Yapeng Fang

Hongbin Zhang
Katsuyoshi Nishinari Editors

Food
Hydrocolloids
Functionalities and Applications
Food Hydrocolloids
Yapeng Fang • Hongbin Zhang •
Katsuyoshi Nishinari
Editors

Food Hydrocolloids
Functionalities and Applications
Editors
Yapeng Fang Hongbin Zhang
School of Agriculture and Biology School of Chemistry and Chemical Engineering
Shanghai Jiao Tong University Shanghai Jiao Tong University
Shanghai, China Shanghai, China

Katsuyoshi Nishinari
School of Food and Biological
Engineering
Hubei University of Technology
Wuhan, China

ISBN 978-981-16-0319-8 ISBN 978-981-16-0320-4 (eBook)

https://doi.org/10.1007/978-981-16-0320-4

© Springer Nature Singapore Pte Ltd. 2021

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We would like to dedicate this book to
Professor Glyn Owen Phillips, who passed
away in July 2020. He has been the Founding
Executive Principal of North East Wales
Institute (presently, the Glyndwr University
Wrexham), the Founding Editor of the journal
Food Hydrocolloids, and the leading
organizer of the Gums and Stabilisers
Conference for the Food Industry and the
International Hydrocolloids Conference. He
established an international network of
scientific communication and collaboration
based on two Hydrocolloids Research
Centres, one in Wrexham and the other in
Wuhan, bearing his name. He was a man of
fidelity and a friend and guide with paternal
affection. It was a pity that the meeting to
celebrate his 90th birthday planned at St
Tropez in 2017 was cancelled, but instead a
special issue of Food Hydrocolloids was
published. His influence on many scientists
was profound and worldwide and will be
greatly missed.
Preface

Food hydrocolloids encompass a wide range of hydrophilic biopolymers such as

proteins, polysaccharides and their derivatives. They are indispensable for the
formulation and processing of our daily food products, e.g., drinks, jams, dressings,
cakes, noodles, hams and more complex functional foods. The importance of
hydrocolloids to the food industry, in our view, has not been fully recognized and
explored. Albeit often used as minor components in foods, hydrocolloids always
perform crucial functions. They not only confer desired structure, stability and
palatability to food products, but also provide with bioactivity and health benefit
that the consumer persistently pursues for. There is a big gap to be connected
between the structures stabilized by food hydrocolloids and the resulting functions,
particularly, the nutritional and health outcomes. The scientific concept of the “food
hydrocolloids” has been consequently expanded from the hydrophilic materials
themselves to the various colloidal structures and functions formed thereby.
In this context, we think it necessary to introduce hydrocolloid knowledge in a
more systematic framework by categorizing into different functionalities and
highlighting the food-related applications. This book aims to be a textbook or
reference tool for undergraduate/graduate students or professionals working in the
field of food science and technology or biopolymer science. It is our purpose that it
can benefit to bridge the divide between fundamental research and industrial appli-
cations. We also hope that it can add to the excellent book Handbook of Hydrocol-
loids edited by Glyn O. Phillips and Peter A. Williams, which has received great
welcome and will be even more popular with its upcoming third edition. The
handbook however differs in organizing knowledge according to each individual
hydrocolloid and aiming to be a technological reference.
This book is the result of long friendship and close collaborations between the
three editors. Y.F. and H.Z. both had stayed in the laboratory of K. N. at Osaka City
University, where Y. F. completed his PhD study and H. Z. conducted his postdoc-
toral research under the kind supervision of K. N. A stronger international collab-
oration network on food hydrocolloids has been set up in China since K. N. joined
the Hubei University of Technology Wuhan in 2013 as a Specially Appointed

vii
viii Preface

Professor. The book is an initiative of the research network and involves many
excellent contributors who are young yet active and creative in the frontline of
hydrocolloid research. Sincere thanks are due to all the chapter contributors whose
efforts make this book a very worthwhile undertaking.

Shanghai, China Yapeng Fang

Shanghai, China Hongbin Zhang
Wuhan, China Katsuyoshi Nishinari
Contents

1 Introduction to Food Hydrocolloids . . . . . . . . . . . . . . . . . . . . . . . . 1

Wei Lu, Xiaobei Li, and Yapeng Fang
2 Solution Properties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
Hongbin Zhang and Ruiqi Li
3 Rheological and Thickening Properties . . . . . . . . . . . . . . . . . . . . . . 75
Katsuyoshi Nishinari
4 Gelling Properties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 119
Katsuyoshi Nishinari
5 Emulsifying Properties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 171
Hui Zhang and Lingli Deng
6 Liquid Foaming Properties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 207
Yongguang Guan
7 Tribological and Sensory Properties . . . . . . . . . . . . . . . . . . . . . . . . 245
Sandip Panda and Jianshe Chen
8 Coating and Film-Forming Properties . . . . . . . . . . . . . . . . . . . . . . . 267
Qian Xiao
9 Self-assembling Properties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 307
Huiyan Zeng
10 Flavour Delivery . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 335
Matthias Schultz
11 Encapsulation and Targeted Release . . . . . . . . . . . . . . . . . . . . . . . . 369
Bin Liu, Lulu Jiao, Jingjing Chai, Cheng Bao, Ping Jiang, and Yuan Li
12 Replacement of Fat or Starch . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 409
Cuixia Sun and Yapeng Fang

ix
x Contents

13 Structuring for Elderly Foods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 445

Makoto Nakauma and Takahiro Funami
14 Bioactivities . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 473
Kang Liu, Xue-Ying Li, Jian-Ping Luo, and Xue-Qiang Zha
15 Dietary Fibers: Structural Aspects and Nutritional Implications . . . 505
Bin Zhang, Shaokang Wang, Santad Wichienchot, Qiang Huang,
and Sushil Dhital
About the Editors

Yapeng Fang is a Distinguished Professor at the School of Agriculture and Biol-

ogy, Shanghai Jiao Tong University. He holds a bachelor and master degree in
polymer physics and chemistry from Shanghai Jiao Tong University, and a PhD
degree in Food Science and Health from Osaka City University. His research interest
is focused on the structure-function relationship of hydrocolloids, and the design of
future foods based on hydrocolloid technologies. He currently severs as an Associate
Editor of the journal Food Hydrocolloids.

Hongbin Zhang is a full professor with tenure at the Department of Polymer

Science and Engineering, School of Chemistry and Chemical Engineering, Shanghai
Jiao Tong University (SJTU). He holds a master degree in applied chemistry and a
PhD degree in polymer materials from SJTU. His research interest is focused on the
interdiscipline of polymer physicochemistry, rheology, colloid chemistry, and food
science, involving polysaccharide, solution, hydrogel and emulsification.

Katsuyoshi Nishinari is currently a specially appointed professor at Glyn

O. Phillips Hydrocolloids Research Centre, Hubei University of Technology. He
worked for National Food Research Institute and then Osaka City University (OCU),
and now is an emeritus professor of OCU. He is also visiting professors at Glyndwr,
ESPCI, SJTU, Osaka, Keio, and other universities. His research interest includes
food hydrocolloids, and rheology applied in food (oral) processing. He currently
serves as the editors of several colloids and rheology related journals.

xi
Chapter 1
Introduction to Food Hydrocolloids

Wei Lu, Xiaobei Li, and Yapeng Fang

Abstract This introductory chapter provides an overview of the definition, classi-

fication, structure, market, regulation, and functional aspects of food hydrocolloids.
The narrow and wide definition of hydrocolloids is compared. A detailed classifica-
tion based on the source of hydrocolloids is summarized and the molecular structure
of typical hydrocolloids, such as polysaccharides and proteins, is introduced. Food
hydrocolloids show diverse potentials in the application of food, nutrition, and
biomedicine industries. They can act as thickening agents, and form gels with
controlled physical properties and functionalities. They can also be employed as
stabilizers for various dispersions, and delivery carriers for bioactive ingredients.
Besides, many hydrocolloids, e.g., whey proteins, or dietary fibers, possess potential
health benefits and can provide basic and essential nutrients for maintaining human
life activity. Furthermore, food hydrocolloids can be tailored into functional mate-
rials with advanced applications in food packaging, biomedical materials,
bionanomaterials, polymer electrolytes, synthesis of inorganic nanoparticles, and
removal of organic pollutants. The market and regulatory aspects of food hydrocol-
loids are also briefly reviewed.

Keywords Hydrocolloids · Definition · Classification · Structure · Functionality ·

Regulation · Safety

W. Lu · Y. Fang (*)
School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China
e-mail: ypfang@sjtu.edu.cn
X. Li
DuPont Nutrition and Health, Shanghai, China

© Springer Nature Singapore Pte Ltd. 2021 1

Y. Fang et al. (eds.), Food Hydrocolloids,
https://doi.org/10.1007/978-981-16-0320-4_1
2 W. Lu et al.

1 History of the Term

The colloidal gels were first investigated by Thomas Graham (1861), who is widely
regarded as the funder of colloid chemistry. He proposed a definition of substances
based on their diffusive behaviors. According to his definition, colloids are slowly
diffusing substances with very high molecular weight and specific structure.
Between these colloidal substances, there exist “feeble forces” that holds them
together in a solution. Graham also defined the colloidal state as a dynamical state
of matter which may change into a crystalloidal status.
About half a century behind the foundation of colloid chemistry, Freundlich,
Ostwald, and Weimarn proposed a new definition of colloids (Mokrushin 1962),
which is different from that given by Graham. They defined colloids as any sub-
stances which are in the dispersed colloidal state. The molecular weight has no
relation to the colloidal state by this definition. For example, the fine suspensions of
gold, silver chloride, or sulfur can also be classified as colloidal solutions according
to this definition. The cognition process on the rationality and limitation of these two
definitions may be also considered as the development process of modern colloid
chemistry.
It was reported that the term “hydrocolloid” derived from the Greek hydro
“water” and Kolla “glue” (Wüstenberg 2015). First known use of hydrocolloid can
be dated back to 1916, which was used to describe a substance that yields a gel with
water. Hydrocolloids are now defined as various long-chain polymers which can
form viscous dispersions and/or gels when dispersed in water. These polymers can
be found in extracts from seaweeds, exudates from trees, flours from grains, products
from fermentations, and many other natural products. They contain many hydroxyl
groups, which significantly increase their hydrophilicity making them hydrophilic
compounds. Furthermore, they can create a dispersion, which is between a true
solution and a suspension and show colloidal properties. In consideration of these
two properties, they are termed as “hydrophilic colloids” or “hydrocolloids.”

2 Definition

A narrow definition of hydrocolloids refers to a series of polysaccharides and pro-

teins that are widely used in different industrial sectors to perform various functions
including thickening and gelling agents, stabilizers of food dispersions (e.g., foams
or emulsions), encapsulation and delivery carriers of functional ingredients and their
controlled release (Phillips and Williams 2009). Besides, all soluble starches, which
chemically belong to the polysaccharide family, can also be considered as a class of
hydrocolloids with many potential applications in the food sector.
Hydrocolloids can also be broadly defined as a variety of biopolymers,
biopolymer-stabilized dispersions, and biopolymer-based gels and particles, such
as polysaccharides, proteins, lipids, protein/polysaccharide aqueous dispersions,
1 Introduction to Food Hydrocolloids 3

emulsions (including Pickering emulsions), foams, bubbles, protein/polysaccharide-

based hydrogels and micro/nanoparticles, solid lipid nanoparticles, liposome, and
oleogels.

3 Classification

Hydrocolloids are widely used in a variety of industries to perform different func-

tions as described above (Williams and Phillips 2009; Lin et al. 2012). The fre-
quently used hydrocolloids and their origins are given in Table 1.1.
Some of the plant hydrocolloids come from trees and tree gum exudates (e.g.,
cellulose, gum Arabic, gum karaya, gum ghatti, gum tragacanth) while some of them
come from plant (cellulose pectin, and starch), seeds (locust bean gum, guar gum,
tara gum, tamarind gum, soybean proteins, zein, rice protein, and gluten), and tubers
(konjac glucomannan). Algal hydrocolloids are mainly derived from red seaweeds
(agar, carrageenan, and furcelleran) and brown seaweeds (alginate).
Microbial hydrocolloids can be produced from simple sugars (glucose or sucrose)
or starch using a microbial fermentation process. Species of bacteria and fungus that
have been used to produce these hydrocolloids by fermentation include Xanthomonas
campestris (xanthan gum), Leuconostoc mesenteroides and Streptococcus mutans
(dextran), Agrobacterium (curdlan), Athelia rolfsii (scleroglucan), Sphingomonas
elodea (gellan), and Aureobasidium pullulans (pullulan).
Animal-sourced hydrocolloids mainly are proteins coming from collagen taken
from bones, skin, and tendons of animals, and skins of fish (gelatin), milk (caseinate
and whey proteins), eggs (egg white proteins), and chitin shells of shrimps and other
Crustaceans (chitosan).

Table 1.1 Source of frequently used hydrocolloids

Source Hydrocolloids
Plant Polysaccharides
Starches, pectins, cellulose, larch gum, guar gum, locust bean gum, tara gum,
tamarind seed gum, acacia gum/gum, Arabic gum, tragacanth, karaya gum, ghatti
gum, inulin, and konjac glucomannan
Proteins
Soy proteins, zein, rice proteins, and gluten (wheat protein)
Algal Alginate, carrageenan, agar, and furcelleran
Animal Gelatin, caseinate, whey proteins, egg white proteins, glycogen, and chitosan
Microbial Dextran, xanthan, scleroglucan, curdlan, pullulan, and gellan
Modified Modified starches, propylene glycol alginate, microcrystalline cellulose (MCC),
amidated pectin, methylcellulose (MC), ethylcellulose (EC),
hydroxypropylmethylcellulose (HPMC), hydroxypropyl cellulose (HPC), and car-
boxymethylcellulose (CMC)
4 W. Lu et al.

4 Structures

4.1 Polysaccharides

The primary molecular chain structure of polysaccharides is monosaccharide units

(Table 1.2) bound together by glycosidic bond, which is formed between the
hemiacetal or hemiketal group of a monosaccharide and the hydroxyl group of
neighboring monosaccharide, known as O-glycosidic bonds. Besides, S-, N-, or
C-glycosidic bonds also exist, where the oxygen of the glycosidic bond is replaced
with sulfur, nitrogen, or carbon, respectively. In addition, α- or β-glycosidic bonds
can be distinguished by the relative stereochemistry (R or S) of the groups of C5 and
C1 in the pyranose ring. If groups connected to these two carbons have the same
stereochemistry, the glycosidic bond is defined as α type, whereas a β-glycosidic
bond is formed when the groups show different stereochemistry.
Therefore, glycosidic bonds in the backbone of polysaccharides mainly include
α-1,4 (e.g., amylose, glycogen, amylopectin), β-1,4 (e.g., pectin, alginate, cellulose,
guar gum, xanthan, konjac glucomannan, xylan), α-1,6 (e.g., dextran), β-1,3 (e.g.,
Arabic gum, laminarin), and β-1,2 (e.g., inulin) glycosidic bonds (Fig. 1.1). The
backbone of some polysaccharides also shows a mixture of the different glycosidic
bonds. For example, carrageenan is composed of repeating sulfated and/or
non-sulfated galactose and 3,6-anhydrogalactose units. The units are linked by
alternating α-1,3 and β-1,4 glycosidic bonds. A similar chain structure can be
observed for gellan gum, which is made up of repeating tetra-saccharides consisting
of two glucose and one of each residue of rhamnose and glucuronic acid.
Polysaccharides composed of the same monosaccharide are called homo-
polysaccharides such as starches and glucan while those composed of more than
one type of monosaccharides are called heteropolysaccharides. Some polysaccha-
rides show a segmented chain structure. Alginic acid (Fig. 1.2), for example, is a
linear biopolymer composed of β-1,4 linked D-mannuronate (M) and L-guluronate
(G) residues. G and M resides are covalently linked together in different sequences
or blocks. They can appear in consecutive M-residues (M-blocks), consecutive
G-residues (G-blocks), or alternating M and G-residues (MG-blocks).
Besides, several polysaccharides show a crystalline zone in their chain structure,
e.g., cellulose, or starch. The crystalline fractions of these polysaccharides with
different structures and properties can be collected by physical or chemical
processing such as microcrystalline cellulose (MCC), microfibrillated cellulose
(MFC), and nanocrystalline cellulose (NCC). Furthermore, the chemical structure
of natural polysaccharide is modified to improve its water solubility or functional-
ities. Examples include modified starch, propylene glycol alginate, amidated pectin,
methylcellulose, ethylcellulose, or carboxymethylcellulose.
In direct analogy with proteins, polysaccharides can also have several levels of
structures. The primary structure of a polysaccharide is defined as the sequence of
monosaccharides. The intramolecular single helices with different persistence
lengths are considered as secondary structures of a polysaccharide while tertiary
1 Introduction to Food Hydrocolloids 5

Table 1.2 Examples of common monosaccharides and corresponding polysaccharides

Monosaccharide Molecular structure Representative polysaccharides
Glucose Starch, cellulose, β-glucan, konjac glucomannan,
gellan, dextran, pullulan, xanthan, xyloglucan

Glucosamine Chitosan

Acetyl-glucose Chitosan
Poly-N-acetyl-β-D-glucosamine

Fructose Inulin

Mannose Konjac glucomannan, guar gum, tara gum, fenu-

greek gum, locust bean gum, xanthan

Galactose Arabic gum, guar gum, carrageenan, agar, tara

gum, fenugreek gum, acacia gum, karaya gum,
xyloglucan, locust bean gum

(continued)
6 W. Lu et al.

Table 1.2 (continued)

Monosaccharide Molecular structure Representative polysaccharides
Arabinose Arabic gum

Galacturonic Pectin
acid

Rhamnose Gellan

Glucuronic acid Gellan, xanthan

Mannuronate Alginate

Guluronate Alginate

structures always exhibit as tight or loosely supercoiled helices. Tightly or loosely

intertwined chains with tertiary structures form the intermolecular quaternary struc-
tures of polysaccharides (Fig. 1.3).
Diener et al (2019) demonstrated several structural levels using carrageenan as
model polysaccharides. In the presence of potassium, a disorder to order the transi-
tion from random coil to single helix (secondary structure) was first observed,
1 Introduction to Food Hydrocolloids 7

Fig. 1.1 Examples of molecular chain structure of polysaccharides

Fig. 1.2 Structural characteristics of alginates: (a) monosaccharide composition (b) chain confor-
mation, (c) block distribution (Draget and Taylor 2011). The permission for reproduction of the
figure was obtained from Elsevier
8 W. Lu et al.

Fig. 1.3 Schematic elucidation of the multilevel structure of polysaccharide chains: An example
from carrageenan (Diener et al. 2019). The permission for reproduction of the figure was obtained
from ACS Publications

followed by intrachain supercoiling events (tertiary structure) and macroscopic

anisotropic domains, which are part of a network (quaternary structure) with tunable
elasticity up to ~103 Pa. Loosely intertwined helices were also observed as tertiary
structure.

4.2 Proteins

Proteins are biopolymers comprised of one or more chains of amino acid residues.
The amino acid residues in protein chains are linked together by peptide bonds,
which is an amide-type of the covalent chemical bond linking C1 of one α-amino
acid and N2 of another adjacent amino acid (Fig. 1.4).
Protein structure mainly refers to four distinct aspects (Finkelstein and Ptitsyn
2016): (1) primary structure, the amino acid sequence; (2) secondary structure,
repeating structures formed by hydrogen bonds, mainly including α-helix, β-sheet,
and β-turns; (3) tertiary structure, the spatial shape of a polypeptide chain or the
spatial arrangement of the secondary structures. Tertiary structure is mainly stabi-
lized by hydrophobic interactions, but also involving disulfide bonds, hydrogen
bonds, salt bridges, or post-translational modifications; and (4) quaternary structures,
1 Introduction to Food Hydrocolloids 9

Fig. 1.4 Diagram of the formation and structure of peptide bond

the structure formed by several polypeptide chains (technically termed as protein

subunits) (Robert et al. 2009).
Proteins are not completely rigid molecules. Proteins may transform between the
above-mentioned structures while they perform their functions. Generally, the ter-
tiary or quaternary structure of proteins is regarded as their conformation, which is
defined as the spatial arrangements of atoms. The conversion between different
conformations generally does not refer to the breakage or formation of covalent
bonds. For example, the structure of protein molecules can be altered by thermal
vibration (thermal denaturation) or the collision with other molecules (Christopher
et al. 2013). Thermal denaturation is also one of the mechanisms that induce the
gelling of protein molecules in solutions. Conformation is different from another
relevant term named configuration, which mainly refers to the fixed three-
dimensional relationship of the atoms in a molecule, defined by the bonds between
them. The change of configuration of proteins may involve the break or formation of
covalent bonds, e.g., the change of primary and/or secondary structure of proteins
(Finkelstein and Ptitsyn 2016).

5 Marketing

The lifestyle changes, the increasing awareness of the association between diet and
health, and emerging new processing technologies contribute to a rapid increase in
the consumption of food hydrocolloids-based ready-made, functional, and healthy
food products, which consequently leads to an increase in the market demand of
hydrocolloids. The market demand for hydrocolloids is estimated at $ 8.8 billion in
2018 and is predicted to grow at a compound annual growth rate (CAGR) of 5.3%
from 2018 to 2023, to reach $ 11.4 billion by 2023 (Singh 2019). North American is
predicted to be the fastest-growing market in these 5 years (Fig. 1.5).
The rising demands for convenient food products in the countries of the Asia
Pacific, South America, and the Middle East & African regions promote the devel-
opment of the processed food industry worldwide, which accordingly leads to a
growth in the hydrocolloids market. In addition, manufacturers always focus on the
10 W. Lu et al.

Hydrocolloids Market by region (USD billion)

5
North American Europe Asia Pacific RoW

0
2016 2017 2018 2019 2020 2021 2022 2023
Year

Fig. 1.5 Hydrocolloids market by region. RoW indicates the rest of the world (Singh 2019). The
data from 2019 to 2023 is predicted based on the estimated growth rate

product innovation and multifunctionality of hydrocolloids with the objective of

offering high-quality hydrocolloids for the end-users. Thus, manufactures perform
various research and development (R&D) activities and these activities accordingly
contribute to the rapid growth of the hydrocolloids market. One of the major factors
that restrict the development of the hydrocolloids market is the international regu-
lations and quality standards of food additives (e.g., stabilizers). The fluctuations in
the price of raw materials for producing hydrocolloids can also affect the market.
In 2018, the thickeners sector, animal sector, food & beverage and gelatin sector
are estimated to account for the largest share based on functions, sources, applica-
tions, and types of hydrocolloids, respectively. Take gelatin as an example, gelatin
has wide applications in food products such as desserts, candies, ice creams, and
marshmallows. In Europe, gelatin is classified as food and is not subjected to food
additive legislation, which creates a lucrative opportunity for manufacturers in the
next few years.

6 Legislation and Safety

6.1 Background

Food hydrocolloids, such as pectin, agar, starch, and gelatin, have been used for
centuries, and consumers are quite familiar with these hydrocolloids. Many hydro-
colloids in use today were developed long before regulatory approvals, and their use
1 Introduction to Food Hydrocolloids 11

level and use in specific applications were restricted. For instance, carrageenan, agar,
or alginate extracted from seaweeds are eaten as basic foods. These edible polymers
from seaweeds are thus considered safe for use in food products. This principle of
“generally recognized as safe” (GRAS) is still in use today but under a more rigorous
review. For example, carrageenan can be classified into λ-, κ, and ι-, carrageenan.
Some of the modified carrageenan, e.g., semi-refined version, found its way of being
used as food texturizing agent only since the 1980s and 1990s while refined
carrageenan has been used for more than 60 years (Imeson 2010). However,
intentional acid-hydrolyzed carrageenan, like poligeenan, cannot form gels and
even have been proved to cause in vivo gastrointestinal ulceration and tumors via
food, water, or oral route. Poligeenan has been also reported to show immune system
toxicity and potentially suppress the immune response (McKim et al. 2018). These
results suggested that degraded fractions of nature hydrocolloids which can be safely
used as food additives need strict safety evaluation before being used in the food
industry.

6.2 Legislation

Regulatory approval of a food ingredient is necessary because the additives would

have no function or market without approval by the governments. Food hydrocol-
loids are generally regulated either as a food additive or as a food ingredient. Except
for gelatin, most of the food hydrocolloids are currently regulated as food additives.
Regulatory authorities strictly control the approval of food additives (Imeson
2010). Chemical modifications are generally not allowed except for cellulose deriv-
atives, starches, and propylene glycol alginate. Physical and enzymatic modifica-
tions are allowed, e.g., physically modified pectin (Slendid™, CP Kelco Division of
JM Huber). Some new hydrocolloids are brought to the market under the “generally
recognized as safe” principle, such as konjac, tara gum, and pullulan. Gellan gum is
the last hydrocolloid that completely goes through a food additive petition in the
global market. It took many years and cost tens of millions of dollars to get the
approval of gellan gum. The total profit earned by gellan gum over the next 20 years
after its approval is still lower than the cost for its approval. These probably suggest
that it is almost impossible for a new hydrocolloid to accomplish a full approval
process in the global market in the foreseeable future. Cassia gum has recently been
approved in France (August 2008) and it is expected to be approved in the EU
market soon.

6.2.1 International System

The Food and Agriculture Organization (FAO) and the World Health Organization
(WHO) established the Codex Alimentarius Commission (CAC) in 1962. CAC is an
intergovernmental organization consisting of delegations from FAO and WHO
12 W. Lu et al.

member countries that take part in developing the food standards. Another organi-
zation, the Joint FAO/WHO Expert Committee on Food Additives (JECFA) was
established before the CAC in 1956 (Magnuson et al. 2013), which is considered as
the oldest and most active body in doing a similar job.
The main task of CAC is to address the safety and nutritional quality of food
products and to develop international standards for promoting trade of foods, such as
setting standards for approved food additives, determining its limits of addition and
safety indexes (e.g., limits for contaminants and toxins, and residue limits for
pesticides), and establishing hygiene and technological function practice codes.
These standards, guidelines, practice codes, and recommendations constitute the
Codex Alimentarius.

6.2.2 European System

European Commission first cleared food hydrocolloids in 1995 based on the Direc-
tive 95/2/EU for Food Additives, which is known as the Miscellaneous Additives
Directive (MAD). MAD authorized a large number of food hydrocolloids (including
alginates, starches, celluloses, and most gums) for their utilization in food products
at suitable levels (Phillips and Williams 2009). Since the adoption of the Directive,
the Commission proposed several modifications by considering the development of
the market. A second modification was made in 1998 and the Member States of the
EU were required to put into effect this updated version in 2000. The new regulation
in this Directive (98/72/EC) came into force in all EU Member States and it affected
many hydrocolloids, mainly including potassium alginate, sodium alginate, konjac,
carrageenan, guar, and pectin.

6.2.3 US and Japan Systems

Since the CAC is the final regulation and can provide for worldwide approvals,
countries (outside the EU) are free to adopt their standards. In United States (USA),
the Food and Nutrition Board of the Institute of Medicine (funded by the United
States Food and Drug Administration, FDA) published the Food Chemicals Codex
(FCC), and the regulation about hydrocolloids can be found in the Fourth Edition
pressed in 1996. Japan also has its own regulation system, including many food
additives specific to Japan (Dai and Chau 2017).

6.2.4 China Systems

In China, except for starch, the usage of hydrocolloids in foods is regulated by

“National standards for food safety: standards for the use of food additives
(GB2760),” which was first established in 2011 (GB2760-2011), and then modified
in 2014 (GB2760-2014). It stipulates the type of hydrocolloids that can be used in
1 Introduction to Food Hydrocolloids 13

China and their levels in the products. Based on GB2760-2014, around 40 hydro-
colloids are allowed to be used in the China market. For example, the most widely
used hydrocolloids in meat and fish products include carrageenan, xanthan gum,
guar gum, agar, gelatin, alginate, locust bean gum, curdlan, and konjac
glucomannan. These frequently used hydrocolloids are all naturally derived.
Generally, these edible hydrocolloids are used at a low level, mainly functioning
as thickening agents, stabilizers, and gelling agents. Many hydrocolloids can also be
considered as dietary fibers, which have many potential health benefits. Thus, a low
level of using edible hydrocolloids in food products is always safe or even good for
health. However, some manufacturers use a high level of hydrocolloids for specific
purposes (cost, stability, etc.), leading to a potential safety issue of the products. In
addition, many hydrocolloids can also be used in industrial products, and industrial-
grade hydrocolloids are much cheaper than food grade, but potentially contain
various harmful ingredients, which can threaten human health. Therefore, it is illegal
to use industrial-grade hydrocolloids in food products but it still can happen that
some producers illegally add them into foods due to their low cost.

6.3 Consumer Concerns

No matter what the scientific facts are, it is the consumers’ approval that decides the
market future of a food ingredient, and hydrocolloids are no exception. In terms of
perception, a label is very important. A friendly label always can reduce people’s
perception of too much chemical connotation, and thus feel they are safe. For
example, label-friendly carbohydrate gum and vegetable gum are used to describe
methylcellulose and hydroxypropylmethylcellulose in the USA, respectively.
Another example, carboxymethyl cellulose (CMC) is a very chemical name, whereas
cellulose gum, a synonym of CMC, sounds much more acceptable. Therefore, EU
authorities allow the use of cellulose gum on the label.
Another main concern is genetic engineering and relevant genetically derived
hydrocolloids. For example, seeds, seaweed, or other plants could be genetically
modified to yield a higher production of relevant hydrocolloids; kelp could be edited
to produce more alginates; the mature time of a carob tree that can produce a
commercial scale of bean gum could be significantly shortened as compared with
the normal 12–15 years. However, the safety of these genetically derived hydrocol-
loids is not conclusive, and their application is still constrained by consumer
concerns (Imeson 2010).
14 W. Lu et al.

Table 1.3 Hydrocolloids commonly used in food applications

Applications Hydrocolloids
Structuring Thickening agents Xanthan, methyl cellulose (MC),
agents carboxymethyl cellulose (CMC), guar
gum, hydroxypropylmethyl cellulose
(HPMC), locust bean gum, konjac
glucomannan, tara gum, and
xyloglucan
Gelling agents Amylose, agar, carrageenan, gellan
gum, pectin, alginate, casein, soybean
proteins, gelatin, xanthan, MC, HPMC,
locust bean gum, konjac glucomannan,
xyloglucan, and curdlan
Stabilizers Emulsifiers Gum Arabic, modified starch, modified
cellulose, crystalline cellulose, pectins,
galactomannans, casein/caseinate,
whey proteins, and soy proteins
Foaming agents Casein, whey proteins, zein, gelatin,
pectins, modified cellulose, chitin, and
chitosan
Delivery Nanoparticles, films, fibrils, microcap- Whey proteins, casein, gelatin, zein/
carriers sules, hydrogels particles gliadin, agar, carrageenan, alginate,
chitosan, guar gum, cellulose, pectin,
xanthan gum, and starch
Bioactive Dietary fibers, antimicrobial, anti-virus, Resistant starch, cellulose, hemicellu-
ingredients anti-coagulant, antioxidant, anti-cancer, lose, guar gum, pectins, β-glucans,
reducing blood lipids, enhancing the psyllium; sulfated polysaccharides
immune system and bone health, anti- from marine algae (e.g., sulfated
cardiovascular disease, lowering blood fucoidan, carrageenan, galactans,
pressure, losing weight, satiating effect fucoidan, sulfated rhamnogalactans,
laminaran, and alginic acid), and whey
proteins
Advanced Food packaging, biomedical materials, Starch, modified cellulose, chitin/
materials scaffolds, biomimetic materials, nano- chitosan, pectin, gluten, soy proteins,
structured membranes, electrolytes, gelatin, zein, crystalline cellulose, and
decontamination materials crystalline starch

7 Functionality and Application

Hydrocolloids can be functionally used as the thickening and gelling agents, stabi-
lizers for emulsions and foams, and delivery carriers for bioactive components. They
can be also tailored into advanced nanomaterials with various specific applications in
both food and biomedical industries. Besides, hydrocolloids and their derivate show
diverse health benefits and are considered as a class of bioactive ingredients. In this
section, these main functionalities of hydrocolloids are summarized (Table 1.3).
1 Introduction to Food Hydrocolloids 15

7.1 Structuring Agents

Hydrocolloids have been used in a variety of food formulas to improve their quality
and shelf-life. The two major applications of hydrocolloids are thickening and
gelling agents. Regarding the thickening, they have been used in many food systems,
such as gravies, soups, sauces, salad dressings, and toppings. In terms of gelling,
hydrocolloids can be used in products like jelly, jam, marmalade, low-calorie gels, or
restructured foods.

7.1.1 Thickening Agents

The process of thickening refers to the entanglement of polymer chains, leading to a

significant increase in the viscosity of the system. Thickening occurs above a critical
polymer concentration (overlap concentration, C*), which refers to the transition
from the “dilute region,” where the polymers can move freely without entanglement
in solution, to the “semi-dilute region,” where molecules crowding promotes the
overlap of polymer coils and thus leads to the entanglement. Below C*, the polymer
dispersions exhibit Newtonian behavior whereas shows a non-Newtonian behavior
above this concentration (Phillips and Williams 2009).
The viscosity of the polymer solution is significantly affected by the molecular
mass of the polymer. The dependency of viscosity on the shear rate increases with
the increasing molecular mass. When the molecular mass of the polymer increases,
the shear thinning of the polymer dispersion occurs at a lower shear rate value. In
addition, the hydrodynamic size of polymers can be obviously influenced by their
molecular structures. Linear and stiff molecules always show a larger hydrodynamic
size than that with a branched and flexible structure and thus yield a much higher
viscosity (Phillips and Williams 2009).
Hydrocolloids that have been applied as thickening agents include starch, gum
Arabic or acacia gum, modified starch, xanthan, gum tragacanth, galactomannans,
gum karaya, and CMC (Saha and Bhattacharya 2010) (Table 1.4). The thickening
effect of hydrocolloids can be influenced by many factors, including the type of
hydrocolloids used, concentration, pH, temperature, or the food system in which
they are used.

7.1.2 Gelling Agents

Gels are a form of matter intermediate between solid and liquid states, showing both
elastic (mechanical rigidity) and flow (liquid) properties (Aguilera 1992). A sol
(liquid) to gel (solid) transition occurs when gels are formed. Gelation is the process
that the polymer chains crosslink to form a three-dimensional (3D) network that
entraps water within it (Oakenfull and Glicksman 2009). Food hydrocolloids are
widely employed as gelling agents to produce food gels with acceptable properties,
16 W. Lu et al.

Table 1.4 Hydrocolloids used as thickeners in foods

Hydrocolloids Properties Applications
Xanthan Highly shear thinning; keeps viscous Soups and gravies, ketchup,
at high temperature and wide pH beverages, toppings, desserts,
ranges and fillings
CMC High viscosity; decreased viscosity Gravies, salad dressings,
by adding electrolytes and at low pH ketchup, and fruit pie fillings
MC and HPMC Viscosity increases with increasing Cake batters, beverages, salad
temperature; viscosity is independent dressings, and whipped
of pH and electrolytes toppings
Gum Arabic Low viscosity; shear thinning dis- Soft drinks and fruit juice based
persion at low shear rates (<10/s); beverages
near Newtonian behavior above the
shear rate of 100/s
Galactomannans (guar High viscosity at low shear rate; Dairy products, i.e., ice cream,
gum, locust bean gum highly shear thinning; the viscosity is ketchup, fruit juices, pudding
and tara gum) independent of electrolytes; powder, and cake batter
decreased viscosity at very high and
very low pH, and high temperatures
Konjac glucomannan Highly viscosity independent of Noodles, jelly desserts, surimi,
salts; forms thermally irreversible meat products
gels in the presence of alkali
Xyloglucan Heat, acid, salt resistant, short texture Sauce for grilled eat, cutlet
(non-thread forming)
Gum tragacanth Rehydrates fast in cold or hot water Salad dressing, bakery emul-
to produce highly viscous dispersion, sion, fruit beverage, and sauce
up to 4000 mPas at 1 w/w%

especially the texture properties (Table 1.5). Food gels are 3D network structures of
high moisture. Food gels can resist flow under pressure and retain their distinct
shape. They are viscoelastic systems with a storage modulus (G0 ) larger than the loss
modulus (G00 ) (de-Vries 2004).
Many classifications can be done for gels (See Chap. 4). Gels formed by covalent
bonds are called chemical gels, and those formed by secondary bonds such as
hydrogen bonds, electrostatic and hydrophobic interactions, and/or van der Waals
forces are called physical gels (Djabourov et al. 2013). Gels can be formed by
physical (e.g., heat or pressure), chemical (e.g., pH), or biological treatments (e.g.,
enzymes). For example, milk gels can be formed by acid or by rennet, while KGM
gelation is induced by alkali. However, some physically induced gels, for example
by heating, can make covalent bonds and thus lead to the formation of chemical gels.
The gel formation always depends on several physicochemical factors, including
temperature, pressure, ionic strength, pH, presence of enzyme, solvent quality, the
concentration of gelling agents, selection of different biopolymers and their molec-
ular weight (Banerjee and Bhattacharya 2012). Selection and control of these factors
result in the formation of different kinds of gels, such as hydrogels, organogels,
xerogels, aerogels, emulsion gels, oleo gels, weak gels, fluid gels, or temperature-
sensitive gels (thermoreversible or thermo-irreversible) (Graham et al. 2019). Gels
1 Introduction to Food Hydrocolloids 17

Table 1.5 Hydrocolloids used as gelling agents

Hydrocolloids as a
gelling agent Properties Applications
Modified starch Cold-set gel; thermal irrevers- Dairy desserts
ible and opaque
Agar Cold-set gels; Bakery products, jellies
thermoreversible
κ-, and ί-Carrageenan Cold-set gels; Puddings, milk shakes, tofu
thermoreversible
Low methoxy pectin Cold-set gels at low pH; Jams, glazes, jellies, milk-based
thermoreversible desserts
High methoxy pectin Cold-set gels at low pH; ther- Jams, jellies
mal irreversible
Gellan gum Cold-set gels; Jellies with different flavors
thermoreversible; highly
transparent
Alginate Thermal irreversible gels; sta- Restructured foods, bakery creams
ble when being heated
Methyl and Thermoreversible gels; insta- Cake batters, beverages, whipped
hydroxypropylmethyl ble when being chilled toppings, salad dressings
cellulose
Curdlan Thermoreversible and thermal Surimi, sausage
irreversible gels

with a wide range of properties can be obtained depending on the conditions that are
employed for gel formation. Some gels are brittle, i.e., fractures at a small deforma-
tion, while the others are deformable and would not break even at very large
deformation. They can be transparent or also be opaque.

7.2 Stabilizers

Hydrocolloids are widely used as stabilizers of food dispersions such as emulsions

and foams, which can be used to design a variety of functional food structures with
desired properties in texture, stability, flavor release, or nutrition.

7.2.1 Emulsifier

Hydrocolloids-based emulsifiers can be seen in many food products include carbon-

ated ice cream, soft drinks, and sauces. Many hydrocolloids are able to act as
stabilizers (stabilizing agents) of oil-in-water (O/W) emulsions, whereas only a
few of them can act as emulsifiers (emulsifying agents). Emulsifying agents are
required to have surface activity at the oil–water interface, and thus the ability to
adsorb to the surface of newly formed oil droplets during homogenization, leading to
18 W. Lu et al.

the formation of stable O/W emulsions (Dickinson 2009). Emulsifying activity and
emulsion stability should be briefly described here as is done for foaming ability
later.
The commonly used polysaccharide-based emulsifiers are Arabic gum, sugar beet
pectin, modified starches, galactomannans, and modified celluloses. The emulsify-
ing ability of these polymers has their chemical structure bases in either (1) the
non-polar chemical groups attached to the molecular backbone (e.g., propylene
glycol alginate, or hydrophobically modified starch/cellulose) or (2) the presence
of a protein component covalently-, or physically linked to the polysaccharide (e.g.,
Arabic gum, sugar beet pectin, or guar gum).
Bovine milk and egg proteins are the most widely used protein-sourced emulsi-
fiers such as casein and whey protein. Casein is a group of amphiphilic proteins
which can be classified into αs1-, αs2-, β-, and κ-casein dependent on their sensibility
to free calcium ion. Whey protein contains a family of globular proteins with rigid
structures, mainly including α-lactalbumin, β-lactoglobulin, bovine serum albumin
(BSA), and immunoglobulins. Animal-sourced gelatins (bovine, pig, or fish) also
have amphiphilic characters, and show surface activity, but they typically exhibit a
poor stabilization effect towards emulsions. Recently, the development of plant-
sourced protein-based emulsifiers is receiving more and more attention, which is
mainly driven by the vegetarian’s will of replacing animal proteins to reduce the
consumption of animal proteins, as well as to promote food sustainability and
security. Some plant proteins are promising emulsifiers, including lupin proteins,
pea proteins, corn germ proteins, and soy proteins (Ozturk and McClements 2016).

7.2.2 Foaming Agent

Foam is a dispersion where a high-volume of gas is dispersed in a liquid. The

stabilization of foams requires an interfacial barrier to prevent coalescence of
neighboring gas bubbles, e.g., the protein layer at the air–water interface. Food
particles at the air–water interface can also stabilize or destabilize foams, depending
on their properties (e.g., wettability or spreading). Foaming properties generally refer
to the foaming capacity and foam stability. Foaming capacity (or foamability) is the
capacity for the continuous phase to embed gas while the foam stability is defined as
the ability to maintain the gas for a certain time. Many food-derived hydrocolloids
have been widely used as stabilizers of foams, including casein, whey proteins, zein,
gelatin, pectin, cellulose, chitin, and chitosan (Dickinson 2017). The foaming
properties of hydrocolloids, e.g., proteins, depend on the intrinsic factors (e.g.,
chemical structure, size, hydrophobicity, or surface chemistry) and external factors
(e.g., concentration, pH, temperature, and the presence of other components).
1 Introduction to Food Hydrocolloids 19

Fig. 1.6 Summarized functional delivery systems based on hydrocolloids

7.3 Delivery Carriers

Proteins and polysaccharides are two important kinds of biopolymers employed to

fabricate food-grade delivery systems. They are biocompatible, biodegradable, and
non-toxic polymers, which makes them attractive building units for delivery systems
in food, biology, and medicine applications. In addition, the diversity of molecular
structures exhibited by proteins or polysaccharides (e.g., molecular weight, branched
or linear structure, charges, polarities, amphiphilicity, dimensions, or reactivities)
offers researchers the opportunity to build delivery carriers with desired specifica-
tions (McClements 2016). These protein- or polysaccharide-based delivery carriers
can be prepared by bottom-up methods that involve self-assembly of individual
molecules into large particles, or top-down methods that involve breaking-down of
large biopolymers into small fragments. Multiple intermolecular forces usually take
part in the formation and stabilization of these carriers for bioactive components
including electrostatic interaction, hydrophobic effect, hydrogen bond, steric repul-
sion, and/or van der Waals attractive force.
Proteins and polysaccharides commonly used to fabricate delivery carriers can be
classified into water-soluble and water-insoluble ones. Water-soluble proteins and
polysaccharides include casein, whey protein, agar, gelatin, carrageenan, alginate,
pectin, guar gum, and xanthan gum while water-insoluble ones have zein, chitosan,
gliadin, starch, or cellulose. Functional delivery carriers developed based on these
biopolymers exhibited different structures, e.g., nanoparticles, films, fibers, hollow
microcapsules, emulsions, or hydrogel particles (Fig. 1.6), through which a variety
of nutraceuticals were delivered, such as polyphenols, vitamins, carotenoids, unsat-
urated fatty acids, fish oils, or essential oils (Abd El-Salam and El-Shibiny 2012;
Ezhilarasi et al. 2012; Fathi et al. 2014).
20 W. Lu et al.

7.4 Bioactive Ingredients

Except for the above-mentioned functionalities, many food hydrocolloids also show
potential health benefits. For example, whey proteins and their derivate show a
variety of health benefits, including antimicrobial and antiviral properties, and
enhance immune defense and bone health, and protect against cancer and cardio-
vascular disease, and reduce oxidative stress and increase levels of glutathione, and
lower blood pressure, and potentially decrease the low-density lipoprotein (LDL)
and blood cholesterol level, moderate blood sugar and increase both the level of
insulin and the sensitivity of its effects, reduce inflammation, have beneficial effects
on inflammatory bowel disease, and induce satiety and reduce hunger, and poten-
tially help to lose weight (Birsen Bulut and Nihat 2012). Many of these health
benefits can be attributed to bioactive peptides released by intestinal hydrolysis of
whey proteins, e.g., antimicrobial peptides, angiotensin-converting enzyme (ACE)
inhibition peptides, antioxidant peptides, or blood-sugar-control peptides.
Polysaccharides have been also reported to possess a variety of health benefits.
One of their major health benefits is being considered as dietary fibers, which can
positively generate a series of biological activity (Fig. 1.7), including but not limited
to eliminating constipation or stimulating colonic muscular activity, encouraging
stool expulsion, promoting the level of short-chain fatty acids (SCFAs), gastroin-
testinal peristalsis, inhibiting the population of pathogenic bacteria (Clostridia),
acting as prebiotics (diet fibers) and antimicrobial agents, and reducing the lipid
absorption (Mudgil and Barak 2013; Makki et al. 2018). Dietary fibers are a family
of polysaccharides, and can be further divided into insoluble and soluble forms
(Deehan et al. 2017). The definition of “soluble” and “insoluble” is different in the
nutritionist community and physicochemical community. This is a serious problem.
KGM is usually classified as a soluble dietary fiber by nutritionists, but
physicochemists struggle to dissolve KGM. It depends on the acetyl content. If the
acetyl group is introduced artificially, the solubility is improved. But some natural
KGM is not so soluble. Water-insoluble dietary fibers such as cellulose and hemi-
cellulose have a fecal bulking effect and are hard to be digested by the gut bacteria.
However, water-soluble fibers can be fermented by the gut bacteria and release
health-beneficial metabolites such as SCFAs. Furthermore, many soluble non-starch
polysaccharides such as guar gum, pectin, β-glucans, and psyllium, can form a gel
structure in the intestinal tract which can delay the absorption of glucose and lipids
(Deehan et al. 2017).
In addition to dietary fibers, polysaccharides also show a variety of potential
health benefits. Sulfated polysaccharides derived from marine algae, for example,
are reported to possess many biological activities, including anti-coagulant, anti-
virus, antioxidant, immunomodulation, inducing osteoblastic cell differentiation,
protecting gastric mucosa against acid and pepsin, inhibiting UV-B induced matrix
metalloproteinase-1 activity in human skin, and reducing cholesterol and triglycer-
ides (TG) levels (Wijesekara et al. 2011). These health benefits of sulfated
1 Introduction to Food Hydrocolloids 21

Fig. 1.7 Influence of low- and high-fiber diet on the composition, diversity, and function of gut
microbiota (Makki et al. 2018). The permission for reproduction of the figure was obtained from
Cell Press

polysaccharides are always correlated to their molecular structure, such as molecular

weight.
Another attractive health benefit of polysaccharides is their great potential in
reducing the risk of cancer. Many studies, mainly observational, have investigated
the correlation between fiber intake and the risk of colon or rectum cancers.
Intervention studies have confirmed the impact of dietary fiber on the recurrence
22 W. Lu et al.

of adenoma, which are often considered an early marker of colorectal cancer.

Nevertheless, there is no hard evidence that can prove the reducing-effect of fiber
intake on the risk of colorectal cancer. Thus, the guideline on dietary fiber intake
aiming to reduce the colorectal cancer risk seems to be lacking enough evidence but
individuals with a low level of fiber intakes potentially have an increased cancer risk
(Mudgil and Barak 2013).

7.5 Functional Materials

In addition to the above-mentioned functionalities, many food hydrocolloids can

also be tailored into a variety of functional materials, such as food-packaging film,
artificial joint prosthesis, bone tissue engineering materials, and cartilage tissue
engineering (Kapoor and Kundu 2016).

7.5.1 Food-Packaging Materials

Foods are generally packaged to extend their shelf-life and to illustrate the compo-
sition and nutrition information to the consumers. With the increasing concern on the
environmental and food safety requirements, the development of renewable biode-
gradable food-packaging materials raises more and more attention over the last few
years, and a class of natural biopolymer-based materials has been developed as
promising food-packaging materials. The food-derived biopolymers that were used
to develop food-packaging materials can be classified into two major groups, poly-
saccharides and proteins. Polysaccharide-based packaging materials include starch,
cellulose, chitin/chitosan, konjac glucomannan, and pectin while protein-based
materials mainly have soy protein, wheat gluten, corn zein, gelatin, whey proteins,
and casein (Tang et al. 2012).
Polysaccharide-based food-packaging films have different characteristics
depending on the materials (Cazón et al. 2017): (1) Starch-based food-packaging
materials always have moderate oxygen barrier properties but poor moisture barrier
and mechanical properties; (2) Chemically modified cellulose derived packaging
materials always show good moisture barrier properties, and are suitable for baked
goods, fresh products (e.g., meat, fish, vegetables, fruits), processed meats, cheese,
and candy. Esterified cellulose-based materials are excellent for high moisture foods
as it allows respiration and reduces fogging; (3) Chitin/chitosan-based materials
have clear, tough, flexible, and good oxygen barriers properties but poor long-term
stability and low water vapor barrier properties; (4) Pectin based packaging material
possesses high water vapor permeability, which limits its use in food packaging.
Different protein-based packaging materials also have their specific properties:
(1) Gluten-based materials show good oxygen barrier properties but poor carbon
dioxide (CO2) barrier properties, which makes them possible to act as packaging
materials for specific applications, e.g., mushrooms; (2) Soy protein-based materials
1 Introduction to Food Hydrocolloids 23

are always brittle and show poor water resistance; (3) Films formed by gelatin do not
have good mechanical properties, even reinforced by several techniques, which thus
limits their application as a packaging material; (4) Zein films always showed similar
tensile strength but higher oxygen permeability as compared with gluten films. This
is attributed to an easier diffusion of oxygen across the helical structure of zein as
compared with the highly cross-linked structure of wheat gluten (Cazón et al. 2017).
Furthermore, nanocomposites based on the above-mentioned polysaccharides
and proteins have also been developed as food-packaging materials, including
starch-kaolin, starch-clay, cellulose acetate-clay, pectin-montmorillonite (MMT),
gluten-MTT, gelatin-MMT, and SPI-MMT. These nanocomposites based packaging
materials show improved tensile strength, thermal stability, and water/oxygen barrier
capacity as compared with those produced by sole polysaccharides or proteins (Tang
et al. 2012).

7.5.2 Biomedical Materials

Inorganic particle based biomedical nanomaterials can cause cytotoxicity owing to

their accumulation, aggregation, decomposition, and/or generating reactive oxygen

Fig. 1.8 Biomedical applications of chitosan-based nanogels (Wang et al. 2017). The permission
for reproduction of the figure was obtained from Royal Society of Chemistry
24 W. Lu et al.

species inside the cell. However, natural biopolymers, such as food polysaccharides
and proteins, always have good biocompatibility and biodegradability, and they can
be developed as functional and eco-friendly advanced materials. Food-derived bio-
polymers can be developed into diverse kinds of biomedical materials (Fig. 1.8). The
biopolymers that have been employed to produce these advanced biomedical mate-
rials include chitin/chitosan, cellulose/nanocrystals, hyaluronan, gelatin, silk pro-
teins, and whey proteins.
For example, polysaccharides nanocrystal can be developed into a variety of
biomedical materials, such as electrolytes, nanoscaffolds, nanosponges, various
biomimetic materials, cellular bioimaging materials, permselective membranes,
and templates for the synthesis of inorganic nanoparticles (Lin et al. 2012). Natural
polysaccharide nanocrystals potentially have prolonged retention in the circulation

Fig. 1.9 Several hydrocolloids-based advanced materials (a) adsorption of toxic metal on gum
fibers (right) and gum (left) (Padil et al. 2018). The permission for reproduction of the figure was
obtained from Elsevier. (b) nanocrystalline cellulose-based biomimetic optical nanomaterials with a
peak reflected wavelength of covering the entire visible spectrum (Shopsowitz et al. 2010). The
permission for reproduction of the figure was obtained from Springer Nature. (c) morphology
observation of inorganic nanoparticles prepared using cellulose nanocrystal as the template (Lin
et al. 2012). The permission for reproduction of the figure was obtained from Royal Society of
Chemistry
1 Introduction to Food Hydrocolloids 25

system, rendering them a promising biomedical material that is biodegradable and

safe for the environment and human health.

7.5.3 Template for Synthesizing Inorganic Nanoparticles

Many hydrocolloids, e.g., cellulose nanocrystal (Fig 1.9c), can act as a template or
scaffold of synthesizing inorganic nanoparticles. For example, mesoporous silica
nanoparticles can be successfully fabricated by using rod-like cellulose nanocrystal
as the template (Dujardin et al. 2003; Gruber et al. 2011). The cellulose crystalline
based materials have also been used as the template for producing gold (Au), silver
(Ag), uranium (U), palladium (Pd), platinum (Pt), selenium (Se), Au-Ag, Ag-Pd,
cadmium sulfide (CdS), zinc sulfide (ZnS), lead sulfide (PbS), and titania (TiO2)
nanoparticles. The reduction reaction between the template and metal ions is the
main mechanism for producing these nanoparticles, and the utilization of cellulose
nanocrystal as the template can produce stable and high-density particles with a
controlled size distribution, and significantly improve their chemical properties.

7.5.4 Other Types of Functional Materials

Food hydrocolloids can also be developed into many other types of advanced
materials, such as mesoporous biomimetic optical nanomaterials with a peak
reflected wavelength of covering the whole visible spectrum or even into the near-
infrared by simple modification (Shopsowitz et al. 2010) (Fig 1.9b), bio-inspired
mechanically adaptive nanomaterials used as adaptive substrates of intracortical
microelectrodes (Shanmuganathan et al. 2010), nanostructured membranes with
potential permselective function to differently charged substances (Thielemans
et al. 2009), reinforced polymer electrolytes with high ionic conductivity and good
stability (My Ahmed Said Azizi Samir et al. 2004), and decontamination materials of
removing organic pollutants (Namazi and Dadkhah 2010) or toxic metal (Padil et al.
2018) (Fig 1.9a).

8 The Future Trends

Over the last half a century, great progress in the science and technology of food
hydrocolloids was gained along with the rapid development of the food industry.
Food hydrocolloids carry the flavor, texture, processing, nutrition, and health char-
acteristics of foods and thus play an essential role in the food industry. In view of an
increasing concern of consumers in a healthy way of life, both scientists and
consumers pay more and more attention to the functionality of food products. The
health benefits of food hydrocolloids have been revealed one after another. Never-
theless, the physiological activities of many emerging natural polysaccharides or
26 W. Lu et al.

proteins have not been clarified. Furthermore, food hydrocolloids can potentially
contribute to the future food structure design due to their ability in acting as
structuring agents, e.g., elderly food, special diet food for diabetics, and low-salt
foods, controlled-textured-flavored-foods for dysphagia. Moreover, excellent plas-
ticity and mechanical properties of food hydrocolloids-based advanced materials
make them possible to have promising applications in food, chemical, and biomed-
ical industries, such as controlled or target delivery carriers of bioactive nutrients and
drugs, meat analogue, artificial muscles, artificial joints or cartilage, or electronic
elements. All in all, the food hydrocolloids market continues to see robust growth
and the prospect of the future is worth to be expected.

References

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Chapter 2
Solution Properties

Hongbin Zhang and Ruiqi Li

Abstract Solution properties are important physicochemical properties of food

hydrocolloids. A number of critical molecular and conformation parameters such
as the molecular weight, intrinsic viscosity, radius of gyration, persistence length,
etc. are deduced from dilute solutions. The main functionalities of food hydrocol-
loids, such as viscoelasticity, physical stabilization, emulsification, suspension,
mouthfeel, texture modification, delivery of actives, essentially rely on their solution
properties. Various hydrocolloids have been widely used in aqueous solution to
provide a variety of structures and functionalities to diverse food or non-food
products. This chapter starts a brief survey of the main features of the chemical
constitution and molecular structures of food hydrocolloids, and then an introduction
into the conformation of single chains in solution, using examples for the explana-
tion. After discussing the single chain behavior, the collective properties of poly-
mers, mainly polysaccharides, in bulk are introduced and discussed based on the
relevant dilute and semi-dilute solution theory. The impact of molecular parameters
and chain conformation of polysaccharides on their solution properties is empha-
sized, while several molecular and conformation parameters are summarized. The
utilization of the solution properties of food hydrocolloids is also described. A series
of common polysaccharide liquid crystals are also briefly discussed.

Keywords Polysaccharide · Thermodynamics · Hydrodynamic parameter · Scaling

law · Chain conformation

H. Zhang (*) · R. Li
Advanced Rheology Institute, Department of Polymer Science and Engineering, School of
Chemistry and Chemical Engineering, Frontiers Science Center for Transformative Molecules,
Shanghai, R. P. China
e-mail: hbzhang@sjtu.edu.cn

© Springer Nature Singapore Pte Ltd. 2021 29

Y. Fang et al. (eds.), Food Hydrocolloids,
https://doi.org/10.1007/978-981-16-0320-4_2
30 H. Zhang and R. Li

1 Introduction

Natural polymers such as polysaccharides and proteins have attracted much attention
because of their great importance as functional food hydrocolloids as well as
renewable resources and biodegradable materials, dealing with sustainable develop-
ment and environmental conservation. These biopolymers have been widely used in
aqueous solutions or in hydrogels to provide structure and function, e.g., viscoelas-
ticity, physical stabilization, delivery of actives, emulsification, mouthfeel, texturiz-
ing, to a variety of food or non-food products.
Since many important properties of polymer solutions, such as viscosity, visco-
elasticity, and phase behavior, are mainly dominated by the polymer chain confor-
mation and dimension, for a comprehensive understanding of molecular interactions,
along with for many technical applications, a knowledge of the polymer chain shape
in solution is essentially important. Only by understanding their chain conformation
and solution properties, establishing new characterization methods, and improving
the theory of natural polymer solutions, it will be possible to clarify the relationship
between their performances and structures, and to design more rational molecules
and build functional materials with good performance to achieve added application
value.
It should be noted at first that the structure of biopolymers, especially natural
polysaccharides is far more complicated than the general synthetic polymers, typi-
cally, such as polyolefins (polyethylene, polypropylene, polystyrene, and so on),
which have only very simple repeating units. However, even for the simplest
polysaccharide family of glucans, their repeating unit is glucose residue sterically
connected by several glycosidic linkages, and only the difference in glycosidic
linkages yields a number of different glucans, for instance, starch (amylose, amylo-
pectin) (α-(1!4) linkage in backbone), cellulose (β-(1!4) linkage), curdlan
(β-(1!3) linkage), and so on. These glucans are of varying chemical structures,
and thereby result in various chain conformations in solution, and diverse solution
properties. Second, very different from small molecule solution, polymer solution
has its unique properties due to the high molecular weight of the polymer (Mw), such
as the slow dissolution process and high viscosity. The properties of the polymer
solution are Mw dependent, and the polymer has the character of molecular weight
polydispersity. And third, a polymer solution refers to a homogeneous system
consisting of two or more components dispersed in a solvent in a molecular state.
Polymer dissolution in solvent needs to go through two stages, first swelling, and
then dissolution. The molecular diffusion in the solvent at first causes the movement
of the polymer chain segment. After that, the molecular diffusion of polymer into the
solvent starts, and the mixture becomes a homogeneous system. Due to the huge Mw
of the polymer, the diffusion rate of the polymer is less than that of small molecules.
The crosslinked polymer can only swell and do not dissolve.
Polymer solutions can be divided into dilute, semi-dilute, and concentrated
solutions. The properties of the solutions can vary greatly with the polymer concen-
tration. When the concentrations are small (generally less than 1%), there is no
2 Solution Properties 31

contact between the polymer chains, and the solution viscosities are very low. Many
important physical quantities are measured in dilute solution conditions. In semi-
dilute solutions, the polymer chains contact and may be entangled with each other,
and the solutions have high viscosities. In concentrated solutions, the polymer chains
display entanglement; meanwhile, the viscosities are very high.
This chapter starts a brief survey of the thermodynamics (mainly free energy and
osmotic pressure) of polymer solutions at equilibrium, including the introduction to
the Flory–Huggins mean-field theory and Flory–Krigbaum dilute theory, to intro-
duce some important physical parameters such as the Flory–Huggins interaction
parameter (χ) and the second virial coefficient (A2). Then, the concentration regimes
of polymer solutions, and the viscosity and scaling of some polysaccharides in
solutions are discussed, and some important molecular and conformation parameters
such as polymer Mw and its distribution (Mw/Mn), intrinsic viscosity [η], the mean
square radius of gyration (Rg), persistence length (Lp), and chain stiffness parameter
(B) are summarized. Finally, some polysaccharide liquid crystals are also briefly
introduced.
Here, the relevant measurement methods of the physical parameters are not
emphasized as can be easily found in textbooks. Rheology, an important aspect of
the polymer solution, which is not included in this chapter, will be discussed in
Chap. 3.

2 Thermodynamics of Polymer Solutions

The thermodynamics of polymer solutions has been well described in the literature.
For a polymer, there are good solvents that dissolve the polymer well and
non-solvents that do not dissolve it. A solvent with an intermediate quality can
dissolve the polymer to some extent.
While amorphous polymers (for example, polystyrene and polymeth-
ylmethacrylate) are readily dissolved in the good solvent, semi-crystalline polymers
(for example, polyethylene, polypropylene, cellulose, chitin, and curdlan) are not
easy to dissolve. Polymer chains within a crystallite are in a thermodynamically
stable state. Heat treatment may help their dissolution. Notably, the dissolution of
cellulose and chitin has long been a very difficult problem, and it has traditionally
been mainly dissolved by organic solvents and high-temperature heating (Kim et al.
2013; Kaliannan et al. 2001). Lina Zhang's group broke through the traditional
method of heating organic solvents to dissolve macromolecules, proposing a new
method for dissolving poorly soluble natural polysaccharides such as cellulose,
chitin, and chitosan in low-temperature alkali/urea aqueous solution system, and
revealed its mechanism (Cai and Zhang 2010).
There are some concentration regimes in the polymer solution, i.e., dilute, semi-
dilute, and concentrated solution. Here, the thermodynamics of dilute solutions is
primarily focused. With the increase of the polymer concentration, the thermody-
namics of the polymer solution will deviate from that of the ideal solution.
32 H. Zhang and R. Li

Thermodynamic properties of the polymer solution depend not only on the polymer
itself but also on how good the solvent is for the polymer. The solvent/polymer
interaction and the degree of polymerization dictate the properties. The Flory–
Huggins mean-field theory describes well the features of polymer solutions, which
has been well verified in literature.

2.1 Flory–Huggins Mean-Field Theory

2.1.1 Lattice Chain Model

The miscibility of the polymer with a given solvent can be well explained in the
mean-field theory. Flory applied this theory to polymer solutions. The simplest
lattice chain theory is generally referred to as Flory–Huggins mean-field theory.
For the polymer solution, the mean-field theory compares the free energy of the
polymer/solvent system before and after mixing.
Figure 2.1 shows a two-dimensional version of the lattice model. The system
consists of nsite sites. Each site can be occupied by either a solvent molecule or a
monomer of the polymer. Double occupancy and vacancy are not allowed. A linear
polymer chain occupies N sites on a string of N  1 bonds. Polymer chains, each
consisting of N monomers, are laid onto empty sites one by one until there are a total
of nP chains. Then, the unoccupied sites are filled with solvent molecules. The
polymer volume fraction ϕ is related to nP by nP ¼ nsiteϕ/N and the number of the
solvent molecules nS is thus given by nS ¼ nsite(1  ϕ).
Before mixing, the polymer molecule and the solvent occupy the volume of
nPNνsite and nSνsite, respectively, where νsite is the volume per site. Upon mixing,
the total volume nsiteνsite does not change. The enthalpy of mixing ΔHmix and the

Fig. 2.1 The Lattice model

for polymer solution. Gray
and white sites are occupied
by polymer chains and
solvent molecules,
respectively. Reproduction
with permission from
Teraoka (2002), Copyright
2002 John Wiley and Sons
2 Solution Properties 33

Gibbs free energy change ΔGmix thus are equal to the change in the internal energy
ΔUmix and the Helmholtz free energy change ΔAmix, respectively.

2.1.2 Flory–Huggins Interaction Parameter χ

The entropy of mixing, especially at low concentrations, is small for polymer-

solvent systems. Therefore, upon mixing (i.e., enthalpy of mixing), the change in
the interactions governs the miscibility. The interactions here are typically van der
Walls interactions, dipole–dipole interactions, and hydrogen bonding.
The lattice fluid model only considers the interactions between nearest neighbors.
The interactions reside in the contacts. Mixing the polymer and solvent changes the
overall interaction energy through the rearrangement of contacts. The Flory–Hug-
gins interaction parameter χ is defined as the product of the lattice coordinate Z and
the energy change reduced by kBT:


ðεPP þ εSS Þ
χ ¼ Z εPS  =kB T ð2:1Þ
2

where kB is the Boltzmann constant, εSS, εPP, and εPS are used to denote the
interactions for a solvent–solvent (S–S) contact, a polymer–polymer (P–P) contact,
and a polymer–solvent (P–S) contact, respectively.
A positive χ suggests that the P–S contacts are less favored compared with the P–
P and S–S contacts. A negative χ means that P–S contacts are preferred, promoting
solvation of the polymer. Generally, χ decreases with an increase of T. In a hydrogen
bonding pair, for example, χ usually changes from negative to positive with
increasing T.
The importance of χ parameter stems from being the only way to consider the
specific chemical nature of polymer and solvent. For many systems, χ has been
reported to increase with increasing the polymer concentration and decrease with
temperature (Fig. 2.2). χ value can be obtained commonly by osmotic pressure
measurements (Lei et al. 2016).

2.1.3 Gibbs Free Energy and Osmotic Pressure

Flory obtained the entropy of mixing ΔSmix per site as

ϕ
ΔSmix =ðkB nsite Þ ¼ ln ϕ þ ð1  ϕÞln ð1  ϕÞ ð2:2Þ
N

ΔSmix given by Eq. (2.2) is greater than the entropy of mixing for an ideal solution of
nP solute molecules and nS solvent molecules.
The change ΔUmix in the internal energy per site is
34 H. Zhang and R. Li

Fig. 2.2 Flory–Huggins

interaction parameters
measured at different
temperatures (◓: 50  C; ○:
37.5  C; and ◒: 25  C) for
the mixing systems of H2O
and pullulan. Reproduction
with permission from Eckelt
et al. (2008), Copyright
2008 ACS

ΔU mix =ðnsite kB T Þ ¼ χϕð1  ϕÞ ð2:3Þ

ΔUmix maximizes at ϕ ¼ 1/2. The sign of ΔUmix is the same as that of χ since
ϕ(1  ϕ) > 0 for 0 < ϕ < 1. ΔUmix depends on the interaction through χ. The system
with the same χ has the same ΔUmix. The solution with χ ¼ 0 is called an athermal
solution. In the athermal solution, ΔUmix ¼ ΔHmix ¼ 0 regardless of ϕ.
From Eqs. (2.2) and (2.3), the Helmholtz free energy of mixing,
ΔAmix ¼ ΔUmix  TΔSmix, per site is given as

ΔAmix ϕ
¼ ln ϕ þ ð1  ϕÞln ð1  ϕÞ þ χϕð1  ϕÞ ð2:4Þ
nsite kB T N

The osmotic pressure Π is given as

Πνsite ΠV ϕ
¼ ¼  ln ð1  ϕÞ  ϕ  χϕ2 ð2:5Þ
kB T nsite kB T N

where V ¼ νsite nsite is the volume of the solution.

The chain connectivity, rigidity, and the shape of the monomer are neglected in
the Flory–Huggins theory. Modifications to this theory are possible by taking these
effects into account.

2.2 Flory–Krigbaum Dilute Theory

For dilute solutions, when ϕ « 1,taking into account ln ð1  ϕÞ ¼ ϕ  12 ϕ2 

3 ϕ  ⋯, then Eq. (2.5) is rewritten to
1 3
2 Solution Properties 35

 
ΠV ϕ 1 1
¼ þ  χ ϕ2 þ ϕ3 ⋯ ð2:6Þ
nsite kB T N 2 3

In the low concentration limit, Eq. (2.6) gives the osmotic pressure Π ideal of the
ideal solution:

nsite ϕ
Π ideal ¼ k T ð2:7Þ
NV B

Thus, the ratio of Π to Π ideal compared at the same concentration is

h  i
Π 1 1
¼1þN  χ ϕ2 þ ϕ3 ⋯ ð2:8Þ
Π ideal 2 3

To compare the theory with experiments, Π is expressed in terms of mass

concentration c. Using the identity c ¼ NMA N νϕsite , where M/(NAN ) is the mass of the
monomer, with NA being the Avogadro’s number (the number of molecules in a
mole of a substance, ca. 6.02  1023/mol).
Π/(NAkBT ) is generally expanded in a power series of c:

Π c
¼ þ A2 c2 þ A3 c3 þ ⋯ ð2:9Þ
N A kB T M

In this virial expansion, A2 is the (osmotic) second virial coefficient. The meaning
of A2 will become clearer when expressing it in the lattice model by comparing
Eqs. (2.8) and (2.9):
   2
1 N
A2 ¼  χ N A νsite ð2:10Þ
2 M

The above-mentioned Flory–Huggins interaction parameter χ and the second

virial coefficient A2 are important parameters representing the interaction between
the polymer and the solvent molecules. A positive A2 (polymer
 in goodsolvent)
Π ideal
deviates χ upward compared to that of the ideal solution ðN A k B TÞ ¼ c=M . When
A2 ¼ 0, the solution in a wide range of concentrations is close to the ideal solution
(the solvent is called as θ solvent). When the solvent quality decreases, χ increases.
For χ > 1/2, A2 is negative. In poor solvent, the polymer chain becomes compacted,
the solvent becomes unable to dissolve the polymer. According to Eq. (2.10),
Table 2.1 summarizes the relationship between A2 and χ. A2 is a measure of the
nonideality of the polymer solution, also indicative of solvent–polymer interaction.
A2 ¼ 0 when the entropy of mixing compensates repulsive polymer–solvent inter-
actions or attractive polymer–polymer interactions. For a given polymer-solvent
system, the light scattering experiment at different concentrations of the polymer
gives an estimate of A2.
36 H. Zhang and R. Li

Table 2.1 Relationship between A2 and χ for polymer in solvent. Reproduction with permission
from Teraoka (2002), Copyright 2002 John Wiley and Sons
A2 χ Solvent quality Chain conformation
+ <1/2 Good Extended
0 ¼1/2 θ
 >1/2 Poor Compacted

Table 2.2 Distinction of the freely jointed chain, freely rotating chain, and real chain. Reproduc-
tion with permission from Teraoka (2002), Copyright 2002 John Wiley and Sons
Model Bond length Bond angle Torsion angle
Freely jointed chain Fixed Free Free
Freely rotating chain Fixed Fixed Free
Real chain Fixed Fixed restricted

3 Chain Model (Teraoka 2002; Rubinstein and Colby 2003;

Strobl 1997)

A chain with no interactions between monomers that are separated by a sufficient

number of bonds along the chain is called an ideal chain. Real chains interact with
both their solvent and themselves. In a real polymer chain, the effect that two
monomers cannot occupy the same space is called an excluded volume effect, and
the excluded volume plays a more important role in polymer solutions than it does in
small molecule solutions. According to the lattice model, an ideal chain has no
excluded volume allowing an overlap of the monomers. A real chain is a regular
chain with an excluded volume. Although the ideal chain does not exist in reality, the
model allows solving many polymer solution problems in a mathematically rigorous
way extensively. In the ideal situation, the real chain behaves like an ideal chain, like
in a dilute solution with a special kind of solvent called a theta solvent.
There are several classical models to describe ideal chains, such as freely jointed
chain model, freely rotating chain model, restricted rotation chain model, equivalent
freely jointed chain (Gaussian chain) model, and wormlike chain model. Each model
makes different assumptions about the allowed values of torsion and bond angles.
However, every model ignores interactions between monomers separated by a large
distance along the chain. One of the simplest models of ideal chains is the freely
jointed chain model with a constant bond length and no correlations between the
directions of different bond vectors, while freely rotating chain model assumes all
bond lengths and bond angles are fixed, and all torsion angles are equally likely and
independent of each other. The distinction of freely jointed chain, freely rotating
chain, and a real chain is described in Table 2.2. A simple unified description of all
ideal polymers is provided by the equivalent freely jointed chain model (Kuhn
model). The equivalent freely jointed chain has NK freely jointed effective bonds
of length LK. The effective bonds constitute a Kuhn segment, and the length LK is
called the Kuhn segment length, characterizing the stiffness of a given polymer
2 Solution Properties 37

Fig. 2.3 Representation of

a random coil (a) as a real
chain; the circles represent
the monomer units which
are linked by the bonds
(short thin lines); (b) in the
Kuhn model (long thin
lines), the long lines indicate
the Kuhn segments; (c) as a
wormlike chain (long
smooth solid line).
Reproduction with
permission from Denkinger
and Burchard (1991),
Copyright 1991 John Wiley
and Sons

chain, NK is the number of Kuhn segments per chain. The Kuhn model is shown in
Fig. 2.3.
As far as the wormlike chain model is concerned, it is a freely rotating chain with
a fixed bond angle but an unrestricted torsion angle, and the chain conformation is
not zigzag but a smooth curve in a three-dimensional space, as shown in Fig. 2.3. A
wormlike chain is specified by the persistence length Lp (in the wormlike chain
model, Lp is equal to the average projection length along the initial direction of the
polymer chain) and the contour length Lc (the length of the fully extended chain).
When Lp  Lc, the chain is either sufficiently short or rigid; in the limit of Lc/Lp!0,
the wormlike chain is a rod; for Lc  Lp, the chain is sufficiently long or flexible; in
the limit of Lc!1, the wormlike chain is the same as the ideal chain. In addition, the
physical feature of the wormlike chain is that its Kuhn length LK is twice the
persistence length Lp.

4 Concentration Regimes: Dilute, Semi-dilute,

and Concentrated

Polymer solutions are broadly classified into three regimes: dilute, semi-dilute, and
concentrated (de Gennes 1980; Graessley 1980). Further, semi-dilute and concen-
trated solutions may be entangled or nonentangled. The polymer chains in the
solution can be regarded as spherical coils. Figure 2.4 depicts three ranges of
polymer concentration c: c « c, c ffi c, and c » c, where c is the so-called overlap
concentration of the polymer.
38 H. Zhang and R. Li

Fig. 2.4 Concentration regimes for solutions of linear flexible polymers: dilute solution, c « c;
solution at the overlap concentration c ffi c; semi-dilute solution, c » c. Reproduction with
permission from de Gennes (1980), Copyright 1980 Cornell University Press

When c is below c, the solution is called dilute. At c « c, the coils are separated
from each other, behaving more or less independently (Fig. 2.4a). The chains interact
primarily with solvent molecules. Thus, the solution is close to an ideal solution.
As the concentration c increases, the coils become congested and eventually
contact each other. At c, the whole volume of the solution is filled with these
spheres (Fig. 2.4b). At c » c, chains are overlapped and entangled. In comparison
with the chains in dilute solutions, their mobility is greatly reduced. The solution in
this regime is called semi-dilute. The thermodynamic properties of the semi-dilute
solutions are very different from those of an ideal solution extrapolated to the same
concentration. In semi-dilute solutions, polymer chains have many other chains
overlapping them though monomers are not congested. As a whole, the chains are
congested, and the interactions between the chains are therefore strong. With a
further increase in c, the overlaps become more serious.
The existence of the semi-dilute regime is characteristic of polymer solutions. The
properties of semi-dilute solutions are different significantly from those of dilute
solutions. With a tenfold increase in the concentration, the Π can easily increase by a
factor of several hundred, whereas in the ideal solution, the osmotic pressure is
proportional to c. Furthermore, in semi-dilute solutions, the overall chain motion is
slow because of the entanglement of the chains. A semi-dilute solution of a high Mw
polymer can barely flow due to its high viscosity. The Π and the time scale of motion
depend heavily on concentration and Mw of the polymer.
The upper limit of the semi-dilute range is sometimes denoted by c. Above c,
the monomers are congested, and sometimes the solution is called concentrated, in
which each segment of the polymer chain does not have sufficient available space.
Typically, the polymer volume fraction at c is between 0.2 and 0.3. For a polymer
with a sufficiently high Mw, there is a broad range of concentrations between c and
c. The semi-dilute regime is often specified by c  c < c.
2 Solution Properties 39

4.1 Relationship Between Overlap Concentration c

and Chain Dimension

In a dilute solution where chains do not overlap, the whole single chain can be
regarded as roughly being confined in a sphere of diameter RF, the root-mean-square
end-to-end distance (RF) of the chain, which follows

RF ’ LK N K ν ð2:11Þ

where ν is an exponent that depends on solvent quality. ν is 0.5 for neutral polymers
in θ solvent, 0.588 for neutral polymers in good solvent and polyelectrolytes with
added salt, and 1 for polyelectrolytes in salt-free solutions (Lopez et al. 2017; Colby
2010).
The overlap concentration c is given by

c ’ ðM w =M 0 Þ=RF 3 ðM w =M 0 Þ13v ð2:12Þ

where M0 is the molar mass of a monomer. The exponent (1-3ν) is 1/2 (for ν ¼ 0.5)
for an ideal chain and -4/5 (i.e., 0.77 for ν ¼ 0.59) for a real chain.
Alternatively, c can be expressed as

c ’ ðM w =N A Þ=Rg 3 ð2:13Þ

where Mw/NA is the mass of each chain, with NA being the Avogadro’s number, Rg is
the chain dimension, i.e., the root-mean-square radius of gyration (the radius of the
sphere in which the chain roughly occupies its space). For an ideal chain, the ratio of
RF to Rg is 60.5’2.45.
Note that different from RF that is defined for linear chains only, Rg is defined for
any chain architecture, including nonlinear chains such as branched chains. Thus,
Eq. (2.14) gives a universal measure for c.
For rigid-chain polymers in solution, the overlap concentration c is given by

c ’ ðM=N A Þ=Lc 3 ð2:14Þ

where Lc is the contour length of a thin rod-like polymer chain. c of a rod chain is
much lower as compared with a flexible linear chain.
Experimentally, c can also be determined as the reciprocal of another character-
istic parameter of a polymer, the intrinsic viscosity [η] as will be discussed in the
following.
40 H. Zhang and R. Li

4.2 Molecular Weight Mw and Molecular Weight Distribution

Mw/Mn

Compared with low molecular weight compounds, besides the difference of very
high molecular weight of the polymer, another significant character of polymer is
that nearly all of the polymer is a molecule mixture with a different degree of
polymerization. This polydispersity is present, especially in plant polysaccharides.
Polymers are usually polydisperse and have distribution in molecular weight. Some
representative values are used as a typical molecular weight of the polymer, i.e.,
different average molecular weights (Lapasin et al. 1992).
Let the polymer consists of ni chains of exact molecular weight Mi, in which the
ith component has a degree of polymerization i. The difference between Mi and
Mi + 1 is the mass of the repeating unit. The number-average molecular weight Mn is
defined as
X X
Mn ¼ nM=
i i i
n
i i
ð2:15Þ

The weight-average molecular weight Mw is defined as

X X
Mw ¼ nM
i i i
2
= nM
i i i
ð2:16Þ

Also used is the z-average molecular weight Mz further weighted with Mi:
X X
Mz ¼ nM
i i i
3
= nM
i i i
2
ð2:17Þ

The viscosity-average molecular weight Mη is defined as

P 1
 ni M i α α
i
Mη ¼ P ð2:18Þ
ni M i
i

where α is the exponent of the Mark–Houwink equation. Generally, α is in the range

of 0.5–1.0, thus Mn < Mη < Mw, and Mη is closer to Mw.
For a perfectly monodisperse polymer, Mn ¼ Mw ¼ Mz. Otherwise, Mn < Mw < Mz.
The ratio of Mw to Mn is used to express the polydisperse of the polymer. The ratio,
often abbreviated as Mw/Mn, is called polydispersity index:

X X X 2
M w =M n ¼ n
i i
nM =
i i i
2
nM
i i i
ð2:19Þ

Unless the polymer is monodisperse, Mw/Mn >1. A sample with a greater Mw/Mn
has a broader molecular weight distribution.
2 Solution Properties 41

One of the most important experimental techniques for determining the dilute
solution behavior of polysaccharides is light scattering. Light scattering has been
often used to characterize polymer chains in a solution, by which Mw and Mw/Mn as
well as Rg, and A2 can be obtained, and furthermore, the information on the
conformation of the polymer chain—whether it is spherical, random-coiled, or
rod-like, can be recognized.
Size exclusion chromatography (SEC) coupled with multiangle laser light scat-
tering (MALLS) has been widely used to characterize the molecular weight and
molecular weight distribution of a polymer. Although SEC uses a flow system, the
separation principle and the analysis are based on a static property of the polymer
chains in solution. SEC sometimes is also called gel permeation chromatography
(GPC) for organic solvent as mobile phase, or gel filtration chromatography (GFC)
or aqueous GPC for aqueous mobile phase. GPC is, by now, one of the most routine
and effective techniques for characterizing the polydispersity of polymeric materials.
Some other techniques include the concentration gradient multiangle laser light
scattering (CG-MALLS), asymmetric flow field flow fractionation-multiangle laser
light scattering (AFlFFF-MALLS), and horizontal agarose gel electrophoresis
(AGE) (Zhang et al. 2015; Striegel and Timpa 1996).

4.3 Intrinsic Viscosity [η]

As it is known, one of the most salient features of macromolecules is the intense

increment in viscosity that they produce when, even in minute amounts, they are
dissolved in ordinary solvents. The viscosity intensifying effect, characterized by the
intrinsic viscosity [η], is extensively used for analysis or characterization of synthetic
polymers, biomacromolecules, nanoparticles, and colloids. The [η] is a quantitative
characteristic of a polymer, representing an increase in the solution viscosity when
the polymer concentration is raised to some level. As expected, a polymer chain with
a greater dimension has a larger [η]. In the Zimm model, it is assumed that as the
polymer moves, it drags the solvent inside its pervaded volume with it (Rubinstein
and Colby 2003). The Zimm model has the correct physics for [η], which provides
information about fundamental properties of the solute and its interaction with the
solvent, and can be precisely related to the conformation of either linear or nonlinear
flexible chains, wormlike macromolecules and micelles, and rigid particles of
arbitrary shape.
The increment in the solution viscosity, η, with respect to that of the pure solvent,
η0, the relative viscosity, is the ratio ηr ¼ η/η0. The specific viscosity of a solution of
concentration c is defined as ηsp ¼ ðηη 0Þ
η0 ¼ ηr  1 . The intrinsic viscosity [η] is
defined as:
42 H. Zhang and R. Li

Fig. 2.5 The specific viscosity ηsp (filled symbols) and the logarithm of relative viscosity lnln ηr
over concentration c (open symbols) plotted and linearly fitted based on Huggins equation and
Kraemer equation, respectively, vs. polymer concentration of hyaluronic acid in 0.1M NaCl at
25  C

ηsp ηr  1
½η ¼ ¼ ð2:20Þ
c c

The [η] value can be obtained as the intercept in a linear least-squares fit of the
reduced specific viscosity, ηsp/c, based on the Huggins equation. Alternatively, it can
be obtained by linear extrapolation of the inherent viscosity, (lnln ηr)/c to zero
concentration, according to the Kraemer equation (Goh et al. 2006a, b).

ηsp
¼ ½η þ K 0 ½η 2 c Huggins equation ð2:21Þ
c
ðln ηr Þ=c ¼ ½η þ K 00 ½η 2 c Kraemer equation ð2:22Þ

where K0 and K00 are the Huggins and Kraemer constants, respectively. For a polymer
in good solvents, K0 value is within 0.3–0.4, and in θ solvents its value is 0.5–0.8;
and if aggregation of the polymers occurs, K0 will be above 0.8. A relationship holds
between the dimensionless constants K0 and K00, for a random coil polymer,
K0  K00 ¼ 0.5.
A classical procedure for the determination of [η] consists of the measurement of
solution viscosities at a series of concentrations, followed by extrapolation of ηsp/c to
zero concentration. The measurements of [η] are usually made with Ubbelohde glass
capillary viscometers that allow in situ dilution. It is often to display the [η]
determination in a dual Huggins–Kraemer plot, as illustrated in Fig. 2.5.
2 Solution Properties 43

Table 2.3 Molecular param- Polysaccharide Mw  105 [η] (dL/g)

eters of hyaluronic acid
CFG 3.33 1.48
(HA) with different molecular
weights and corn fiber gum HA-1 3.56 8.78
(CFG) HA-2 12.9 12.89
HA-3 22.3 33.98

Table 2.4 Molecular param- Polysaccharide Mw  105 [η] (dL/g)

eters of some branched
CFG 2.0 1.12
polysaccharides
SSPS 8.5 0.34
GA 10.2 0.18
OSA-s 144 0.39

Fig. 2.6 Comparison of [η] (left) and Rg (right) of HA measured by different characterization
techniques as a function of molecular weight. Mark–Houwink relationship for Mw is included as a
reference in dashed line using published K and α parameters (K ¼ 0.0336 ml/g and α ¼ 0.79) (Milas
et al. 2001) (left figure). The dashed line (right figure) represents a linear fit of the data points. The
solvent is 0.1 M NaCl solution. Reproduction with permission from Luan et al. (2011), Copyright
2011 Elsevier

The reciprocal of the intrinsic viscosity 1/[η] can be used to represent the overlap
concentration cof a given polymer.
The intrinsic viscosity [η] is a characteristic property of polysaccharide solutions.
Table 2.3 compares the different molecular features of hyaluronic acid (HA) and
corn fiber gum (CFG) with different Mw. Since HA macromolecules possess a linear
chain structure, the [η] values of their solutions are high and increase significantly
with Mw, for instance, from 8.78 dL/g for Mw ¼ 3.56  105 (HA-1) to 33.98 dL/g for
Mw ¼ 2.23  106 (HA-3). Furthermore, when comparable Mw is considered, the [η]
of HA-1 (8.78 dL/g) is much bigger than that of the highly branched CFG macro-
molecules (1.48 dL/g) (Zhang et al. 2015; Luan et al. 2011). In comparison with a
linear chain of HA, like CFG, the [η] values of some other branched polysaccharides
such as gum arabic (GA), octenyl succinate anhydride-modified starch (OSA-s), and
soluble soybean polysaccharides (SSPS) that have densely branched structures are
also very small, although their Mw can be quite high, e.g., Mw ¼ 1.02  106 for GA
and even Mw ¼ 1.44  107 for OSA-s (Table 2.4) (Jin et al. 2017).
44 H. Zhang and R. Li

Table 2.5 Polydispersity Sample SEC-MALLS AFlFFF-MALLS AGE

index Mw/Mn of several
HA-1 1.2 1.2 1.2
hyaluronic acid (HA) samples
measured by different HA-2 1.1 1.1 1.8
techniques HA-3 1.5 1.5 1.4
HA-4 1.0 1.1 1.9

A scaling law of [η] with Mw known as the Mark–Houwink equation has been
established for a number of polysaccharides, which will be discussed in the
following.
Figure 2.6 and Table 2.5 display an example of measured results of hyaluronic
acid by using different abovementioned characterization techniques (Goh et al.
2006a). This work compares the application of different techniques that provide
consistent results on the conformational properties of HA in solutions (Luan et al.
2011).
The [η] is determined by dilute solution viscometry. When light scattering
techniques and viscometry measurements are combined, Mw and Mw/Mn, Rg, A2,
and [η] can be determined, which provides a basis for understanding the solution
properties of polysaccharides.

4.4 Scaling of Viscosity

The scaling of various properties such as viscosity with polymer concentration as

well as molecular weight governs much of the practical use of polymer in industry. A
power-law type correlation between the zero-shear specific viscosity ηsp, 0 and the
polysaccharide concentration c (Eq. 3.13) with different values of b in the two
regimes can be applied (Lapasin et al. 1992). The use of zero-shear specific viscosity
ηsp, 0 instead of zero-shear viscosity η0 removes the contribution of solvent viscosity.

ηsp,0 ¼ acb ð2:23Þ

For polymers with different primary structures but similar conformational char-
acteristics, the range of b values in both regimes is relatively narrow. For the dilute
regime, the exponent b is 1.0–1.5, and for the semi-dilute regime b is 3.5–4.5. Many
polysaccharides such as hyaluronic acid, guar gum, carboxymethyl cellulose,
carboxymethyl amylose, and λ-carrageenan can be fitted to this form of the master
curve (Lapasin et al. 1992; Morris et al. 1981; Ren et al. 2003). Table 2.6 shows the
exponent b values in the dilute and concentrated regimes. For most polysaccharides
with the random coil conformation in concentrated solutions, typically, ηsp, 0 ~ c3.3,
similar to the theoretical prediction of ηsp, 0~c3.75 by de Gennes (de Gennes 1980).
This strong dependence of ηsp, 0 on c reflects the dominant role played by the
intermolecular interactions.
2 Solution Properties 45

Table 2.6 Values of the exponent b in the dilute and concentrated regimes, and of c[η] for some
polysaccharides in aqueous solution at 25  C
b b
Polymer (dil.) (conc.) c[η] Notes Ref.
Hyaluronic acid 1.4 3.3 4 0.015 M NaCl, Morris et al. (1980, 1981)
pH ¼ 7
1.4 3.9 2.5 0.15 M NaCl, Morris et al. (1980, 1981)
pH ¼ 2.5
1.0 3.5 – Salt-free Yu et al. (2014)
1.5 4.2 – 0.15 M NaCl Yu et al. (2014)
Alginate 1.4 3.3 4 0.2 M NaCl, Morris et al. (1981)
pH ¼ 7
1.1 2.0 1.5 0.1 M NaCl Krause et al. (2001)
1.0 4.15 6 0.1 M NaCl Milas (1996)
Xanthan 1.1 4.0 3.2 0.1 M NaCl Gravanis et al. (1987)
1 0.1 M NaCl Esquenet and Buhler (2002),
Brunchi et al. (2014)
1.4 3.3 1.4 1 M KCl Stokke et al. (1992)
Cellulose 0.7 2.2 1.4 [amim]Cl Kuang et al. (2008)
Carboxymethyl 1.3 3.9 – 0.1 M NaCl, Ellis and Ring (1985)
cellulose pH ¼ 7
0.7– 3.5 1 Water ~ 2 M Lopez et al. (2017)
1.3 NaCl
Amylose 1.05 1.9 1 H2O (65  C) Ellis and Ring (1985)
Carboxymethyl 1.4 3.3 4 0.5 M NaCl, Morris et al. (1981)
amylose pH ¼ 7
L-carrageenan 1.25 3.2 3.0 0.75 M NaCl Pereira et al. (1982)
Guar gum 1.3 5.1 3.3 0.1 M urea Robinson et al. (1982)
Pectin 1.2 3.3 – 0.1 M NaCl Axelos et al. (1989)
Scleroglucan 1.4 3.3 1.4 1 M KCl Stokke et al. (1992)
Schizophyllan 1.5 4.3 1.2 DMSO, Fang et al. (2010)
Mw ¼ 8  104
1.6 2.8 4.3 DMSO, Fang et al. (2010)
Mw ¼ 14.8  104
Xyloglucan 1.3 4.0 1 H2O Wang et al. (1997)
Colanic acid 1 3.6 – 0.2 M NaCl Ren et al. (2003)
or 0.2 M CaCl2

It is customary to refer to the generalized representation as suggested by Morris

et al. (1981), consisting of a master curve of reference coordinates ηsp, 0 and c[η], of
which an example is shown in Fig. 2.7 for a series of polysaccharides. Moreover, the
ηsp, 0~c[η] plot associates the transition from a dilute to a concentrated regime to the
coil overlap parameter c[η]. Thereby the approximate value of c can be obtained
from [η] data. Experimental c[η] values for many polysaccharides such as
hyaluronic acid, alginate, guar gum, carboxymethyl amylose, λ-carrageenan, and
so on fall in the range 2.5–4, or even a wider range 1–6 (see Table 2.6), much higher
46 H. Zhang and R. Li

Fig. 2.7 Zero-shear specific viscosity as a function of the coil overlap parameter, c[η], for a range
of random coil polysaccharides in salt solution at pH ¼ 7. (open circle) dextran; (filled circle)
carboxymethyl amylose (0.5M NaCl); (open triangle) high mannuronate alginate (0.2M NaCl);
(filled triangle) high guluronate alginate (0.2M NaCl); (open square) λ-carrageenan (0.075M KCl);
and (filled square) hyaluronic acid (0.015M NaCl). Reproduction with permission from Morris et al.
(1981), Copyright 1981 Elsevier

than those calculated theoretically from the Simha critical condition of coil
overlapping (Weissberg et al. 1951; Simha and Utracki 1975).
The addition of salt can strongly influence the correlation between ηsp, 0 and the
polyelectrolyte polysaccharide concentration c. Figure 2.8 shows the specific vis-
cosity of semi-flexible polyelectrolytes of CMC (Mw ¼ 3.2  105, DS ’ 1.2) (Lopez
et al. 2017) and hyaluronic acid (Mw ¼ 1.8  106) (Yu et al. 2014) solutions as a
function of polymer and added salt concentration, respectively. As seen in Fig. 2.8
(left), the solution viscosities of CMC decrease with increasing the salt concentra-
tion, the effect being more pronounced at lower CMC concentrations. In diluted
regime, the scaling relationship between specific viscosity and CMC concentration
changes from ηsp~c1.3 for csalt¼2 M to ηsp~c0.68 for the salt-free solution. However, a
similar scaling relationship of ηsp c3.5 is observed at high CMC concentration. In
2 Solution Properties 47

Fig. 2.8 Double-logarithmic plot of specific zero-shear viscosity versus concentration of CMC
(left). (Reproduction with permission from Lopez et al. (2017), Copyright 2017 ACS) and
hyaluronic acid solution (right) (Reproduction with permission from Yu et al. (2014), Copyright
2014 Elsevier)

addition, the scaling of entanglement concentration in 0.01M and 0.1M NaCl shows
a behavior of neutral polymer in good solvent, characteristic of highly screened
polyelectrolyte solutions. For HA, salt-free HA solutions present a scaling relation-
ship as ηsp~c1.0 (in dilute concentration region) and ηsp~c3.5 (in semi-dilute concen-
tration region); for saline HA solutions (0.15M NaCl), the scaling exponents are 1.5
and 4.2, corresponding to dilute and semi-dilute concentration region, respectively
(Fig. 2.8, right plot) (Yu et al. 2014).

5 Chain Conformational Analysis of Polysaccharides

in Solution
5.1 Hydrodynamic Radius Rh, Radius of Gyration Rg,
and Shape-Factor ρ

The size of polysaccharides in solution can be measured by a number of techniques,

typically, the dynamic and static light scattering method. The root mean square
radius of gyration Rg, obtained by static scattering experiments, serves as a practical
overall parameter for coil dimensions. The effective hydrodynamic radius Rh of a
macromolecule represents another useful quantity for polymer dimensions.
Based on the dynamic light scattering method, Rh can be calculated using the
Einstein–Stokes equation:

kB T
Rh ¼ ð2:24Þ
6πηD

where D is the diffusion coefficient, kB, T, and η are the Boltzmann constant, the
absolute temperature, and the solvent viscosity, respectively.
48 H. Zhang and R. Li

Since the definitions of Rg and Rh are different, they do not normally coincide
with a given polymer chain, although every single dimension can indicate the size of
the coil. It has been recognized that chains cannot be properly described by just one
of these radii. A combination of both quantities, the shape-factor or asymmetry-
factor ρ, enables detailed insight into the chain architecture:

Rg
ρ¼ ð2:25Þ
Rh

For rigid chains ρ 2, for flexible random coils ρ is 1.5–1.8 (Kok and Rudin
1981; Harding et al. 1992) in a good solvent and 1.3 in a θ solvent (Kok and Rudin
1981), and for compact spheres ρ is about 0.775 (Brant 1981). For instance, the ρ
value of a Na+-type deacetylated gellan gum in NaCl solution at 25  C, existing as
rigid double-stranded chains, is ca. 3, whereas it is ca. 2 at 40  C at which gellan gum
molecules take the conformation of single-stranded chains (Takahashi et al. 2004;
Takahashi 1999). For high acyl gellan gum in DMSO solution, ρ value is 1.67,
indicating a flexible structure of the chain (Kang et al. 2017).
Different from the hydrodynamic radius Rh defined from the diffusion coefficient
D, another hydrodynamics-related radius, the viscometric radius Rη determined from
the intrinsic viscosity (Flory 1953), is also an indicator of the coil size.
 1=3
3½η M
Rη ¼ ð2:26Þ
10πN

In comparison with Rh, theory predicts Rη/Rh values of ca. 1.2 (Lewis et al. 1991;
Oono 1983; Pyun and Fixman 1964; Chong and Fixman 1965). For hyperbranched
chains, experimentally Rη slightly smaller than Rh and their ratio Rη/Rh roughly
remaining 0.95 have been reported (Li et al. 2013).

5.2 Scaling of [η], Rg, and A2 with Mw

5.2.1 Mark–Houwink Equation

Intrinsic viscosity [η] is a measure of the capacity of a polymer molecule to enhance

the viscosity. It depends on the size and the shape of the polymer molecule.
Experimentally, the intrinsic viscosity [η] of a polymer in solution is related to
molecular weight M by the Mark–Houwink equation

½η ¼ KM α ð2:27Þ

where K is a constant of the unit of L/g, and α is called a Mark–Houwink exponent.

Figure 2.9 is an example of the molecular weight dependence of [η] for amylose in
DMSO.
It is known that the value of parameters K and is different from polymer to
polymer, related to the shape and chain structure of the macromolecule, and can
2 Solution Properties 49

Fig. 2.9 Molecular weight dependence of [η] for amylose in DMSO reported in the literature.
Reproduction with permission from Norisuye (1996). Copyright 1996 Elsevier

Table 2.7 Mark–Houwink Chain conformation Solvent α

exponents
Linear flexible θ solvent 0.5
Linear flexible Good solvent 0.7–0.8
Rigid – >1

depend on the nature of solvent as well. In ordinary good solvents, the constants
K and α obtained are valid only within a rather limited range of M.
The polymer conformation parameter α decreases with increasing molecular
compactness. Apparently, α is greater for a more extended conformation. The values
of α are 0–0.5, 0.5–0.8, and 0.8–1.8 for spherical, random coil, and rod conforma-
tions, respectively (Burchard 1996). Generally, α is in the range of 0.5–0.8 for linear
flexible polymer in good solvent (Miyaki et al. 1980; Lewis et al. 1991; Kniewske
and Kulicke 1983). When the polymer chain is more extend, or presents as semi-stiff
conformation, the α value will be above 0.8 and even beyond. In the θ solvent, the
flexible chain has α ¼ 0.5. Note that the apparent α exponent for hyperbranched
polymers in a good solvent can be less than 0.5 (Lu et al. 2013). Values of α are listed
in Table 2.7 for some typical shapes and conformations of the polysaccharides.
The Mark–Houwink equation has been established as summarized in the follow-
ing Table 2.8 for the given polysaccharide of hyaluronic acid and Table 2.9 for many
other polysaccharides.
50 H. Zhang and R. Li

Table 2.8 The reported K and α of the Mark–Houwink equation for hyaluronic acid in literature
K (mL/ T
Mw  104 g) α Solvent ( C) Ref.
>10 0.0279 0.763 0.1 M HCl 25 Cleland and Wang (1970)
0.0228 0.816 0.2 M NaCl 25
0.0318 0.777 0.5 M NaCl 25
7.70–170 0.036 0.78 0.2 M NaCl 25 Laurent et al. (1960)
80–220 0.033 0.79 0.1 M NaCl 25 Milas et al. (2001)
1–100 0.033 0.79 0.1 M NaCl 25 Luan et al. (2011)
10–100 0.057 0.75 0.15 M NaCl 25 Terbojevich et al. (1986)
<10 0.0013 1.056 0.15 M NaCl 37 Mendichi et al. (2003)
10–100 0.0339 0.778 0.15 M NaCl 37
>100 0.395 0.604 0.15 M NaCl 37
10–300 0.029 0.8 0.15 M NaCl 25 Li et al. (1997)
<100 0.00346 0.779 0.15 M NaCl 25 Bothner et al. (1988)
>100 0.0397 0.601 0.15 M NaCl 25
– 0.029 0.8 0.15 M NaCl 25 Wedlock et al. (1983)
– 0.039 0.77 0.15 M NaCl 30 Ueno et al. (1988)
180–1250 (hylan 0.033 0.77 0.15 M NaCl 25 Al-assaf et al. (1995)
form)
40–200 0.0508 0.716 0.2 M NaCl 20 Gura et al. (1998)
40–270 0.0199 0.829 0.2 M NaCl 25 Yanaki and Yamaguchi
(1994)
42–140 0.0278 0.78 0.1 M NaNO3 25 Soltes et al. (2002)
1–7.2 0.0003 1.2 0.2 M PBS pH 37 Shimada and Matsumura
7.3 (1975)
31–150 0.057 0.76 0.2 M PBS pH 37
7.3

5.2.2 Scaling of Rg with Mw

Table 2.10 shows the molecular weight dependence of Rg of hyaluronic acid in 0.1M
NaCl and a comparison of the magnitude of Rg for this linear polymer and the
branched polysaccharide of CFG with a similar Mw. Similar to the impact of Mw on
the [η] (Table 2.4), the Rg values of the linear hyaluronic acid increase significantly
with Mw, from 81 nm for Mw ¼ 3.56  105 (HA-1) to 167 nm for Mw ¼ 2.23  106
(HA-3), and for a comparable Mw, the Rg of HA-1 (81 nm) is much bigger than that
of the high branched CFG macromolecules (24 nm) (Zhang et al. 2015).
Rg can also be related to Mw by the relation:
0
Rg ¼ kM αw ð2:28Þ

where k and α0 are constants for a given polymer at a given temperature in a given
solvent. Usually, α0 of ~0.7 indicates a relatively stiff rod-like conformation, ~0.6 for
2 Solution Properties 51

Table 2.9 The parameters of the Mark–Houwink equation for various polysaccharides in different
solutions at different temperatures
K
104
T Mw  (mL/
Polysaccharide Solvent ( C) 10-4 g) α Ref.
Amylose 1 M KOH 25 22–310 118 0.89 Cowie (1960)
DMSO 25 22–310 125 0.87
EDA 25 22–310 1550 0.70
Cellulose DMAc-LiCl 30 12.5–70 1.278 1.19 McCormick
et al. (1985b)
EMIMAc 0– 3–100 – 0.4– Gericke et al.
100 0.6 (2009)
Carboxymethyl cellulose 0.2 M NaCl 30 0.21– 43.0 0.74 da Silva et al.
0.52 (2018)
0.01 M NaCl 25 21–229 87.76 0.93 Clasen and
Kulicke
(2001)
Methyl cellulose 0.1 M NaNO3 25 – 800 0.92 Schittenhelm
Sulfoethyl cellulose 0.1 M NaNO3 25 56 1.74 1.19 and Kulicke
Carboxymethylsulfoethyl 0.1 M NaNO3 25 135.5 65.8 0.91 (2000)
cellulose
Hydroxyethylsulfoethyl 0.1 M NaNO3 25 29.4– 230 0.80
cellulose 30.3
Hydroxyethyl cellulose 0.1 M NaNO3 25 – 410 0.73
Hydroxypropyl cellulose 0.1 M NaNO3 25 – 420 0.68
Hydroxyethylmethyl 0.1 M NaNO3 25 – 1700 0.60
cellulose
Hydroxypropylmethyl 0.1 M NaNO3 25 – 3600 0.53
cellulose H2O 25 49–380 850 0.51 Clasen and
Kulicke
(2001)
Cellulose acetate DMAc 25 4.55– 1900 0.60 Kamide et al.
14.5 (1981)
DMSO 25 4.55– 1700 0.61
14.5
H2O 25 4.55– 2000 0.60
14.5
Formamide 25 4.55– 2000 0.60
14.5
0.01 M NaCl 25 21–229 87.76 0.93
Pullulan H2O 25 0.5–240 221 0.66 Kato et al.
(1984)
H2O 25 2.08– 236 0.658 Kawahara et
123.8 al. (1984)
0.1 M NaCl 25 4.8–150 1.79 0.67 Kato et al.
(1983)
H2O 25 0.53– – 0.675 Nishinari et
101.5 al. (1991)
(continued)
52 H. Zhang and R. Li

Table 2.9 (continued)

K
104
T Mw  (mL/
Polysaccharide Solvent ( C) 10-4 g) α Ref.
Chitosan 0.2 M 30 94–251 1.04– 0.81– Wang et al.
CH3COOH/ 168 1.12 (1991)
0.1 M
CH3COONa
0.1 M AcOH/ 25 4.78– 18.1 0.93 Roberts and
0.2 M NaCl 63.01 Domszy
0.1 M AcOH/ 25 4.78– 30.4 1.26 (1982)
0.02 M NaCl 63.01
0.2 M AcOH/ 25 – 893 0.71 Lee (1975)
0.1 M
AcONa/4 M
urea
0.3 M AcOH/ 25 20 820 0.76 Rinaudo et al.
0.2 M AcONa (1993)
(DA = 2%)
0.3 M AcOH/ 25 6–28.5 790 0.79 Brugnerotto
0.2 M AcONa et al. (2001)
(DA = 0–3%)
0.3 M AcOH/ 25 6–28.5 740 0.80
0.2 M AcONa
(DA = 12%)
0.3 M AcOH/ 25 6–28.5 700 0.81
0.2 M AcONa
(DA = 22–
24%)
0.3 M AcOH/ 25 6–28.5 630 0.83
0.2 M AcONa
(DA = 40%)
0.3 M AcOH/ 25 6–28.5 570 0.825
0.2 M AcONa
(DA = 56–
61%)
0.02 M ace- 25 9.5–18 843 0.92 Berth and
tate buffer/0.1 Dautzenberg
M NaCl (2002)
Chitin 2.77 M 20 10–120 1000 0.68 Einbu et al.
NaOH (2004)
DMAc/5% 25 9–51 24 0.69 Terbojevich
LiCl et al. (1988)
DMAc/5% 30 8–71 76 0.95 Poirier and
LiCl Charlet
(2002)
Agarose 0.75 M 35 8–14 700 0.72 Rochas and
NaSCN Lahaye
(1989)
(continued)
2 Solution Properties 53

Table 2.9 (continued)

K
104
T Mw  (mL/
Polysaccharide Solvent ( C) 10-4 g) α Ref.
Alginate 0.01 M NaCl 20 10–270 0.048 1.15 Smidsrød
0.1 M NaCl 20 10–270 0.20 1.0 (1970)
1 M NaCl 20 10–270 0.91 0.87
I~1 20 10–270 1.20 0.84
Lentinan 0.1 M NaOH 25 14.5– 51 0.81 Zhang et al.
113 (2005)
Curdlan DMSO 25 5–100 1.60 0.74 Futatsuyama
et al. (1999)
DMSO 25 5.1–192 4.80 0.65 Zhang and
Nishinari
(2009)
H2O:cadoxen 25 7–68 2.50 0.65 Hirano et al.
(1:1) (1979)
0.3 M NaOH 25 5.3–200 79.0 0.78 Nakata et al.
(1998)
Konjac glucomannan H2O 25 4.9–120 5.30 0.78 Prawitwong
et al. (2007)
Cadoxen 25 26–69 3.55 0.69 Kohyama
(1993)
Methyl-konjac H2O 30 16–120 6.37 0.74 Kishida et al.
glucomann (1978)
Gellan gum 0.025 M 25 1.3–6.2 74.8 0.91 Dreveton et
TMACl al. (1996)
0.01 M NaCl 25 6.8 1660 0.89 Masuelli
(2014)
Na+ high acyl gellan gum DMSO 25 42–101 11.6 0.67 Kang et al.
(2017)
Xanthan gum 0.01 M NaCl 25 170 0.046 1.41 Masuelli
(2014)
0.1 M NaCl 25 30–1000 1.70 1.14 Milas et al.
(1985)
Petcin PBS 20– 8.45– 0.234 0.8224 Kar and
60 10.28 Arslan (1999)
0.0155 M 25 2.3–7.1 1.4 1.34 Owens et al.
NaCl (1946)
κ-carrageenan 0.028 NaCl 20 0.58–46 77.8 0.90 Vreeman et
0.118 NaCl 20 0.58–46 88.4 0.86 al. (1980)
0.218 NaCl 20 0.58–46 209 0.78
I~1 20 0.58–46 520 0.67
0.1 M NaCl 25 25–40 30 0.95 Kalitnik et al.
(2013)
λ-carrageenan 0.1 M NaCl 20 34–87 – 0.6 Almutairi et
al. (2013)
(continued)
54 H. Zhang and R. Li

Table 2.9 (continued)

K
104
T Mw  (mL/
Polysaccharide Solvent ( C) 10-4 g) α Ref.
Guar gum 0.1 M urea 25 44–165 3.8 0.72 Robinson et
al. (1982)
H2O 25 38–140 7.76 0.98 Doublier and
Launay
(1981)
Dextran 0.1 M NaCl 25 0.94– 13.6 0.47 Kato et al.
519.3 (1983)
Schizophyllan 0.01 M 25 10.7– – 1.1, Kashiwagi et
NaOH 568 1.8 al. (1981b)
DMSO 25 10.7– 231.4 0.69
568

Table 2.10 The values of Rg Polysaccharide Mw  105 Rg (nm)

of hyaluronic acid (HA) with
CFG 3.33 24
different molecular weight
and corn fiber gum (CFG) HA-1 3.56 81
HA-2 12.9 120
HA-3 22.3 167

flexible random coils in a good solvent, and ~0.5 for flexible random coils and ~0.33
for compact coils in a θ solvent. For instance, for pullulan in water, α0 ¼ 0.568
(De Nooy et al. 1996), and in the θ solvent of ethylene glycol, α0 ¼ 0.5 (Nordmeier
1993).
Table 2.11 summarizes the parameters in the relationship between the radius of
gyration and the molecular weight for a series of polysaccharides and their
derivatives.

5.2.3 Scaling of A2 with Mw

The A2 value directly reflects the degree of interactions between polymers and
solvents. For polymers in a good solvent the value of A2 is positive; and A2
0 for polymers in a θ solvent or aggregation. Usually, A2 decreases with increasing
Mw as well as Rg, and obeys the following scaling rule:

A2 ¼ KM n
w ð2:29Þ

where K and n are constants for a given polymer at a given temperature in a given
solvent. The n value is normally 0.2–0.3 at a good solvent limit (Takahashi et al.
2 Solution Properties 55

Table 2.11 The parameters of Eq. (3.4) for the selected list of polysaccharides
T k
Polysaccharide Solvent ( C) Mw  10-4 104 α0 Ref.
Hyaluronic acid 0.1 M NaCl 25 1–100 – 0.5 Luan et al.
(2011)
0.2 M NaCl 25 66.7–423 – 0.5 Takahashi
et al. (2003)
0.15 M NaCl 25 600–800 – 0.57– Cowman and
0.60 Matsuoka
(2005)
Amylose 1 M KOH 70 141–223 – 0.80 Ahmad et al.
(1999)
0.1 M KOH 25 1.2–120 – 0.62 Roger et al.
H2O 25 24–120 – 0.52 (2000)
Amylopectin 1 M KOH 70 141–223 – 0.58 Ahmad et al.
(1999)
Cellulose DMAc-LiCl 30 12.5–70 – – McCormick
et al. (1985b)
Cellulose diacetate DMAc 25 19.4 680 0.53 Muroga et al.
THF 25 19.4 299 0.56 (1987)
Acetone 25 19.4 7390 0.31
Cellulose triacetate DMAc 25 8.0 463 0.55
Methyl cellulose 0.1 M NaNO3 25 – 440 0.55 Schittenhelm
Sulfoethyl cellulose 0.1 M NaNO3 25 56 120 0.65 and Kulicke
Carboxymethyl 0.1 M NaNO3 25 – 66 0.70 (2000)
cellulose
Hydroxyethylsulfoethyl 0.1 M NaNO3 25 29.4–30.3 380 0.58
cellulose
Hydroxyethyl cellulose 0.1 M NaNO3 25 – 260 0.59
0.1 M NaNO3 25 – 330 0.55
Hydroxypropyl 0.1 M NaNO3 25 – 250 0.56
cellulose 0.1 M NaNO3 25 – 660 0.51
Hydroxyethylmethyl 0.1 M NaNO3 25 – 380 0.53
cellulose 0.1 M NaNO3 25 – 260 0.59
hydroxyethylethyl 0.1 M NaNO3 25 – 140 0.63
cellulose
Hydroxypropylmethyl 0.1 M NaNO3 25 – 470 0.51
cellulose
Pullulan H2O 25 6.5 164 0.57 Muroga et al.
(1987)
H2O 25 0.5–240 147 0.58 Kato et al.
(1984)
0.1 M KOH 25 0.62–130 – 0.62 Roger et al.
H2O 25 0.6–130 – 0.58 (2000)
Chitin 2.77 M NaOH 20 10–120 1700 0.46 Einbu et al.
(2004)
DMAc-LiCl 25 9–51 240 0.69 Terbojevich
et al. (1988)
(continued)
56 H. Zhang and R. Li

Table 2.11 (continued)

T k
Polysaccharide Solvent ( C) Mw  10-4 104 α0 Ref.
Chitosan 0.2 M 25 3.6–46 299 0.59 Christensen
NH4OAc et al. (2008)
0.02 M acetate 25 9.5–18 750 0.55 Cowie (1960)
buffer/0.1 M
NaCl
Lentinan 0.1 M NaOH 25 14.5–113 2300 0.58 Zhang et al.
(2005)
Konjac glucomannan H2O 25 4.9–120 277 0.60 Prawitwong
et al. (2007)
Na+ high acyl gellan DMSO 25 42–101 206 0.61 Kang et al.
(2017)
DMAc dimethylacetamide, THF Tetrahydrofuran

Fig. 2.10 Relationship between A2 and Mw for hyaluronic acid in 0.1M NaCl. Reproduction with
permission from Luan et al. (2011), Copyright 2011 Elsevier

2003; Cowman and Matsuoka 2005); and a higher value (>0.5) is a characteristic of
branched structures (Tao et al. 2007). For, example, n ¼ 0.61, 0.80, 0.66 for
amylopectin, glycogen and dextran, resepectively (Burchard 2001). The scaling
rule holds only in the semi-dilute regime.
A2 is in the range of 1.0  103 to 3.5  103 mol ml g2 for hyaluronic acid in
0.1M NaCl, depending on its Mw (Luan et al. 2011). Figure 2.10 shows the power-
law relationship linking A2 and Mw for a series of hyaluronic acid samples. The
obtained exponent of n ¼ 0.2 is in very good agreement with the predicted value of
2 Solution Properties 57

0.2 (Cowman and Matsuoka 2005) as well as the experimental value of 0.19
(Takahashi et al. 2003).
A famous relation between intrinsic viscosity [η], coil size Rg, and molar mass M,
for linear chains, is known as the Fox–Flory equation (Rubinstein and Colby 2003)
 
Rg 3
½η ¼ Φ ð2:30Þ
M

where Φ is the Flory hydrodynamic constant, which is roughly related to the chain
draining in solution. Φ ¼ 2.5  1023/mol is a universal constant for all polymer-
solvent systems, and in theta solvent Φ ¼ 2.84  1023/mol.
In the Zimm model, the following relationship among these physical parameters
can be established (Lopez et al. 2017):

N A R2g Rh
½η ð2:31Þ
Mw

The main property of the above relationship is that the ratio of

ð½η M w Þ=ðN A Rh R2g Þ is a constant. For high acyl gellan gum chains in the good
solvent of DMSO, a value of ca. 4 for the ratio of ð½η M w Þ=ðN A Rh R2g Þ is found
(Kang et al. 2017).

5.2.4 Persistence length (Lp)

Polysaccharides may exhibit different conformations in solutions. The wormlike

chain model is famous for describing the chain flexibility in solutions.
The persistence length (Lp) is a key parameter characterizing the flexibility of
linear polymers and therefore their conformation. The value of Lp for a given
polymer can be taken as a measure of the chain stiffness. It is notable that the values
of Lp of various polysaccharides may greatly vary as a function of Mw, solvent
conditions, and characterization techniques, also strongly depending on the calcula-
tion methods. There are various methods to determine Lp. Lp can be derived from the
light scattering data by Benoit–Doty treatment of the wormlike chains (Benoit and
Doty 1953). Lp may also be obtained by using the Mw dependence of the [η] from the
slope in the Bushin–Bohdanecky plot based on the Yamakawa–Fujii–Yoshizaki
(YFY) theory (Bohdanecky 1983).
A summary and comparison of Lp for various polysaccharides are given in
Table 2.12. The well-known Lp values for DNA are also shown for comparison.
It is shown in Table 2.12 that the Lp values of the stiff-chains are much larger than
those of the flexible random chains. Generally speaking, as the values of Lp increase,
the macromolecules become stiffer, more extended, indicating a stretching semi-
flexible chain. The quality of solvent undoubtedly has a significant effect on the
polymer conformation. While an Lp value is 5.2 nm for K+ acetylated gellan in pure
58 H. Zhang and R. Li

Table 2.12 Persistence length, Lp, for various polysaccharides

T Mw 
Sample Solvent ( C) 104 Lp (nm) Ref.
Na+ high acyl 50 mM NaNO3/ 25 42.0– 8–10 Kang et al. (2017)
gellan DMSO 100.7
K+ acetylated DMSO 25 220 5.2 Brownsey et al. (1984)
gellan 90% DMSO 25 160 11.64
K+ deacetylated 90% DMSO 25 88–96 13.14–
gellan 15.54
Na+ 25 mM NaCl 40 3.47– 9.4 Takahashi et al. (2004)
deacetylated 11.5
gellan – 9.47 17 Takahashi (1999)
75 mM NaCl 25 23.8 98 Okamoto et al. (1993)
– 43.4 102 Dentini et al. (1988)
Konjac Phosphate buffer 20 24–74 13 Kök et al. (2009)
glucomannan
Amylose DMAc-3% LiCl 25 50–200 3.26 Cao et al. (2000)
H2O 25 – 1.5 Roger et al. (2000)
0.1 M KOH 25 – 1.6
D2O 25 30–80 1.9–
3.2
KOD 25 – 4.2
DMSO 25 0.56–170 1.2 Nakanishi et al. (2002)
Cellulose 9%, 10% LiCl/ 25, 1.65–70 11–25 Terbojevich et al. (1985);
DMAc 30 McCormick et al. (1985a)
Carboxymethyl 0.02 M, 0.1 M 25 12–120 12–16 Hoogendam et al. (1998)
cellulose NaNO3
High salt – – 5.0
concentrations
water/cadoxen – – 8.5
Cellulose DMAc 25 19.4 139 Muroga et al. (1987)
diacetate
Cellulose THF 25 19.4 39
diacetate
Cellulose Acetone 25 19.4 77
diacetate
Cellulose DMAc 25 8.0 59
triacetate
Galactomannan H2O 20 – 8–10 Patel et al. (2006), Morris et
al. (2008a)
Pectin 0.1 M NaCl 20 15–18 10–13 Morris et al. (2008b)
Hyaluronic acid 0.15 M NaCl 37 4–550 6–8 Robinson et al. (1982)
Curdlan DMSO 25 5.1–192 5.81 Zhang and Nishinari (2009)
Lentinan 0.2 M LiCl/ 25 21.7– 6.0 Wang et al. (2012)
DMSO 84.7
DMSO 25 – 5.1
DMSO 25 19.1– 4.8 Wang and Zhang (2009)
53.8
(continued)
2 Solution Properties 59

Table 2.12 (continued)

T Mw 
Sample Solvent ( C) 104 Lp (nm) Ref.
Chitin 8% NaOH/4% 25 215.6 4 Li et al. (2010)
urea
2.77 M NaOH 20 10–120 11.5– Einbu et al. (2004)
13
5% LiCl/DMAc 25 9–51 15–50 Terbojevich et al. (1988)
Chitosan 265 mM HAc/ 25 3.27– 9.5 Kang et al. (2018)
200 mM NH4Ac 42.66
Pullulan H2O 25 6.5 (Mn) 1.2–1.9 Muroga et al. (1987)
H2O 25 10–100 1.4–3.1 Adolphi and Kulicke
(1997)
Alginate H2O 25 – 91 Stokke and Brant (1990)
0.001–0.1 M 22.5 95.9 4–6 Muroga et al. (1987)
NaNO3
0.2 M NH4OAc 25 3.6–46 12 Christensen et al. (2008)
Single-stranded 0.02 mM NaCl 25 – 30  4 Stokke and Brant (1990)
xanthan
Double- 0.02 mM NaCl 25 – 68 
stranded 7
xanthan
Double-helix 0.1 M NaCl 25 740– 120 Sato et al. (1984)
xanthan 7400
ι-carrageenan 0.1 M NaCl 25 – 22.6 Schefer et al. (2014)
(random coil)
ι-carrageenan 0.1 M NaCl 25 – 26.4
(helix)
Succinoglycan 0.01 M NaCl 25 35.4– 50 Nakanishi and Norisuye
71.4 (2003)
Scleroglucan H2O 25 – 80  Stokke and Brant (1990)
10
Schizophyllan 0.01 M NaOH 25 10.7–568 180  Kashiwagi et al. (1981a)
30
H2O 25 50–570 100– Chun and Park (1994)
200
DNA: single- 8 M urea 21 2800– 4 Tinland et al. (1997)
stranded 53860
bases
Double- 0.1 M NaCl 20, – 58–68 Norisuye (1993)
stranded 25

DMSO (Brownsey et al. 1984), it is 11.64 nm for this polysaccharide in 90% DMSO
aqueous solution at 25  C. Significantly different values of Lp are also reported for
chitin in different solvents as 4 nm in 8% NaOH/4% urea (Li et al. 2010), 11.5–
13 nm in 2.77M NaOH (Einbu et al. 2004), and 15–50 nm in 5% LiCl/DMAc
(Terbojevich et al. 1988).
60 H. Zhang and R. Li

5.2.5 Chain Stiffness Parameter B

Smidsrød and Haug (1971) proposed an empirical method to determine the empirical
chain stiffness parameter B for polyelectrolyte polysaccharides:

B ¼ S=½η v0:1 ð2:32Þ

where S is a constant, [η]0.1 is the [η] at 0.1M NaCl ionic strength, and the exponent ν
usually ranges from 1.2 to 1.4.
A number of polysaccharides contain acidic groups (typically –COOH), which
are essentially polyelectrolytes. Their [η] value decreases with increasing ionic
strengths of the solution according to the following relation:

½η ¼ ½η 1 þ SI 1=2 ð2:33Þ

where [η]1 is the [η] at infinite ionic strength, and I is the ionic strength of the
solution. The [η]1 and S values can be derived from the intercept and the slope of the
linear functions by the measurement of the [η] at different I, of which Figs. 2.11 and
2.12 are examples for hyaluronic acid (Luan et al. 2011) and chitosan (Gooday et al.
1986), respectively.

Fig. 2.11 Plots of intrinsic viscosity [η] versus I1/2 for hyaluronic acid in aqueous solution (NaCl
as a supporting electrolyte). Reproduction with permission from Luan et al. (2011), Copyright 2011
Elsevier
2 Solution Properties 61

Fig. 2.12 Plot of intrinsic

viscosity [η] versus I1/2 for
chitosan with the degree of
acetylation of 0.42.
Reproduction with
permission from Gooday
et al. (1986), Copyright
1986 Springer Nature

A list of stiffness B parameters of different polysaccharides has been compiled

(Lapasin et al. 1992) (Table 2.13). A lower value of B is associated with stiffer chain
conformation. For example, a small B value of 0.00525 for xanthan gum indicates
that the polymer adopts a stiff conformation (Tinland and Rinaudo 1989), whereas
the B value is 0.045–0.065 for flexible carboxymethyl cellulose (Smidsrød and Haug
1971), 0.10 for k-carrageenan (Vreeman et al. 1980), and is 0.0147–0.0201 for
hyaluronic acid with intermediate stiffness (Luan et al. 2011). Variation in structure
composition, type, DS, the molecular weight of the polysaccharide as well as
chemical modification to the molecular structure can strongly influence the magni-
tude of B value, i.e., the stiffness of the polymer chain. Alginate with different ratios
of guluronate/mannuronate and alternate block ranks shows different chain stiffness
(Smidsrød et al. 1973). Similar situation can be observed for xanthan with different
ratios of Ac/Pyr substitution (Shatwell et al. 1990). The flexibility of the chitosan
chain increases with increasing the degree of deacetylation (Gooday et al. 1986).
With the increase in DS, the CMC chain becomes more flexible (Smidsrød and Haug
1971). For pectin, high DS content makes the chain much stiffer (Smidsrød and
Haug 1971). The high Mw hyaluronic acid chain is more flexible than low Mw one
(Luan et al. 2011). λ-carrageenan chain has a smaller B than k-carrageenan chain, and
62 H. Zhang and R. Li

Table 2.13 Stiffness parameter B for various polysaccharides (B value of DNA is also shown for
comparison)
Polymer B Notes Ref.
Alginate 0.031 High guluronate Smidsrød et al.
0.040 High mannuronate (1973)
0.065 Alternate block copolymer of
guluronate and manuronate
0.02 High guluronate Abodinar et al.
0.022 Low guluronate (2014)
Amylose 0.22 – Smidsrød and Haug
xanthate (1971)
Carboxymethyl 0.20 DS = 0.80
amylose
Carboxymethyl 0.044 DS = 0.40
cellulose 0.045 DS = 0.56
0.045 DS = 0.73
0.065 DS = 1.00
0.078 DS = 1.00 Trivedi and Patel
0.075 DS = 1.08 (1982)
0.065 DS = 1.35
λ-carrageenan 0.053 – Morris et al. (1978)
κ-carrageenan 0.10 – Vreeman et al.
(1980)
Chitosan 0.043 DD = 0.48 Gooday et al.
0.061 DD = 0.58 (1986)
0.091 DD = 0.88
0.1 DD = 1.00
0.03 – Abodinar et al.
(2014)
0.02–0.1 – Anthonsen et al.
(1993)
0.08 – Kienzle-Sterzer
(1984)
0.043 DA = 0.52 Muzzarelli et al.
0.061 DA = 0.42 (1986)
0.060 DA = 0.20
0.091 DA = 0.12
Hyaluronic acid 0.0147– Mw = 1  104–1  106 Luan et al. (2011)
0.0201
0.065 – Smidsrød and Haug
Pectin 0.034– DS = 0 (1971)
0.044
0.044 DS = 0.27
0.052 DS = 0.58
0.026 DS = 0.78
(continued)
2 Solution Properties 63

Table 2.13 (continued)

Polymer B Notes Ref.
0.005 DS = 0.89
0.015 HM + 60% sucrose Michel et al. (1984)
0.020 Without sucrose
0.03 LM Abodinar et al.
0.03 LM (2014)
Xanthan 0.12 0.1 M NaCl Ac:Pyr = 1:1 Shatwell et al.
0.00044 0.1 M NaCl Ac:Pyr = 9:2 (1990)
0.00615 0.1 M NaCl Ac:Pyr = 48:1
0.00662 0.1 M NaCl Ac:Pyr = 1:4
0.00545 0.1 M NaCl Brunchi et al.
(2014)
DNA 0.0055 – Sharp and Bloom-
field (1968)
DS degree of substitution, DD degree of deacetylation, DA degree of acetylation, HM high-
methoxyl, LM low-methoxyl, Ac/Pyr the ratio of degrees of acetyl (Ac) and pyruvic acid (Pyr)
substitution

chemical modification can make it less stiff (Morris et al. 1978; Vreeman et al.
1980).
Note that in comparison with the method to deduce conformation parameters of
the polymer chain in solution by using the well-known Mark–Houwink equation, the
main advantage of this method is that the stiffness can be determined solely from the
intrinsic viscosity data without any knowledge of Mw.

6 Polysaccharide Liquid Crystal

Liquid crystals (LCs)—alternatively named anisotropic fluids—are intermediate

between ordinary fluids. LCs are endowed with only short-range order or structure,
and crystalline solids.
As concerns the formation of polysaccharide liquid crystals, a lot of polysaccha-
rides have been found to give mesomorphic phases in the aqueous environment and
other solvents, e.g., cellulose and some cellulose derivatives (Gilbert 1990),
schizophyllan (Van and Teramoto 1982), scleroglucan (Yanaki et al. 1984), xanthan
gum (Salamone et al. 1982; Lim et al. 1984), curdlan (Dobashi et al. 2004, 2005),
konjac glucomannan (Dave et al. 1998).
Cellulose and many cellulose derivatives can form lyotropic LCs in suitable
solvents, whereas other cellulose derivatives give thermotropic LCs. For instance,
cellulose acetate and cellulose triacetate form anisotropic solutions in different
64 H. Zhang and R. Li

solvents such as mixtures of trifluoroacetic acid and chlorinated liquids (Gilbert

1990). Hydroxypropyl cellulose (HPC) is reported to form stable and easy-to-handle
mesophases in water and in dimethylacetamide (Werbowyj and Gray 1976). HPC
also forms thermotropic LCs (Suto et al. 1982). Cellulose tricarbanilate yields liquid
crystalline solutions when dissolved in methyl ethyl ketone (Vogt and Zugenmaier
1983) and cellulose itself is found to form LCs in dimethylacetamide/LiCl solvent
system. Nanocellulose suspensions can form a liquid crystalline structure at a critical
concentration due to its inherent properties from the attractive van der Waals forces
and repulsive electrostatic forces (Moon et al. 2011; Habibi et al. 2010) Aqueous
suspensions of nanocrystalline cellulose colloidal rods can be used to establish a new
anisotropic soft material—a liquid crystal “hydroglass” (Xu et al. 2019).
Aqueous xanthan gum solution presents excellent viscosity, also exhibits liquid
crystalline behavior with high stability in a wide range of concentrations (Rwei and
Nguyen 2014b) The viscosity of the xanthan gum solution in the liquid crystal
region follows a power law with an index n of roughly 0.08, independent of the
xanthan gum concentration (Rwei and Nguyen 2014a).
Liquid crystal behavior is also found in the biphasic systems of single-walled
carbon nanotubes dispersed in hyaluronic acid solutions, which exhibits liquid
crystalline nematic phases in equilibrium with the isotropic domains (Moulton
et al. 2007).

7 Conclusions

In this chapter, we concisely introduced the thermodynamics of polymer solutions,

which has been well described in the literature, especially the Flory–Huggins mean-
field theory and the concept of two vital physical parameters of χ and A2. While the
concentration regimes and the scaling law of polymer solutions are detailed, several
key molecular and conformation parameters are summarized.
However, the determination of molecular parameters and conformation charac-
teristics has been a particularly challenge because of some factors such as the
extremely low water-solubility, high molecular weight and polydispersity, sensitiv-
ity to the sider substituents, and especially the polyelectrolyte nature and the strong
tendency of the polymer chain to aggregate even in a very dilute solution. These
parameter values may significantly vary depending on the molecular weight, solvent
conditions, and characterization techniques, as well as the calculation methods. The
conformation parameters may also be sensitive to variations of the polydispersity of
the polymer. To deduce a global conformation of the polymer chain, the theory of
scaling functions presupposes homogeneous monodisperse fractions in molar mass
and dimension of the polymers. However, in comparison with the relatively readily
available synthetic polymers with narrow molecular weight distribution, natural
biopolymer samples, especially plant polysaccharides and algal polysaccharides,
2 Solution Properties 65

generally possess a broad molecular weight distribution. Therefore, it is necessary

and essential to make conformation analysis by using samples with pertinent Mw and
narrow polydispersity.
The full understanding of the molecular structures, chain conformation, function-
alities, and solution properties of typical food hydrocolloids and their relationships
are prerequisite for food researchers to efficiently utilize them on food product
innovation due to the complexity of these diverse natural biopolymers with varied
molecular structures and physicochemical properties, manufacturing processes, and
consumption requirements. With the emergence of different characterization
methods, recently, the determination of molecular chain conformation and the
illumination of its importance on solution properties remain the pressing issues in
the food hydrocolloids field. To design and develop more rational macromolecule
structures and build functional food materials with good performance based on the
relationship between chain conformation and solution properties will still be an
acclaimed methodology to achieve deeper application value.

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Chapter 3
Rheological and Thickening Properties

Katsuyoshi Nishinari

Abstract Rheology in food studies all the mechanical properties relating deforma-
tion and flow. Since the texture is known to be important to govern the palatability
with taste and aroma, and also recognized as a key factor in designing foods for
persons with difficulty in mastication and deglutition, hydrocolloids controlling
rheological and thickening properties have been attracting more attention than
before. This chapter describes the necessity and importance of rheology and then,
fundamental concept of elasticity, viscosity, static and dynamic viscoelasticity, and
the measuring methods in relation with the molecular structure. Although yield stress
and thixotropy have been recognized and studied, they are both not so well under-
stood, and recent developments are described in relation with thickening properties.
Recent development of fractional calculus and microrheology is described. Appli-
cation of thickening properties is also described.

Keywords Viscosity · Elasticity · Viscoelasticity · Yield stress · Thixotropy

1 Introduction

Rheology is a science on the deformation and flow of matters. The word rheology is
the combination of “rheo”—flow and “logy”—science in Greek. In the early stage of
rheology, British Rheologists Club (1942) proposed a classification of deformation
which is divided into elastic deformation and flow. Between the two most simple
extremities, Hookean elasticity and Newtonian viscosity, viscoelastic deformation
has been most well studied in relation with plastic industry and food industry.
Rheology has been proved to be useful to understand quantitatively the gelation
process (treated in the next chapter), the transformation of a liquid to solid, dispersed
systems, the mixture of different materials because most foods consist of many

K. Nishinari (*)
Glyn O. Phillips Hydrocolloids Research Centre, School of Food and Biological Engineering,
Hubei University of Technology, Wuhan, PR China
e-mail: katsuyoshi.nishinari@hbut.edu.cn

© Springer Nature Singapore Pte Ltd. 2021 75

Y. Fang et al. (eds.), Food Hydrocolloids,
https://doi.org/10.1007/978-981-16-0320-4_3
76 K. Nishinari

ingredients. Rheology is developing steadily influenced by the development of other

related science. The new measurement techniques such as atomic force microscopy,
optical tweezers in microrheology make possible to detect the nano/micro scale
distance, and the length scale studied in rheology is widened to nano, micro, meso,
and macroscale. Femtosecond spectroscopy and time-resolved X-ray diffraction
photoelectron spectroscopy study the very fast change of rheological properties
accompanying the structural changes, and the timescale is also widened. Rheology
is also closely related with dielectric (e.g. broadband dielectric measurement) and
related studies, and thus can be treated from the common background such as
stimulus-response theory. Stimulus-response approach has been proved useful to
understand systematically the transient phenomena in statistical physics, but now it
seems to be used also in psychology. Remember the conditioned reflexes of Pavlov’s
dog. It is expected to be applied to sensory evaluation of texture and flavor of foods.
Soft matters such as foods, biological tissues, polymers, colloids are now understood
based on common background (de Gennes 1979; Doi 2013). Rheology in food has
been studied extensively because the perceived texture is closely correlated with
rheology (Bourne 2002; Chen and Engelen 2012; Nishinari and Fang 2018). As is
widely recognized, texture and flavor are most important attributes governing the
palatability of foods. With the advent of aged society, the number of persons who
have the difficulty in the mastication and swallowing is increasing, which increases
the number of cases of pneumonia and other symptoms. Rheology is not yet
completed for food science, and more developments are expected to solve these
problems and to create better foods, better storage, and distribution methods in
collaboration with other disciplines. It is widely recognized that instrumental mea-
surements give the objective evaluation and basis of the perceived texture of foods
which are described by subjective terms. Interpretation of sensory terms such as firm,
hard, stiff, rigid, etc. is often confusing and depends on individuals.
Although the elastic modulus obtained from the initial slope of force-deformation
curves of the agar gel is larger than that of the gelatin gel (Fig. 3.1a), the breaking
strength of the agar gel turns out to be smaller than that of the gelatin gel even after
the calibration of the cross-sectional area. Therefore, it is impossible to answer to a
question “which of the two gels, an agar gel or a gelatin gel, is ‘firmer’ or ‘harder’?”
if the definition of the hardness or the firmness is not given.
In Fig. 3.1b, how we could answer to the question “Which of the two liquids,
sugar syrup or mayonnaise, is thicker?” Most people reply that sugar syrup is thicker
than mayonnaise. This indicates that most people sense the viscosity at a higher
shear rate than ca. 1 s
1 in the mouth. As shown in this example, daily language such
as “thick” and “thin” is vague.
Scott Blair studied the relation between the instrumental measurement and
sensory evaluation of texture using fractional calculus which has recently
re-attracted more attention. Although the theory of viscoelasticity is well developed
in the linear region where the stress and strain or strain rate are in the linear relation,
the large deformation and fracture are not so well established. In this chapter, after
describing the basic concepts in the linear viscoelasticity, large amplitude oscillatory
shear (LAOS), yield stress, thixotropy, microrheology are described. Application of
thickening agents in food production is also described. Then, recent application of
3 Rheological and Thickening Properties 77

Fig. 3.1 (a) Force-deformation curves of cylindrical (20 mm diameter and 30mm height) gels of
4% agar and 25% gelatin at a compression velocity of 10 mm/min. The curve GI and GII for gelatin
gels were obtained with approximately equal probability (Nishinari et al. 1980). (b) The viscosity as
a function of shear rate for sugar syrup (closed circle, flow behavior similar to honey) and
mayonnaise (open circle). Reproduction with permission from Nishinari et al. (1980). Copyright
1980 JSFST

fractional calculus and also the relation between the molecular structure and
viscosifying function are described.

2 Elasticity

Theory of elasticity treats the deformation of solids. Solids do not flow and maintain
a certain shape. A solid which does not deform at all even under a force is called a
“rigid body.” The distance between two points in a rigid body is regarded as
constant. The concept of a rigid body is an ideal model. In reality, even a diamond
or iron deforms when it is subjected to a large force.
The simplest model of a deformable solid is an elastic body. An elastic body
shows a constant strain (ratio of deformation) instantaneously, that is without delay,
when it is subjected to a constant stress (force per unit area). This solid is called a
Hookean body. Imagine a rod of a radius r and a length l which is subjected to a force
f. Hooke’s law for this rod is written as f/(πr2) ¼ E (δl/l), where the left-hand side is
the force per unit area, which is called (elongational or compressional) stress, while
the right-hand side is the product of the elastic constant E and the (elongational or
compressional) strain. Elongational strain indicates the ratio of the deformation δl to
the initial length l, and it is often written as ε ¼ δl/l. Elastic constant E defined for the
elongational or compressional deformation is called Young’s modulus, which rep-
resents the stress required to produce the unit elongation or unit contraction. It
indicates the resistance to the deformation. The unit of E is force over area, that is
Pa ¼ N/m2, in SI.
When the force f is exerted on the upper surface of the parallelepiped body to the
parallel direction of the surface, then it shows a uniform deformation which is
78 K. Nishinari

Table 3.1 The relation between 4 elastic parameters

G,E G, κ E, κ G, μ E, μ κ, μ
G G G 3κE G E 3κ ð1
2μÞ
9κ
E 2ð1þμÞ 2ð1þμÞ
E E 9Gκ
Gþ3κ
E 2(1 + μ)G E 3κ(1
2μ)
κ GE
9G
3E
κ κ 2ð1þμÞG E
3ð1
2μÞ
κ
3ð1
2μÞ
μ E
2G
2G
3κ
2G
2ð3κþGÞ
3κ
E
6κ
μ μ μ

characterized by a small angle γ. For a small angle, tan γ can be approximated by γ,

which is called the shear strain. Then, Hooke’s law is written as f/A ¼ Gγ, where G is
called shear modulus or rigidity.
Next, let us consider the volume change of a spherical elastic body (volume v)
which is surrounded by liquid. When the liquid pressure is raised from p to p+Δp by
a pump, the volume of the sphere will decrease from v to v-δv. Then, Hooke’s law is
written as δp ¼
κ δv/v. Since it is the custom to represent the sign of the
compressional pressure as positive, the negative sign in the right-hand side origi-
nates from the decrease of the volume under compression. Here, κ is called bulk
modulus, and its inverse 1/κ is called compressibility.
When a cylinder of length l and the radius r is extended to its axial direction
without any force at side surface, the radius will be decreased from r to r-δr. The
ratio of the transverse strain
δr/r to the longitudinal strain δl/l is called Poisson’s
ratio μ: μ ¼ -(δr/r)/(δl/l). The negative sign originates from the fact that when the
length of cylinder is increased (δl > 0), the radius is decreased (δr < 0). Generally,
the value of Poisson ratio ranges from 0.5 for incompressible material like rubber to
0 for porous material like cork.
The Relation Among Four Elastic Parameters E, G, κ, and μ
Commonly used four elastic constants E, G, κ, and μ are not independent, and are
related with each other as shown in Table 3.1. Only two constants are independent,
and therefore, when two constants are determined the other two can be calculated.
On the other hand, the determination of only one constant is not complete to
understand the whole elastic property of that material.
As mentioned above, the Poisson ratio μ ranges from 0 to 0.5. In Table 3.1,
the bulk modulus κ is given as a function of E and μ. When μ approaches 0.5, the
denominator (1
2μ) approaches to zero and thus the bulk modulus approaches to
infinity, therefore, the bulk modulus is much larger than the other elastic parameter
G and E. This is just the case for rubber. The fact that the bulk modulus is much
larger than G and E means that this material needs a very high pressure to reduce the
volume, hence this is called an incompressible material. In such a case, Young’s
modulus is about three times of the shear modulus: E ¼ 3G.
Shear deformation appears in the torsion, and can be determined by the measure-
ment of the moment. The deformation mode is the same as in rotational viscometer.
When a beam is bent by an applied couple, an upper part is stretched and the
lower part is compressed. There is a part near the middle layer which retains the
3 Rheological and Thickening Properties 79

original length, and is called a neutral layer. Then, Young’s modulus is involved in
the bending, and is obtained from the determination of the sag.
Since measurement of bending is easy, it is often used to study rheological
properties of solids. To apply three point bending test to a soft matter such as agarose
gels, the points were replaced by cylindrical rods (Bonn et al. 1998).
Strain energy of elastic materials is given by the elastic modulus  (strain)2/2 for
all deformation modes, elongation, shear, and expansion.
Poisson’s ratio of most engineering solids, metals, plastics, and glasses is known to
be about 0.3 while those of many soft foods such as cheese and jellies are reported as
0.5 just like a rubber. Cork and flexible foams show no lateral expansion when
compressed thus their Poisson’s ratio is very close to zero. Poisson’s ratios of
vegetables and fruits are reported higher than those of metals and plastics but lower
than those for cheese and jellies, for example, 0.37 for an apple tissue (Powel 1994). It
is also important to take into account the deformation rate (in most cases compression
is selected), since the real food is not purely elastic but rather viscoelastic, and shows a
time dependence. When a 1.33% gel of gellan was compressed slowly at 0.005 mm/
min, no lateral expansion was observed (Nakamura et al. 2001).

3 Viscosity: Newtonian Fluid

There are many models for viscous fluids, and a Newtonian fluid is the simplest
model. In this model, the tangential stress is written by a linear equation of the
shear rate.
Imagine the fluid is filled between the lower and the upper plates (Fig. 3.2). When
the upper plate is moved to the right direction parallel to the x-axis, the fluid in the
neighborhood of the upper plate is dragged with the plate, and the fluid far from the
upper plate is dragged with a slower velocity.

Fig. 3.2 Fluid flow in a narrow gap δ between two very large planes. Upper plate (y = δ) moves
parallel to x-axis. Lower plate (x-axis) is fixed at y = 0. Velocity gradient ∂u/∂y is given by shear
rate dγ/dt because the shear γ is dx/dy, and the velocity u is dx/dt
80 K. Nishinari

The velocity vector is written as u ¼ [u( y), 0, 0]. In the Newtonian fluid, the
tangential stress τ is proportional to the velocity gradient, ∂u/∂y, the change in the
velocity of the fluid flow in the direction perpendicular to the direction of fluid flow,
τ ¼ η∂u/∂y, where the proportional coefficient η is called viscosity. This is called a
Newton’s law of viscosity. When a constant shear stress is given, the velocity
gradient ∂u/∂y is small for a fluid of which the viscosity is large. The viscosity
represents the tangential stress which is required to produce the unit velocity
gradient, thus it is the resistance to the flow.
The viscosity of the fluid can be determined by various methods as described
below.

3.1 Capillary Viscometer

Let us imagine a fluid flow in a capillary. Symmetrical considerations indicate that

the velocity distribution in the liquid will be symmetrical around the axis of the
capillary, and have its maximum value on this axis. Flow rate Q of a Newtonian fluid
(viscosity η) in a capillary (radius a and length l) under a pressure difference Δp is
given by

Za
Q¼ 2πru dr ¼ πΔpa4 =8ηl
0

taking into account that u is the flow velocity at r (the distance from the axis r ¼ 0),
and 2πrdr is the area of the circular ring from r to r+dr. The flow velocity u ¼ (Δp/
4ηl) (a2
r2) takes maximum at the central line r ¼ 0, and becomes slower with
increasing distance from the central line. The viscosity of the liquid can be deter-
mined by measuring Q, volume V of liquid flowing through the capillary in time t,
i.e. Q ¼ V/t. This is called Hagen–Poiseuille equation.

3.2 Falling Ball Viscometer

This is the simplest evaluation of the sedimentation of food particles in the suspen-
sion and creaming in the emulsion. Imagine a solid sphere (radius a and density ρ) is
falling slowly at a velocity v in a surrounding infinite medium (viscosity η, density
ρ0). The frictional force exerting on the falling sphere is given by Stokes law 6πaηv
when the velocity is slow. When the falling velocity is constant (acceleration is
zero), the upward force and downward force should be balanced, and thus the falling
velocity v is given by

2a2
v¼ ðρ
ρ0 Þg
9η
3 Rheological and Thickening Properties 81

where g is the gravitational acceleration. When the density of the sphere ρ is lower
than that of the medium ρ0, the sphere will float up, and the floating velocity can be
calculated in the same way. If the spherical material is not a solid but immiscible
liquid such as in the case of creaming up oil droplet in water, the coefficient in the
Stokes formula is given by 4πaη instead of 6πaη. If the sphere is a gas bubble, this
coefficient is known to be closer to the solid sphere 6πaη rather than to that of the
liquid sphere because the surface of the bubble is usually covered with a hard layer.
Various rotational viscometers are used: liquid samples are put between the cone
and plate in cone plate viscometers, and samples are put between an inner cylinder and
an outer cylinder in coaxial cylindrical viscometers (called also Couette viscometers).

3.3 Reynolds Number

In the above-mentioned viscometers, viscosity can be determined only for a laminar

flow in which the fluid moves in parallel layers. In the laminar flow, the streamlines
show regular shapes and do not intersect each other. In fluid flow of high velocity, the
flow becomes turbulent where the fluid shows a non-regular motion and the velocity at
any point varies in both direction and magnitude with time. Turbulent motion is
accompanied by the formation of eddies and the rapid interchange of momentum in
the fluid. The change from the laminar flow to turbulent flow occurs at a critical value
of Reynolds number Re: Re ¼ ρvl/η, where ρ is the density of the fluid, η is the
viscosity of the fluid, v is the velocity of the fluid in motion relative to some solid body
characterized by a linear dimension l (called characteristic dimension).

ρvl ρl3 ðv2 =lÞ mass  acceleration inertial force

Re ¼ ¼ ¼ ¼
η ηðv=lÞl 2 viscosity  shear rate  area viscous force

Thus, the physical meaning of Reynolds number is the ratio of the inertial force to
the viscous force. Characteristic length represents the radius or the diameter
for spheres, and is an equivalent diameter (or radius) for non-spherical objects. The
equivalent radius is defined for non-spherical objects based on Stokes’ law. The
Reynolds number is used to predict the transition from laminar flow to turbulent flow.

4 Non-Newtonian Flow: Shear Thinning and Shear

Thickening

Most liquid foods, except water, alcohol, oil, are not Newtonian fluids. Simple
models frequently used to analyze the flow behavior of liquid foods are shown in
Fig. 3.3.
82 K. Nishinari

Fig. 3.3 (a) Shear stress plotted against shear rate for commonly used simple models of fluids.
(1) Newtonian fluid σ ¼ η (dγ/dt), (2) Power law fluid (shear thinning) σ ¼ η (dγ/dt)n, n <
1, (3) Power law fluid (shear thickening) σ ¼ η (dγ/dt)n, n > 1, (4) Bingham fluid σ ¼ η (dγ/dt) +
σ y, (5) Herschel–Bulkley fluid σ ¼ η (dγ/dt)n + σ y. (b) Viscosity as a function of shear rate for
Newtonian (1), shear thinning (2) and shear thickening (3) fluids

Newtonian fluid shows a linear relation between shear stress and shear strain, and
the straight line for shear stress vs shear strain passes through the origin. The
viscosity defined by the shear stress divided by shear strain for a power law fluid
σ ¼ η (dγ/dt)n with n < 1 decreases with increasing shear rate, and the behavior is
called shear thinning (Model 2 in Fig. 3.3b). Most common liquids show such a
behavior. Shear thinning may be attributed to many reasons depending on the sample
structure: alignment of rod-like particles or molecules in the flow direction, disinte-
gration of aggregated particles. The viscosity for a power law fluid with n > 1
increases with increasing shear rate, and the behavior is called shear thickening
(Model 3 in Fig. 3.3b). There are not so many examples, but starch paste and some
other examples are shown later.
Liquid foods filled in tubes such as mayonnaise or tomato ketchup or tooth paste
do not begin to flow if it is not subjected to a certain stress. The minimum stress
required to cause a flow is defined as yield stress. A fluid which has a yield stress and
shows a linear stress–strain rate as in Newtonian flow above the yield stress is called
a Bingham liquid (Model 4 in Fig. 3.3a). Hershel–Bulkley liquid is a fluid which has
a yield stress and shows a non-linear stress–strain rate above the yield stress (Model
5 in Fig. 3.3a). Spreadability of butter, cream, jam, paste (legume, fish, meat, nuts,
seed, spice, herb, etc.) is closely related with yield stress, and is discussed later.
In addition to these five models, the following Casson equation is also frequently
used.

σ 1=2 ¼ σ c 1=2 þ ηc 1=2 ðdγ=dt Þ1=2

where σ c and ηc are called the Casson yield stress and the Casson viscosity coeffi-
cient, respectively. The intercept of fitting line is square root of Casson yield stress.
Casson yield stress is the minimum stress that is needed for the fluid to flow.
Casson’s equation has been widely used in the evaluation of chocolate.
3 Rheological and Thickening Properties 83

Fig. 3.4 (a) Shear rate dependence of the viscosity of xanthan and guar solutions showing the
crossover at 50 s
1. The shear rate was changed stepwise from 0.03 to 1000 s
1 for 10 min. Closed
square G1, 2.0 wt% guar (Mw ¼ 1.4  106) solution; closed circle G2, 2.6 wt% guar (Mw ¼ 6.3 
105). closed triangle, X2, 4.1 wt% xanthan (Mw ¼ 3.4  106) solution. Reproduction with
permission from Nishinari et al. (2011), Copyright 2011 Elsevier. (b) Cox–Merz plot for aqueous
solution of hydroxypropylmethyl cellulose (Mw ¼ 2.6  106) with different concentrations.
Complex viscosity η* is defined in Sect. 5. Reproduction with permission from Clasen and Kulicke
(2001). Copyright 2001 Elsevier

4.1 Steady Shear Viscosity of Polymer Solutions

Polysaccharide thickening agents show shear thinning behavior. Xanthan is a micro-

bial polysaccharide consisting of a linear (1–4) linked β-D-glucose backbone with a
trisaccharide chain on every other glucose at C-3, containing a glucuronic acid
residue linked (1–4) to a terminal mannose unit and (1–2) to a second mannose
that connects the backbone, and has also been used widely in food industry as a
thickener and texture modifier because its solution is stable over a wide pH and
temperature range (Sworn 2021). The persistence length of xanthan was determined
as 120 nm (Sato et al. 1984) and it explains well why this polysaccharide is an
excellent thickener. Guar is one of galactomannans consisting of a mannan backbone
and galactose side chains (Nishinari et al. 2007). The ratio of mannose to galactose is
approximately 2:1. Rheological and related characteristics of guar have been exten-
sively studied, and Mark–Houwink–Sakurada exponent and the persistence length
are reported as 0.7 and 4 nm, respectively (Picout and Ross-Murphy 2007; Picout
et al. 2001). In comparison with corresponding data 0.65 and 2.5 nm for a standard
flexible polysaccharide, pullulan (Nishinari et al. 1991; Shingel 2004) it is evident
that guar is slightly stiffer and its solution with the same molar mass and concentra-
tion shows higher viscosity than that of pullulan.
Steady shear viscosity of aqueous solutions of guar gum and xanthan gum
solutions as a function of shear rate is shown in Fig. 3.4a. Xanthan solution shows
pronounced shear thinning as reported by many previous workers. An apparent shear
thickening behavior at lower shear rate is caused by structural formation after the
84 K. Nishinari

preparation of solution and it does not appear in the curve observed by lowering the
shear rate. Wagner et al. (2016) recently reexamined this problem.
The lower molar mass guar shows less shear thinning and an apparent Newtonian
plateau at lower shear rate as expected (Clasen and Kulicke 2001). This is consistent
with previous data for guar gum fractions degraded enzymatically by β-mannanase
for different times (Cheng and Prud’homme 2000).
When the complex viscosity η* ¼ G*/iω (Sect. 5) as a function of angular
frequency ω, and the steady shear viscosity η as a function of shear rate dγ/dt are
plotted, the so-called Cox–Merz plot, both viscosities coincide as shown in Fig. 3.4b.
When the complex viscosity and the steady shear viscosity do not coincide, it may
mean that the microstructure at the relaxed state is destroyed by shear force. Then,
the steady shear viscosity at higher shear rate may be smaller than the corresponding
complex viscosity. However, it should be reminded that more fine local structure
cannot be detected by this method. It is yet useful to see the validity of Cox–Merz
rule for solutions of thickening agents.
Richardson and Ross-Murphy (1987a, b) reported that 3% guar gum solutions
obeyed the Cox–Merz rule, but xanthan solutions deviated from it, and they ascribed it
to some microstructure, “weak gel” (although senior author of this paper regretted to
use this term 20 years later and prefers to use “structured liquid” because it is not a gel)
formation in xanthan solutions. While guar gum solutions can be “thickeners,” xanthan
solutions are “stabilizers” in which fine particles can be suspended without sedimen-
tation. This suggests that some “structure” exists in xanthan solutions. Wientjes et al.
(2000) did not find the validity of Cox–Merz rule for guar gum solutions. It is difficult
to know the cause of the discrepancy between papers of these two groups.
Steady shear viscosity of xanthan solution has been studied extensively. Since
xanthan chain is very stiff (persistence length 120 nm as mentioned above), the
solution shows a shear thinning behavior. Yet, Einaga et al. (1977) show a Newto-
nian plateau at very low shear rates for a solution of much stiffer schizophyllan
(persistence length 180 nm, M ¼ 4.3  106) in water at 30  C using a Zimm–
Crothers type viscometer.
It was shown that globular protein solutions do not obey the Cox–Merz rule
(Ikeda and Nishinari 2001a, b).
One important reason why xanthan is widely used as a thickener and stabilizer is
that its remarkable shear thinning behavior in comparison to galactomannans such as
guar and locust bean gum (LBG) in addition to its insensitivity to the change in
temperature and pH. The high viscosity at low shear rates makes xanthan a good
stabilizer which suspends fine particles of herbs at rest, and the low viscosity at
higher shear rate confers it a flowability in motion. Recently, the steady shear
viscosity of bacterial cellulose (BC) was compared with that of xanthan and LBG,
and it was found the BC solution was more shear thinning than xanthan solution
even at a lower concentration (Paximada et al. 2016). BC is also reported to be a
good suspending agent for chocolate drink preventing the precipitation of cocoa
particles, and in addition BC has a great heating stability after sterilization and thus
the viscosity remains unchanged after heat treatment (Shi et al. 2014). This potential
of BC was not exploited so much although it has attracted more attention as a unique
jelly dessert known as nata de coco or stabilizer of ice cream (Azeredo et al. 2019).
3 Rheological and Thickening Properties 85

The occurrence of shear thickening depends on the phase volume, particle size
(distribution), particle shape, as well as those of the suspending phase (Barnes 1989,
2000). The increase of the viscosity above a critical shear rate is ascribed to the
transition from a two- to a three-dimensional spatial arrangement of the particles.
After this transition, the viscosity decreases again with increasing further shear rate.
Shear thickening behavior in food systems has been overviewed (Bagley and
Dintzis 1999). Suchkov et al. (1997) reported a shear thickening behavior of 11S
broad bean globulin (legumin) in 0.6 mol/dm3 NaCl at pH 4.8 which was shown to
have an upper critical temperature of 21  C and a critical protein concentration of
18%. Dintzis et al. (1996) reported that waxy starches (corn (maize), rice, barley, and
potato) showed shear thickening to a greater extent than did wheat, normal rice or
normal corn starches when dissolved gently and dispersed at 3.0% concentration in
0.2N NaOH.
Shear thickening was observed for a 17.22% schizophyllan (Mw ¼ 450 kDa)
solution around a shear rate of 0.01–0.1 s
1 (Fang et al. 2004b). At the same shear
rate range, a steep change in the birefringence was observed indicating an abrupt
increase in molecular orientation. This is due to the extreme stiffness of
schizophyllan chain as mentioned above.
Recently, the classical most well-known corn starch suspension was revisited,
and shown that the so-called walking on water effect could be observed only above
52.5 wt% (42 vol%); a 2.1 kg rock laid on starch suspension with different concen-
trations fell, but a falling rock was bounced when it hit the surface of the starch slurry
of 52.5 wt% (Crawford et al. 2013).
Solutions of mamaku polysaccharide extracted from black tree fern were shown
to show the shear thickening behavior (Goh et al. 2007). Another example of shear
thickening behavior was reported for sulfated polysaccharides from seaweeds (Shao
et al. 2014).

4.2 Concentration Dependence of Viscosity

To understand the concentration dependence of the viscosity of polymer solutions, it

is necessary to take into account the molar mass, molecular conformation, and shear
rate dependence as is described in Chap. 2. The concentration dependence of
viscosity of flexible polymers is usually represented by the relation between zero
shear specific viscosity and the coil-overlap parameter defined by the product (C[η])
of the concentration (C) with the intrinsic viscosity [η]. Morris et al. 1981 Here, the
intrinsic viscosity is determined by the extrapolation of the specific viscosity to zero
concentration. The specific viscosity ηsp is defined as ηr
1, where the relative
viscosity ηr is defined as the ratio of the viscosity of the solution to that of the
solvent ηs: ηr ¼ η/ηs.
As shown above, most polymer solutions are non-Newtonian fluids and show a
shear thinning behavior. At sufficiently low shear rate, the viscosity does not depend
on the shear rate and shows a Newtonian plateau, so the viscosity observed at the
86 K. Nishinari

shear rate extrapolated to zero can be obtained, and it is called simply a zero shear
viscosity (η0).
As described above, some models have been proposed to interpret the shear rate
dependence of the viscosity of polymer solutions most of which are non-Newtonian
liquids. Cross model is widely used to fit the observed shear rate dependence where
both the zero shear viscosity η0 and the infinite shear viscosity η1 are observed:

η
η1 1
¼ ,
η0
η1 1 þ ðK γ_ Þm

where K has the dimension of time, and m is dimensionless. The value of m ranges
from 0 to 1, and represents the degree of shear thinning. When m is closer to zero it
tends to more Newtonian liquids, while the most shear thinning liquids have a value
of m tending to unity.
Some other models Carreau model, Sisko model, etc. are also used (see more
different models in Barnes (2000) and Lapasin and Pricl (1999)). Risica et al. (2010)
used Cross eq to analyze the shear rate dependence of guar gum and
hydroxypropylmethyl guar aqueous solutions.
The double logarithmic plot of the zero shear specific viscosity (ηsp0) of polymer
solutions and polymer concentration shows two straight lines. The slope of these
straight lines is smaller at lower concentrations than at higher concentrations, and the
crossover point C* of these straight lines shifts to lower concentrations with increas-
ing molar mass of a polymer.
Irrespective of chain flexibility, the slope at lower concentrations is reported as
ca.1 and the slope at higher concentrations as 3.3 for many polysaccharides, but
some other slopes are also reported and tabulated in Lapasin and Pricl 1999. When
the zero shear specific viscosity is represented by a power law (ηsp,0 ~ Cn), the
exponent (n) takes different values below and above C*. The exponent n for most
polysaccharide solutions ranges from 1.1 to 1.6 below C*, and from 1.9 to 5.6 above
C* (tabulated in Lapasin and Pricl 1999).

4.3 Salt Effect

The viscosity of polyelectrolyte solutions is influenced strongly by the presence of

salt. In contrast to the common procedure to determine the intrinsic viscosity of
non-electrolytic polymers, the reduced viscosity (ηsp/C) of polyelectrolyte solutions
shows a turn up in the extrapolation of polymer concentration. It is caused by the
polymer coil expansion by electrostatic repulsion. By adding salt, the reduced
viscosity decreases and the extrapolation to zero concentration becomes possible.
See, for example, the zero shear rate reduced viscosity vs. equivalent monomer
concentration of sodium pectate at various counterion concentrations (Lapasin and
Pricl (1999)). Smidsrød and Haug (1971) proposed an empirical method to deter-
mine the stiffness parameter B by determining the intrinsic viscosity at 0.1M ionic
strength (See Chap. 2). While the viscosity of flexible polyelectrolye solutions is
3 Rheological and Thickening Properties 87

decreased by the addition of salt, that of stiff chains such as xanthan is comparatively
insensitive to the addition of salt because the volume occupied by the stiff chain in
the solution is not so much reduced as in the case of flexible chains. This compar-
ative insensitivity/stability of the viscosity is one of the reasons why xanthan is used
widely in food processing as mentioned before.
Wyatt et al. (2011) reported that the zero shear viscosity of xanthan showed
different behaviors in the presence of salt depending on the polymer concentration.
They observed that the zero shear viscosity of dilute xanthan solutions (500 ppm and
50 ppm) decreased while that of concentrated solution (4000 ppm) increased signif-
icantly by the addition of salt (50 mM NaCl). They also observed a similar behavior
for anionic polysaccharides carrageenan and welan and also for a cationic polysac-
charide chitosan. They stated that these results could be understood by scaling
theories. Wyatt et al. (2011) found that Cox–Merz rule was valid for xanthan
solutions, which is contradictory with previous reports by Lee and Brant (2002)
and Richardson and Ross-Murphy (1987b), and they attributed this disagreement to
the difference in the concentration.

4.4 Viscosity of Suspensions

The viscosity of dilute suspensions is well described by Einstein’s equation η ¼

η0 (1 + 2.5φ), where η0 is the viscosity of a suspending medium, and φ represents the
volume fraction of the dispersed phase. Figure 3.5 shows the normalized viscosity

Fig. 3.5 Verification of

Einstein’s equation.
Reproduction with
permission from Hunter
(1998). Copyright 1998
Oxford University Press
88 K. Nishinari

(the ratio of the viscosity of the suspension to that of the solvent) vs. the volume
fraction for various materials. It is striking that this equation holds well irrespective
of the different nature of materials tested, spores, glass, and yeast. It is dependent
only on the volume fraction.
This equation does not hold, however, for concentrated suspensions, and various
modified equations have been proposed (Hunter 1998).

4.5 Extensional Viscosity

There have been many studies on the extensional viscosity but less than those on
shear viscosity because of the difficulty in the measurements. Methods of measure-
ments are stretching liquid between two flat plates, cylindrical viscoelastic filament
uniaxially extended by rotating clamp, crossed slot devices, and opposed jets
(Barnes 2000, p. 160; Larson 1999, p. 19). While the shear viscosity is defined by
ηs ¼ τ=_γ where τ is the shear stress and γ_ is the shear strain rate, the extensional
viscosity is defined by ηe ¼ σ=_ε where ε_ is the extensional strain rate. In Newtonian
fluids, it is known that ηe ¼ 3ηs. However, most food fluids are non-Newtonian, and
the ratio of ηe/ηs, which is called Trouton ratio, is generally greater than 3. When a
stick or a plate in a viscous liquid such as sugar syrup or honey is pulled up, the
liquid flows downward and does not flow upward because they are Newtonian fluids.
But, imagine the suspending of egg white just after cracking an eggshell or drooling
from the mouth of a baby. Both of these liquids egg white/ saliva flow downward but
then flow upward because of their elasticity. Egg white plays a role of cushion for an
egg yolk and saliva and other mucins in humans have their own biological function
which cannot be achieved if they are Newtonian liquids.
Chan et al. (2007) performed a comparative study of shear and extensional
viscosity of casein, waxy maize, and their mixtures. They measured the extensional
viscosity by observing the filament thinning when the liquid filament was extended.
Chan et al. (2007) found that the maximum stretchable length of casein, waxy maize,
and their mixtures were well described by a master curve using a capillary number
(the product of the viscosity and the stretching speed divided by the surface tension)
for viscous fluids, but the more concentrated samples showed a deviation from this
master curve indicating the important effect of viscoelasticity.
Extensional viscosity of thickening agents has attracted much attention recently
in relation with developing dysphagic treatments. Thickening agents are widely used
in hospitals to prevent the aspiration. Thickening of a thin liquid such as tea, juice, or
soup is effective (Cichero 2013). In addition to the increase of the viscosity, the
cohesiveness of the liquid is believed to be necessary for safe swallowing, and the
liquid extension test is suitable for this (Nishinari et al. 2019).
3 Rheological and Thickening Properties 89

5 Viscoelasticity

Simple mechanical models consisting of a spring which obeys the Hooke’s elasticity
law σ ¼ G ε (σ stress; G elastic modulus; ε strain) and a dashpot which obeys the
Newtonian viscosity law σ ¼ ηdε/dt (η viscosity) are used widely. A spring repre-
sents an ideal elastic body which deforms instantaneously (without delay) and
recovers to the initial state when the external force is removed. A dashpot is an
ideal viscous material where the shear stress σ is proportional to the shear rate dε/dt.
In the shear deformation, τ and γ are used for stress and strain in most textbooks.
Viscoelasticity is a relaxational phenomenon which depends on the time scale of
the measurement. Typical viscoelastic phenomena are stress relaxation and creep. In
the former, when the constant strain is given to a viscoelastic material, the induced
stress decreases with the lapse of time, and it is called stress relaxation. In the latter,
when a viscoelastic material is subjected to a constant stress, the induced strain
increases with the lapse of time, and it is called creep. In these experiments, the
stimulus causing the response is step-like and kept constant (does not depend on the
time), and then, this is called static viscoelasticity.
A simple model consisting of a spring and a dashpot combined in series is called a
Maxwell element. When a stress σ is applied to this model, the total strain ε is the
sum of strains of each element, spring, and dashpot, and the total stress is equal to the
stress of each element σ ¼ σ 1 ¼ σ 2 because they are combined in series. Therefore, σ
¼ Gε1 ¼ ηdε2/dt, where the suffixes 1 and 2 refer to the spring and dashpot,
respectively. Substituting these into ε ¼ ε1+ε2, the relation between stress and strain
(called constitutive equation) for a Maxwell element is obtained: dε/dt ¼ (1/G)(dσ/
dt) + (σ/η). A Maxwell model represents a liquid-like viscoelasticity because it
shows an infinite deformation because of the dashpot. When this viscoelastic
material is subjected to a step strain at time t ¼ 0, the stress is decreased with the
lapse of time. This phenomenon is called stress relaxation. The above Maxwell eq
gives the solution for this phenomenon, σ(t) ¼ ε 0 Ge-t/τ, where τ ¼ η/G is called the
relaxation time, indicating that the stress decreases exponentially.
A simple model consisting of a spring and a dashpot combined in parallel is called
a Kelvin–Voigt element. When a stress σ is applied to this model, the total stress σ is
the sum of stresses of each element, and the total strain is equal to the strain of each
element, ε ¼ ε1 ¼ ε2 because they are combined in parallel. Therefore, σ 1 ¼ Gε1 and
σ 2 ¼ ηdε2/dt, respectively. Substituting these into σ ¼σ 1+σ 2, the constitutive equa-
tion for a Kelvin–Voigt element is obtained: σ ¼ Gε + ηdε/dt. A Kelvin–Voigt
model represents a solid-like viscoelasticity in spite of the presence of a dashpot
because it is combined in parallel with the spring which shows only a finite
deformation. When a viscoelastic material is subjected to a step stress σ0 at time
t ¼ 0, the strain is increased with the lapse of time. This phenomenon is called creep.
The above Kelvin–Voigt eq gives the solution, γ(t) ¼ (σ 0/G) [1-exp(-t/τr)], where τr
¼ η/G is called retardation time. The retardation time is the time at which the strain
reaches (1-1/e) times the final strain (σ 0/G) after an infinite time.
90 K. Nishinari

Fig. 3.6 Strain vector ε*

and stress vector σ* on the
complex plane. ε* rotates
around the origin with
constant angular velocity ω,
while σ* also rotates with
the same ω with a leading
phase δ. σ// and σ┴ show
stress components with the
same phase and the leading
phase to ε*

In real material, the two element model such as Maxwell element and Kelvin–
Voigt element cannot describe well the rheological behavior. Assume that the strain
ε(t) of the material subjected to the constant stress σ0 at time t ¼ 0 is proportional to
σ 0, then ε(t) ¼ σ0 J(t) where J(t) is called creep compliance. When J(t) does not
depend on σ 0, this material behavior is called linear. If σ 0 is doubled, ε(t) induced by
the stress is also doubled in this case.
J(t) consists of three parts:

J ðt Þ ¼ J 0 þ J d φðt Þ þ t=η ðÞ

where J0 is the instantaneous compliance which obeys Hooke’s law, t/η is the
viscous flow following the Newton’s law, and the Jd φ(t) is the retarded elasticity,
and φ(t) is called creep function, φ(0) ¼ 0, φ(1) ¼ 1. This is generally valid
irrespective of the model. The creep function is given by φ(t) ¼ 1
exp(
t/τr).
These parameters can be determined graphically.
Dynamic Viscoelasticity
In the dynamic viscoelastic measurements, the viscoelastic material is subjected to
the sinusoidal strain or stress, and the induced sinusoidal stress or strain is observed.
The oscillational stress and strain can be represented in complex plane as vectors
rotating at an angular velocity ω as shown in Fig. 3.6, where δ is the phase
difference.
The shear strain is written as γ* ¼ γ 0 exp(iωt) ¼ γ 0 eiωt, and the shear stress is σ*
¼ σ 0 ei(ω t + δ) . The complex elastic modulus is defined by G* ¼ σ*/γ*. Therefore,
G* ¼ (σ0/γ0)eiδ ¼ (σ0/γ0) (cosδ + i sinδ).
When G* is written as G0 + iG00 , the real part G0 is called storage modulus, and the
imaginary part G00 is called loss modulus.

G0 ¼ ðσ 0 =γ 0 Þ cos δ, G00 ¼ ðσ 0 =γ 0 Þ sin δ

The ratio G00 /G0 is called mechanical loss tangent tan δ ¼ G00 /G0 , which is related
to the energy loss as shown below.
Complex viscosity is defined by
3 Rheological and Thickening Properties 91

η* ¼ G*=iω ¼ η0
i η00

And its absolute value is |η* | ¼ [(G0 )2 + (G00 )2]1/2/ω

η0 ¼ G00 /ω, η00 ¼ G0 /ω
For some polysaccharide solutions, the so-called Cox–Merz rule, which states
that the complex viscosity plotted vs angular frequency agrees with the steady shear
viscosity plotted vs shear rate, holds as is shown in Fig. 3.4b. But, this rule is an
empirical rule with no sound theoretical validation, and many deviations are also
reported.
Dynamic Behavior of Maxwell Element and Kelvin–Voigt Element
When a sinusoidal strain ε0eiωt is given to a Maxwell element, the frequency
dependence of complex modulus is given by

iωτG
GM *ðiωÞ ¼
1 þ iωτ

and the storage and loss moduli are

ω2 τ 2
G M 0 ð ωÞ ¼ G
1 þ ω2 τ 2
00 ωτ
G M ð ωÞ ¼ G
1 þ ω2 τ 2

When a sinusoidal stress σ 0eiωt is given to a Kelvin–Voigt element, the frequency

dependence of complex compliance is given by

1 1
J KV *ðiωÞ ¼
G 1 þ iωτ

Loss Modulus is Related to the Energy Loss

Question: Calculate the energy dissipated in a viscoelastic material during one
period of oscillation, and show that it is proportional to G00
Answer

2π 2π 2π
Zω Zω Zω
dγ π
η γ_ dt ¼ η γ_ 2 dt ¼ ηðγ 0 ω cos ωt Þ2 dt ¼ ηω2 γ 0 2 ¼ πγ 0 2 ωη ¼ constG00
dt ω
0 0 0

Mechanical Spectra: Frequency Dependence of Storage and Loss Moduli

Frequency dependence of storage and loss moduli for polymer solutions or colloid
dispersions can be classified into four categories as shown in Fig. 3.7. In the dilute
solutions where polymers dissolve with little or no overlap with each other, G0 < G00
92 K. Nishinari

Fig. 3.7 Typical angular frequency ω dependence of G0 and G00 of commonly used thickening
agents. It should be noted that values of G0 and G00 depend not only on the concentration but also on
the molar mass and conformation of hydrocolloids or the coil-overlap parameter

and at very low frequencies it is seen that G0 ~ ω2, G00 ~ ω. At higher concentrated
solutions G0 < G00 at lower frequencies, but at higher frequencies G0 > G00 and thus at
an intermediate frequency the crossover G0 ¼ G00 is observed. This is interpreted as
follows: at higher frequencies entangled polymer chains play a knot of temporary
network and thus somewhat a solid-like behavior is observed, but at lower frequen-
cies, the time of oscillational period is long enough for chains to disentangle, and
thus it shows a liquid-like behavior G0 < G00 . The transition from the dilute region to
entangled region is seen by the coil-overlap parameter ¼ intrinsic viscosity 
polymer concentration as mentioned before. For networks or elastic gels, G0 > G00
at a wider frequency range and tan δ < 0.1. Between an elastic gel and concentrated
(entangled) polymer solutions, another frequency dependence is observed, and it is
called a structured liquid. This frequency dependence is also seen in globular protein
solutions. Previously, it was called a weak gel. But, as will be discussed in detail in
the next chapter, it is essentially a liquid and flows above a yield stress, which is
discussed later in this chapter.
Time–Temperature Superposition
As is well known a material behaves like a liquid for a slow deformation, and shows
a solid-like behavior for a fast deformation (see e.g. https://en.wikipedia.org/wiki/
Pitch_drop_experiment). We also know that many solids become soft or firm when
heated or cooled. For many synthetic polymers, the data obtained at low (high)
temperatures are equivalent to those obtained at a high (low) frequency or a short
(long) time scale, and then it was found that the data can be superposed by shifting
horizontally (parallel to the time or temperature axis). When such a superposition
between time and temperature is possible, it is called thermorheologically simple,
and the resulted curve is called a master curve. The time–temperature superposition
was successfully applied to many synthetic polymers, but great caution is needed to
apply to food biopolymers.
3 Rheological and Thickening Properties 93

Lissajous–Bowditch Figure and LAOS (Large Amplitude Oscillatory Shear)

When the sinusoidal stress and strain, σ ¼ σ 0sinωt and γ ¼ γ0sinωt, are taken as
x-axis and y-axis in the orthogonal coordinate system, the Lissajous–Bowditch
figure is obtained. In the elastic material, the stress and strain are in phase, and the
Lissajous–Bowditch figure is a straight line. In the viscous material, the stress and
strain are π/2 out phase, and the Lissajous–Bowditch figure is a circle. In the
viscoelastic material, the stress and strain are δ out of phase, and the Lissajous–
Bowditch figure is an ellipse.
Generally, the linear viscoelastic range is wider for flexible linear polymer
solutions than for structured liquids. Beyond the linear viscoelastic range, the
resulted Lissajous–Bowditch figure is skewed. While the deviation from the ellipti-
cal shape is very small for a concentrated polysaccharide solution (hydroxyethyl
guar gum), a pronounced distortion was recognized for a structured liquid
(sc1eroglucan) particularly at low frequency (Lapasin and Pricl 1999).
To understand quantitatively the non-linear viscoelastic behavior of food mate-
rials, large amplitude oscillatory shear (LAOS) has recently been attracting much
attention. LAOS behaviors are often shown by the strain dependence of G0 and G00 at
a constant angular frequency ω, and are classified into four types: (1) strain thinning
(G0 and G00 decreasing); (2) strain hardening (G0 and G00 increasing); (3) weak strain
overshoot (G0 decreasing, G00 increasing followed by decreasing); (4) strong strain
overshoot (G0 and G00 increasing followed by decreasing) (Hyun et al. 2011).
However, this presentation cannot capture and clarify the whole characteristics of
non-linear rheology. Chebyshev polynomial presentation has been proposed to get
more physical meaning from LAOS data (Cho et al. 2005; Ewoldt et al. 2008).
The elastic (σ 0 ) and viscous (σ") stress are given by

σ 0 ðxÞ ¼ γ 0 ∑ en ðω, γ0 Þ T n ðxÞ

n: odd

σ 00 ðxÞ ¼ γ 0 ∑ vn ðω, γ0 Þ T n ðyÞ

n: odd

where Tn (x) is the nth order Chebyshev polynomial of the first kind, T1(x) ¼ x, T3(x)
¼ 4x3
3x, T5(x) ¼ 16x5
20x3 + 5x, ⋯
and x ¼ γ/γ 0, y ¼ γ_ =γ_ 0 , provide the appropriate domains of [
1, +1] for
orthogonality, en (ω, γ0) and vn (ω, γ0) are elastic and viscous Chebyshev
coefficients.
In the Fourier representation of LAOS, the stress response σ (t; ω, γ0) to the strain
stimulus γ (t) ¼ γ_ 0 sin ωt or strain rate stimulus γ_ ðtÞ ¼ γ_ 0 sin ωt is given by
X
σ ðt; ω, γ 0 Þ ¼ γ 0 G0n ðω, γ 0 Þ sin nωt þ G00n ðω, γ 0 Þ cos nωt g,
n odd

X
σ ðt; ω, γ 0 Þ ¼ γ_ 0 η00n ðω, γ 0 Þ sin nωt þ η0n ðω, γ 0 Þ cos nωt g:
n odd
94 K. Nishinari

The relationships between the Chebyshev coefficients in the strain or strain rate
domain and the Fourier coefficients in the time domain are given by

en ¼ G0n ð
1Þðn
1Þ=2 n : odd,

vn ¼ G} n =ω ¼ η0n n : odd,

In the linear regime at small strains, e3/e1 <<1 and v3/v1<<1, and the linear
viscoelastic relations

e1 ! G0 and v1 ! η0 ¼ G00 =ω:

are obtained. The strain-stiffening ratio S and the shear thinning ratio T are defined as
follows:
 
S ¼ G0L
G0M =G0L ¼ ð4e3 þ . . . :Þ=ðe1 þ e3 þ Þ
 
T ¼ η0L
η0M =η0L ¼ ð4v3 þ . . . :Þ=ðv1 þ v3 þ Þ

where G0 L and G0 M are large strain modulus and minimum strain modulus
respectively;

dσ  X
G0M   ¼ nG0 ¼ e1
3e3 þ ⋯,
dγ γ¼0 n odd n

σ X
G0L   ¼ G0n ð
1Þðn
1Þ=2 ¼ e1 þ e3 þ ⋯,
γ γ¼γ n odd
0

For a linear elastic response, S ¼ 0 and S > 0 means intracycle strain stiffening.
For a linear elastic response, T ¼ 0 and T > 0 means intracycle shear thickening.
LAOS studies on dough and gluten gels have been done using this method
(Ng et al. 2011; Yazar et al. 2017a).
The characteristic shapes of Lissajous–Bowditch figures for a gluten gel over a
range of strain amplitudes from γ 0 ¼ 0.02 to 6.0 at a fixed angular frequency at ω ¼
1.0 rad s
1 are plotted in Fig. 3.8 (Ng et al. 2011).
At smaller strain amplitudes, the relation between the stress and strain appears as
an ellipse:
 
σ 2
2σγG0 þ γ 2 G0 þ G00 ¼ G00 γ 20 ,
2 2 2

This quadratic form can be diagonalized, and the major and minor axes of
the ellipse are centered at the origin. The enclosed area is given by πγ 02G", indicating
the energy lost during one cycle of oscillation. As the strain amplitude is increased,
the material response and shape of the Lissajous–Bowditch curves shown in Fig. 3.8
deviate significantly from this simple behavior. First a gradual “softening” of the
3 Rheological and Thickening Properties 95

Fig. 3.8 Lissajous–Bowditch curves after 12 oscillatory cycles for a gluten gel at a fixed angular
frequency ω ¼ 1.0 rad s
1 with (a) γ 0 ¼ 0.02, 0.05, 0.10; (b) γ 0 ¼ 0.10, 0.20, 0.50; the curve for
γ 0 ¼ 0.10 from (a) is repeated. (c) γ 0 ¼ 0.50, 1.00, 2.00; (d) γ 0 ¼ 2.0 and 6.00. As the imposed strain
amplitude γ 0 increases from (a) to (d), the magnitude of the maximum stress grows and the axes are
rescaled. Experimental data are plotted as open symbols. The decomposed elastic stresses are shown
in the lower panel as a dotted line for γ 0 ¼ 6. Predictions from the nonlinear generalized gel model
are plotted as a solid line for each strain amplitude. Reproduction with permission from Ng et al.
(2011). Copyright 2011 AIP

material is indicated by the clockwise rotation of the major axis toward the strain-
axis. Second, a distinct “stiffening,” indicated by the upturn of the shear stress, is
observed at large strains (Fig. 3.8d). The magnitude of the enclosed area also
increases with increasing amplitude, indicating an increasingly dissipative response.
These non-linear features cannot be fully captured by simply reporting G0 and G00 as
a function of strain amplitude, as is usually done in linear viscoelasticity analysis
(Ng et al. 2011).
Duvarci et al. (2017) applied this method to suspensions, emulsions, and elastic
networks (tomato juice, mayonnaise, soft and hard dough). In the strain dependence
of G0 and G00 at a constant angular frequency, tomato paste showed the type (I),
strain thinning (G0 and G00 decreasing) while mayonnaise showed the type (III), weak
96 K. Nishinari

strain overshoot (G0 decreasing, G00 increasing followed by decreasing). A slight

strain softening (S < 0, e3/e1 < 0) at smaller strains and strain hardening at larger
strains were observed for tomato paste and mayonnaise. Strain hardening (e3/e1 > 0)
and shear thinning (v3/v1 < 0) were observed for soft dough and hard dough.
Biaxial deformation has been widely used in baking investigation, and gluten
doughs showing high resistance to biaxial deformation and pronounced strain
hardening behavior were found to be good for baking, i.e., larger bread volume
(Kokelaar et al. 1996). Bread volume has been used as an index of the bread forming
performance of dough. Recently, bread volume of gluten-free flour doughs were
plotted as a function of large strain modulus G0 L or minimum strain modulus G0 M,
and it was found that the bread volume increased with increasing G0 L and or G0 M
(Yazar et al. 2017b).

6 Yield Stress

The concept of yield stress is intuitively evident: most fluid foods filled in tubes such
as mayonnaise, mustard, ketchup, pastes, etc. begin to flow only when they are
subjected to a certain value of stress. The minimum stress required to cause flow is
the yield stress. The first simplest model is a Bingham fluid: σ ¼ ηB (dγ/dt) + σ y,
where σ y is the yield stress, ηB is the Bingham plastic viscosity. Another simple
model, Herschel–Bulkley fluid σ ¼ ηHB (dγ/dt)n + σ y has also been used for better
describing the behavior.
Yield stress governs the thickness of coated chocolate on the biscuit as shown
below (Fig. 3.9).

Fig. 3.9 Chocolate coating

on biscuit. The yield stress
of chocolate dictates the
thickness of chocolate
3 Rheological and Thickening Properties 97

The gravitational force exerting on the melted chocolate f is given by f ¼ mg¼

Vρg, where m is the mass, V¼ a  b  c is the volume, ρ is the density of the
chocolate. From the definition of the yield stress, this gravitational force should
balance with the yield stress  surface area ¼ τy  a  b
Therefore, a  b  c  ρ  g ¼ τy  a b
The thickness c of the chocolate is c ¼ τy/ρg
For a chocolate with τy ¼ 5 Pa, ρ ¼ 1.06  103 kg/m3, the thickness of the
chocolate is given by
 
c ¼ 5 N m
2 = 1:06  103 kg=m3  9:8 m s
2 ¼ 0:00048 m ¼ 0:48 mm:

Yield stress can in principle be determined by decreasing the shear rate in the plot
of the shear stress vs shear rate. Then, the shear stress obtained at zero shear rate is
the yield stress. Rigorously speaking, it is impossible to determine the absolute yield
stress which is conceptually the idealization. In a very long time scale everything
flows even at a very low stress (Barnes 1999).
Although it is not easy to measure, the concept of the yield stress is meaningful in
food science and technology, and it should be reminded that the yield stress values
obtained by different methods can be compared only over the same range of shear
rates (Barnes 1999; Bonn et al. 2017).
Many papers have been published on the existence of the true yield stress. Using
four model yield stress materials, 0.2% carbopol, a commercial hair gel (containing
carbopol, water triethanolamine as a stabilizing agent), a commercial shaving foam,
and a cosmetic water-in-oil emulsion (composed of 80% water with carbopol
940 (1%) in Vaseline oil (6%) and Polyethylene glycol 600 (6%) with several
additives, so that it remains stable under shear when used), Mϕller et al. (2009)
showed that the yield stress really exists and it marks a transition between the fluid
which flows and the solid which does not flow. These four samples are called
“simple” yield stress materials because the viscosity depends only on the shear
rate, and the yield stress is a material property while for thixotropic yield stress
materials the viscosity depends also on the shear history, which is discussed in the
next Sect. 7. “Apparent” viscosity ¼ stress/(instantaneous shear rate) which is time
dependent and therefore not “true” viscosity, of all the four materials as a function of
stress showed a steep rise with decreasing stress and then leveled off to show a
plateau value below a certain stress. The plateau value of the apparent viscosity
increased with the measurement time up to ~103 s. Fig. 3.10a shows the apparent
viscosity of 0.2% carbopol as a function of time for different stress values.
Above the stress 27 Pa, the apparent viscosity quickly reaches the steady value
and then it stays constant up to ~103 s, while below the stress 25 Pa, the apparent
viscosity continues to increase even at 104 s. Therefore, above the stress 27 Pa, the
material flows at a constant viscosity which is a decreasing function of the stress,
while the apparent viscosity continues to increase with time as η ~ t0.6 as indicated by
the dashed straight line, indicating that the system behaves almost as a solid having a
very large viscosity within the experimentally accessible observation time. Thus, this
stress 27 Pa can be defined as a yield stress. The same phenomenon was also
observed for other three different materials. Below a critical stress, the viscosity
98 K. Nishinari

Fig. 3.10 (a) Apparent viscosity of 0.2% carbopol as a function of time at different stresses. At and
above the stress 27 Pa, the apparent viscosity η of the sample shows a constant value (time
independent) while at and below the stress 25 Pa, η increases with increasing time η ~ t0.6 as
indicated by the dashed straight line. (b) A creep compliance J(t) of the same data as in left figure.
At large t a fluid state will have a slope of 1 (indicated by the dotted line), while a solid state will
have a slope of 0. At stresses of 27 Pa and above, slopes are nearly 1, while they are nearly 0 at 25 Pa
and below—data for six stresses are collapsed and hardly discernable. Reproduction with permis-
sion from Mϕller et al. (2009). Copyright 2009 IOP

increases in time; the material eventually stops flowing. Above the critical stress, the
viscosity decreases continuously in time; the flow accelerates. Thus the viscosity
jumps discontinuously to infinity at the critical stress. This phenomenon is thus
called viscosity bifurcation (Coussot et al. 2002a, b)
As shown clearly in Fig. 3.10b, at and below the stress 25 Pa the deformation was
zero indicating the solid behavior below this critical stress (yield stress), while the
compliance increases with increasing time with the slope 1 indicating the fluid
behavior at and above the stress 27 Pa. Both Fig. 3.10a, b show clearly the existence
of the yield stress, and this is also recognized also in the other three different yield
stress materials, a hair gel, a shaving foam, and a water-in-oil emulsion.
Many methods have been proposed to determine the yield stress. Dinkgreve et al.
(2016) using the same four model simple yield stress materials compared the yield
stress values, and concluded that the Herschel–Bulkley fit to the stress-shear rate
gives the lowest and the most reliable value for the yield stress. Vane geometry is
sometimes useful to determine the yield stress because it is free from slippage
(Stokes and Telford 2004).

7 Thixotropy

The viscosity of many structured liquids at a constant shear rate decreases with the
lapse of time, which is called thixotropy. It is different from shear thinning behavior
which is defined as the decreasing of the viscosity when the shear rate is increased.
3 Rheological and Thickening Properties 99

Fig. 3.11 (a) Shear stress transients in step shear rate tests for 6 wt% gum arabic in water. The
shear rates of different steps were shown in the panel (b) Time evolution of the transient shear stress
in sudden reduction of shear rate from 1000 s
1 to different lower shear rates for 6 wt% gum arabic
solution. The different lower shear rates were shown in the panel. Reproduction with permission
from Li et al. (2011). Copyright 2011 Elsevier

All the shear thinning liquid show a thixotropic behavior since it takes a time to
recover the initial microstructure even if it is short.
Thixotropy is a time dependent rheological phenomenon, and it is understood as
the microstructural formation at rest and the breakdown of that structure during
shearing. The thixotropy is explained by the competing two processes aging (struc-
ture formation) and rejuvenation (structure breakdown).
Delayed failure of fish myosin gels was studied, and the time to failure as a
function of scaled applied stress showed two regimes (Brenner et al. 2013). This time
depends on the bond reformation rate; the time is short when the stress is high and
bond reformation is not sufficient while it is long when the stress is low and bond is
reformed (Sprakel et al. 2011; Lindstrom et al. 2012).
A hysteresis loop test which records a stress–shear rate curve is a simple dem-
onstration of the thixotropic behavior and has been used widely for many foodstuffs
such as yogurt. Although a loop test, shear stress vs shear rate, has been frequently
used to study thixotropic behavior because it is easy to do, Barnes (1997) depreciated
this method. Though it gives some qualitative information, he recommends the
stepwise experiments where the shear rate or stress is changed from one constant
value to another with a carefully controlled prehistory. The advantages of the
stepwise change of shear rate in comparison with the hysteresis loop method are
that not only the initial condition can be well-defined and reproducible, but also the
shear rate during the actual test remains constant. Hence, the effect on an established
structure of shearing at a fixed shear rate can be measured as a function of time
(Mewis and Wagner 2012).
Figure 3.11 shows an example of such an experiment for solutions of gum Arabic
which is widely used as emulsifiers and stabilizers in food industry. When the shear
rate is increased/decreased in a step-like manner, the induced stress shows an
overshoot/undershoot, and then decreases/increases to a stationary value. The tran-
sient stress shown in Fig. 3.11 can be written
100 K. Nishinari

1
σ¼h   i2
p1ffiffiffi þ p1ffiffiffi
p1ffiffiffi
σs σi σ s exp ð
kt Þ

where σ i and σ s represent the initial and stationary values of the stress, respectively,
and k is a kinetic constant with k ¼ kf + kb. Here, kf and kb are rate constants, which
modulate the kinetics of structural buildup and breakdown processes, respectively.
The rate constant kf depends on temperature, whereas kb is a function of the applied
shear rate, and t is the testing time. Although old textbooks describe a gum Arabic
solution as a Newtonian solution, the recent improvement of the rheometry makes it
possible the measurement at low shear rates. The deviation from the Newtonian
behavior at low shear rates (<10 s
1) suggests the existence of the molecular
aggregation which is broken down by shear flow. The above equation was also
used by Grassi et al. (1996) for scleroglucan and by Fang et al. (2004a) for
schizophyllan. Oleogel consisting of rapeseed oil and shellac (a resin secreted by
an insect) also showed a thixotropic behavior (Patel et al. 2013). This indicates that
an oleo gel is a structured liquid and not a gel as defined in the next chapter. Actually,
this oleo gel shows a flow behavior without fracturing when it is subjected to shear
which is widely observed for structured liquids with yield stress such as xanthan,
welan, rhamsan, schizophyllan, and scleroglucan.
The stepwise experiment to examine the thixotropic nature of low-fat mayon-
naise-like emulsion gels, containing egg yolk, rapeseed oil (30% instead of 80% in
full-fat mayonnaise), salt, sugar, vinegar, and 4.0% alginate solution and KGM
powder, was performed (Yang et al. 2020). Structural recoverability was found for
the low-fat mayonnaise-like emulsion gels in the stepwise experiment, and the
stackability was enhanced by adding small amount of calcium ions. Stackability is
required for mayonnaise, and is related with yield stress.
The opposite behavior of thixotropy is called antithixotropy where start-up flow
or a sudden increase in shear rate causes an increase in viscosity over time (Mewis
and Wagner 2012).
The dilute solution of globular protein shows another unique aspect, an
antithixotropic rheological behavior; the viscosity of beta-lactoglobulin solutions
at a fixed low shear rate increased with the lapse of time (Renard et al. 1996).

8 Microrheology

In the conventional or bulk rheology, the sample size is generally mm order and
1 cm3 in volume, and therefore it is difficult to get the information of local region of
inhomogeneous sample. Microrheology is useful to examine the rheological prop-
erties of a local region in inhomogeneous materials such as cytoplasm, thick egg
white, and emulsion gels. The idea to determine the elasticity and viscosity in
inhomogeneous materials dates back to Freundlich and Seifriz, and Heilbronn in
1920s, and development of theories on Brownian motion and experimental
3 Rheological and Thickening Properties 101

techniques, electronics, spectroscopy, light scattering, thereafter made the method

more quantitative.
There are two types of measurements in microrheology: passive and active
methods. The microrheological information obtained by these two methods are
equivalent by virtue of the fluctuation–dissipation theorem (FDT) in the equilibrium
state (Doi 2013; Waigh 2016; Yang et al. 2017).
In the passive microrheology, the mean square displacement (MSD) of the probe
particle is detected and analyzed by video tracking, laser tracking, and diffusing
wave spectroscopy.
Multi-particle tracking microrheology was performed to obtain local elasticity
and heterogeneities of gellan microgels using fluorescent colloidal particles with
diameter of 0.5 μm (Caggioni et al. 2007).

9 Fractional Calculus Bridging the Instrumental

Measurement and Sensory Evaluation

9.1 Scott Blair’s Approach to Understand the Firmness

Judged by Humans

Intermediate materials between elastic solid and viscous fluid such as cheese cannot
be characterized only by modulus or by viscosity. In most food rheology, the
intermediate state of the elasticity and viscosity has been studied as viscoelasticity
where the rheological behavior of such materials is analyzed by static or dynamic
viscoelastic measurement such as creep, stress relaxation or oscillation measure-
ment, and large deformation and fracture mechanics have also been studied. Scott
Blair proposed alternative method to analyze the rheological behavior of foods and
other industrial materials by virtue of the “principle of intermediacy” (Scott Blair
1969). Instead of the complex plane representation of the storage modulus and loss
modulus, he proposes a Cartesian coordinate consisting of the modulus G ¼ σ/ε and
the viscosity η ¼ σ/(dε/dt) to explain the “principle of intermediacy.” An interme-
diate material between the elastic solid and the viscous liquid can be represented in
this plane as a vector φ ¼ dkε/dtk, where k is a fraction and dkε/dtk is a fractional
derivative which Scott Blair introduced into rheology to understand the relation
between the sensory assessment and the instrumental measurement. The elastic
material with k ¼ 0 is judged by the strain ε and the viscous material with k ¼ 1 is
judged by the strain rate, that is d1ε/dt1, and therefore, it is reasonable to think that
the fractional differentiation of the strain dkε/dtk should be used for the judgment of
the firmness for the materials with 0 < k < 1 intermediate between the elastic
material and the viscous material.
To represent the intermediate state of the elasticity and viscosity, the quasi-
property χ was defined as (Scott Blair 1947)
102 K. Nishinari

 
χ ¼ σ= d k ε=dt k

where dnε/dtn¼ψ
1 σ k(k-1) (k-2)⋯(k-n+1) t(k-n) ¼ ψ
1 σ t(k-n) Γ(k+1)/ Γ(k-n+1),

where, Γ(k-n+1) is a gamma function

Z1
Γ ð k þ 1Þ ¼ e
z zk dz ¼ k ΓðkÞ ¼ kðk
1Þ ðk
2Þ⋯ðk
n þ 1Þ
0

Although most physicists had not agreed to use “quasi-properties,” Reiner

supported Scott Blair’s concept.
The fractional calculus has been used to discuss the rheological properties of
various soft materials (Koeller 1984; Jaishankar and McKinley 2012; Rogosin and
Mainardi 2014). The mechanical element having an intermediate behavior of the
spring and dashpot is called Scott Blair element or springpot model having a
constitutive equation

dβ γ
σ¼G ð 0 < β < 1Þ
dt β

Therefore, from the experimental observation of J(t), or G(t), or G0 (ω) and G00 (ω),
parameters G and β of springpot model can be determined.

9.2 Recent Development of the Application of Fractional

Calculus to Liquid Foods

Wagner et al. (2017) based on fractional calculus studied the rheological properties
of thickening agents used in dysphagia. Fractional Maxwell model (FMM) and
Fractional Jeffereys model (FJM) were used. FMM is a series combination of two
springpots with quasi-properties V, α and G, β, and it is reduced to a spring or
dashpot when α ¼ 0 or α ¼ 1, respectively. FJM is a parallel combination of FMM
and a dashpot with the viscosity ηs.
The steady shear viscosity can be also written using these parameters. Wagner
et al. (2017) used the Cox–Merz rule to correlate the small amplitude oscillational
data of G0 and G00 with the steady shear viscosity, and they found generally a good
agreement although they needed some corrections taking into account the viscosity
at higher shear rate. They fitted the experimental data of polysaccharide thickening
solutions used for dysphagic treatment, a commercial thickener TUC and saliva,
using the above equations, and obtained the parameters V, α, G, β and ηs of fractional
models. They replaced G with K, and β with n, where K is the so-called consistency
index of the fluid in the power law fluids (Curves 2 and 3 in Fig. 3.3a). Instead of η in
power law models (2) and (3) in Fig. 3.3a, K is often used and called consistency
index (with the unit of Pa sn), while n is called flow behavior index (dimensionless).
3 Rheological and Thickening Properties 103

Fig. 3.12 Fractional parameter phase space for food consistency. The markers correspond to the
values of the quasi-property K and fractional exponent n best describing the shear thinning in the
steady shear viscosity of the TUC solutions, polysaccharides, and whole saliva at γ_ ¼50 s
1. The
dashed diagonal lines bordering the shaded regions denote isoviscosity curves at γ_ ¼50 s
1
corresponding to the “thin” (0.001–0.05 Pa s) “nectar-like” (0.05–0.35 Pa s), “honey-like”
(0.35–1.75 Pa s), and “pudding-like” (>1.75 Pa s), preparations of starch solutions by the National
Dysphagia Diet Task Force. Reproduction with permission from Wagner et al. (2017). Copyright
2017 Elsevier

They analyzed the thickening behavior of various polysaccharides and a com-

mercially available thickening agent TUC using fractional calculus power law model
using the Cox–Merz rule

nðγ_ Þjγ_ ¼ω ¼ jη*ðωÞj ¼ Kωn‐1

In the fractional calculus treatment of viscoelasticity, K is called quasi-property.

They found an excellent fitting using this equation for thickeners and plotted the data
in the K-n plane (Fig. 3.12). K is taken as an abscissa and n is taken as an ordinate.
The exponent n ¼ 1 corresponds to the purely Newtonian viscous liquid, and n ¼
0 corresponds to a Hookean elastic solid. Each point in the interior of this parameter
space in the range 0 < n <1 represents viscoelastic power law fluids.
In this K-n plane, equations of the dashed diagonal straight lines are

log η log K
n¼1þ

log 50 log 50

where η ¼ 0.001, 0.05, 0.35, and 1.75 from the left to right, and the slope of the
diagonal straight lines is
1/log50.
As is seen clearly from Fig. 3.12, it is evident that the characterization of the
“thickness” of these fluids only by the viscosity is insufficient. Tara gum is close to
104 K. Nishinari

the n ¼ 1 Newtonian line, but the other fluids flax seed, whole saliva, guar gum, and
TUC are all below n ¼ 0.5, and elasto-plastic or yield stress like response is very
important. The importance of the elastic contribution for the dysphagia treatment is
in good agreement with the concept that cohesiveness of bolus is important for safe
swallowing (Nishinari et al. 2019). The advantage of the analysis based on fractional
analysis is that it contains only a few parameters which can be determined by the
well-defined rheological measurements.
Faber et al. (2017a, b) studied the texture of cheese using the fractional calculus.
They favored their approach based on the springpot model over the more undirected
approach of statistically correlating large amounts of rheological data to the results
from quantitative descriptive analysis (QDA) of multiple texture attributes. Their
reservations to the latter approach are for two reasons. First the deformations and
observations in QDA that lead to the texture judgment are ill-defined and may vary
from subject to subject. This a priori weakens correlations with the measurements
obtained from the carefully designed rheological experiment. Their second argument
to favor the fractional calculus approach is that material properties are intrinsic
properties whereas texture attributes are extrinsic in nature (Reiner 1971).

10 Further Developing of Thickening Properties

10.1 Viscosity of Mixed Hydrocolloids

The viscosity of the mixture of hydrocolloids has been studied from a long time ago.
The so-called synergistic interaction between xanthan and guar, the viscosity of
mixed solution as a function of mixing ratio showed a maximum, was reported
(Sanderson 1981). The word synergism is used when the viscosity of the mixture is
higher than the sum of the viscosities of the individual gum dispersions. However, in
most cases, the viscosity of the mixture is lower than the sum, for example, the
viscosity of mixtures of konjac glucomannan with xanthan, guar gum, carrageenan,
sodium alginate, methyl cellulose, hydroxymethyl cellulose, sodium carboxymethyl
cellulose, all the mixture except with xanthan showed the lower viscosity than the
sum of the individual polysaccharide (Liang et al. 2011).
Recently, Zhang et al. (2015) examined the interaction of corn fiber gum (CFG)
with various polysaccharides, hyaluronan (HA), guar gum (GG), carboxymethylcel-
lulose (CMC), hydroxyethyl cellulose (HEC), konjac glucomannan (KGM), pectin,
and chitosan using a shear stress synergy index Is, which is defined by

σ iþj
Is ¼ ,,
σi þ σ j

where σ iþj, σ i and σ j represent the shear stress for the mixture of i and j, i alone and
j alone in the flow curve σ  γ_ written as follows:
3 Rheological and Thickening Properties 105

Z γ_ 2
1
σ¼ σdγ_ ,
γ_ 2
γ_ 1 γ_ 1

(Pellicer et al. 2000). This index Is was used to judge whether the interaction is
synergistic or antagonistic. When Is > 1 it is judged as synergistic while when Is <
1 it is judged as antagonistic or lack of synergism. Zhang et al. (2015) found the
synergistic interaction, i.e. Is > 1 for CFG with HA, HEC, GG, KGM, pectin,
chitosan and concluded that CFG has a great potential as a viscosifying polysaccha-
ride. CFG itself shows a very low viscosity and almost a Newtonian behavior like
gum Arabic, while the interaction with other polysaccharides induces a greater
viscosity.
As for the temperature dependence of viscosity of thickening agents, xanthan
shows low temperature dependence and so stable, but in heat processing the viscos-
ity lowering is sometimes required so that the stirring force can be weak. Xyloglucan
(XG) is suitable for such requirement. Viscosity of tapioca starch (TS) suspension is
reported to be more heat resistant when xyloglucan was added (Pongsawatmanit
et al. 2006). The influence of temperature on the apparent viscosity of TS paste was
estimated by an Arrhenius plot ln (viscosity) vs 1/T. The activation energy Ea
determined from the slope of this plot for TS mixed with XG was found much
smaller than for TS alone, indicating that the addition of XG improved the heat
stability. It was also found that the water separation from freeze-thaw samples was
much reduced by XG.
Exactly the same reasoning was used in the same year 2006, to show the thermal
stability of the mixture of hydroxypropyl-substituted guar (HPG) and carboxymethyl
hydroxypropyl- substituted guar (CMHPG) in comparison with each individual
HPG and CMHPG (Zhang and Zhou 2006). Zhang and Zhou found that the
activation energy was smaller for the mixed CMHPG/HPG solution indicating that
the mixture showed the improved temperature tolerance with respect to the viscosity.
They also found that the viscosity of the 1:1 mixed solution was higher than that of
each individual solution.

10.2 Molecular Structure and Viscosifying Function

The beneficial effect of dietary fibers is recognized well, and this function is
generally believed to be conferred by high viscosity. However, in the supplement
utilization, too high viscosity hinders the sufficient intake of dietary fibers, and
therefore enzymatically degraded guar gums, degraded sodium alginate, dietary
fiber from psyllium seed husk, and other polysaccharides have been commercialized
(www.mhlw.go.jp/english/topics/foodsafety/fhc/02.html).
Cellulose chemists have modified the structure of cellulose by adding some
residues at C2, C3, and C6 thus controlling the solubility and viscosifying properties.
106 K. Nishinari

Similar trials have been made for various polysaccharides using chemical and
enzymatic modifications.
Recently, new variants of xanthan were created using genetic engineering
(Wu et al. 2019). Since the shear rate dependence of these variants shows quite a
different behavior, the further developments are expected.
Zhao et al. (2017) found that a new exopolysaccharide from a strain Klebsiella
sp. is very resistant against the extreme pH. Although solutions of this new poly-
saccharide show a commonly observed shear thinning behavior at pH range from
2 to 12, this polysaccharide is degraded outside of this pH range. However, when the
alkaline degraded polysaccharide is acidified into neutral pH, both moduli increased
and the crossover was observed at pH 10.5. Below that pH, the solution showed a
structured liquid behavior. It suggests that the backbone structure of this polysac-
charide was not degraded, and some conformational change was reversible as
observed in scleroglucan, which showed a rheological recovery induced by the
partial recovery of triple helices from coil conformation after the degradation at
high pH (Aasprong et al. 2003). The advantage of this new polysaccharide is that the
viscosifying properties are stable and not so much changed by the addition of
sodium, potassium, magnesium, and calcium ions as in xanthan.

11 Application of Hydrocolloids as a Thickener

11.1 Controlling the Viscosity and Stabilizing of Liquid

Foods, Acidified Milk, Sauces

High methoxyl pectin (HMP) has been used widely to stabilize acidified milk drinks
for over 60 years. Stabilization mechanism of HMP was attributed to the steric
repulsion among HMP adsorbed casein aggregates and not by a weak gel network,
and the adsorption of HMP to the surface of casein aggregates was by electrostatic
attraction (Parker et al. 1994; Jensen et al. 2010). Kiani et al. (2010) studied the
stabilization of a traditional Iranian yogurt drink by gellan/HMP, and suggested that
gellan confers the additional stabilization effect. Recently, another pectic polysac-
charide extracted from Ulmus davidiana was found effective to stabilize yogurt
preventing syneresis and also promoting the growth of lactic acid bacteria (Yeung
et al. 2019). Many other hydrocolloids with low viscosity such as soluble soybean
polysaccharides (Maeda and Nakamura 2021), gum tragacanth, and Persian gum
(Azarikia and Abbasi 2016) were also shown effective for the stabilization of
acidified milk drinks. While low viscosity acidified yogurt drinks are liked by
healthy young consumers, high viscosity ones are preferred by traditionalists and
also believed to be safer for dysphagic patients in hospitals.
Since starch is the most widely used hydrocolloids, it is important to know the
effect of seasonings on rheological properties. Hirashima et al. (2005, 2012) found
3 Rheological and Thickening Properties 107

the great difference of the viscosities of 3 wt% maize starch suspensions/pastes to

which sucrose, acids, and salt were added before and after the heating.
Xanthan is used frequently to thicken the starch paste. Caramel sauces made from
potato starch containing glucose syrup were thickened by xanthan (Krystyjan et al.
2012). Xanthan was also added to tapioca starch paste to increase the viscosity and
the stability against heating (Chantaro et al. 2013).
Low calorie mayonnaise using xanthan gum has been commercialized in Japan. A
body mouthfeel should be maintained while reducing oil content in mayonnaise. The
viscosifying function of xanthan surely plays an important role here, but its stability
in acidic pH and the texture constancy in the presence of salt are also required. A
good flavor release is also required and it is related with shear thinning behavior of
xanthan. A mixture of xanthan, locust bean, guar and tamarind gums is used to make
a tofu of higher water holding capacity and with a good mouthfeel. Xanthan is used
to prevent retrogradation of rice-cake (mochi) and also to improve the texture. A
thickener consisting of xanthan and low methoxyl pectin is proposed. Japanese
noodles containing xanthan are reported to have an elastic texture. Xanthan is also
used for glaze in order to protect frozen fish and vegetables preventing syneresis
(Nishinari 1988).
Hanpen is a traditional fish product in Japan. It contains many air bubbles and has
a sponge-like texture. Meat of sharks and cod are mashed with yams and then boiled
in hot water. Sharks and yams are necessary for producing air bubbles. A traditional
sponge cake containing many foams and with a moist mouthfeel, Karukan, also
relies on yam polysaccharides. The foam producing and stabilizing capacity of yam
polysaccharides is believed to originate from high viscosity but also from some
elasticity.
Spinnability is related with the breakup length when the liquid is subjected to
extension. When a glass rod is immersed into a liquid and then pulled up vertically,
even sugar syrup which is very close to a Newtonian fluid shows a thread because a
liquid adhering on the glass rod is flowing down, and when all the liquid flows down,
the thread breaks. However, when the fluid has a certain elasticity, the fluid extends
by rubber elasticity and when the fluid breaks, the suspending fluid jumps up
because of the elastic recovery. Therefore, the maximum extendable length of the
fluid depends on the ratio of the viscosity and elasticity, which is known as the
relaxation time (See Sect. 5). This property of fluid is closely related with cohesive-
ness which plays an important role together with viscosity in the safe swallowing
(Nishinari et al. 2019). Elongational measurement is important for evaluating the
spinnability and thickened fluids used in dysphagic treatments (Turcanu et al. 2018)
because it is related with not only viscosity but also with elasticity.
In some thickening application, the short texture is required rather than long
texture. Here, the “short” means neat and tidy mouthfeel and the draining and
dripping off in the processing, which can be quantified by the break up length
when the liquid is subjected to extension. In the operation of spreading honey on
the bread, long texture hampers it. Some patent proposes to mix agar to make the
texture short. This short texture is also preferred in thick sauce for breaded cutlets,
and xyloglucan is often used for such a requirement. This short texture is also
108 K. Nishinari

suitable in sauce for barbecued /grilled meat and for a round dumpling made from
powdered rice.
Many polysaccharides such as arabinoxylans, cereal beta-glucans, chitin and
chitosan, dextran, mesquite gum, pullulan as well as vegetable proteins and milk
proteins are used as thickening as well as stabilizing and emulsifying agents in
various foods (Nussinovitch and Hirashima 2019).
Although the detailed mechanism of aspiration in dysphagic patients is not yet
understood, thickening of liquid foods has been found effective to reduce the risk of
aspiration. Thickening slows the liquid flow so that the epiglottis may be able to
close the airway to prevent the penetration of the food bolus, although this scenario
was not demonstrated so well (Cichero 2013; Steele et al. 2015; Nishinari et al.
2016). Many kinds of thickening agents are commercially produced and used in
hospitals and organizations for disadvantaged persons (IDDSI, International Dys-
phagia Diet Standardisation Initiative: https://iddsi.org/; Japan Care Food Confer-
ence: https://www.udf.jp/).

11.2 Rheological Control of Texture by Polysaccharide

Thickeners: Noodles/Pasta and Breads

Noodles are popular foods especially in Asia, and pasta, spaghetti, and macaroni,
etc. are also popular in Western countries. Rice noodles are popular in Vietnam,
Thailand, and other Asian countries, and high amylose rice (long grain indica type)
is suitable for noodles. Recently, to reduce the excessive inventory and promote the
utilization of low amylose rice (short grain japonica type), hydrocolloids are added
to powdered rice to improve the texture in Japan. Noodles from low amylose rice are
evaluated too sticky and not enough firm. Nitta et al. (2018) compared the texture of
rice noodles treated by Ca2+-induced setting of alginate or LM pectin, and found that
the stickiness was reduced and the firmness was increased by the addition of alginate
or pectin. Kita et al. (2009) reported the similar improvement of the texture of
japonica rice noodles by adding tamarind seed gum (TSG). They found that the
decrease in hardness of noodles by frozen storage was reduced by the addition of
TSG. Chitosan was also used for rice noodles (Klinmalai et al. 2017). Esterified
tapioca starch was also used for salted noodles (Eguchi et al. 2014). Silva et al.
(2013) examined the effect of adding various polysaccharides, guar gum, LBG,
KGM, HPMC, and xanthan on the mechanical properties of sweet potato noodles
containing broccoli powders. They found that shear modulus of noodles was lowest
in noodles with hydroxypropyl methylcellulose (HPMC) and xanthan, which was
attributed to the restriction of swelling of starch granules by these polysaccharides
with high water binding capacity.
Dough rheology has been a very important applied rheology since the birth of
rheology, and the relation between the bread making property and the rheological
properties of dough has been extensively studied. Viscosifying hydrocolloids have
3 Rheological and Thickening Properties 109

been widely used for this, but the relation between the viscosifying property and the
final bread property is not so well understood. Gluten plays a crucial role in the
baking to form and keep “cell” walls, and many hydrocolloids are expected to
improve the baking performance, but it was only partially successful. Doughs
showing a strain hardening behavior in extension test were affirmed to have a
good bread making property (Kokelaar et al. 1996). Since glutenin was identified
as a main gluten fraction which plays an important role in elastic and strain
hardening properties of dough, instead of using gluten whey protein particles are
mixed with wheat starch to make a dough showing a similar strain hardening
behavior to a normal wheat dough (van Riemsdijk et al. 2011).
Application of hydrocolloids for rice bread has also been reported. It has attracted
much attention to make the gluten-free bread since the celiac disease was recognized
to be caused by gluten. Hydrocolloids at 1% and 2% w/w, pectin, carboxymethyl-
cellulose (CMC), agarose, xanthan or oat β-glucan were added to rice flour for
gluten-free bread (Lazaridou et al. 2007). The loaf volume is one of most important
index for bread making quality and it was found that the incorporation of xanthan at
1% into the gluten-free breads did not change the loaf volume and at 2% supple-
mentation level even decreased the volume; this formulation exhibited the lowest
volume among all preparations. This was consistent with previous findings.
Recently, however, in comparison of locust bean gum, guar gum, xanthan gum,
tamarind seed gum, native gellan gum, dextrin, LM pectin, fermented cellulose
CMC, konjac glucomannan, HM pectin, κ-carrageenan, ι-carrageenan, and
λ-carrageenan in baking, the highest loaf volume was found when xanthan was
used (Morimoto et al. 2015). HPMC, guar gum, pectin, xanthan gum, TSG were
added to rice flour to make a gluten-free bread (Jang et al. 2018), and the authors
concluded TSG gave the best bread among these hydrocolloids. The inconsistency
among reported results may be due to the difference in the hydrocolloids because
most commercially available polysaccharides have many variations: molar mass
may be different, degree of substitutions or branching if any, cation type if it is
charged polymers, etc. Therefore, unfortunately, it is impossible to compare directly
the papers without confirming that the hydrocolloids used are the same or not in
addition to the other factors such as mixing ratios and mixing conditions. This
situation led Japanese workers to form a collaborative research group using the
common samples of gellan to study the gelling and viscosifying properties as
described in the next chapter for gelling properties.
Because of the issue of celiac disease, gluten-free pasta and bread have been
investigated by many research groups which tried to use various hydrocolloids to
improve the quality and especially textural properties (Padalino et al. 2016; Pahwa
et al. 2016; Rai et al. 2018). Lynch et al. (2018) reviewed potential application of
exopolysaccharides for gluten-free bread, and showed that the added dextran
increased the specific volume and reduced the crumb hardness and extend the
shelf life of the bread better than HPMC, which was attributed mainly to water
binding ability of this high molar mass and branched dextran (Rühmkorf et al. 2012).
110 K. Nishinari

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Chapter 4
Gelling Properties

Katsuyoshi Nishinari

Abstract Except beverages, soups, and dried foods, most foods are eaten in gel
state, and thus it is important to understand the mechanisms of gel formation to
improve the food quality. Starting from the definition and classification of gels, this
chapter describes the molecular forces responsible for gel formation, how the
network structure is formed, rheological determination of gel point, critical molar
mass and concentration below which no gelation occurs. Then, characterization
methods, gelation kinetics, mechanical spectra, and thermal scanning rheology are
described based on studies performed for gelation of polysaccharides and proteins.
Various factors influencing structure and properties of polysaccharide and protein
gels, in particular, gelation rate, temperature, molar mass, concentration, interaction
between long chains and short chains, small molecules such as sugars, acids, salts,
and polyphenols are described. Release of molecular chains followed by erosion of
gels induced by immersion in solvents is also described. Special categories of gels,
microgels, mixed gels, cryogels, and oleogels are also described.

Keywords Definition · Classification · Transition · Polysaccharides · Proteins

1 Introduction

Gels are everywhere around us. In the breakfast, boiled or fried eggs are served.
Tofu, yogurt or cheese is also served. They all are gels. Jams on the toast are also
gels. There are many food gels, dessert jellies, pudding, marshmallow, etc. When
water used to boil fish or meat is concentrated and cooled, a gel is formed. Though
raw rice grains are not gels, cooked rice grains, and cooked pasta and noodles might
be family members of gels. When dried foods with very low water content are
masticated and crushed into small fragments, they are mixed with saliva forming a

K. Nishinari (*)
Glyn O. Phillips Hydrocolloids Research Centre, School of Food and Biological Engineering,
Hubei University of Technology, Wuhan, People’s Republic of China
e-mail: katsuyoshi.nishinari@hbut.edu.cn

© Springer Nature Singapore Pte Ltd. 2021 119

Y. Fang et al. (eds.), Food Hydrocolloids,
https://doi.org/10.1007/978-981-16-0320-4_4
120 K. Nishinari

bolus and then swallowed. Before swallowing, the solid foods are transformed to
gels before forming a swallowable bolus. Our gastric juice is in acidic pH ca. 1–1.5.
Why the stomach wall is not injured? It is protected by mucin gels. Acid-induced
gelling property of alginate has been used for treating gastroesophageal reflux
disorder. When some part of our body is injured, blood comes out and it clots
(gels). When we catch a cold, sputum is formed. It is also a gel. They all are formed
from a liquid which is solidified or a solid becomes a gel by absorbing liquid.
Therefore, apart from two extremes, very dried foods and liquids, most semi-solid
foods can be called gels although mainly polysaccharide and protein gels are
discussed in this chapter. Some shampoos are called gels although they are not
gels in a scientific word. It is necessary to define a gel.

2 What Is a Gel?

The definition of gels has been a problem discussed for a long time. Jordan Lloyd
said “The colloidal condition, the ‘gel’, is one which it is easier to recognize than to
define”. In the review on structure of gels, she classified gels into (1) heat-reversible
gels such as gelatin or agar in water, cellulose acetate in benzoyl alcohol and
(2) heat-irreversible gels such as silica in water, colloid in chloroform and alcohol,
many metallic sulfides and oxides in water. Since then, many definitions have been
done for polymer gels (Djabourov et al. 2013). Recently, low molar mass gelator has
been attracting attention in relation to pharmaceutical and cosmetic industry but not
so much applications are found in food industry except oleogel. Gels formed from
synthetic and biological polymers have many things in common and it is useful to
discuss together comparing each other (Djabourov et al. 2013; Nishinari et al. 2016a;
Tokita and Nishinari 2009) although the present chapter focuses mainly on polysac-
charide and protein gels. Gels can be defined both from a rheological behavior and
from a structural feature.
Rheological definition of a gel is that the system does not flow, and it can be
characterized by the presence of a plateau region of storage modulus and the low
tanδ (<0.1) at an angular frequency range from 10
3 to 102 rad/s, which is accessible
by many commercially available rheometers. This should be called an operational
definition and it cannot exclude the possibility to find the violation of this definition
if a material obeying this definition shows a liquid-like rheological behavior at
further lower frequencies. We should remember a famous saying of a prophetess
Deborah “Even the mountains flow before the Lord” or everything flows. We could
enjoy to visit the Website of a pitch drop experiment. A drop of the pitch flows (falls)
every 9 years or 10 years or 11 years. It shows clearly that the distinction of a liquid
and a solid is not simple, and depends on the “patience” of the observer. Since it is
difficult to set the reasonable time-scale for both patient and impatient persons, it is
more practical to clarify the fluidity by the yield stress concept (discussed in the
previous Chap. 3) and the large deformation behavior above the yield stress. In this
sense, a commonly used tube tilting method is not a good method because of its
4 Gelling Properties 121

dependence on the patience of the observer. Impatient observers will not see whether
it flows after a long time under the gravity which does not induce flow during a short
time (see the discussion on delayed failure in Chap. 3). If it flows above the yield
stress, it is a structured liquid and should not be called a gel as mentioned below. If it
is a gel, it will be broken down into separate parts and will not flow. If divided parts
recover the initial continuity immediately on contact, such materials could be used in
various areas in adhesive, coating, electric conductivity, and tissue engineering.
These are called self-healing gels (Cordier et al. 2008; Halake and Lee 2017), and
it is expected that the principle may be also applied in encapsulation of bioactives
and endurable packaging materials in the future.
Structural definition of a gel is based on the connectivity of the system. Gel is a
system consisting of molecules, particles, chains, or other structural elements which
are partially connected to each other in a fluid medium by crosslinks to the macro-
scopic dimensions. According to this structural definition, the loss of fluidity is the
result of connectivity. Entanglement of long object may be regarded as connected by
delocalized crosslinks. The sol-gel transition can be treated by a percolation theory
(Tokita 1989). This is also another operational definition, and it cannot exclude the
possibility of finding gels whose constituents are not directly connected.
There are two material groups having a name “gel,” but it can cause a confusion.
These two are a weak gel and a fluid gel. Weak gel was named for structured liquids
which show a weak frequency dependence for storage and loss modulus at a
commonly accessible angular frequency range from 10
3 to 102 rad/s, and the
mechanical loss tangent is higher than 0.1. These materials are essentially liquid
but it does not flow below its yield stress, thus it is apparently different from other
liquids with no yield stress. Since yield stress has attracted further attention recently,
it was discussed in the previous chapter for rheology. It may be better to mention
here that it is not a good question to ask “which is more viscous, mayonnaise or whip
cream and sugar syrup or honey?” The former has a yield stress and the latter not. In
the present chapter, this is not called a weak gel but called structured liquid because it
is essentially a liquid with non-zero yield stress.
The second class material, fluid gel is against the definition of gels mentioned
above in the sense that it flows; fluid includes the meaning of flow. In this case, we
should define the length scale of the “phase.” Strictly speaking, the so-called fluid
gels should be called microgels. Everybody knows that sands “flow.” And there is an
expression “flowing sands.” Everybody also knows that a grain of sand is a solid
although a mass of sands “flows.” These granular materials like sands, powders, and
other solid particles flow, but this flow is different from the liquid flow which has
been studied in fluid dynamics. The flow of granular materials stops at the angle of
repose of these granular solids when put on the horizontal plane while liquids with
no yield stress (described in the previous chapter) flatten showing no angle of repose.
Dried sands and powders don’t stick each other, and therefore contradict with the
connectivity in the above definition, but this situation can be modified by increasing
moisture content or adding liquids. Such a material has, however, been called a paste
rather than a gel. They are similar to slurries. But, it is difficult to make a clear
122 K. Nishinari

distinction between a gel and a paste. It may be impossible to define the highest and
the lowest liquid content for gels. This is related to the following problem.
Emulsion gel and oleogel are different from food polymer gels consisting of
polysaccharides and proteins. In fats such as butter or margarine, the solid fat content
is an index to characterize the state of the material. This index decreases with
increasing the temperature. The difference between the gel-sol transition and the
melting of fat is that the latter is more crystalline than the former. What is the border
of the crystallinity or the orderliness of the structure when we define the gel? There
may be gels very close to crystalline state and also other gels very close to
amorphous state. It may be impossible to define a borderline which depends on
many factors, which is discussed later.
It is therefore difficult to propose the simple definitive definition of gels, and the
present author has a similar feeling with his good friend, Nijenhuis who says A gel is
a gel, as long as one cannot prove that it is not a gel. The difficulty arises when we
define a border between a gel and non-gel as pointed above such as liquid content,
and the degree of the order, and glass. If the border can be found as a discontinuous
transition, it can be clearly defined, but if the border is not discontinuous and vague,
it might be difficult to distinguish the materials on both sides, gel and non-gel. The
above tentatively proposed definition of gels could be operationally practical, but the
author will be happy if more persuasive definition is proposed because the science is
the never-ending endeavor of human beings.

3 Classification of Food Gels

Gels can be classified from various points of views based on mechanical properties,
molecular forces, constituting ingredients, temperature dependence of elastic mod-
ulus, transparence, electric charges, etc.

3.1 Classification of Food Gels Based on Mechanical

Properties

When we hear that it is easier to recognize than to define a gel, we expect that a gel
may wobble which is easy to recognize. The amplitude we observe is related to the
elastic modulus (Nishinari 1976) or simply we can push a gel with a finger. Gels may
not have the elastic modulus of common solids such as metals, stones, glass, or
plastics which have the moduli >109~11 Pa. Moduli of most rubbers are of the order
of 106 Pa. What is the maximum and the minimum moduli for gels? There is no
consensus about this because there may be a very firm gel which is very close to
solids, and a very tenuous or sloppy gel very close to liquids. The minimum modulus
4 Gelling Properties 123

necessary for a gel to be self-standing is ca103 Pa, and this and yield stress are
recognized important in 3D printing (Nan et al. 2020).
Beyond the small deformation range, gels show a failure or fracture. Gels close to
liquids show failure but not clear fracture. The strain at failure or fracture is an
important property in the application of gels. Generally, gels consisting of long
chains show a deformable behavior while gels consisting of short chains or small
molecules show a failure at small strain and are brittle. Obviously, textures of
deformable gels and brittle gels are very different (Nishinari 2000a).
Soft gels are not chewed by teeth but compressed/squeezed between the tongue
and the hard palate. The transition of the mastication mode from the tongue-palate
squeezing to the chewing by teeth could be well predicted by a simulating compres-
sion experiment of a gel between an artificial tongue and the base plate of the
uniaxial compression machine, and depended on the deformability of gels (Ishihara
et al. 2013, 2014).

3.2 Classification of Food Gels Based on Molecular Forces

Molecular forces which form gel network are covalent and non-covalent bonds such
as hydrogen bonds, hydrophobic interactions, ionic bonds. Gels formed by
non-covalent weak secondary forces such as hydrogen bonds, hydrophobic interac-
tions, van der Waals forces are called physical gels, while gels formed by covalent
bonds are called chemical gels. In many food protein gels, physical and chemical
bonds coexist (Djabourov et al. 2013).
It seems that it is still not possible to understand the structure-property relation of
all different gels in common language which is aimed in this chapter. Researchers
studying polysaccharide gels consisting of linear chain molecules use the network
theory derived from rubber elasticity and the ordering process is related with coil-
helix transition and aggregation processes while globular protein gels are studied
from the viewpoint of molecular rearrangement (Nishinari and Takahashi 2003;
Nishinari et al. 2000). Gelatin is a fibrous molecule and its gelation has been studied
extensively, but in food area, particle gels formed from globular proteins are another
important material. Helix-coil transition is also a molecular rearrangement, but this
term is not so much used among researchers interested in polysaccharide gels or
gelatin gels which are jokingly called an honorary polysaccharide. Both of these gels
are common in a sense that structural elements are connected or in modern parlance,
after de Gennes’ introduction, “percolated” to span the space. The term percolation
originates from a coffee percolator (Djabourov et al. 2013).
124 K. Nishinari

3.3 Classification of Food Gels Based on Molar Mass

of Network Elements

Most food gels are made from polysaccharides or proteins. When they are degraded
into lower mass compounds, they cease to form a gel. Everybody knows that sucrose
solutions don’t form a gel even if they are concentrated. They precipitate above a
certain concentration at each temperature. An important difference between lower
molar mass compounds such as sugar or salt and higher molar mass compounds
(polymer) is that the former solutions precipitate when the temperature or pressure or
concentration is changed so that the extrinsic condition is outside of the condition of
saturated solution while the latter materials have plural conformations such as helix
and coil and their aggregates which lead to network structure called commonly gels.
Precipitated lower molar mass materials such as butter or margarine are usually not
called gels. However, lower molar mass gelators (Weiss and Terech 2006) have been
attracting much attention and especially oleogels in food area (Singh et al. 2017).
Pullulan is one of most soluble polysaccharides and its films are used in food
packaging, but its gels are not studied in gel technology so much, because materials
called gelling agents form a gel at lower concentrations.

3.4 Classification of Food Gels Based on Ingredients

In foods, many polysaccharide gels and protein gels have been known, and gels
consisting of mixed polysaccharides and proteins have also been studied. Emulsion
gels which consist of polysaccharide or protein gels containing oil droplet have been
attracting much attention in the past two decades. Aerated gels have been also
studied recently because it can reduce the calorie inducing satiety thus can be a
convenient tool to prevent the obesity. Recently, gelation of oils has attracted much
attention, and is described later.

3.5 Classification of Food Gels Based on Origin

Gels can be also classified by the origin of materials, plant or animals, sea weed,
seed, bean, legume, fish, meat, egg, milk, and microbial origin such as curdlan,
gellan, and so on. This viewpoint is important in designing food products (Phillips
and Williams 2020).
4 Gelling Properties 125

3.6 Classification of Food Gels Based on Temperature

Dependence of Elastic Modulus

Some biopolymer solutions of agar, carrageenan, gellan, and gelatin form a gel on
cooling, but egg white, milk proteins, solutions of methylcellulose and other cellu-
lose derivatives, and curdlan form a gel on heating (Nishinari 2000b; Nishinari and
Zhang 2004). They are often called cold-set gels and heat-set gels, respectively.
Some of them are thermoreversible and the others are thermo-irreversible; gels of
agar, carrageenan, gellan, gelatin are formed on cooling, return to sol state on
heating, thus called thermoreversible gels, while boiled egg white or tofu or konjac
is a typical thermo-irreversible gel because they do not show gel-sol transition after
gel formation. However, some thermoreversible gelation of ovalbumin (OVA), one
of the main proteins in egg white, is well-known and studied extensively (Doi 1993).

3.7 Classification of Food Gels Based on Optical Properties

Gels of gellan and gelatin are transparent while egg white gels, starch gels, and many
other gels are opaque. But it is well established to make a transparent egg white
protein gel (Doi 1993). Most pigments are added artificially but some gels have a
color in gels where naturally binding pigments exist.

3.8 Classification of Food Gels Based on Shape of Network

Elements or Crystallinity

Fibrous gel, particle gels, or mixture of these may exist, since rearrangements or
ageing always occurs and these may be related to syneresis. Partially ordered
structured gels or liquid crystalline gels also exist. Even in apparently homogeneous
gels, an internal structure consists of ordered domains and amorphous domains. It
may be impossible to define the upper and lower limit of the degree of order.

3.9 Classification of Food Gels Based on Electric Charges

Most gelling polysaccharides have anionic groups such as carboxyl group (gellan
(Morris et al. 2012)) or sulfate group (carrageenan) while chitosan molecule has a
positive charge. Gels of agarose, curdlan, and starch are electrically neutral without
electric charges although most commercially available samples contain some ions.
Since amino acids are positively or negatively charged, gelling behavior of proteins
is strongly influenced by pH and ionic strength.
126 K. Nishinari

3.10 Classification Based on Other Criteria

It is also possible to classify gels based on size (microgels and bulk gel), pH
dependence, water holding capacity, thermal conductivity, liquid (water, oil, alco-
hol), or gas content. This viewpoint is useful in food processing and preservation.
Macroscopic shape such as spherical, rod, tetrahedron, rectangular can be controlled
if it is moldable, but these geometrically regular shaped gels may not be naturally
occurring gels. It is an important problem to keep a shape in food processing and
flavor release.

4 Gel-Sol Transition

4.1 Molecular Forces

Suzuki et al. (1972) examined whether the pressure would prompt the gel formation
in which hydrogen bonds play a main role, while the pressure would retard it in
which hydrophobic interactions play a main role. Investigating the dependence of
equilibrium constant K in the gel-sol phase transition on the temperature and
pressure (∂1nK /∂T ) p ¼ ΔH/ RT2 and (∂1nK/∂P) T ¼ -ΔV/RT, respectively,
they studied the effect of pressure and temperature on the gelation of gelatin and
methylcellulose (MC). They found that the pressure prompted the gel formation of
gelatin, while the pressure retarded that of MC, and therefore, it is concluded that
hydrogen bonds and hydrophobic interactions are dominating molecular forces for
gelatin and MC, respectively. However, they noticed that ΔV changes the sign above
6000 atm, and stated that both hydrophobic interaction and hydrogen bonds concern
in the gelation of MC.
Kometani et al. (2015) studied the pressure effect on the gelation of agarose and
MC (Fig. 4.1). The cloud-point temperature, Tcloud, was determined from cooling
curves by the temperature of 75% transmittance because it corresponds to the
steepest point of decreasing transmittance in all curves. In the same way, the
“transparency temperature,” Ttrans, at which the solution returns to a transparent
sol state was determined from heating curves by the temperature of 75% transmit-
tance. From the Clapeyron-Clausius relation dT/dP ¼ TΔV/ΔH together with the
enthalpy change determined by thermal measurements, the volume change for
agarose was determined as ΔV ¼
4.57  10
6 m3/kg which is an order of
magnitude larger than that for gelatin (Fig. 4.1a). This suggests that the gels of
agarose and gelatin are stabilized under high pressure, but their stabilization mech-
anism may be different. In addition, it should be mentioned that both κ- and
ι-carrageenan gels formed also by hydrogen bonds are destabilized by pressure
(Gekko and Kasuya 1985).
The slope dT/dP ¼ 2.97  10
2 K/MPa determined from Fig. 4.1b for MC is
consistent with the reported values ΔV ¼ 0.80  10
6 m3/kg and ΔH ¼ 11.8 kJ/kg
4 Gelling Properties 127

Fig. 4.1 Plots of Tcloud (square) and Ttrans (diamond) as a function of pressure for 1 wt % solutions
of agarose (Fig. 4.1a) and methylcellulose (Fig. 4.1b). Triangles and cross show the gelling
temperature Tgel and melting temperature Tmelt, respectively, measured by the falling-ball method.
Reproduced with permission from Kometani et al. (2015), Copyright 2015ACS

taken from literatures, and indicates that MC gel is destabilized by compression as

contrasted with agarose (Kometani et al. 2015).
The main molecular forces responsible for gel formation in agarose and MC are
hydrogen bonds and hydrophobic interaction, respectively, and both appear as an
endothermic peak on heating in DSC. In the former, gel-to-sol transition occurs,
while in the latter sol-to-gel transition occurs. However, in the latter, at some
intermediate concentration range, an exothermic peak appears on heating which is
attributed to the appearance of anisotropic phase (Yin et al. 2006). This anisotropic
phase formation at higher concentration for stiff molecules is also known for xanthan
(Lee and Brant 2002), gellan (Nitta et al. 2010), schizophyllan (Fang et al. 2004),
and other biopolymers (Djabourov et al. 2013).
Shirakawa et al. (1998a, b) found that xyloglucan from which galactose residues
were removed formed a gel on heating and returned into sol state on further heating
and that this transition was thermally reversible (Fig. 4.2). Enthalpy change ΔH
accompanying gel to sol transition on heating was reported 24.2 J/g for gelatin
(Gekko et al. 1992), 33.2 J/g for κ-carrageenan (Watase and Nishinari 1987a), 40 J/g
for agarose (Watase and Nishinari 1987b) while ΔH accompanying sol to gel
transition on heating is 16.0 J/g (Haque and Morris 1993) or 10–17 J/g (Funami
et al. 2007) for methylcellulose, 6.9 J/g for methylhydroxypropylcellulose (Yuguchi
et al. 1995). ΔH found for the degalactosylated xyloglucan was 4.4 J/g, which is the
same order of magnitude to the latter group where hydrophobic interaction plays a
main role. The gelation of xyloglucan by galactose removal is still studied recently
(Sakakibara et al. 2017).
128 K. Nishinari

Fig. 4.2 Sol-gel transition 140

temperature for xyloglucan
as a function of galactose
removal ratio. ■ lower 120 Sol

Transition temperature (°C)

temperature transition point;
□ higher temperature 100
transition point. Reproduced
with permission from 80
Shirakawa et al. (1998a),
Copyright 1998 Elsevier
60

40 Gel

20

0
20 30 40 50 60 70 80
Removal–ratio of Galactose (%)

As is seen in the gelation of MC and xyloglucan, the chemical modification

changes the solubility or gel forming ability. Gel forming ability is the intermediate
between the insolubility and solubility. Carboxymethylation of curdlan confers
curdlan water solubility but makes lose its gelling ability (Jin et al. 2006; Nishinari
and Zhang 2004).
On the other hand, molecular forces in the gelation of globular protein gels are
more complicated (Djabourov et al. 2013; Walstra 2003).

4.2 Rheological Determination of Sol-Gel Transition

One of the most widely used methods for the determination of sol-gel transition is
that proposed by Winter and Chambon (1986). These authors examined the gelation
process of polydimethylsiloxane (PDMS) to determine the gel point. PDMS was
chosen for its well-defined chemical nature, the need of catalysis, the availability of
prepolymers with different molecular weights, and the elastomeric nature of the
samples after crosslinking. The advantage of this method was to stop the catalyst
action instantaneously to prevent the further crosslinking reaction. Since the mea-
surement of the frequency dependence of G0 and G00 takes a time, it was examined
before and after the crossover of G0 and G00 , which was possible by stopping the
crosslinking reaction by stopping the catalytic action by stopping agent (sulfur). By
this method, they could observe the storage modulus G0 and loss modulus G00 as a
function of frequency near the crossover point of G0 and G00 observed as a function of
time at a constant frequency. Before the gelation point GP, G0 < G00 and after GP,
G0 > G00 . The frequency dependence of G0 and G00 was observed from 6 min before
4 Gelling Properties 129

the time of crossover of G0 and G00 , t0, to the time after 6 min of t0. At an earlier stage
of the crosslinking reaction, both G0 and G00 are strongly frequency dependent and
G0 < G00 , and at time t0, both G0 and G00 show a similar frequency dependence, and
after the time t0, G0 tends to show a plateau which is a characteristic mechanical
behavior for rubbers. They proposed the following criterion for the critical gelation
point.

G0  G00  ωn ð1Þ
00 0
tanδ ¼ G =G ¼ tan ðnπ=2Þ ð2Þ

which is now called Winter-Chambon criterion. At the critical point, both of these
two equations should be satisfied simultaneously. The exponent n in the two
equations should be the same. These equations are valid at times longer than a
certain characteristic time t0 or at lower frequencies than 1/t0. First, this criterion was
proposed for such a chemical gel, but later, Nijenhuis and Winter (1989) found that
this criterion is valid also for physical gels of polyvinyl chloride (PVC). Since then
many papers have been published to study the critical gelation point, and although
initially the exponent was proposed as 0.5, other values between 0.3 and 0.7 have
been reported for various gelation processes (Winter 2016). Figure 4.3a shows the
mechanical spectra, i.e., the frequency dependence of G0 and G00 of a 2.5 wt %

Fig. 4.3 (a) Angular frequency dependence of G0 (open symbols) and G00 (filled) for a 2.5% gellan
gum solution at various temperatures shown on the right side of each curve. The data are shifted
along both the horizontal and the vertical axes by shift factors “a” and “b,” respectively, to avoid
overlapping. Numbers on the left side of each curves represent “a” and “b,” respectively.
Reproduced with permission from Miyoshi and Nishinari (1999), Copyright 1999 Springer. (b)
Angular frequency dependence of G0 (filled symbols) and G00 (open) for a 50mg cm-3 OVA
solutions heated at 80  C for 60 min and then reheated after addition of 60 mM NaCl for various
times Δt ¼3, 5, 15, 45, and 240 min. The data are shifted along both the horizontal and the vertical
axes by shift factor “a” to avoid overlapping. Reproduced with permission from Koike et al. (1998),
Copyright 1998 Elsevier
130 K. Nishinari

sodium type gellan solution at temperatures from 40  C to 10  C (Miyoshi and

Nishinari 1999). At higher temperatures, G0 and G00 of the solution shows strong
frequency dependence and G0 < G00 while at low temperatures G0 > G00 and both
moduli tend to show a plateau. At 26  C, the solution satisfies the above equations,
and is in the critical state.
Figure 4.3b shows the mechanical spectra of G0 and G00 of a 50 mg cm
3 OVA
solution prepared by a two-step heating method; first heating at 80  C for 60 min and
then cooled, and then reheated at the same temperature for various heating time
periods Δt (Koike et al. 1998). Critical gels satisfying the eqs. (1) and (2) were
observed for Δt from 3 to 30 min indicating the fractal structure persisted in this
range of the heating time. Koike et al. (1998) had reported that OVA formed linear
aggregates, worm-like cylinder, diameter 12 nm, persistence length 23 nm, and the
molar mass per contour length 16,000 nm
1 after heating the solution at 80  C by
dynamic laser scattering (Nemoto et al. 1993) (See Fig. 4.13). Transparent gels were
formed by both one-step and two-step heating in a certain condition of the salt
concentration and heating time, and these gels were found to satisfy the critical
condition eqs. (1) and (2). However, such a critical gel behavior was not observed for
translucent gels. Although these authors ascribed the main molecular forces for the
gel formation are hydrophobic interaction, they found the necessity of further
structural study based on neutron scattering and SEM, and this is discussed again
in Sect. 7.
Hossain et al. (1997) studied the gelation of iota-carrageenan solutions, and found
that G0 ¼ G00 at 57 C and n ¼0.42. The value of n was found to decrease with
increasing concentration of iota- carrageenan. The crossover of G0 and G00 as a
function of time or temperature has often been taken as an indication for the gelation
point, but this is only right for a critical gel with the relaxation exponent n ¼ 0.5.
Very small values 0.l for the relaxation exponent were reported for bacterial poly-
ester, poly(β -hydroxyoctanoate), and gelatin. The critical gelation point at which G0
and G00 show the same frequency dependence may lie in between the entangled
solution behavior and a weak gel behavior described before. To determine the
gelation point by Winter-Chambon criterion, it is convenient to plot the tan δ
observed at different frequencies as a function of time.
Figure 4.4 shows gelation process of carboxymethyl cellulose crosslinked with
polyfunctional glycidyl ether.
However, this method of determination of gel point was found not applicable in
some systems. In the gelation of pluronic (called also poloxamer, PEO-PPO-PEO
triblock copolymer, where PEO ¼ poly(ethylene oxide), PPO ¼ poly(propylene
oxide)) used in drug delivery, it was found that at lower temperatures (33  C) a
liquid-like behavior G0 <G00 was found, on further heating gel is formed at >34  C,
however, at higher temperatures it was found that G0 >G00 at high frequencies and
G0 <G00 at low frequencies, which is characteristic for entangled polymer solutions
(Nyström and Walderhaug 1996).
Globular protein solutions show mechanical spectra similar to those of structured
liquids such as xanthan solutions, that is, G0 >G00 at the angular frequency range from
10-1 to 102 rad s-1 and tan δ  0.1 (Ikeda and Nishinari 2000; Ikeda and Nishinari
4 Gelling Properties 131

Fig. 4.4 Loss tangent tan δ

as a function of time for the
hydrogels at various fixed
frequencies (0.5, 1, 5,
10 Hz) measured at constant
temperature 60  C. tgel is the
gel point. Reproduced with
permission from Lawal et al.
(2011), Copyright 2011
Springer

2001a, b; Matsumoto and Inoue 1993). During isothermal heating at 70 C, 5% w/w
β-lactoglobulin in a 0.1 mol/dm3 NaCl aqueous solution shows a gelation behavior
(Ikeda and Nishinari 2001a). It was observed that G0 >G00 all the time, and the
gelation occurred at around 3000 s, which was also confirmed by tilting tube.
Thus, the Winter-Chambon criterion cannot be used. For such a case, the tempera-
ture or the time at which G0 begins to increase steeply is generally taken as the
gelation point (Tobitani and Ross-Murphy 1997).

4.3 Spinodal Decomposition or Nucleation and Growth?

Gelation of agarose was thought to progress through spinodal transition based on

Cahn’s theory (Feke and Prins 1974). San Biagio et al. (1996a) recognized the
characteristic or “signature” behavior of spinodal decomposition in the gelation of
agarose (concentration < 1%); the occurrence of a low-angle scattering ring, the
exponential increase of scattered light, its typical dependence upon the scattering
vector represented by linear Cahn’s plot, the occurrence of a transient viscosity peak.
Summarizing their data, they constructed a C-T phase diagram according to which at
higher concentrations (>1.5%), only direct gelation occurs at 60  10  C, and below
this temperature range both demixing mediated and direct gelation are thought to be
possible. They conclude that this approach is useful to understand the gelation
beginning from the break of symmetry, the formation of inhomogeneous structure
from a homogeneous sol state through the spinodal decomposition, and the possi-
bility to control the final gel structure. San Biagio et al. (1996b) studied the gelation
of bovine serum albumin (BSA) by the same approach and found the occurrence of
spinodal demixing of sol after the unfolding of the native BSA.
Morita et al. (2013) constructed a C-T phase diagram of agarose, and found that
an agarose solution formed a gel on cooling and then the phase separation occurred,
which was detected as a cloud point. The spinodal points are found below these
132 K. Nishinari

temperatures. They also recognize characteristic features of spinodal decomposition

mentioned above. They attributed the opacity of agarose gels to the frozen concen-
tration fluctuation within the gel already formed. They pointed out that the sol-gel
transition and the phase separation are independent phenomena, and showed a
porous structure of a quenched gel. They finally emphasize the importance to take
into account this porous structure induced by spinodal decomposition in addition to
the formation of the crosslinking points (junction zones) to understand the macro-
scopic properties of gels.
MC gelation has been studied by many research groups. Group of Lodge
(Arvidson et al. 2013) studied gelation and phase separation of MC with three
different Mw using Winter-Chambon criteria and light transmittance. The gelation
point was determined by frequency independent tan δ for concentrated solutions
(C  10C*, where C* is the coil-overlap concentration). They found a high corre-
lation between the Tgel and the cloud point which is different from previous papers
reporting the phase separation and the gelation are distinct events. In addition, they
showed that gelation of MC has strong dependence on heating rate while the melting
of the gel has little dependence on cooling rate, and thus suggested that
thermogelation of MC proceeded by a nucleation and growth mechanism
(McAllister et al. 2015) rather than spinodal decomposition although they cited
several papers which propose the different mechanism that the gelation of MC is
induced by spinodal decomposition.

4.4 Jamming Transition: Another Molecular Rearrangement

Induced by Shear

In relation to shear thickening of suspensions, shear induced close packing formation

has been attracting much attention. Jamming transition is functionally defined to
occur when, with increasing packing fraction or decreasing applied shear stress, a
yield stress is first observed (Liu and Nagel 1998). Tanaka (2011) classifies jamming
gels with delocalized crosslinks as viscoelastic fluids with long relaxation times.
The minimum packing fraction where a yield stress is observed depends on a
number of suspension properties, including shape, polydispersity, friction coeffi-
cient, roughness, density matching, temperature, and attractive forces (Brown and
Jaeger 2014).

4.5 Zippering and the Size of Junction Zone

To correlate the microscopic states of a macroscopic system with thermodynamic

state variables such as the temperature, volume, and pressure, it is necessary to
determine the partition function in the statistical mechanics. A simple zipper model
4 Gelling Properties 133

for thermoreversible gels consists of aggregated ordered structures such as helices

and stiff chains which can be opened from both ends (Nishinari et al. 1990). From
the partition function including the number of parallel links N, the number of zippers
N, binding energy ε and the degeneracy (rotational freedom) g, the heat capacity
C can be calculated.
In the heating DSC measurements, dT/dt is positive, then the endothermic peak is
equivalent to the maximum of the heat capacity C. In the cooling DSC measure-
ments, dT/dt is negative, then the exothermic peak is again equivalent to the
maximum of C. When the concentration of gels increases, the mobility of chain
molecules decreases, and then the degeneracy g will decrease. As a result, the peak
of the heat capacity shifts to higher temperatures. This is in accordance with
Eldridge-Ferry’s empirical formula. This zipper model approach can also explain
that the transition is sharper in cooling than in heating as has been observed for many
polysaccharide gels such as agarose, κ-carrageenan, and gellan (Watase and
Nishinari 1987b; Miyoshi and Nishinari 1999).
A single DSC endothermic peak for a κ-carrageenan gel on heating was success-
fully fitted by choosing appropriate structural parameters. This treatment was extended
to multi-component systems in which junction zones consist of associations with
different kinds of zippers. A broader DSC endothermic peak for 0.42% κ-carrageenan
/0.186% konjac glucomannan gel was thus fitted (Nishinari et al. 1990).
Using van’t Hoff’s law, Eldridge and Ferry (1954) proposed a method to deter-
mine the heat ΔHm absorbed on forming a mole of crosslinks—dlnC /dTm ¼ ΔHm /
RTm2, where R is the gas constant, Tm (gel melting/K) is the transition temperature,
and C is the gel concentration. Integration of this equation leads to lnC ¼ ΔHm /
RTm + const. This EF equation is based on the following chemical equilibrium:
2 moles crosslinking loci ! 1 mole crosslinks.
Tanaka and Nishinari (1996) modified the EF equation taking into account the
molar mass dependence and proposed a method to estimate the bonding energy ε
(i.e., the enthalpy change for binding a single repeat unit into the network junction),
the number ζ of repeating unit in the molecular chain constituting a crosslink, and the
junction multiplicity s (the number of polymer chains combined in a single junction)
(Nishinari et al. 1996). It is expected that this method is applied for food polymers
although it is not so easy to get food polymers with different molar masses (Nishinari
and Fang 2021).

4.6 Critical Molar Mass and Concentration for Gelation

For thermoreversible gels, the term “junction zone” has been frequently used to
describe the crosslink because each crosslink involves aggregates of ordered molec-
ular chains like helices or extended stiff chains. It is required to know the lower limit
of the molar mass below which a helix is not formed for each polymers. The
minimum concentration necessary for gelation might be related to a persistent length
of the chain. It has never been found that gels are formed in a dilute solution of
flexible polymers such as polyethylene oxide or pullulan which only entangle each
134 K. Nishinari

other but don’t form junction zones. Such a system cannot retain water solvent, and
the system flows if the concentration is not high enough. Koga and Tanaka reported
that the critical concentration for gelation shifted to lower concentrations with
increasing persistence length of linear chain molecules by Monte Carlo simulation.
Solutions of flexible chains, like pullulan, polyethylene oxide, don’t form a gel even
at quite a high concentration. However, since it is known that these polymer
solutions produce a film when the solvent water is evaporated they should form a
gel before becoming a solid film. It is evident that solutions of monomers of these
polymers, glucose, or ethylene glycol do not form a gel even if solvents are
evaporated. These substances with low molar mass are known to form a crystal
when the solution is concentrated. Therefore, there should be a critical molar mass of
polymers below which no gelation occurs.
Ogawa et al. (2006) prepared 6 gellan gum samples with different molar masses
converted to sodium form and examined the helix-coil transition. The weight
average molar mass for higher molar mass samples at 25  C (Mw,25) determined
by DLS was two times higher than that determined at 40  C (Mw,40), indicating the
double helix formation while the ratio Mw,25/Mw,40 is becoming smaller with
decreasing molar mass, and was 1 for the lowest molar mass sample
(Mw ¼ 17,000) indicating that the lowest molar mass sample does not form a double
helix. This is consistent with results obtained by circular dichroism and DSC. The
minimum molar mass for gelation of pectin, κ-carrageenan, and alginate has been
discussed recently (Nishinari and Fang 2021).
The critical gelation concentration has been determined by scaling in which the
shear modulus G0 near the gelation threshold is plotted against the distance of the
concentration from the critical concentration: G0  (C – Ccr)t. The critical exponent
t  2 has been reported by many workers (Djabourov et al. 2013). Joly-Duhamel
et al. (2002) reported that the G0 of gelatin gels could be represented well by the helix
concentration for all the gelatin extracted from various fish although gelatin
extracted from warm water fish begins to form a gel at a higher temperature than
that from cold water fish.
Concentration dependence of the elastic modulus of gels has been studied for a
long time, and it has been known empirically that the modulus is represented by a
power law. The exponent for agarose has been known larger ~4 at lower concen-
trations and smaller ~2 at higher concentrations. A cascade treatment and a modified
rubber elasticity theory were applied to fit the concentration dependence of moduli
for agarose, amylose, pectin, κ-carrageenan, etc. Both these theories predict that the
exponent is larger at lower concentration region and smaller at higher concentration
region. However, an opposite tendency was found in heated ovalbumin gels: the
exponent is about 4 at lower concentration region and about 10 at higher concen-
tration region (Koike et al. 1996).
Generally, the minimum concentration required for gelation is found lower for
linear polymers but gelatin needs higher concentration than agarose or gellan
(Fig. 4.5). This is not a rigorous statement because it may be possible to make the
opposite situation by preparing gels with short chain gellan samples which need
4 Gelling Properties 135

β-lactoglobulin
particulate
fine stranded
amyloid strands

Fig. 4.5 Storage modulus of linear chain polymers and globular protein gels as a function of
concentration. This comparison is not strict because the molar masses and other molecular param-
eters are not the same. Taken from Djabourov et al. (2013) collecting the data from Clark et al.
(1983); Milas, Rinaudo (1996); Kavanagh et al. (2000); Veerman et al. (2002). Reproduced with
permission from Djabourov et al. (2013), Copyright 2013 Cambridge University Press

higher concentration than agarose. Djabourov et al. (1988, 2013) plotted G0 of

gelatin gels as a function of helix content χ and found G0 ¼ 0 at χ < 8%.

4.7 Rigid Network Chains

Common characteristics in physical gels are that network chains are very stiff. Stiff
chains tend to align to form a liquid crystal. Structure formation has been discussed
in relation to Flory’s phase diagram (1956) for rod-like polymers; agarose (Hayashi
et al. 1978), MC (McAllister et al. 2015), xanthan (Lee and Brant 2002). Since stiff
chains are expected to form a gel at a low concentration, it has been attracting much
attention.
Fibrils have been attracting much attention in relation to amyloidosis diseases
such as Alzheimer’s, Creutzfeldt-Jakob disease (CJD), and type II diabetes (van der
Linden 2012). Recently, β-lactoglobulin (β-lg) fibrils formed after heating at 80  C
and pH 2 for 10 h were observed by atomic force microscopy (AFM) and it was
reported that the fibrils have a multi-stranded helical shape with twisted ribbon-like
structures having persistence length from 1000 to 3000 nm (Adamcik et al. 2010).
Since the fibrils of globular proteins have a similar structure to those causing
amyloidosis diseases, it is necessary to confirm the safety of these materials.
Bateman et al. (2010) reported that β-lg fibrils could be digested in simulated gastric
fluid. They found that the fibrils were digested completely by pepsin within 2 min.
Fibrils of β-lg are known to form a gel above a certain concentration depending on
pH, ionic strength (I ), and temperature. A phase diagram of solutions of β-lg fibrils at
pH 2 was constructed as a function of concentration (0–2 wt%) and ionic strength
(0–800 mM NaCl) at 20  C (Bolisetty et al. 2012). Gel phase is found in the region
C ¼ 0.3–2.0 wt% and I ¼ ca 40–100 mM. Recently, Cao et al. (2018) found that the
136 K. Nishinari

G0 /C 2.2, G0 /C 2.4 (Pa)

G0 /I 4.4 (Pa)
I (mM NaCl) C(wt%)

Fig. 4.6 Normalized G0 of BLG fibril networks as a function of (a) ionic strength I (mM NaCl) and
(b) polymer concentration C (wt%). Reproduced with permission from Cao et al. (2018), Copyright
2018 APS

dependence of plateau modulus G0 on the polymer concentration C scales as G0 ~ C 2.2

for semiflexible fibrils or G0 ~ C 2.4 for rigid fibrils (Fig. 4.6b).
Based on the recent theoretical treatment on the stiff chain networks by groups of
MacKintosh, Janmey, Weitz, Cao et al. deduced G0 ~ C 11/5 for semiflexible fibrils
network and G0 ~ C 5/2 for rigid fibrils network, which coincided well with their
experimental observation (Fig. 4.6b). As for the dependence of plateau modulus G0
on the ionic strength I, Cao et al. (2018) found G0 ~ I 4.4 for both semiflexible fibrils
and rigid fibrils (Fig. 4.6a). They explained this high exponent taking into account
the DLVO theory which led to the exponent 4.1.
It was shown recently that β-lg fibrils in the presence of transglutaminase (TGase)
form a gel at a much lower β-lg concentration (Wu et al. 2016) because TGase has a
crosslinking ability to form isopeptide bonds between glutamine and lysine residues
in proteins, thus introducing both inter- and intramolecular covalent crosslinks.

4.8 Interaction of Short Chains and Long Chains

Fang et al. (2007) re-examined the so-called egg-box model which has been pro-
posed for the gelation of alginates and pectins, where two or more chains are
involved in cooperative binding, forming the egg-box structure, by isothermal
titration calorimetry (ITC). Three distinct and successive steps in the binding of
calcium to alginate were identified with increasing concentration of Ca ions. They
were assigned to (1) interaction of Ca2+ with a single guluronate unit forming
monocomplexes; (2) propagation and formation of egg-box dimers via pairing of
4 Gelling Properties 137

Fig. 4.7 Coexistence of oligoguluronate (GB), short chain guluronate, with alginates promotes the
aggregation and strengthens the gel in the presence of sufficient calcium while at low Ca2+ it inhibits
the network growth by binding with calcium. The blue dots represent Ca2+ and the green dots
represent Na+ and H2O etc. Reproduced with permission from Liao et al. (2015), Copyright 2015
Elsevier

these monocomplexes; and (3) lateral association of the egg-box dimers, generating
multimers.
Since alginates and pectins both are known to have a similar structure and form
gels in the presence of calcium, it is expected to see the similar multiple step gelation
process also in pectin. The different behaviors in alginates and pectin gelation were
attributed to the structural difference; alginates are block-copolymer while arrange-
ment of copolymer units in pectins is more random (Fang et al. 2008).
Recently, effects of the addition of oligoguluronate, guluronate block (GB) to
alginates with and without salt on rheological properties were studied, and the ratio
of Ca to guluronate G, R(Ca/G) was found to play an important role (Liao et al.
2015). The addition of GB was inhibitive in the range of 0.25 < R (Ca/G) < 0.60 and
promotive in the range of R > 0.60. These two effects were shown to be associated
with the different molecular events that dominate the gelation of alginate, namely,
egg-box dimerization and lateral aggregation. Quantitative analysis indicates a
competitive binding rather than a screening binding during egg-box dimerization,
which led to the inhibitory effect in the lower Ca concentration regime. On the other
hand, in the higher Ca concentration regime where alginate gelation is predominated
by chain lateral aggregation, the dimers formed by GB could act as a binder to
enhance the aggregation of alginate dimers, resulting in a promotive effect on
alginate gelation (Fig. 4.7). The results are consistent with the microstructures
observed by AFM.
138 K. Nishinari

44 (a) Control 1.2 (b)

Iysine
S3
43 S2 0.9
S1
β-Ig
To(°C)

42 0.6

φ
41 0.3

40 0
0 2 4 6 8 10 0 2 4 6 8 10
r r

Fig. 4.8 The effect of β-lg or its hydrolysates on the onset DSC temperature (T0) of helix formation
of κ-car (a) and relative extent (φ) of the conformational transition of κ-car (b) as a function of the
mixing ratio r ¼ β-lg/κ-car. The concentration of κ-car 0.15%; pH ¼ 4.7. Reproduced with
permission from Cao et al. (2016b), Copyright 2016 ACS

Cao et al. (2016a) studied the mixture of κ-carrageenan (κ-car) and β-lg with
different mixing ratios r ¼ β-lg/κ-car, and found that the exothermic peak enthalpy
of κ-car accompanying coil to helix transition detected by DSC decreased with
increasing r. It was analyzed quantitatively based on a theory of McGhee and von
Hippel (1974) which was used to explain the binding of protein to DNA. The relative
extent of conformational transition ϕ (r) could be experimentally measured as the
ratio of the enthalpy change of conformational transition of protein/polyelectrolyte
mixture (ΔH (r)) to that of pure polyelectrolyte (ΔH (r ¼ 0)). The inhibiting degree ϕ
of κ-car helix formation by β-lg was quantified by the enthalpy change as a function
of r. Electrostatic complexation in soluble state had subtle effect on the coil-to-helix
transition of κ-car, which is due to the relatively high freedom of κ-car at the initial
stage of protein binding. Electrostatic complexation in insoluble state however
greatly suppressed the conformational transition, which is due to the high physical
hindrance imposed by β-lg upon extensive protein binding. The effect of protein/
polyelectrolyte electrostatic complexation on the conformation transition of poly-
electrolyte was described quantitatively. The effect was closely associated with the
molar mass of β-lg or its hydrolysates: Larger hydrolysates S1 (>2000 Da) had an
inhibitory effect on the conformational transition of κ-car, smaller hydrolysates S3
(<1000 Da) tended to promote it (Fig. 4.8).

4.9 Cation- or Acid-Induced Gelation

Gelation of globular proteins can be induced either by heating (heat induced

gelation) or by cold gelation. In the cold gelation, a low concentration solution of
native proteins is heated or subjected to high pressure and soluble aggregates are
formed. The aggregates remain soluble on cooling. Then, gelation is induced either
by adding salt or by changing the pH toward the isoelectric point of the proteins in
order to reduce the electrostatic repulsion. Cold-set gels have been used as a vehicle
4 Gelling Properties 139

to deliver thermally labile and water insoluble functional ingredients (Chen et al.
2006), and recently cold-set emulsion gels are also attracting much attention
(McClements 2017; Khalesi et al. 2019).
Ion-induced gelation of alginate has been used to produce an imitation ikra
(salmon roe). When a droplet of non-gelling sodium alginate falls into calcium
lactate solution, a thin film is formed on the surface of droplet. This process was
patented by a Japanese chemical industry (Nishinari 1988). This very fast gelation
can be used to produce artificial berries and onion rings (Draget 2021). Since this
gelation of sodium alginate is too fast and leads to inhomogeneous gel formation,
slow release of calcium ions from insoluble calcium salts is used by chelating agents
EDTA, citrate) or glucono-delta-lactone to control the gelation rate and to form
homogeneous gels (Draget 2021). Alginate is also used to treat heart burn (acid
reflux) since alginate solution forms a gel raft on the top surface in the stomach
because of its low pH (ca 1.5) (Gaviscon has been used).

5 Characterization Methods

5.1 Gelation Kinetics: Time Dependence of Storage and Loss

Moduli G0 and G00 of a Solution at a Constant
Temperature and a Frequency

When a solution prepared at a non-gelling temperature is kept at a certain gelling

temperature, G* begins to increase with lapse of time. Generally employed condition
of the measurements are as follows: the frequency and the amplitude should be as
low as possible so that the structure being formed might not be broken, however, the
frequency ca l Hz has been chosen in most cases. The geometry of the apparatus for
viscoelastic measurements should be chosen so that the injected sample solution can
be quenched (cooled rapidly) or can be heated rapidly. This can be satisfied generally
if the required volume of sample solution is small.
Generally, the gelation proceeds faster at higher temperatures for a heat-setting
system whilst it proceeds faster at lower temperatures for a cold-setting system. It is
easier to carry out the rheological measurement at the temperature where the gelation
proceeds slowly for the analysis of kinetics. In the first order kinetics, the storage
modulus is written as follows:

G0 ðt Þ ¼ G0sat ½1
exp ð
k ðt
t 0 ÞÞ ,

where t0 is the latent time (gelation time), k is the rate constant, G0 sat is the saturated
value of the storage modulus after a long time. Solutions of some biopolymers such
as methyl cellulose, degalactosylated xyloglucan, curdlan, glycinin, and
β-conglycinin form a gel on heating, while solutions of other biopolymers such as
agarose, carrageenan, gelatin, and gellan form a gel on cooling (All these were cited
140 K. Nishinari

in Nishinari 1997). When the gelling polymer is composed of fast gelling and slow
gelling components, the above kinetic equation can be modified using two rate
constants and two latent times for each components. More kinetic models have
been proposed, see p. 386 in Lapasin and Pricl (1999). Recently, the gelation of
β-lactoglobulin was analyzed by a simple eq. G0 (t) ¼ G0 sat exp(
β/t) proposed by
Scott Blair in 1963.
Lapasin et al. (1990) reported that both moduli increased first and then decreased
during gelation of calcium pectate. The decrease of the moduli was attributed to the
structure breakdown into smaller flow units.
Rennet-induced gels have been studied extensively by many groups as a basis for
cheese production. The dependence of casein micelle size on the gelation has been
studied using micelles with different sizes prepared by differential centrifugation,
and it was found that smaller casein micelles form a stronger gel faster (Niki et al.
1994). It was interpreted on the basis that the κ-casein, which plays a main role in
rennet-induced casein gelation, is abundant on the surface of casein micelles.
As in the enzymatic modification of casein, partial hydrolysis has a great potential
to control the structure and property of proteins. Doi et al. (1987) performed a limited
proteolysis of ovalbumin (OVA) (Mw ¼ 45,000) using a pepsin, and obtained an
intermediate OVA (Mw ¼ 42,500), called p-OVA having a similar physicochemical
properties with native OVA, and compared the gelation. This will be described later
in Sect. 7.
To control the rheological and structural properties of proteins, transglutaminase
(TGase) was used by several research groups. Jaros et al. (2010) reported G0 max in
GDL-induced gelation process of TGase-treated casein solution at 30 min after the
GDL addition, and G0 max depended strongly on the incubation time by TGase. G0 max
increased and then deceased with the incubation time, and was attributed to
rearrangement and not due to the slippage frequently observed in gelation experi-
ments. Syneresis and tanδ showed the minimum at 30 min after the GDL addition
when G0 showed the maximum (Fig. 4.9). In the processing of yogurt, various
polysaccharides or microparticulated whey proteins are added to increase G0 ,
which reduces the syneresis and increases the creaminess (Jørgensen et al. 2019).
Soy protein gels are used not only for tofu but also attracted attention as raw
materials for cheese and yogurt to replace animal milk, and the optimization
condition has been studied by many research groups (Nishinari et al. 2018).

5.2 Mechanical Spectra: Frequency Dependence of G0

and G00 at a Constant Temperature

Figure 4.10 shows the frequency dependence of gellan solutions with a small and
large amount of NaCl. Most commonly available commercial gellan samples contain
salts to enhance the gelling ability. This figure shows the behavior of gellan which is
converted into pure sodium type to see the effect of salts on the gelation behavior.
4 Gelling Properties 141

Fig. 4.9 (a) Averaged gelation curves of three enzyme treated casein solutions (27 g kg
1). TGase
treatment was with 3U TGase g
1 protein at 40  C, the numbers refer to incubation times in hours.
Inactivation was with 1 g L
1 N-Ethylmaleimide, and gelation was induced by 40 g L
1 GDL at
30  C. Dotted line shows the corresponding pH decay. (b) G0 max (□) with the corresponding tan δ
(■) and syneresis (○) of cross-linked casein gels as affected by enzyme incubation time. Data are
mean values  standard deviations of three individual experiments. Reproduced with permission
from Jaros et al. (2010), Copyright 2010 Elsevier

Fig. 4.10 Frequency dependence of G0 (open) and G00 (closed) of 2 wt% sodium type gellan with
30 mM NaCl at 5  C (a) and 0.3 wt% sodium type gellan with 100 mM NaCl at 35  C (b)
Reproduced with permission from Nitta et al. (2010), Copyright 2010 ACS

Both storage modulus G0 and loss modulus G00 show a weak frequency dependence
in Fig. 4.10a, while both G0 and G00 show a plateau in Fig. 4.10b although the
polymer concentration is much lower in the latter.
The behavior in Fig. 4.10a is seen in a structured liquid (often called a “weak gel,”
but this term was not to be used because it is essentially a liquid), and is different
142 K. Nishinari

from a “true gel” or “elastic gel” behavior shown in Fig. 4.10b. The difference
between a structured liquid and an elastic gel appears in a larger value of tangent
delta for the former because of its liquid nature. The large deformation behavior
shows a more clear difference between a structured fluid and an elastic gel; the
former flows while the latter fractures above the yield stress. For example, see the
strain dependence of G0 and G00 in Fig. 4.4 and the frequency dependence G0 and G00
in Fig. 4.3 for κ-carrageenan in a paper by Ikeda and Nishinari (2001d).
In the globular protein such as soybean β-conglycinin, the gelation commences
only at higher temperatures than the denaturation temperature. The frequency
dependence for G0 and G00 of β-conglycinin or glycinin solution begins to show a
plateau at 68  C or 80  C, respectively (Nagano et al. 1994a; Nishinari et al. 2014).

5.3 Thermal Scanning Rheology

Temperature dependence of storage and loss moduli G0 and G00 of a solution at a

constant frequency.
Figure 4.11 shows the gelation process of a 15% soybean β-conglycinin solution.
At a constant temperature of 80  C, G0 increased, and continued to increase when the
temperature was lowered from 80  C to 20  C at the rate of 2  C/min. G0 decreased
with increasing temperature from 20  C to 80  C at the same rate. Values of G0 are
symmetrical about the vertical line at t ¼ 3600 s. The increase in G0 after 1800 s
when heating is stopped and the temperature is lowered is attributed to the further
formation of network structure by hydrogen bonding which may be broken by
subsequent heating from 20  C to 80  C. Although it is almost impossible to evaluate
the contribution of hydrophobic interactions, hydrogen bonding, ionic interactions,
and covalent bonding quantitatively because these interactions operate

Fig. 4.11 Gelation process 4000 100

of 15% soybean temperature
β-conglycinin solution at
pH 7.6. The solution was 80
heated at 80  C for 30 min 3000
(¼1800 s), and then cooled
temperature/°C

to 20  C at 2  C /min, and 60
heated again to 80  C at the
G’ / Pa

same rate. Reproduced with 2000

permission from Nagano 40
et al. (1994b), Copyright G’
1994 ACS
1000
20

0 0
0 1000 2000 3000 4000 5000 6000
time/s
4 Gelling Properties 143

simultaneously, Fig. 4.11 clearly shows the important contribution of hydrogen

bonding to the gel formation of β-conglycinin.
At higher temperatures for a 2 wt% solution of sodium type gellan, G0 < G00 while
on cooling moduli show a steep increase at ca 35  C (Tch) which is attributed to coil-
to-helix transition, which coincides with the temperature at which the molecular
ellipticity shows a transition. On further cooling, G0 and G00 show a crossover at 8  C
(Tsg) which is ascribed to sol-gel transition (Miyoshi and Nishinari 1999). This
sol-gel transition was found not the formation of a true gel but a structured liquid.
Strictly speaking, this procedure to determine the gelation point is not precise
because the crossover point depends on the frequency of measurement in addition
to scan rate.
For gelling polysaccharides, it is necessary to pay attention to the slippage which
sometimes leads to a serious misinterpretation. The cooling curve of G0 and G00 using
a normal cone and plate geometry showed a peak for both G0 and G00 in the cooling
process of carrageenan solutions, while that using perforated cylinders which pre-
vent the slippage, no peak of G0 and G00 was observed (Richardson and Goycoolea
1994). Zhang et al. (2001) also found the peak of G0 and G00 as a function of time for
2% dispersions of KGM at higher temperatures at ω ¼ 1rad/s. It is known that
gelation of KGM is faster at higher temperatures. Zhang et al. (2001) measured the
compressive force during gelation of KGM which is free from slippage, and then
they did not observe the peak even at higher temperatures confirming that the peak of
G0 and G00 observed was caused by the slippage.

6 Physical Properties of Gels

6.1 Effect of Gelling Rate on Gel Properties

The elastic modulus of gelatin gels increased with time at lower temperatures not
reaching an equilibrium value even after 100 h (Nijenhuis 1981, 1997). The thermal
stability of gelatin gels increased with increasing storage time at the temperature
which is a little higher than gelation temperature (Michon et al. 1997). For agarose
gels, which are other helix-forming thermoreversible gels, the effect of the storage
temperature near the gelation temperature on rheological and structural properties
has been studied and it was found to differ from the behavior of gelatin gels. Aymard
et al. (2001) found that holding solutions of agarose for long times at high temper-
atures decreased the strength of the gels formed on cooling. They interpreted this by
structural observation that during the holding period the un-gelled solutions resolved
progressively into regions of high and low polymer concentration, and that the
resulting heterogeneity gave weaker networks when the solutions were gelled by
cooling. On the other hand, Mohammed et al. (1998) reported that agarose gels
showed larger elastic modulus and were more thermally stabilized by cooling more
slowly. Recently, it was shown that storage Young’s modulus and the fracture stress
and strain of gellan gels increased with decreasing cooling rate (Nitta et al. 2014).
144 K. Nishinari

As mentioned before, a gel may be formed before a solid film is formed when the
solvent is evaporated from, for example, solutions of pullulan which is usually not
called a gelling polysaccharide because it forms a gel only at higher concentrations.
The evaporation speed influences the length scale of the structure being formed, but
this problem has only been tackled recently (Schaefer et al. 2015) mainly in relation
to spin coating. This may be called a kind of concentration quench, as temperature
quench influenced the formation of gel structure.

6.2 Temperature Dependence of Elasticity of Gels

Sol-gel transition in polysaccharides can be classified into four groups: (1) cold-set
gels like agarose, kappa- and iota-carrageenans, and gellans which form a gel on
cooling the solution; (2) heat-set gels like some cellulose derivatives such as methyl
cellulose (MC), curdlan, konjac glucomannan (KGM) which form a gel on heating
the solution; (3) inverse reentrant gels like a mixed solution of methyl cellulose and
gelatin, which forms a gel at higher and lower temperatures and stays a sol state at an
intermediate temperature range. Gelatin may be replaced by some po1ysaccharides
which form a gel on cooling; and (4) reentrant gels like xyloglucan from which some
galactose residues are removed. It forms a gel at an intermediate specific temperature
range and remains in the sol state at higher and lower temperatures outside of this
specific temperature range. Some copolymer gels show a complex temperature
dependence: Sol-gel transition is observed at two or three different temperatures,
i.e. they show reentrant transition (Nishinari 2000a, b, 2001, 2009).
There have been many investigations and debate on the temperature dependence
of the elastic modulus or gels relating with the entropic and energetic contribution.
Since it was difficult to observe the elastic modulus of thermoreversible gels as a
function of temperature from a low temperature, the increase in modulus with
increasing temperature was not observed. It was concluded that the gel elasticity is
energetic elasticity in previous studies while the opposite was also asserted. To
understand this problem it is effective by dividing the elastic modulus into entropic
and energetic contribution from the temperature dependence of the elastic modulus
of gels of agarose with different molar masses (Nishinari et al. 1984).
Reel-chain model was proposed to explain the temperature dependence of
thermoreversible gels (Nishinari et al. 1985). In this model, network consists of
Langevin chains which allow to treat large deformation. On heating, some segments
are released from junction zones consisting of aggregated helices. There should be a
limit for number of segments released before gel-to-sol transition occurs. Some
statistical calculations gave the elastic modulus as a function of the binding energy,
upper limit number of segments released from the junction zone, average distance
between junction zones. It predicts the monotonic increase of the modulus for a high
binding energy as expected from rubber elasticity theory, and the monotonic
decrease for a low binding energy, and an intermediate behavior showing the
4 Gelling Properties 145

maximum of the modulus as a function of temperature. This theory was applied to

many thermoreversible gels to explain the temperature dependence of the modulus.

6.3 Molar Mass Dependence of Elastic Modulus

Since it is difficult to prepare samples with different molar masses with a narrow
molar mass distribution, there have not been so many studies on the molar mass
dependence of elastic modulus of gels. Saunders and Ward (1955) studied rheolog-
ical properties of gelatin gels with different molar masses and showed that the elastic
modulus increased with increasing molar mass up to a certain value and then leveled
off whilst the breaking stress continued to increase with increasing molar mass.
Similar tendency was observed for gels of alginate (Smidsrød 1974) and
κ-carrageenan (Rochas et al. 1990). The gelation kinetics of dispersions of konjac
glucomannan with different molar masses has been studied by measurements of
storage shear modulus and it was shown that the storage modulus of the dispersion of
the same concentration increased with increasing molar mass (Yoshimura and
Nishinari 1999).
Effects of molar mass on the stress-strain curve and on the temperature depen-
dence of the modulus of agarose gels are shown in Fig. 4.12. Both the stress and
strain at break increased (Fig. 4.12a), and the entropic behavior of elastic moduli was
enhanced (Fig. 4.12b) with increasing molar mass of agarose.

Fig. 4.12 (a) Stress-strain relation of 2.4% (w/w) gels of agarose with different molar masses
(• < □ < r < ○) at 25  C. Reproduced with permission from Watase and Nishinari (1983),
Copyright 1983 Springer. (b) Temperature dependence of storage modulus E0 for five agarose
fractions with Mw from 3.4  104 g mol
1 (F1) to 48.5  104 g mol
1 (F5) (Nishinari and Watase
1993)
146 K. Nishinari

Molar mass dependence of the moduli was found sometimes contradictory

(Nishinari and Fang 2021). While from a figure of G0 as a function of concentration
in amylose gels showed a greater G0 for a higher molar mass amylose (Clark et al.
1989), a similar plot for oat β-glucan showed a greater G0 for a lower molar mass
(Lazaridou et al. 2003). This apparent inconsistency is originated from the
non-equilibrium nature of these physical gels. In both amylose and oat β-glucan,
the gelation proceeded faster for lower molar mass because of the higher mobility.
This slower pace of gelation is different in amylose and β-glucan, and therefore,
great care is required to compare the molar mass dependence of the modulus. Gel
formation of higher molar mass fractions is so slow that the modulus at a time not
long enough is smaller than that for lower mass fractions. Gelation of gelatin is also
known to take a long time as mentioned before (Nijenhuis 1981, 1997).

6.4 Molecular Motion (Rearrangement) in Gels

Agarose forms a gel at a very low concentration < 0.1 wt% depending on the molar
mass, sulfate content (purified agarose or idealized agarose contains no sulfate
groups but most commercially available agarose contains a small amount of sulfate
groups). Even after gelation it is not in the equilibrium state. Syneresis, which is an
exudation process of water from the network, is also observed in globular protein
gels such as cheese (syneresis is necessary) and yogurt (syneresis is not preferable).
This process is not so fast, and therefore gel engineers study small and large
deformation rheological behavior of these gels in pseudo-equilibrium state. Agarose
gels as well as other polysaccharide gels are believed to be formed by aggregated
double helices connected by flexible chains. Since the concentration is low, there are
free spaces where molecular motion is occurring.
A step-like decrease of tan δ was observed while the endothermic peak in DSC
and the steep change in the specific ellipticity in CD were observed at 25  C on
heating a 1.6% gellan gel, and these changes are attributed to helix-coil transition
(Nitta et al. 2001; Nishinari et al. 2001). A molecular motion or molecular
rearrangement such as the helix-coil transition occurs in an apparently solid material
with the elastic modulus of ca 105 Pa. This was a surprise for authors because of the
appearance of a solid of the gellan gels. From the viewpoint of the solid content,
most part of the gel consists of water and thus there must be much enough space
where large-scale molecular motion or rearrangement can occur.
Helix-coil transition can occur in different ways. Segments can be released from
junction zones on heating as was discussed in reel-chain model approach (See Sect.
6.2). On cooling, these released segments are reeled into junction zones so that
junction zones become longer and/or thicker. If the elastic modulus is mainly
determined by the number of elastically active chains which connect junction
zones as in the theory of rubber elasticity, the increase in elastic modulus should
be attributed to the increase in the number of elastically active chains rather than to
the thickening or lengthening of junction zones. It is quite possible that long
4 Gelling Properties 147

segments released from junction zones form a new junction by helix formation.
Helix-coil transition does not necessarily occur only at the chain ends, but it also may
occur at the intermediate point of long chains.
There may be different junction zones with different thermal stabilities. Weak
junction zones with low thermal stabilities may disintegrate at lower temperatures
which arc directly related to helix-coil transitions, and these chains produced from
the disintegration of helices may form helices on cooling only at lower temperatures.
The distribution of thermal stability may cause a broad endo- and exo-thermic peak
in DSC heating and cooling curves as well as gradual increase and decrease of the
specific ellipticity in heating and cooling CD measurements. According to a fibrous
model (Morris et al. 2000), the elasticity of gels arises mainly from stretching and
bending of fibers that consist of aggregated helices. When some ends of these fibers
arc converted into flexible chains by helix-coil transition, these ends will cease to
contribute to the elasticity and then the elastic modulus will decrease. Even if one of
these conformational changes occurs, it does not necessarily lead to the gel-to-sol
transition. When these changes are only local, only less ordered helices are
converted into coils, and the whole network structure is not broken down, and
keeps the size and shape of the self-supporting gel.

6.5 Chain Release and Erosion of Gels

Most gel technologists using chemical gels formed by covalent bonds believe that a
polymer gel consists of a three-dimensional crosslinked network and swells in a
solvent to a certain finite extent, but does not dissolve even in a good solvent. It was
found that some molecular chains in a gellan gel release out from the gel when it is
immersed in a solvent such as water or salt solutions (Tanaka and Nishinari 2007;
Hossain and Nishinari 2009). After a long time, the gel swells infinitely, and
disperses completely in a great amount of water. On immersion of gels in water,
cations and non-crosslinked chains diffuse out from the gel into the surrounding
medium, and the gel structure is weakened. Salt diffusion from the gels into the
surrounding medium is faster than chain release; chains which lose condensed or
bound ions cannot retain a helical conformation, and so they then diffuse out.
Analysis of the released chains revealed that the shorter chains are released predom-
inantly. The modulus increase by immersion was observed at first, and it was
suggested by stiffening of elastically active chains. Potassium ions in the external
solvent were found to strengthen the gel structure of gellan, yet
tetramethylammonium (TMA) ions in the external solvent, which are known to
inhibit the gelation though not inhibiting helix formation, were also found to
increase the modulus.
A mass loss in gellan gels prepared by calcium ions were also found, and after
28 days these gels maintained constant composition without further mass loss during
the remainder of the test period between 28 and 164 days (De Silva et al. 2013). This
shows the difference between monovalent cations and divalent cations as will be
discussed in Sect. 6.7.
148 K. Nishinari

6.6 Syneresis/Water Holding Capacity (WHC)

Syneresis is a phenomenon in which water oozes out from the gel network, and is
known from long time ago but its mechanism is not so well understood although the
importance has been recognized because of its close relation with flavor release and
juiciness perception. Many methods of measurements have been proposed: centri-
fugation, compression, wiping the surface, measuring water on the top of a test tube
containing the gel, etc. As Mizrahi described (Mizrahi 2010), the syneresis may be
governed by the swelling pressure which is the difference between the osmotic
pressure and the network pressure Πsw ¼ Πos –Πnet, where the osmotic pressure is
produced by polymer chains in the gel and it drives water into the gel, thus causing
the swelling and stretching of polymer network. Stretched chains tend to contract to
compensate the decrease of the entropy caused by the chain stretching, thus giving
rise to an internal pressure which is called a network pressure. Mizrahi (2010)
suggested several methods to prevent the syneresis by increasing the osmotic
pressure, controlling network pressure and crosslinking.
Miwa et al. (1994) compared the syneresis of 3–4 wt % gels of curdlan, carra-
geenan, agar, konjac after kept at 4  C for 20 h, and the syneresis of both curdlan and
konjac was 10.3% which was much larger than that of carrageenan (1.4%) and agar
(0.6%). However, after freezing and thawing, syneresis of both curdlan and konjac
increased to ca 21% while it was much more than that for carrageenan and agar
because these gels became sponge-like and water ran out through larger pores. This
encouraged food industry to use curdlan and konjac in frozen foods such as surimi to
improve the texture. Syneresis of agar gels was reported to decrease with increasing
agar concentration/added sucrose/sulfate residue. It should be reminded that the
order of the addition of sucrose to make polysaccharide gels, before or after gelation,
changes the structure/mechanical properties and syneresis. For example, the addition
of a higher sucrose content (>55 wt%) in 1 wt% agar gelation before heating was
found to make the gel inhomogeneous and led to a weaker gel with more syneresis
(Yang et al. 2015). It was found that the elastic modulus decreased after syneresis
although the concentration was increased by syneresis, and it was attributed to the
loosening of the network structure caused by syneresis (Nagasaka and Taneya 2000;
Nishinari and Fang 2016, 2017). It was also shown that syneresis was reduced by
adding sucrose or starch to curdlan though it also reduced the gel strength. While this
practical manipulation may be sometimes useful, the basic understanding of syner-
esis in polysaccharide gels is not yet reached (Ako 2017; Divoux et al. 2015).
Syneresis in globular protein gels may be understood in the first approximation by
permeability B which is proportional to the area and the fraction of pores in a cross
section of the gel (Walstra 2003). Walstra explains the great difference in the
syneresis in ground coffee (B ~ 10
8 m2), rennet milk gel (B ~ 10
12 m2), and
polysaccharide gel (B ~ 10
17 m2) by the approximate value of the permeability. The
difference of B values roughly explains that some polysaccharide gels retain water
while globular protein gels show a much more syneresis. While syneresis is unde-
sirable in most gels, it is an essential process in cheese making, where most of the
liquid (whey) in the rennet gel has to be removed (Walstra 2003). In these protein
4 Gelling Properties 149

gels, rearrangements of strands consisting of globular protein aggregates lead to

loose ends which in turn form junctions thus becoming more compact expelling
liquids. When junctions are broken, a pressure is built up, which is called “endog-
enous syneresis pressure,” that is causing syneresis even without external pressure
such as gravity. However, the experimental determination of the endogenous syn-
eresis pressure seems to be difficult, and thus the complete understanding of the
syneresis of protein gels is still a challenge.
Nieuwland et al. (2016) found a good correlation between the structure, mechan-
ical properties, and WHC for ovalbumin (OVA) gels at a pH range from 5.8 to 6.8:
the microstructure of OVA gels became less dense based on SEM and CLSM, and
the Young’s modulus decreased, in parallel with the decrease in WHC with
decreasing pH.
Urbonaite et al. (2016) prepared 14 w/w% WPI gels at pH 7.2 with different pore
sizes ranging from 10
2 to 100 μm by changing NaCl content from 0 to 300 mM to
examine the effect of coarseness (pore size) and the mechanical properties, Young’s
modulus on WHC. They found that, depending on the lower or higher than 100 mM
NaCl, fine stranded gels (length scale <0.1μm) or coarse stranded gels (> 0.1μm)
were formed. They found the Young’s modulus showed the maximum at 100 mM
NaCl, while the WHC decreased with increasing concentration of added NaCl, and
concluded that coarseness was more dominant than stiffness (Young’s modulus) for
water holding.
Dai et al. (2016) examined the effects of the addition of 0.5% KGM on the
physicochemical properties of yogurt, and found that the syneresis was decreased
while retaining the structural stability and the firmness of full-fat yogurt. They found
the suppression of syneresis was equivalent to previous reports in which inulin or
orange fiber was added to skimmed yogurt and noticed that total solid content was
also responsible for the reduction of syneresis.
To improve the texture and water holding capacity of myofibrillar protein gels,
dietary fibers are widely used. Effects of the addition of insoluble fibers (microcrys-
talline cellulose, oat fiber, walnut shell flour) and native and modified starches
(potato, tapioca) on liquid loss, hardness and resilience (recoverable deformation)
of chicken breast myofibrillar proteins were measured (Gravelle et al. 2017). Since
fat and connective tissues were removed, the samples were myofibrillar protein gels,
and liquid loss was mainly from water. Among the fillers tested, native potato starch
was most effective to reduce the liquid loss, which was attributed to the difference in
hydration estimated by T2 relaxation time of water.

6.7 Effect of Sugars, Salt, Acid, and Polyphenols

on Rheological Behavior of Gels

It has been empirically known that the addition of sucrose to polysaccharide gels
make them transparent, less brittle and reduce the syneresis. The Young’s modulus,
150 K. Nishinari

fracture stress, and melting enthalpy increased and then decreased with increasing
sucrose or glucose in agarose or κ-carrageenan gels (Nishinari and Fang 2016). The
different strengthening effect of sugars has been correlated with a number of
equatorial hydroxyl residues in sugars. Kawai et al. (2007, 2008) extended the
reel-chain model in which flexible chains are released from junction zones consti-
tuting aggregated helices to explain the observed strain hardening and then softening
in uniaxial compression. As the end-to-end distance increases due to the deforma-
tion, more and more segments are reeled out from the junction zone. Finally, one end
of the chain is liberated from the junction and the chain becomes dangling. The
appearance of dangling chains causes the strain softening because they cease to
contribute to the elasticity.
Drying of gels has been studied from various viewpoints, and recently the effect
of sugars on drying kinetics of agarose gels was studied (Mao et al. 2017).
Epiga1locatechin gallate (EGCG), a polyphenol abundant in tea leaves was found
to induce gelation of non-gelling xyloglucan (Nitta et al. 2004). Yan et al. (2016)
further reported that other gallate analogs also induce gelation of xyloglucan.
Generally, salt does not influence so much the gelation of neutral polysaccharides
such as agarose but sometimes enhances the gelation of KGM or xyloglucan. Effects
of salts on gelation of charged polysaccharides have been extensively studied. While
the monovalent cations shield the electrostatic repulsion of carboxyl groups in
gellan, divalent cations bind different molecular chains of gellan, and therefore
once the gel is formed it does not return to the sol state on heating up to 90  C
(Miyoshi et al. 1994; Djabourov et al. 2013).

7 Particulate Disordered Structure: Globular Protein Gels

When globular proteins are denatured by heating, high pressure or denaturants such
as urea or guanidine hydrochloride, hydrophobic portion folded and buried inside
were exposed outside, which is called unfolding, and then unfolded polypeptide
chains form a three-dimensional network (Doi 1993). In comparison with network
structure formed from long chain molecules which are predominantly entropic and
have been successfully elucidated to some extent by developing rubber elasticity
theory, particulate disordered gels are less well understood. Rheology and structure
of rennet-induced or acid-induced casein or whey protein gels (Nicolai et al. 2011),
ovalbumin gels (Nemoto 2000; Nieuwland et al. 2016) as well as soybean proteins
(Nishinari et al. 2014), and other plant proteins have been extensively studied.
A great difference from thermoreversible polysaccharide gels such as agarose,
kappa-carrageenan, gellan is that the minimum concentration required is much
higher for globular protein gels, but recently, gelation of fibrils made from globular
protein changed this common knowledge. This was described in “4.7 Rigid network
chains.” Most polysaccharide gels except cellulose derivatives such as methyl
cellulose form a gel by hydrogen bonds and thus form a gel on cooling while
globular protein gels are formed on heating and it is thought that the denaturation
4 Gelling Properties 151

of the native protein structure is prerequisite for gel formation. The main force
responsible for gelation is thought to be hydrophobic interaction, but other second-
ary forces are also contributing.
Dutch school employed a fractal model and measured the permeability and
rheology to correlate the structure and property of milk protein gels. Bremer et al.
(1989) concluded that acid casein gels can be described very well by a fractal model
with a fractal dimension 2.3, while it was not applicable for rennet-induced casein
gels because of microsyneresis. The modulus decreased with increasing temperature
which contradicts some earlier publications affirming that the casein gels are entro-
pic like rubber elasticity (van Vliet and Walstra 1985). The increase of the aging
temperature to 50  C and extending the aging time to 157 h changed the gel to a more
solid like material; the storage modulus tends to show a rubber like plateau and the
loss tangent decreased. Storage modulus G0 as a function of measuring temperature
Tm for acid sodium caseinate gels aged at different temperatures increased with
increasing aging temperature, but decreased with the temperature of measurement
(Roefs and van Vliet 1990). It is generally accepted that in casein gels, an ultimate
effect of structural rearrangement is syneresis: expulsion of liquid. Syneresis is
desired for manufacturing of cheese but not for yogurt.
Doi (1993) classified the gelation of globular protein into random aggregates and
string beads network. His group studied the gelation of ovalbumin (OVA) by
rheology, dynamic light scattering (DLS), CD, TEM, SEM, CLSM based on the
conformational studies reporting that OVA molecules are worm-like chains, linear
aggregates, with cylindrical diameter 12 nm and the persistence length 23 nm (Doi
et al. 1987; Nemoto 2000). As a result of heat denaturation, hydrophobic regions are
exposed on the surface, and linear polymer or aggregate is formed depending on the
balance of hydrophobic interaction and electrostatic repulsion (Fig. 4.13).
They found that the heat induced gelation of 39–59% OVA solution showed a
critical structure obeying the Winter-Chambon criteria (Koike et al. 1996), and
found a very low exponent 0.09–0.14. Gels formed with concentrations lower than
25% and higher than 59% did not show critical structure. The exponent of concen-
tration dependence was found to change from 4 below 59% to 10 above the
concentration range from 59% to 89%, where the modulus an order of 109 Pa
close to the glass modulus was found. This is totally different from common
concentration dependence of modulus for polysaccharide gels where the exponent
is ca 4 at lower concentration range, but ca 2 at a higher concentration range as
described in “4.6 Critical molar mass and concentration for gelation.”
A two-step heating was employed in addition to one-step heating; the OVA
solution was first heated without NaCl for 60 min, and quickly quenched to room
temperature where linear aggregates molar mass higher than several millions were
formed. After addition of NaCl (20, 60, and 100 mM) to the viscous solutions, they
were reheated for various time periods from 2 to 240 min and then immediately
cooled (Koike et al. 1998). Transparent gels were obtained by both the one-step and
the two-step heating. Doi (1993) described methods to obtain transparent egg white
or milk whey protein gels controlling the pH and ionic strength by two-step heating.
152 K. Nishinari

Fig. 4.13 Schematic representation of the heat denaturation of OVA, leading to the formation of
linear polymer or aggregate (Doi et al. 1987). Reproduced with permission from Doi et al. (1987),
Copyright 1987 John Wiley & Sons

More recently, gelling behavior of OVA and p-OVA (formed by a limited

hydrolysis with pepsin) was analyzed based on SANS (small angle neutron scatter-
ing) in addition to DLS (Hiroi et al. 2016). In spite of the similarity of physico-
chemical properties, intrinsic viscosity, secondary structure, denaturation
temperature, OVA and p-OVA showed significantly different gelling behaviors on
heating. Heating 6 wt% solutions at neutral pH without salt led to a transparent gel
for OVA while a turbid gel was formed for p-OVA. On heating OVA solution, the
resulted aggregate size was found proportional to the protein concentration judging
from the independence of the peak position in the SANS intensity as a function of
scattering angle. On the contrary, an increase in the amount of large clusters formed
on heating was thought to disturb the correlation of p-OVA dimers resulting in wide
distribution of the distance between each solute and thus a two-phase separated
structure was formed. The fact that the cleavage of only 22 N-terminal residues from
OVA led to a very different gelling behavior is expected to be a hint to develop
further utilization of OVA.
Globular protein solutions show a “solid-like” mechanical spectra (shown in
Chap. 3) even before gelation probably because they form a colloidal crystalline
structure (Matsumoto and Inoue 1993; Ikeda and Nishinari 2001a, b, c). Tobitani and
Ross-Murphy (1997) made a temperature-concentration diagram constructed by the
gelation temperature for each concentration of globular protein which is determined
4 Gelling Properties 153

by interpolation and extrapolation of the data set, a locus of nominally infinite

gelation time.

8 Mixed Gels

Just as in alloys of metals and plastics, many mixed gels have been prepared to
respond to the demand of various functionalities (Morris 1986). Many papers have
been published on the mixed gels of xanthan, locust bean gum, konjac mannan, etc.,
and some common features are recognized for synergistic interaction, which means
that the only one polysaccharide does not form a gel at that concentration, but forms
a gel by mixing with other polysaccharide. 1) One polysaccharide undergoes coil-
helix conformational transition, such as agarose, carrageenan, furcellaran, gellan,
and xanthan gum. 2) The backbone linkage of the other polysaccharide in the
mixture is β(1,4) linkage, such as in konjac glucomannan or galactomannan, or
xyloglucan (Morris 1994).
Polysaccharide mixture gels have been extensively studied, and Unilever model
and Norwich model have been proposed and examined. Gels of the mixture of
xanthan and galactomannan or konjac glucomannan have been studied, and it was
proposed that helical xanthan associates with the smooth region of galactomannan
devoid of galactose side chains (Unilever model, Dea et al. 1977) while Norwich
model proposed that the mixed gel was formed between coiled xanthan and
galactomannan or KGM because gels were formed only after the helical xanthan
was denatured into coil state (Brownsey et al. 1988). The latter statement was
affirmed by adding helix enhancing cations to raise the helix-coil transition and in
such a condition the mixture did not form a gel on cooling. However, the gelation in
the mixture of pyruvate free xanthan (PFX) and KGM was observed to start about
40  C more than 60  C lower than coil-to-helix transition temperature of xanthan at
acidic pH (3.5 and 4) higher than 100  C, and that the synergistic gelation of KGM
with PFX and commercial xanthan was not so different. Thus, the above-mentioned
prerequisite of Norwich group for the gel formation of the mixture, the conformation
of xanthan should be in coil state, was refuted (Agoub et al. 2007).
The effect of salt is important to study the mixtures. Annable et al. (1994) studied
the mixture of xanthan and KGM in the presence of different salts using dynamic
viscoelasticity, DSC, and ESR. They showed that the gelation temperature shifted to
lower temperatures when electrolyte is present, with divalent cations having a
greater effect than monovalent cations. These observations are explained by the
fact that electrolyte promotes xanthan self-association at the expense of xanthan/
KGM interaction, and thus the temperature for gelation on cooling shifted to lower
temperatures with increasing xanthan self-association. The effect of salt follows the
Hofmeister series. The strength of the cationic effect was as follows:
K+ ~ Cs+ < Na+ ~ NH4+ ~ Ba2+ < <Mg2+ ~ Ca2+ (Annable et al. 1994).
Recently, Takemasa and Nishinari (2016) found the nuclear Overhauser effect
(NOE) between low molar mass galactomannan and xanthan, which is a direct
154 K. Nishinari

evidence for the synergistic interaction because NOE is seen only for nuclei closer
than about 0.5 nm.
Although synergistic interaction leading to an industrially interesting phenome-
non such as enhancement of thickening and gelling ability has been attracting great
interest, the most common phenomenon occurring in the mixing of different bio-
polymers is phase separation. Fang et al. (2006) constructed a state diagram of
gelatin/κ-carrageenan aqueous mixtures based on turbidity measurement, confocal
scanning laser microscopy (CSLM), DSC, and zeta potential measurements. They
found a coexistence of associative and segregative (associative-co-segregative)
phase separations at low temperature and low NaCl concentration in addition to
compatible, associative, and segregative phase separation behaviors. A coexistence
of associative and segregative phase separations was observed and it was attributed
to a kinetically trapped state by gelation.
The effect of the addition of insoluble fibers and starches (discussed in Sect. 6.6
Syneresis) on mechanical properties of myofibrillar protein gels (Gravelle et al.
2017) is discussed here. When the addition of fiber was low (mass fraction mf of
filler <0.1–0.15), the hardness increased with increasing mf. While the resilience,
estimated from the immediately recovered deformation, was decreased with increas-
ing mf in myofibrillar protein gels with insoluble fibers, that of starch-filled protein
gels was almost independent of mf. This was interpreted that swelling of starch
granules made the filler more flexible and diminished the stress concentration at the
interface. The stress concentration Sc as a function of mf estimated for completely
bonded interface was shown to decrease, and thus the relative strength Rs which is an
inverse of the stress concentration, Rs ¼ 1/Sc, increased with increasing mf (Gao and
Lelievre 1994).
The fundamental problems and many examples of mixed gels are discussed in
Chap. 10 of Djabourov et al. (2013) Cao et al. (2016a, b) and in Nicolai (2019).

9 Fracture of Food Gels

Fracture of gels can be divided into brittle fracture and ductile fracture. Fracture
occurs at the structural defects. To avoid the uncertain distribution of structural
defects, the fracture test is sometimes done for notched samples (van Vliet and
Walstra 1995). While Young’s modulus obtained at the small deformation is not so
influenced by the depth of the notch l, the fracture stress decreased with increasing l.
In the extension test, the gel tends to fracture at the clamps or slips if the gel is not
clamped tightly or too loosely, therefore it is not easy to obtain reproducible results.
A memorable record in food gels was reported by van Kleef (1986) who could use
224 experiments which were successfully performed free from slippage or fracture of
the gels at the clamps among more than several hundreds of experiments for OVA
gels. He got the relation between the fracture stress and the protein concentration C:
σ b ¼ 102C2.6 for the pH 10 gels and σ b ¼ 5.89C3.2 for the pH 5 gels.
Fragment size distribution of masticated fish sausages was analyzed, and it was
found that the distribution was fitted well by combination of the lognormal
4 Gelling Properties 155

distribution with exponential tail indicating that the fragmentation process has a size-
segregation-structure between large and small parts. For large fragments population,
incorporation of exponential distribution N(s) ¼ Ae-s/B in addition to lognormal
distribution was effective. In the mastication, the segregation of fragments into larger
and smaller fragments occurs, but the number of large fragments decreases with the
progress of mastication and the value of B decreases with the increasing number of
mastication (Kobayashi et al. 2010). Recently, the fragmentation process in the
mastication was studied using agarose gels with different sizes and with different
molar masses. The effects of gel size and molar mass were recognized in the early
stage of mastication. The average particle size showed a high correlation with the
hardness (Moritaka et al. 2019).
Soft gels are known to be crushed between the tongue and the hard palate
(Nishinari et al. 2020). Many such food gels have recently been produced for
disadvantaged persons. Ishihara et al. (2013, 2014) studied the compression of
agar gels and gellan gels using an artificial tongue made from silicone gel, and the
transition from the tongue only breaking to chewing by teeth was found to be
predicted from the instrumental uniaxial compression.

10 Microgels (Fluid Gels)

Microgels can be produced by shearing the gelling polymer solutions undergoing

sol-gel transitions (Norton et al. 1999) or other method using emulsion in which
gelling polysaccharides are dispersed. These microgels have been used as texture
modifiers, stabilizers and especially for reduced fat foods. With a finite yield stress,
microgels behave as a solid at rest but flow when subjected to a stress above the yield
stress. In addition to many kind of polysaccharides from agar (Farres and Norton
2015), carrageenan (Garrec et al. 2013), and other origins, whey protein isolate
(Moakes et al. 2015) was also used to make microgels.
Microgels of alginates were studied to encapsulate probiotics such as lactobacil-
lus which are labile in severe gastrointestinal environments. In comparison with
external gelation where alginate solution is dripping into calcium chloride solution,
internal gelation of alginate is performed in alginate emulsion containing insoluble
calcium carbonate particles by slowly lowering pH using GDL, which can produce
small alginate beads ca 1μm or even smaller than 200 nm (Paques et al. 2013).
Smaller gel beads are advantageous because larger particles than 25μm (this value
depends on the shape of particles, hard and irregular particles are perceived as gritty
than soft and round particles of similar size; Engelen et al. 2005) are detected on the
palate, and thus can extend the application. Cai et al. (2014) compared alginate beads
prepared using calcium carbonate and calcium disodium ethylenediamine- tetra-
acetate and found that encapsulation using the former protected L. acidophilus
more effectively in the survival test, which was related to the mechanical strength
of the microcapsules.
Protein microgels can be also produced by ultrasonication of heated β-lg at low
pH (Murphy et al. 2017, 2018). Produced microgels of β-lg with a diameter ca
156 K. Nishinari

230 nm were shown to be stable without coalescence over 6 weeks storage; however,
emulsions containing limonen stabilized by the microgels were susceptible to
creaming, flocculation, and limonen was lost during storage, which remain as future
problems.

11 Cryogels

A cryogel is formed by the cryogenic treatment, freezing–frozen storage–thawing of

the precursor system. Scientific research became active since the discovery of
cryogels of poly(vinyl alcohol) (Lozinsky and Okay 2014). According to this
definition, traditional Japanese foods such as koori-tofu (dried tofu after freezing),
bo kanten (agar-stick, resulting from freezing–thawing) are cryogels but since they
are xerogels and are eaten after absorbing solvent (water) by cooking, the charac-
teristics of these xerogels are quite different from those of swollen food gels. Most
gels lose their elastic characteristics once they are frozen and thawed. Most of them
lose their solvents and only dried framework remains like agar-stick and koori-tofu.
Cryogels keeping the rubber-like texture and water even after freeze–thaw cycles
have been studied extensively including PVA and hyaluronan (Djabourov et al.
2013; Zhang et al. 2013). Other cryogels of amylopectin, gelatin, maltodextrin,
potato starch, oat β-glucan have also been studied, but most of them are sponge-
like cryogels (Lozinsky and Okay 2014). LBG solution was found to form a gel after
the repeating cycles of freezing and thawing (Tanaka et al. 1998). It was found that
LBG does gel from solutions containing in excess of 1% solids at room temperature
on a time-scale of several months. The gelation mechanism of LBG in concentrated
sucrose solutions was shown to be governed by frustrated crystallization process
with nucleation and growth stages rather than reversible pairwise crosslinking and
the gelation rate became maximum at
5  C (Richardson and Norton 1998). This
experimental finding may give a clue to explain the well-known fact that LBG forms
a gel via a freeze–thaw cycle.
Giannouli and Morris (2003) reported that ca. 0.5% xanthan solution forms a
cryogel when subjected to a freeze–thaw cycle. As has been known, xanthan
solution is a structured liquid and can suspend solid particles. This network is
known to be strengthened by calcium ions. Giannouli and Morris reported that the
cryogelation was abolished by adding sucrose higher than 30% and also by adding
0.4 mM calcium ions. The inhibiting role of sucrose was attributed to the freezing
point depression, and thus ice crystallization was inhibited leading to the insufficient
alignment and association of xanthan chains. The weakening of cryogelation of
xanthan by calcium ions was suggested that the strengthening the structure of
xanthan by calcium restricts the alignment during freezing, thus hampering the
cryogel formation.
Doyle et al. (2006) studied the effect of sugars (sucrose, glucose and fructose) and
sorbitol on the cryogelation of galactomannans. They prepared galactomannans with
different M/G ratios from 2.65 to 4.16 by enzymatic modification of guar gum
4 Gelling Properties 157

(M/G ~ 1.6), and in addition they used LBG with much higher molar mass. They
found that Young’s modulus determined from the initial slope of the compression
curve, the stress at break, increased up to ca 50% sugar and then decreased with
increasing concentration of sugars. On the other hand, the strain at break was found
to decrease monotonically with increasing sugar concentration. In addition, they
found that Young’s modulus, stress at break as a function of M/G increased steeply
around M/G ¼ 4, and the chain length did not affect so much for these behaviors.
They raise again the freezing point depression as one of the reasons. Since the
concentration of sugars used in this study was 40–60% and was lower to make the
system glassy state where conformational ordering of polysaccharide chains was
inhibited. Then, the inhibition of polymer–polymer association by binding of sugar
molecules to polysaccharide chains was thought to be the main reason for the
decrease in Young’s modulus, and stress at break after reaching the maximum.
It was found that cereal β-glucans from oat, barley, and wheat (molar mass
ca. 200  103) form cryogels, and both elastic modulus and the fracture stress
increased with increasing freeze–thaw cycles (Lazaridou and Biliaderis 2004).
They further examined the effects of sugars and polyols (fructose, glucose, sorbitol,
sucrose, and xylose) on rheological and thermal properties of cryogels of barley
β-glucans (Lazaridou et al. 2008). While both G0 and G00 decreased by adding
fructose, glucose, sucrose, and xylose, both modulus increased by adding sorbitol.
On heating, G0 of cryogels slightly decreased up to 50–60  C, then decreased steeply
indicating the melting. The melting temperature shifted to lower temperatures except
cryogels with sorbitol. Heating DSC curves also showed the endothermic peak
accompanying the melting, and the melting peak temperature shifted to lower
temperatures in the presence of polyols. However, the endothermic enthalpy was
increased by the addition of polyols. They interpreted these experimental results by
the freezing point depression of the β-glucan solutions by adding of sugars that may
decrease the ice crystallization leading to the weaker structural formation of
cryogels. They attributed the difference between rheological behavior and thermal
behavior to the difference in the global structure and local structure.
Though some trial cryogel production of food hydrocolloids such as
hydroxyethyl cellulose irradiated with UV-visible light at room temperature and in
the frozen state or chitosan cryogels formed in moderately frozen aqueous solutions
using glutaraldehyde as a crosslinker (Okay 2014), they are not still acceptable in
food use. It is expected that other polysaccharide or protein cryogels are developed.

12 Oleogels

Since the findings of adverse effect of trans fatty acids on blood lipids and coronary
heart disease risk in the early 1990s, food industry tries to find a better way to modify
the texture-property of oil/fat products without using a large amount of crystalline
triacylglycerol molecules which are rich in saturated and or trans fatty acids. Solid
fat foods such as butter and margarine keep a shape but are spreadable when
158 K. Nishinari

subjected to shear stress higher than their yield stress. One of the most important
differences between polysaccharide and protein gels and solid fat is that the linear
elastic range is very narrow in the latter in comparison with the former because of the
crystalline nature in the latter. The most widely employed method of producing
oleogels is dispersing a gelator into liquid oil. The well-known gelators include
waxes such as rice bran waxes, beeswax, shellac, and ethylcellulose (Singh et al.
2017; Patel and Dewettinck 2016). All the edible oleogels reported show a structured
liquid behavior as described earlier, that is, both G0 and G00 are almost independent of
frequency and G0 > G00 , tan δ > 0.1 (de Vries et al. 2017; Moschakis et al. 2016;
Zetzl et al. 2014; Patel et al. 2013).
The finding that the slowly formed gels have a stronger structure is widely
observed for polymer gels but the different behavior is reported for oleogels made
by dispersing gelator in edible oil. Oleogels of shellac, a natural resin secreted by lac
insect, were prepared in the following way: the dispersion of shellac in the rapeseed
oil was heated above melting temperature of shellac (>85  C), and then cooled to
room temperature resulting in oleogels. The onset of nucleation and crystal forma-
tion was delayed on slower cooling resulting in larger and less dense crystals. The
storage modulus was found to decrease with lowering the cooling rate (Patel et al.
2013). This was explained by the increase in total effective area for smaller crystals
which leads to the stronger network formation due to the higher crystal–crystal
interactions.

13 Applications of Gelling Agents in Food Industry

Nata de coco, a kind of bacterial cellulose produced from coconut water, has been a
favorite dessert jelly, and boomed in the 1990s especially in Japan, and still
continued to be consumed all over the world, may be because of its health benefits,
lowering cholesterol, and effective for diabetes and obesity in addition to its peculiar
textural characteristics (Fontana et al. 2017). It has an anisotropic fibrous texture and
thus expected to be a suitable matrix as vegetarian “meat,” since it can have various
colors and meat-like flavors (Fontana et al. 2017). The addition of small amount of
bacterial cellulose increased the gel strength of tofu and kamaboko and thus
improved the sensory evaluation (Okiyama et al. 1992, 1993; Shi et al. 2014).
In the preparation of mixed gels of κ-carrageenan, KGM, and LBG, the small
deformation rheology was governed by κ-carrageenan, and Young’s modulus
decreased with decreasing κ-carrageenan concentration. The large deformation
behavior examined by gel ring extension test, however, was dominated by KGM,
and the rupture strain increased with increasing KGM concentration (Brenner et al.
2013). These two opposing trends led to a maximum in rupture stress. It should be
noted that only κ-carrageenan has a gelling ability in this combination at neutral or
lower pH, and KGM gels only at higher pH. This result may be generalized to design
texture of food gels consisting of a deformable component and a brittle component.
The instantaneous gelation ability of alginate is applied to make an imitation
ikura (salmon roe). A drop of salad oil is introduced into a sol of carrageenan or
4 Gelling Properties 159

xanthan which does not form a gel. This sol is wrapped by a film of alginate or
pectin. The appearance resembles a natural salmon roe so even a fisherman cannot
distinguish between them (Ueda 1985; Kishi 1977). This is quite often used for sushi
and some other snacks. This became a popular demonstration in open campus event
in universities.
Fish paste gels (surimi or kamaboko) show rubber-like behavior and frequently
added starch plays a role of filler reducing the entropic nature of elasticity (Nishinari
1988). Many trials to improve the texture of surimi or meat products have been done
by adding cellulose, KGM, curdlan, and other polysaccharides (Kaur and Sharma
2019; Zhuang et al. 2020). Anisotropy is an important factor controlling the fibrous
texture, and has been successfully introduced in softened meat and string cheese
(Nishinari 2020). A Japanese invention, crab stick (kani-kama) is consumed inter-
nationally, and the fibrousness is conferred by making the structure anisotropic
sometimes adding alginates or other polysaccharides.
Frozen tofu containing curdlan keeps the smooth texture after thawing (Nakao
et al. 1994) whilst when usual tofu is frozen, the texture becomes rough, which has
been used as koori-tofu in Japanese and Chinese dishes.
Agar has been used widely in Japanese cuisine, and higher molar mass agar
(Mw ~ 5 105) is used for tokoroten (noodle-shaped agar gel served with vinegar and
soy sauce or brown sugar syrup), while medium molar mass agar (Mw ~ 3 105) is
used for sweets and yogurt, and lower molar mass agar (Mw < 105) gels with low gel
strength break down into microgels by a weak force, and thus suitable for mixing
with honey to make it less sticky and easy to spread on toasted bread (Nishinari and
Shiba 2007). Strictly speaking, agar is not a molecule but a mixture of agarose and
agaropectin, and thus the molar mass is a kind of the average. This concept of the
average molar mass has also been used for alginate, starch, and other biopolymers.
Melt-in-the mouth sensation of foods are liked in chocolate, ice cream and jellies.
This means that most humans like the sensation or feeling when chocolate, ice cream
and jellies are melted. Fish gelatin exploitation was pursued by many hydrocolloids
research groups to overcome the BSE issue or religious issues. Xanthan cryogel
(0.5%) (Giannouli and Morris 2003) and mixed gels of xanthan and KGM at acidic
pH (3.5 and 4) (Agoub et al. 2007) were found to melt in the mouth temperature
range, which might be useful for fruit jellies.
Aerogels have attracted much attention because of the light mouth feel and low
calorie. Marshmallow, soufflé, and hanpen are typical examples and have been
enjoyed before a short note with colloids science view appeared in Nature (Kistler
1931). Endeavor to develop aerogels more systematically to apply them in delivery
carrier of active compounds using starch-based aerogels containing agar or micro-
crystalline cellulose (Dogenski et al. 2020) or in food packaging as absorber matrix
(da Silva et al. 2020) is in progress.
Gelling polysaccharides agar, alginate, KGM, curdlan, carrageenan, gellan, pec-
tin, starch are used in controlling the texture, water holding capacity of meat and fish
products, and are also used to make dessert jellies (Nussinovitch and Hirashima
2014; Nishinari 2020).
160 K. Nishinari

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Chapter 5
Emulsifying Properties

Hui Zhang and Lingli Deng

Abstract To date, food hydrocolloids have been extensively studied as a surface-

active ingredient adsorbing at the oil–water interface to stabilize emulsions through
the interfacial properties. This chapter is aimed to review the formation, preparation
methods, colloidal properties, and stability mechanisms of food emulsions. The
effects of the absorbing and non-absorbing food hydrocolloids on the flocculation,
coalescence, creaming/sedimentation, and Ostwald ripening of emulsions are
discussed. Three classes of food hydrocolloids as emulsifiers are categorized: pro-
teins (e.g., gelatin, zein, whey protein, casein, and β-lactoglobulin), polysaccharides
(e.g., gum arabic, pectin, galactomannan, microcrystalline cellulose, and starch), and
protein-polysaccharide conjugates, which can be covalently or electrostatically
formed to provide the improved emulsifying and stabilizing properties. Due to the
unique emulsifying properties, these food hydrocolloids have found a number of
applications in beverages, dairy and meat products in the food industry. The future
prospects of food hydrocolloids with emulsifying properties are proposed at the end
of this chapter.

Keywords Emulsion stability · Emulsifier · Protein · Polysaccharide · Conjugates

1 Introduction

An emulsion is a colloid of two or more immiscible liquids where one liquid acts as a
continuous phase and other liquids discontinuously disperse in it. During emulsion
preparation, an emulsifying agent actively adsorbs at the newly formed oil–water
interface to protect the newly formed droplets from immediate coalescence. Except

H. Zhang (*)
College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou, China
e-mail: hubert0513@zju.edu.cn
L. Deng
College of Biological Science and Technology, Hubei Minzu University, Enshi, China

© Springer Nature Singapore Pte Ltd. 2021 171

Y. Fang et al. (eds.), Food Hydrocolloids,
https://doi.org/10.1007/978-981-16-0320-4_5
172 H. Zhang and L. Deng

Fig. 5.1 Structures of conventional emulsion (a), Pickering emulsion (b), multilayer emulsion (c),
multiple emulsion (d), emulsion gel (e), and nanoemulsion (f)

for small molecular surfactants, food hydrocolloids are traditionally associated with
thickening and gelling behaviors, which result in enhanced emulsion viscosity and
retard the movement of oil droplets. On the other hand, there are some hydrocolloids
which can influence the emulsion stability through the interfacial properties. Hence,
a food hydrocolloid may serve as an emulsifier, as a stabilizer, or in both of these
roles.
Based on the dispersed and continuous phase, food emulsions are commonly
divided into two conventional emulsions, which are water-in-oil (W/O) and oil-in-
water (W/O) emulsions. Additionally, the preparation of several sophisticated emul-
sions has been of wide interest to the food scientists (Fig. 5.1). Examples are
Pickering emulsions, multilayer emulsions, multiple emulsions, emulsion gels,
nanoemulsions, etc. (Sivapratha and Sarkar 2018). Nanoemulsions are dispersions
of nanoscale droplets with a mean droplet diameter between 20 and 100 nm (Solans
et al. 2005). The mean droplet diameter of nanoemulsions sometimes is the same as
microemulsions, but the two systems are quite different, as nanoemulsions are
thermodynamically unstable while microemulsions are thermodynamically stable
(McClements 2012). In Pickering emulsions, solid particles are partly wetted by oil
and water, and act as a physical stabilizer based on a steric mechanism (Chevalier
and Bolzinger 2013). A densely packed layer composed of particles at the oil–water
interface contributes to the formation of Pickering emulsions. Multilayer emulsions
are usually fabricated by layer-by-layer electrostatic attraction. Water-in-oil-in-water
(W/O/W) and oil-in-water-in-oil (O/W/O) emulsions are referred to as “emulsions of
emulsions,” where the droplets of one dispersed liquid are further dispersed in
another liquid (Benichou et al. 2007). Emulsions containing hydrogels have recently
emerged as a new class of functional materials, which offer the advantage of
5 Emulsifying Properties 173

improved mechanical resistance for easier handling, and the opportunity for hydro-
philic bioactive delivery (Dickinson 2012) as well as healthier fat replacers (Freire
et al. 2018).

2 Methods for Creating Emulsion Systems

The homogenization process usually contains a single step or some consecutive

steps, which is of great necessity to create an emulsion system from two immiscible
liquids. During homogenization, a variety of methods for preparing emulsions
greatly affect the flow conditions (i.e., laminar, turbulent, or cavitational) of emul-
sions, which determine the nature of the forces for disrupting a droplet. The methods
such as high-speed mixers, colloid mills, high-pressure valve homogenization,
ultrasonication, microfluidization, and membrane and microchannel homogenization
are high-energy ones, while phase inversion and spontaneous emulsification tech-
niques belong to low-energy methods (Fig. 5.2). Comparatively, low-energy
methods are more applicable in the preparation of nanoemulsions, in which fine
oil droplets are inclined to the spontaneous formation under specific environmental
conditions (e.g., composition, stirring, temperature) (Rao and McClements 2010).
Owing to less equipment requirement and energy costs, low-energy methods are
attractive in the food and beverage industries. However, in low-energy methods,
only a few types of oils and emulsifiers are suitable for preparing stable ultrafine
droplets, and relatively high emulsifier-to-oil ratios are required. High-energy

Fig. 5.2 Illustration of the commonly used emulsification methods

174 H. Zhang and L. Deng

methods are often effective for creating other emulsion systems. High-speed mixers
and colloid mills are only available to form coarse emulsions, in which emulsion
droplets have relatively large droplet sizes (r > 1μm), but the other methods can be
employed to form submicron droplets. In addition, different types of homogenizers
are recommended according to the rheological characteristics of the materials. For
example, the homogenization of highly viscous fluids needs the use of some high-
speed mixers and colloid mills, while low or intermediate viscous materials can be
handled by most other types of homogenizers. Moreover, the different size distribu-
tion of droplets can be accomplished by different methods. For the preparation of
emulsions with narrow droplet size distributions, membrane and microchannel
homogenizers are more effective than other methods. Consequently, a variety of
factors (e.g., the rheological characteristics of materials, the desired droplet size
distribution) should be considered for selecting an appropriate emulsification
method, with a purpose of a particular application (Zhang et al. 2018).

3 Colloidal Properties

The appearance, texture, and shelf life of emulsion products are associated with the
droplet size in emulsion systems. In general, the size of the droplets in a monodis-
perse emulsion is expressed as the droplet diameter or radius, while a particle size
distribution is used to denote the fraction of droplets within a range of discrete size
classes in a polydisperse emulsion. For low-energy methods, the dominant factors
affecting the size distribution of droplets are emulsifier type, emulsifier-oil-water
ratio, ionic strength, etc. The size distribution generated by high-energy methods
mainly depends on emulsifier type and concentration, oil-water interfacial tension,
viscosities of oil and water phases, and the intensity and duration of energy input
(Jafari and McClements 2018). A static light scattering instrument is mainly used as
a particle size analyzer on condition that the emulsion droplets can scatter incident
light in a well-defined manner. But for concentrated emulsions, the extensive
dilution and ultrasonic treatment are of necessity before the light scattering mea-
surements to avoid inaccuracy of results. Alternatively, a light microscope is avail-
able to achieve the assessment of the microstructure and droplet size distribution of
concentrated emulsions.
The interface of each emulsion droplet is a narrow region formed by accumula-
tion of surface-active substances. The type and concentration of surface-active
species determine the composition and structure of the interfacial region, which
may influence the intermolecular distance and local concentration of reactive mol-
ecules, and then accelerate certain types of chemical reactions (e.g., oxidation)
(McClements and Decker 2000). Typically, for monolayer emulsions stabilized by
food-grade emulsifiers (e.g. proteins, polysaccharides, surfactants), the thickness of
the interfacial layer is in the range of 1–10 nm, which is considered as a vital factor
for emulsion stability.
5 Emulsifying Properties 175

Table 5.1 The major colloidal interactions between oil droplets in emulsions (Piorkowski and
McClements 2014)
Interactions Sign Magnitude Range
Electrostatic Repulsive Strong to weak Long to short
Steric Repulsive Strong Short
Depletion Attractive Weak to medium Short
Bridging Attractive Strong Short
Van der Waals Attractive Intermediate Intermediate
Hydrophobic Attractive Strong Long

The electric properties of emulsion droplets, usually characterized in terms of

zeta-potential (ζ), mainly depend on the solution conditions and the adsorbed
emulsifier molecules that are ionized or ionizable (McClements 2010). For example,
ionic surfactants in emulsion systems can be neutral, positively charged, or nega-
tively charged. The electrical charge of proteins is associated with the isoelectric
point and solution pH. Depending on the type of functional groups along the
backbone, surface-active polysaccharides may also have an electrical charge. In
general, the emulsion droplets are covered by the same type of emulsifier. Then,
the electrostatic repulsion caused by the same electrical charge protects the droplets
from aggregation, and significantly affects the interactions between emulsion drop-
lets and other charged species (e.g., surfactants, hydrocolloids, flavors, antioxidants).
For instance, the catalyst electrostatically absorbed on the droplet surface may
promote the lipid oxidation of oil droplets, in comparison to free one (Mei et al.
1998).
In addition to electrostatic interactions, colloidal interactions such as van der
Waals, steric, hydrophobic interactions, depletion, and salt-induced attraction can
also play a significant role in emulsion systems (Table 5.1). Their magnitude (strong
to weak), sign (attractive to repulsive), and range (long to short) determine the nature
of interdroplet interactions, which strongly affect the overall characteristics (e.g.,
stability, rheology, and appearance) of a particular emulsion system. When attractive
forces dominate the interactions, the droplets are inclined to associate with each
other. While the dominant forces are repulsive, the droplets tend to retain their
individual integrity (Zhang et al. 2018).

4 Emulsion Stability

When considering the stability of an emulsion, the distinguishment between its

thermodynamic and kinetic stability is significant. From a thermodynamic point of
view, an emulsion tends to separate due to the reduced interfacial energy. Therefore,
all food emulsions are thermodynamic instability and will eventually break down as
long as they are left long enough (McClements 2015).
176 H. Zhang and L. Deng

The comparison of the free energy of a liquid–liquid system before and after
homogenization can be used to explain the origin of such thermodynamically
unstable systems. The free energies of an oil phase and a water phase keep constant
before and after emulsification, so the difference in the free energy between the
non-emulsified and emulsified states is analyzed as follow:
 
ΔGformation ¼ G f  Gi ¼ GIf  GiI  TSconfig
f
 TSiconfig ¼ ΔGI  TΔSconfig

By definition, the difference in interfacial free energy (ΔGI) between the initial
and final states is equal to the increase in contact area (ΔA) between the oil and water
phases, which is multiplied by the interfacial tension (γ): ΔGI ¼ γΔA. Hence,

ΔGformation ¼ γΔA  TΔSconfig

The increased contact area after emulsification always results in the positive
change in interfacial free energy, which thus opposes emulsion formation. Addi-
tionally, due to the greater number of arrangements accessible to the droplets in the
initial state than the final state, the configurational entropy term (TΔSconfig) is
always negative, which therefore contributes to emulsion formation. The overall
free energy change concerned with the formation of a food emulsion can thus be
expressed by

ΔGformation ¼ γΔA

Therefore, the creation of a food emulsion should be always thermodynamically

unfavorable, because of the increased interfacial area after emulsification.
Compared with the thermodynamic stability, the kinetic stability of food emul-
sions is more attractive and interesting for food scientists and engineers, because of
the particular importance to create food products with desirable properties. Between
the two different free energy states (Glow and Ghigh) probably occupied by plenty of
molecules in a system, the molecules are most likely to occupy the state with the
lowest free energy. At thermodynamic equilibrium, the two states are populated
according to the Boltzmann distribution:


nhigh Ghigh  Glow
¼ exp 
nlow kT

where nlow and nhigh are the number of molecules that occupy the energy levels Glow
and Ghigh, k is Boltzmann’s constant (k ¼ 1.38  1023 J K1), and T is the absolute
temperature.
Compared with the thermal energy of the system (kT), the fraction of molecules in
the lower free energy state becomes greater as the difference between the two free
energy levels is larger. In practice, due to the presence of a free energy barrier, ΔG*,
between the two states (Fig. 5.3), a free energy needs to be acquired to cross the
5 Emulsifying Properties 177

Fig. 5.3 Emulsions are a thermodynamically unstable system, but may exist in a metastable state,
therefore being kinetically stable

barrier if a high free energy state of a system moves into a low one, and the
transformation rate decreases with the increasing barrier height. In general, the
long-term stability of most emulsions needs an activation energy of about 20kT.
Actually, an emulsion system is in different metastable states, probably because of
the sufficiently large barrier associated with a thermodynamically unstable state for a
long time. Nevertheless, a single free energy barrier concerned with a particular
physicochemical process is considered as a most important factor to determine the
overall kinetic stability of an emulsion.

5 Stability Mechanisms

The most important phenomena affecting the long-term stability of an emulsion are
gravitational separation, coalescence, flocculation, phase inversion, and Ostwald
ripening (Fig. 5.4). Creaming, as one type of gravitational separation, describes
that droplets in a system move upward owing to their lower density than the
surrounding liquid, while sedimentation is the other type of gravitational separation,
which describes that droplets move downward because of their higher density than
the surrounding liquid. Flocculation and coalescence are both types of droplet
aggregation. Flocculation describes the process that two or more droplets come
178 H. Zhang and L. Deng

Fig. 5.4 Mechanism of destabilization in a colloidal system. Reproduction with permission from
(Kuroiwa et al. 2015), Copyright 2015 Elsevier

together but remain as individual entities, while coalescence occurs when two or
more droplets merge together, resulting in the formation of a single larger droplet
and eventually a separate layer of oil on the top of a system, namely, “oiling-off.”
Phase inversion describes the conversion from an O/W emulsion to a W/O emulsion
or vice versa. Ostwald ripening is the growth of one emulsion droplet at the expense
of a smaller one as a result of the difference in chemical potential of the material
within droplets. Small-sized unstable particles are being dissolved and re-attached on
the surface of big particles, reaching a more stable thermodynamic state.

5.1 Flocculation

Flocculation is a reversible process of the sticking together of droplets upon colli-

sion, leading to an apparent increase in the droplet size and then an acceleration of
creaming or sedimentation. As droplet collisions reduce or electrostatic and steric
repulsion increase, flocculation can be decreased between droplets. Once droplets
collide, the interface separating two droplets has a naturally thinner tendency and
eventually ruptures. Then, coalescence occurs in a form of one larger droplet.
Emulsions stabilized with ionic hydrocolloids are more likely sensitive to pH and
ionic strength, which would induce flocculation by pH changes or specific types of
ions. The conformation of the hydrocolloids is also affected by pH and ionic
5 Emulsifying Properties 179

Fig. 5.5 Schematic illustration of depletion and bridging flocculation

strength, especially the protein part. The addition of simple salts into the aqueous
phase reduces the thickness of the electrical double-layer, and so diminishes the
range of the electrostatic repulsion.
When a polymer is introduced into a colloidal system, flocculation is often
observed, due to one of two distinct mechanisms—bridging or depletion flocculation
(Fig. 5.5). Bridging flocculation is a process that the insufficient emulsifier cannot
achieve the saturation of freshly formed surfaces and share the adsorbed layer among
neighboring droplets. When the emulsifier adsorbs too slowly or is present at a very
low concentration, most of the individual droplets tend to form an aggregate and
cannot remain as their individual integrity as a result of coalescence or bridging
flocculation.
If the amount of non-adsorbing hydrocolloids in an emulsion is not enough to
immediately cover all the interface, depletion flocculation may occur. McClements
(2000) defined four regions concerned with the instability of an emulsion system
containing polysaccharides, and determined a critical flocculation concentration
(CFC), which is the polysaccharide concentration producing depletion flocculation.
When the polysaccharide concentration (c) in an emulsion exceeds a particular
value, termed as the critical viscosity concentration (CVC), creaming would be
assumed to completely retard: (I) Unstable. No-flocculation (c < CFC) and relatively
low viscosity (c < CVC). The droplets move upward at a rate that is proportional to
the square of their diameter and the reciprocal of the aqueous phase viscosity.
(II) Highly unstable. Flocculation (c > CFC) and relatively low viscosity
(c < CVC). The effective size of the particles in the emulsion increases because of
flocculation, but the viscosity is not high enough to prevent the flocculation. (III)
Stable. No-flocculation (c < CFC) and high viscosity (c > CVC). The droplets are
180 H. Zhang and L. Deng

not flocculated, but the viscosity of the aqueous phase is so large that they cannot
move. (IV) Stable. Flocculation (c > CFC) and high viscosity (c > CVC). The
droplets are flocculated, but the viscosity of the aqueous phase is so large that they
cannot move.

5.2 Coalescence

Coalescence describes the merging of two or more emulsion droplets to form a larger
single droplet. For an emulsion product, coalescence is perceived as highly unfa-
vorable during storage. The shifting of the overall distribution towards larger droplet
sizes may result in the enhanced creaming. However, partial coalescence for the
development of structures is of great significance for some food products, such as
whipped cream and ice cream, because of the conversion of a viscous liquid to a
viscoelastic solid. For many of food triglyceride emulsions, particularly dairy emul-
sions, fat crystallization occurs in such emulsion droplets at storage temperature.
Crystals are both radial and tangential to the surface, and actually growing or
protruding through the membrane. The incomplete coalescence caused by crystals
present in the oil phase results in the formation of irregularly aggregated droplets
with the original identity (Fig. 5.6). Two important additional factors influencing
partial coalescence are: (i) the temperature cycling of cream layers, which causes
growth and melting of fat crystals, and (ii) agitation or stirring, which greatly
increases the likelihood of collision-induced crystal penetration between droplets
(Dickinson 2009).

Fat droplets Competitive adsorption Partial coalescence

and fat crystallization of fat globules

milk protein milk protein milk protein

oil
fat crystals
fat crystals
emulsifier emulsifier emulsifier

Fig. 5.6 Schematic illustration of fat crystallization and partial coalescence. Reproduction with
permission from Cheng et al. (2020) Copyright 2020 Elsevier
5 Emulsifying Properties 181

5.3 Creaming/Sedimentation

Creaming is a process that oil droplets driven by gravity move upward to form a
concentrated cream layer at the top of an emulsion. The decreased droplet size of
colloidal particles can retard creaming, which can be modified by the hydrocolloid
concentration, emulsifying conditions (pH, ionic strength), and emulsification
method. Beverage emulsions are sensitive to creaming due to the fact that the dilute
emulsions are typically made up of a flavor oil blend containing a weighting agent,
and a polymeric emulsifier or stabilizer (Given Jr 2009). Sedimentation also happens
when the weighting agent is overused or supersaturated in the oil phase. Various
hydrocolloids have been applied for manufacture of beverage flavor emulsions, such
as octenyl succinylation starch, whey protein, arabinogalactan, etc. (Klein et al.
2010; Mora-Gutierrez et al. 2018). Electrostatic interactions between proteins and
polysaccharides are used to fabricate sequential multilayers to build droplet shell
integrity and stability against various environmental stresses (ionic strength, pH,
presence of oxidation catalysts, heat, sunlight, etc.), because of the formation of a
more robust interfacial complexes.

5.4 Ostward Ripening

The difference in the radius of droplet curvature results in the chemical potential of
the materials within droplets and then Ostwald ripening, i.e., one emulsion droplet
grows at the expense of a smaller one. Alternatively, the process may be purely
viewed from a perspective of the reduced free energy in the system by means of the
destructed interfacial area. Overall, the effect is an increase in the average radius of
the emulsion droplets with time, as the smaller droplets dissolve and redeposit their
materials onto the larger droplets (Taylor 1998). In theory, Ostward ripening can be
decreased as the monodispersity of an emulsion increases, due to the fact that the
thermodynamic driving force is associated with the size difference of droplets. In
practice, the coarsening tendency can be reduced by mixing of another insoluble
component into the dispersed phase, due to a large counteracting thermodynamic
driving force provided in direct opposition to the Ostwald ripening effect
(Kabal’Nov et al. 1987; Davis et al. 1981).

6 Proteins

6.1 Gelatin

Gelatin is a natural protein hydrolyzed from animal collagen. It is widely used for
foaming, emulsifying, and wetting purposes in food industry. Previous studies have
182 H. Zhang and L. Deng

shown that gelatin acts as an emulsifier with surface activity in O/W emulsions
(Table 5.2) (Lobo 2002). Its emulsification and foamability derive from the hydro-
phobic area of peptide chains on gelatin. Compared with other surface-active sub-
stances, such as globular proteins and gum arabic, gelatin is a weaker emulsifier,
which often produces larger droplet sizes during homogenization when used alone.
Hence, its effectiveness can be improved by either hydrophobically modification
with the attachment of non-polar side-groups or used in combination with other
emulsifiers as complex emulsifiers.
Gelatin from marine sources (warm- and cold-water fish skins, bones, and fins) is
a possible alternative to bovine gelatin, with an advantage of being not associated
with the risk of bovine spongiform encephalopathy. Moreover, fish gelatin is
acceptable in Islam and can be used with minimal restrictions in Judaism and
Hinduism. Surh et al. (2006) studied the emulsifying property of fish gelatin with
various molecular weights, and the effects of pH, salt, and thermal processing on the
stability of the gelatin-stabilized emulsions. It was observed that a higher ratio of
large droplets led to an easier destabilization of the emulsions stabilized by low
molecular weight fish gelatin than high molecular weight ones. Dickinson and Lopez
(2001) compared the emulsifying property of fish gelatin with that of commercial
milk proteins, and found that an optimization of the protein/oil ratio was expected to
prevent coalescence caused by large droplets, especially at high ionic strength.
Due to the limited emulsifying property of gelatin, its complexes with other
emulsifiers have been studied. In a reported study (Surh et al. 2005), the multilayer
emulsions with SDS as the first layer and SDS-fish gelatin complex as the second
layer were fabricated, the droplets in the secondary emulsions exhibited good
stability against droplet aggregation in a water bath for 30 min at different temper-
atures ranging from 30 to 90  C. It was also observed that the emulsions prepared
with a complex of whey protein with fish gelatin using layer-by-layer interfacial
deposition technique were more physiochemically stable than those with individual
proteins (Taherian et al. 2011). Zeeb et al. (2011) studied the stabilizing effect of
enzymatic crosslinking on the beet pectin-fish gelatin double coated emulsions, and
found that the freeze-thaw stability and creaming behavior of the secondary emul-
sions treated by laccase were significantly improved, compared to the individual
emulsions. Lately, the fish gelatin-gum arabic complexes were used to fabricate
concentrated emulsions, and greater intermolecular connectivity between
the adsorbed layers of adjacent oil droplets for a gel network extension was observed
at lower pH (Anvari and Joyner 2017).

6.2 Zein

Zein, as a food-grade abundant material extracted from corn, has been attempted to
accomplish extensive applications in various industries. The hydrophobic property
of zein depends on its high proportion of hydrophobic amino acids, while the degree
of ionization of basic and acid amino acid groups affects the degree of
5 Emulsifying Properties 183

Table 5.2 Typical hydrocolloid emulsifiers and the related emulsions

Emulsifier Emulsion type References
Fish gelatin Oil-in-water emulsion Surh et al. (2006)
Fish gelatin/SDS Oil-in-water emulsion Surh et al. (2005)
Fish gelatin/whey protein isolate Oil-in-water emulsion Taherian et al.
(2011)
Fish gelatin/sugar beet pectin Multilayer emulsion Zeeb et al. (2011)
Fish gelatin/gum arabic Oil-in-water emulsion Anvari and Joyner
(2017)
Zein Pickering emulsion de Folter et al.
(2012)
Zein/tannic acid complex particles Pickering emulsion Zou et al. (2015)
Zein/chitosan complex particles Pickering emulsion Wang et al. (2015)
Zein/propylene glycol alginate particles Pickering emulsion Dai et al. (2018)
Zein/gum arabic nanoparticles Pickering emulsion Li et al. (2018)
Zein/corn fiber gum Pickering emulsion Zhu et al. (2019)
Zein/propylene glycol alginate/rhamnolipid Pickering emulsion Dai et al. (2019)
complex particles (high internal phase)
Zein/sodium caseinate/propylene glycol alginate Pickering emulsion Sun et al. (2018)
(high internal phase)
Zein/tannic acid complex particles High internal phase Zou et al. (2019)
emulsion gel
Whey protein/polysaccharides (locust bean gum, Water-in-oil-in-water Benichou et al.
guar gum, xanthan gum, konjac gum) emulsion (2007)
Whey protein microgels/casein Pickering emulsion Chevallier et al.
(2019)
Whey protein nanoparticles Pickering emulsion Wu et al. (2015)
Whey protein Emulsion gel Mantovani et al.
(2016)
Whey protein microgel particles Pickering emulsion Sarkar et al. (2016)
Whey protein Emulsion gel Luo et al. (2019)
Sodium caseinate, whey protein isolate or iso- Emulsion gel Freire et al. (2018)
lated soy protein
Casein Emulsion gel McIntyre et al.
(2017)
Casein gel particles Pickering emulsion Wang et al. (2018a)
Casein/soy protein or casein/pea protein Emulsion gel Silva et al. (2019)
Casein/whey protein isolate Emulsion gel Balakrishnan et al.
(2017)
β-lactoglobulin Oil-in-water emulsion Purwanti et al.
(2016)
β-lactoglobulin Nanoemulsion Ali et al. (2016)
OSA modified quinoa starch granules Water-in-oil-in-water Marefati et al.
Pickering emulsion (2015)
Guar gum Water-in-oil-in-water de Almeida Paula
emulsion et al. (2018)
(continued)
184 H. Zhang and L. Deng

Table 5.2 (continued)

Emulsifier Emulsion type References
Microcrystalline cellulose/soybean protein Oil-in-water emulsion Xu et al. (2016)
hydrolysate
Nanofibrillated cellulose/guar gum/carboxy Oil-in-water emulsion Golchoobi et al.
methyl cellulose (2016)
Locust bean gum/sodium caseinate Water-in-water Moschakis et al.
emulsion (2018)
Ball-milling and OSA modified areca taro starch Pickering emulsion Liu et al. (2018)
Gelatinized native rice starch, waxy maize starch Pickering emulsion Kasprzak et al.
(2018)
Pectin/zein complex Oil-in-water emulsion Piriyaprasarth et al.
(2016)
Waxy corn starch/locust bean gum Water-in-water Murray and
emulsion Phisarnchananan
(2016)
Octenylsuccinate quinoa starch Pickering emulsion Li et al. (2019)
gel
Quinoa starch Pickering emulsion Saari et al. (2019)
waxy maize starch
oat starch
Native corn (NCS), rice (NRS), wheat (NWS), Oil-in-water emulsion Gómez-Luría et al.
and waxy corn (WCS) starch (2019)
Rice, wheat, potato, OSA potato starch Pickering emulsion Chen et al. (2019)
Wheat native starch and OSA waxy maize starch Emulsion gel Torres et al. (2017)

hydrophobicity. It is classified into four classes with different solubility behaviors,

which are known as α-, β-, γ-, and δ-zein (Anderson and Lamsal 2011). Zein cannot
be easily dissolved in either water or oil, but it is expected to fabricate colloidal
particles by anti-solvent precipitation.
Zein colloidal particles have been extensively studied for preparation of Pickering
emulsions. de Folter et al. (2012) found that the Pickering emulsions stabilized by
unmodified zein colloidal particles were unstable to creaming. However, the
improved stability of emulsions may be obtained by the stabilization of complex
particles of zein and other biopolymers (e.g., proteins and polysaccharides) against
coalescence and creaming through the modified wettability (Wang et al. 2015;
Piriyaprasarth et al. 2016; Li et al. 2018). Zou et al. (2015) fabricated the zein/tannic
acid complex colloidal particles based on hydrogen bonding to stabilize emulsions,
which were extensively crosslinked to form a continuous network among and around
the oil droplets and protein particles, leading to the formation of stable Pickering
emulsion gels. Recently, the zein-propylene glycol alginate complex particles with a
neutral wettability were fabricated to stabilize emulsions with a higher stability
against coalescence (Dai et al. 2018). Zhu et al. (2019) fabricated the zein/corn
fiber gum complex colloidal particle-stabilized Pickering emulsions, which were
observed to have a gel-like network behavior. A neutral wettability was obtained
5 Emulsifying Properties 185

when the mass ratio of zein to corn fiber gum was 2:1, and the prepared Pickering
emulsions exhibited better physical stability.
Zein-based complex particles have been also applied for stabilization of high
internal phase emulsions or emulsion gels. Zou et al. (2019) fabricated the high
internal phase emulsion gels with an oil content ranging between 72 and 87% (w/w)
stabilized with the zein/tannic acid complexes at a particle concentration range of
0.7–1.4%. Lately, the zein-propylene glycol alginate-rhamnolipid complex particles
were prepared with suitable three-phase contact angles to stabilize O/W Pickering
high internal phase emulsions with an oil phase ratio of 75% (Dai et al. 2019).
Similarly, Sun et al. (2018) fabricated the high internal phase Pickering emulsions
stabilized by the ternary zein/sodium caseinate/propylene glycol alginate complexes
at a volume concentration of as much as 80% oil. In a recent report, the Pickering
emulsions at a high oil volume ratio (60%) were fabricated, which were effectively
stabilized by the zein/gum arabic nanoparticles through the formation of a stable and
thick oil–water interfacial layer to hinder agglomeration and Ostwald ripening
(Li et al. 2018).

6.3 Casein

As a heterogeneous phosphorylated protein, casein comprises the major proteina-

ceous component of mammalian milk. Owing to the self-assembly and surface-
active characteristics, casein is expected to stabilize O/W emulsions as an emulsi-
fying agent based on electrostatic and steric stabilization mechanisms. The emulsi-
fying capacity of casein products differs from their form: acid casein > micellar
casein > rennet casein (Roman and Sgarbieri 2006). Tan and McGrath (2012)
studied the emulsion morphology diagrams of the sodium caseinate/oil/water sys-
tem. For the lowest emulsifier concentration, bridging flocculation was evident and
the emulsions were very unstable. Increasing the sodium caseinate concentration
enhanced the emulsion stability and promoted the existence of distinct individual oil
droplets. The further increase in the sodium caseinate concentration caused a
reduced stability, which was ascribed to depletion flocculation. Then the sodium
caseinate self-assembly was initiated. The emulsion stability was again enhanced
due to the formation of a three-dimensional protein network at sufficiently high
sodium caseinate and/or oil concentrations.
In addition, the thermo-reversible gelation behavior was observed in the sodium
caseinate-stabilized emulsions (Dickinson and Casanova 1999). Once heated to
30–40  C, a concentrated liquid-like emulsion is converted into a flocculated
emulsion gel that supports its own weight. The emulsion gel “melts” again slowly
upon cooling, and the return to the original low-viscosity state is accelerated by
stirring. Aggregated casein networks are commonly encountered in making yogurt
and other fermented dairy products. The addition of lactic acid bacteria lowers the
pH from 6.7 to lower than 4.5, resulting in a liquid-like dispersion of casein particles
into a soft solid-like aggregated network. When the pH of sodium caseinate-
186 H. Zhang and L. Deng

stabilized emulsions is lowered towards the protein’s isoelectric point, a transition

from the net repulsion to net attraction occurs, leading to droplet flocculation and
then soft solid-like emulsions. Normally, the stiffness of the casein stabilized
emulsion gels increases with the increasing oil content, as the oil droplets act as
active fillers (i.e. the oil droplets are covered with proteins which are bound to the
surrounding protein network) in the emulsion gels (Silva et al. 2019; Balakrishnan
et al. 2017). It was reported that the sodium caseinate-stabilized emulsions showed
a shear-thinning behavior as a result of a more structured system at pH 7.0, while at
pH 3.0 the addition of laccase improved the emulsion stability by narrowing the size
distribution and increasing the viscosity (Sato et al. 2015). Radford et al. (2004)
studied the effects of alcohol and calcium on the sodium caseinate-stabilized emul-
sions, and found that the addition of calcium ions and/or ethanol resulted in a
pronounced reduction in viscosity and the onset of Newtonian flow. Balakrishnan
et al. (2017) fabricated the emulsion gels by heating the homogenized suspensions of
micellar casein mixed with sunflower oil at various pH (5.8, 6.0, and 6.3) and oil
weight fractions (5, 10, and 15%). The gel stiffness increased with the decreasing pH
and the increasing oil ratio. The gel stiffness did not change significantly after
replacing up to 40% casein by whey protein at pH 5.8 or 6.0, but decreased
significantly at pH 6.3. However, the addition of a small amount of casein into
whey protein microgel stabilized Pickering emulsions effectively improved the heat
stability of the emulsions by competitive adsorption at the interface (Chevallier et al.
2019). McIntyre et al. (2017) prepared the casein-based emulsion gels with milk fat
or rapeseed oil at high (774 mg/100 g) or low (357 mg/100 g) calcium levels.
Compared with the low-calcium emulsion gels, the high-calcium gels were signif-
icantly softer and showed the highest disintegrated rate during the simulated gastric
digestion. The fatty acid releases were similar for all the emulsion gels made from
milk fat, while a higher lipolysis was found in the high-calcium emulsion gels made
from rapeseed oil. Silva et al. (2019) reported that micellar casein could be replaced
by plant proteins (e.g. soy protein and pea protein) while maintaining the same
emulsion gel stiffness.
Sodium caseinate has been found to facilitate the formation of the high internal
phase Pickering emulsions stabilized by the zein/sodium caseinate/propylene glycol
alginate complexes (Sun et al. 2018). The casein gel particles were also fabricated by
covalently genipin crosslinking of a protein network with the self-associated
sub-micelles or calcium induced casein micelles to stabilize Pickering emulsions
(Wang et al. 2018a). The oil droplets stabilized by these particles exhibited higher
stability against flocculation and creaming, compared to those stabilized only by
sodium caseinate.

6.4 Whey Protein

Whey protein has been widely used in food industry as an emulsifier, which consists
of β-lactoglobulin (~65%), α-lactalbumin (~25%), bovine serum albumin (~8%),
5 Emulsifying Properties 187

Fig. 5.7 Schematic presentation of (a) an emulsion-filled protein gel and (b) a protein-stabilized
emulsion gel. Yellow circle, oil; small red circle, protein; light blue, water; green curve, protein
network

and immunoglobulins (Morr and Ha 1993). Compared to casein, whey protein can
be used over a wider pH range since its solubility goes through a minimum at the
isoelectric point at pH close to 5. This makes whey protein applicable to acidic
environments where caseins cannot be used, especially at pH below 4.5. However,
heat treatment has a significant effect on the properties of the whey protein-stabilized
emulsions, since the non-covalent interactions maintaining the secondary and ter-
tiary structures of the globular protein would be broken by thermal energy. It has
been extensively investigated about the stability of the whey protein-stabilized
emulsions in terms of the effect of heating. Euston et al. (2000) found that the
interactions between the adsorbed protein at the emulsion droplet surface and the
non-adsorbed heat denatured protein in the continuous phase were the main cause of
aggregation, and the non-adsorbed protein acted as a “glue” holding the aggregated
emulsion droplets together.
Whey protein has been used for preparation of emulsion gels as well, due to its
characteristic heat-induced gelling property (Dickinson 2012). The major whey
protein in cow’s milk is β-lactoglobulin, which denatures and aggregates upon
heating to a temperature of 70  C, like many other globular proteins. At the
molecular level, this aggregation and subsequent gelation are a result of the devel-
oped hydrophobic interactions between the exposed non-polar regions of the
unfolded β-lactoglobulin molecules. The sulfhydryl-disulfide interchange leading
to covalent crosslinking may also improve the evolving network structure. There-
fore, this complex colloidal system may exist as both an emulsion and a gel, which
can be expressed as “emulsion gel” for understanding. To specify more precisely the
properties of such protein-based systems, two structural arrangements can be distin-
guished: (a) the emulsion-filled protein gel and (b) the protein-stabilized emulsion
gel. As shown in Fig. 5.7a, the emulsion-filled protein gel is a protein gel matrix
within which emulsion droplets are embedded. It is a kind of particle-filled solid due
to the network properties of the spatially continuous matrix in charge of its solid-like
rheological properties. The protein-stabilized emulsion gel is shown schematically in
188 H. Zhang and L. Deng

Fig. 5.7b. It is a type of particulate gel, and its rheological properties are mainly
determined by the properties of the network of aggregated emulsion droplets.
Above a critical concentration, whey protein can be crosslinked to form a three-
dimensional gel network by heating, acidification or enzymatic treatment. The whey
protein emulsion gel structure can be modified by varying pH, droplet size, ionic
strength, and temperature. Mantovani et al. (2016) studied the effect of pH on cold-
set gel formation by acidification of emulsion gel (30% oil and 5% non-heated whey
protein isolate (WPI)) and emulsion-filled gel (emulsion dispersed in heat-treated
whey protein solution), and found that stronger gel strength was obtained at a pH
near the isoelectric point of whey protein because of protein aggregation promoted
by lower acidification rate and electrostatic repulsion. Guo et al. (2017) investigated
the effect of gel structure and rheology on the intestinal digestion of canola oil
dispersed within O/W emulsions gelled with WPI. The softer and microstructurally
homogeneous gels were obtained at lower salt concentrations. The rate of lipid
digestion increased due to the looser, less spatially heterogeneous protein matrix,
in comparison to the firmer gels. Overall, the extent and rate of lipid digestion were
modified by the strength and microstructure of the WPI-stabilized emulsion gels,
which may be used to produce emulsion-based food products. For example, the
emulsion gels containing ω-3 fatty acids and condensed tannins were fabricated with
WPI for potential use of healthier fat replacers (Freire et al. 2018). Luo et al. (2019)
encapsulated capsaicinoids in whey protein emulsion gels, and studied the structural
relationship with the in-mouth breakdown behavior. A greater hardness led to a
smaller bolus particle size, but a higher degree of fragmentation caused greater
surface exposure during mastication.
Besides, soft whey protein microgel particles used as an emulsion stabilizer can
show the combined advantages of biocompatibility and an increased stability against
coalescence. Wu et al. (2015) prepared the Pickering O/W emulsions stabilized by
WPI nanoparticles in the size range of 200–500 nm, which exhibited good stability
against coalescence. Sarkar et al. (2016) designed the Pickering O/W emulsions
using whey protein microgels by a facile route of heat-set gel formation followed by
mechanical shear, and studied the influence of heat treatment on the emulsions
stabilized by these particles. In addition, the whey protein microgel particles could
also act as a stabilizer for waxy corn starch/locust bean gum water-in-water emul-
sions (Murray and Phisarnchananan 2016).

6.5 β-Lactoglobulin

β-Lactoglobulin, a dominant globular protein found in whey fraction, possesses a

remarkable emulsifying property, which is commonly used in the formulation of
food emulsions. β-lactoglobulin has showed great potentials as a transport vehicle
for hydrophobic compounds, since it can bind hydrophobic vitamins and fatty acids
in inner cavities (Kimpel and Schmitt 2015). Moro et al. (2013) studied the effects of
heat treatment on the emulsifying properties of β-lactoglobulin. For a shorter time
5 Emulsifying Properties 189

period of heating, both the foamability and foam stability were improved, but the
emulsifying property diminished. However, after 10 min of heating at 85  C, both
the foaming and emulsifying properties were impaired. Purwanti et al. (2016)
studied the unheated and heat-aggregated β-lactoglobulin stabilized clove oil-in-
water emulsions and limonene-in-water emulsions by microchannel emulsification.
The monodisperse emulsion droplets were consistently produced using unheated or
heat-aggregated β-lactoglobulin with concentrations from 0.5 to 3% (w/w) at pH
7. Ali et al. (2016) fabricated the β-lactoglobulin stabilized biocompatible
nanoemulsions prepared by high-pressure homogenization. The nanoemulsions
with 1 wt% β-lactoglobulin and with 5 wt% Miglyol 812 (a mixture of medium
chain triglycerides) had a relatively small particle size (~200 nm) and a low
polydispersity, when a homogenization pressure of 100 MPa was applied for
4 cycles. These nanoemulsions were made stable for at least 30 days. However,
the emulsification capacity of β-lactoglobulin was reduced at higher homogenization
pressures (200 MPa and 300 MPa).

7 Polysaccharides
7.1 Gum Arabic

Gum arabic was once a kind of hydrocolloids widely used in food industry, and now
there is still 40–50 thousand tons per year consumed in the worldwide market. It
helps to stabilize the flavor and essential oils in production of soft drinks or
concentrated juices. Gum arabic (A. senegal) is a complex branched hetero-
polyelectrolyte with a backbone of 1,3-linked β-galactopyranose units and side-
chains of 1,6-linked galactopyranose units terminating in glucuronic acid or 4-O-
methylglucuronic acid residues. There are three different fractions separated from
gum arabic, namely, arabinogalactan (~90% of total mass), arabinogalactan protein
(~10%), and glycoprotein (~1%) (Randall et al. 1989). Gum arabic is a polysaccha-
ride emulsifier naturally conjugated with protein. The structure of this protein-
polysaccharide complex is known as the “wattle blossom” model (Fig. 5.8), in
which the hydrophobic protein moieties adsorb onto the oil droplet surface, and
the covalently attached hydrophilic carbohydrate blocks protrude into the aqueous
phase against droplet flocculation and coalescence (Dickinson 2008). Though it
serves as a stabilizer to form a thick steric stabilizing layer and protect the flavor
and essential oils in emulsion products at intermediate pH values, high ionic
strengths or high temperatures, gum arabic shows a lower affinity for the oil-water
interface than most other surface-active biopolymers. The α-tocopherol encapsulated
nanoemulsions with gum arabic as an emulsifying and stabilizing agent were
fabricated through solvent-displacement technique (Moradi and Anarjan 2018).
The monomodal size distribution was successfully obtained with a mean particle
size of 10.01 nm, 49.46 nm, and 171.2 nm for the emulsions prepared with 0.05%,
0.1%, and 0.15% gum arabic, respectively. Hu et al. (2019) reported that gum arabic
190 H. Zhang and L. Deng

Fig. 5.8 Schematic

representation of the wattle
blossom model of gum
arabic (a) in solution and (b)
at the oil-water interface.
Hydrophilic carbohydrate
blocks and the backbone
chain of hydrophobic
protein were illustrated

enriched with trace elements (Zn2+, Fe3+, Fe2+) had good emulsion stability as the
molecular weight and arabinogalactan protein content increased, in comparison to
the control gum arabic. Atgié et al. (2018) found that the protein-rich species of gum
arabic irreversibly adsorbed as monolayers at the oil–water interface, and the
absorbed amount drastically increased with both the decreasing ionic repulsions
and the increasing gum concentration. However, those changes corresponded to only
minor composition changes in the adsorbed layer, suggesting a significant role of
controlling the interfacial density in a rational design of gum arabic-based
formulations.

7.2 Pectin

Pectin is another class of hydrocolloid with a fascinating emulsifying character,

while its emulsifying property differs depending on the varieties of plant sources.
For example, citrus and apple pectin can form gels at low pH and serve as thickening
agents but not effective emulsifying agents, while sugar beet pectin is normally used
5 Emulsifying Properties 191

as an excellent emulsifying agent owing to the protein moiety, acetyl groups, and
highly branched polysaccharide structures, and it can form a thick hydrated layer that
may prevent droplets from flocculation and coalescence through electrostatic and
steric repulsive forces. Williams et al. (2005) found that the emulsifying property of
sugar beet pectin was affected by the proportion of ester groups, the accessibility of
the protein and ferulic acid groups to oil droplet surface, and the molecular mass
distribution of the fractions. However, no simple relationship existed between the
emulsifying property and the protein or ferulic acid content. Funami et al. (2011)
reported that the emulsifying property of the deproteinized sugar beet pectin with a
protein content ranging from 5 to 0.5% became worse than that of the untreated
pectin. Siew et al. (2008) reported that an increase in the protein content (12% higher
than the average) of sugar beet pectin would help adsorb onto the oil droplets. Chen
et al. (2016a) observed a significant decrease in the droplet size of the sugar beet
pectin stabilized emulsions, as the protein content of sugar beet pectin increased
from 0.5 to 3%.

7.3 Galactomannan

As the most commonly used plant polysaccharides, galactomannans belong to the

legume family consisting of β-(1-4-) linked D-mannan backbone by substitution of
α-(1-6-) linked D-galactose stubs for certain mannose residues. Locust bean gum and
guar gum are high molecular weight galactomannans used as food additives, since
no significant proportion of hydrophobic groups presents in the carbohydrate struc-
ture. This type of hydrocolloid can be assumed to be used for modifying the
rheological properties of the continuous aqueous phase between dispersed particles
or droplets. To date, the surface and interfacial properties of guar gum and locust
bean gum have been extensively studied (Reichman and Garti 1991).
It was reported that the emulsions with similar droplet size distributions showed
similar stability when crude, purified and bipurified guar gums were used to prepare
emulsions under similar conditions (Garti and Reichman 1993). However, the
emulsions made from the guar gum with a proteinaceous-rich fraction had larger
droplets and relatively low stability with the fastest coalescence rate upon dilution.
Later, Garti and Reichman (1994) found a similar degree of surface activity and
emulsification ability when the guar gum was purified down to 0.8% protein. Thus, it
is conceivable that the slight hydrophobicity of the polymannose backbone may
contribute to some emulsion stabilizing properties.
In a recent study, guar gum was used to fabricate W/O/W double emulsions for
improving the heat stability of anthocyanins at pH 4.0 (de Almeida Paula et al.
2018). In addition, various levels of guar gum (0.5, 1.0, and 1.5%) were used as a fat
substitute to prepare low-fat meat emulsions. The reduction of fat by incorporation
of guar gum was found to increase the emulsion stability and cooking yield but
decrease the penetration force (Rather et al. 2017). To develop a new low-fat
mayonnaise formulation, nine mayonnaise samples containing different
192 H. Zhang and L. Deng

compositions of nanofibrillated cellulose, guar gum, and carboxymethyl cellulose

were compared to the commercial low-fat mayonnaise with 30% fat (Golchoobi
et al. 2016). In addition, the water-in-water emulsions containing sodium caseinate
and locust bean gum have been also fabricated as potential functional ingredients to
mimic fats (Moschakis et al. 2018).

7.4 Microcrystalline Cellulose

Microcrystalline cellulose (MCC) is among the most common cellulose derivatives

in food industry (Nsor-Atindana et al. 2017). It is a hydrocolloid without solubility in
water but adsorbs mechanically at the oil–water interface (Garti and Reichman
1993). The cellulose crystallites are claimed to build a network made from the
majority of the particles being less than 0.2μm. The formed colloidal network of
the free MCC thickens the water phase between oil globules and provides effective
stabilization against their subsequent coalescence (Milani and Maleki 2012).
When colloidal MCC is dispersed in water, it can be expected to simulate the
fat-induced rheology owing to the inherent properties in food applications such as
baked products, frozen desserts, mayonnaise, gravies, and sauces. For example, an
emulsion with 60% soybean oil showed similar stability and rheology characteris-
tics, compared to the emulsion containing 1–1.5% colloidal MCC and 20% soybean
oil (Imeson 2011). The use of MCC as a fat substitute gives a rich creamy texture in
low-fat sauces and dressings, because of the insoluble material to mimic fat. MCC
may be used alone or in combination with other polysaccharides in ice cream, which
is expected to increase the solid content and improve the stability and ice rheology
(Nsor-Atindana et al. 2017).
In general, crystals of cellulose within micro/nanoscale dimension (MCC/NCC)
have been commonly applied for preparation of Pickering emulsions. Kalashnikova
et al. (2011) confirmed that Pickering O/W emulsions could be effectively stabilized
by bacterial cellulose nanocrystals for several months if the particles were properly
dispersed. The colloidal MCC particles (11% colloidal MCC combined with 1%
sodium carboxymethylcellulose) could also form a network around the emulsified
oils and then act as a stabilizer of O/W emulsions and W/O/W multiple emulsions
(Jia et al. 2014), in which MCC not only thickened the continuous phase between the
droplets, but also provided a mechanical barrier to prevent oil droplet coalescence
(Dickinson 2013). In another study, the microrheological property of curcumin
emulsions was changed by the addition of MCC, as evidenced by a transition from
the viscous to viscoelastic behavior of the emulsions. The freeze-thaw stability of the
emulsions was significantly improved by MCC, which was attributed to the
enhanced repulsive steric forces between the curcumin droplets. In addition,
stearoylated microcrystalline cellulose was recently reported to fabricate high inter-
nal phase Pickering emulsions with the highest internal phase ratio of 89% (Pang
et al. 2018).
5 Emulsifying Properties 193

7.5 Starch

Starch (including hydrophobically modified starch) is an abundant, inexpensive, and

natural food ingredient, while the natural variation regarding the size, shape, and
composition of starch granules normally occurs among its numerous botanical
sources. For native starch, it cannot achieve adsorption to the oil–water interface
as an emulsifying agent. Therefore, starch has been modified to become more
suitable for emulsion stabilization. Physical modifications, including milling,
non-solvent precipitation, ultrasonication, high-pressure treatment may be applied
to reduce the size of starch. Decreasing the particle size of starch often relates to the
increased storage stability of its Pickering emulsions. Octenyl succinic anhydride
(OSA) modification is the most commonly used chemical modification method,
which improves the hydrophobicity of starch. OSA modified starch with a degree
of modification <3%, E1450, is a well-established food ingredient with no specific
limitations on its use (Timgren et al. 2011). The Pickering emulsion gels were
formed by stabilization of octenylsuccinate quinoa starch granules at an oil fraction
ranging from 50 to 70% (Li et al. 2019). Liu et al. (2018) compared the stability of
soybean oil-in-water Pickering emulsions stabilized by different areca taro starches
(native starch, OSA esterified starch, ball-milled starch, and compound modified
starch with ball-milling and OSA), and found that the compound modified starch
showed strong surface activity and high emulsion viscosity, leading to the best
emulsifying capacity and stability. Nevertheless, it should be noted that the amy-
lose/amylopectin ratio also affects the emulsion properties. Lu et al. (2018) reported
that the milled high-amylose maize starch particles had the best stabilization ability,
followed by milled normal maize starch particles. Kasprzak et al. (2018) screened a
series of commercially available food starches, and found that a waxy rice starch
showed the O/W emulsifying ability following gelatinization. In addition, the OSA
modified quinoa starch granules were also reported to be used to make W/O/W
double emulsions with solid or liquid shea oil (Marefati et al. 2015). The double
emulsions were freeze-dried and then rehydrated for emulsion reformation, which
had a similar droplet size to that of the initial emulsion, suggesting a high process
stability.

8 Protein-Polysaccharide Conjugates

Numerous studies on the emulsifying properties of protein-polysaccharide conju-

gates have been published in the last few years. A conjugate is the material made
from a protein covalently linked by a polysaccharide via Maillard reaction, in which
temperature, humidity, and concentration of ingredients need to be carefully con-
trolled. Maillard reaction is a series of non-enzymatic browning reactions that occur
naturally between the reducing end of a sugar and amino acids. The functional
properties of protein and polysaccharide can be combined through Maillard reaction
194 H. Zhang and L. Deng

from the production of a novel protein-polysaccharide conjugate, which may be

expected to result in an enhanced protein functionality both as an emulsifier and
a stabilizer.
Several methods including wet heating, dry heating, and molecular crowding are
often used to induce Maillard reaction between proteins and polysaccharides. For
dry heating, to ensure sufficient contact, the powders of the protein and polysaccha-
ride are obtained by freeze-drying the mixture firstly dispersed in water, and then
heated in an apparatus at the controlled time, temperature, and relative humidity.
Wet heating can also be used to prepare protein-polysaccharide conjugates by
heating the ingredients present in a buffer solution for hours. It is considered as a
more efficient method with the improved control over the reaction advancement, in
comparison to the dry heating technique. However, the concentration of reactants
needs to be increased for a higher glycation level, as molecular crowding is of
necessity for Maillard reaction under less adverse treatments in aqueous solutions
(Perusko et al. 2015; Weng et al. 2016). In the presence of high concentrations of
biological macromolecules, the reaction follows the excluded volume theory, and
the crowded environment promotes the conjugation between protein and polysac-
charide. In addition, the extent of protein aggregation can potentially be minimized
in the macromolecular crowding environment. Recently, ultrasound has been
applied to facilitate the Maillard reaction. It was reported that the conjugates
obtained by the ultrasound treatment had better emulsifying properties, compared
to those prepared by classical heating (Xue et al. 2017; Chen et al. 2016b; Liu et al.
2016). Stanic-Vucinic et al. (2013) found that high-intensity ultrasound efficiently
promoted the glycation of β-lactoglobulin by Maillard reaction, and the obtained
conjugates possessed the improved antioxidant capacity, with a minor influence on
protein’s secondary and tertiary structures.
Table 5.3 illustrates some studies involving the conjugates from proteins (e.g.,
whey protein, casein, β-lactoglobulin, and soy protein), and polysaccharides (e.g.,
guar gum, pectin, and dextran). High molecular weight conjugates possess the
properties of protein, strongly adsorbing at the surface of oil droplets, as well as
the hydrophilic properties of polysaccharide, being highly solvated by the aqueous
medium. The conjugation between proteins and polysaccharides may provide much
more improved steric stabilization for emulsion droplets, as illustrated in Fig. 5.9. As
the molecular weight of the polysaccharide moiety sufficiently increases, a thicker
stabilizing layer can be formed by the conjugate in an emulsion. Therefore, the well-
prepared protein-polysaccharide conjugates may show the substantially improved
emulsifying and stabilizing properties compared to native proteins (Sivapratha and
Sarkar 2018).
Great concerns have been also raised by the presence of a protein and polysac-
charide as naturally occurring substances with the emulsifying capacity produced by
Maillard reaction. Cirre et al. (2014) conducted a heat-induced maturation treatment
for corn fiber gum in the solid state, which induced a reduction in the solubility and
incensement in the aggregation of the proteinaceous component. The emulsification
performance and stability of the matured samples were greatly improved, in com-
parison with the control gum. A 3-fold increase in the proportion of the adsorbed
Table 5.3 Formation, characterization, and functionality of protein-polysaccharide conjugates obtained via Maillard reaction
Protein Polysaccharide Reaction condition Characterization methods Main results References
Whey protein Guar gum Wet heating, 50 or 70  C, SDS-PAGE (sodium dodecyl Cumin seed oil (CSO) Farshi
15 min sulfate polyacrylamide gel nanoemulsions were prepared using et al.
electrophoresis) preheated 10% WPI and 0.2% guar (2019)
gum in aqueous phase, and the mix-
ture of CSO and corn oil (30:70) as
oil phase.
5 Emulsifying Properties

Deamidated Citrus pectin Dry heating, 80  C, RH (rela- OPA (o-phthaldialdehyde), Conjugates prepared by dry heating Wang
wheat gluten or tive humidity) 79%, 24 h SDS-PAGE, FTIR (Fourier deamidated wheat gluten/maltodex- et al.
maltodextrin transform infrared), CD (circular trin (1/1) for 9 h showed the best (2019)
dichroism) emulsifying properties.
Whey protein Pectin Dry heating, 80  C, RH 79%, SDS-PAGE, HPSEC (high- Solubility and emulsion stability Wefers
6–168 h pressure size exclusion were improved. et al.
chromatography) (2018)
Casein Carrageenan Dry heating, 60–80  C, RH OPA, Color, FTIR, fluorescence Conjugates obtained at 60  C for Qiu et al.
79%, 6–72 h measurements 24 h showed the best emulsifying (2018)
activity and stability.
Bovine serum Sugar beet Dry heating, 85  C, RH 79%, SDS-PAGE, Color, GPC (gel Covalent conjugates exhibited good Chen et al.
albumin pectin 5h permeation chromatography) emulsion stability under the extreme (2018)
environmental conditions.
Rice protein Dextran Wet heating, pH 12, 80– TNBS (2,4,6-trinitrobenzene sul- Emulsifying activity and stability Cheng
100  C, 10–30 min fonic acid), HPSEC, SDS-PAGE, were increased by factors of 1.79 and et al.
FTIR 2.2, respectively. (2018)
Egg-white Guar gum Dry heating, 60  C, RH 80%, SDS-PAGE, Color, GPC, FTIR Emulsion capacity and stability were Hamdani
lysozyme 10 days significantly improved. et al.
(2018)
Sodium Locust bean Dry heating, 80  C, RH 79%, SDS-PAGE, Color, FTIR Conjugates produced at 80  C after Barbosa
caseinate gum 4 days 24 h showed good emulsifying et al.
properties. (2018)
(continued)
195
Table 5.3 (continued)
196

Protein Polysaccharide Reaction condition Characterization methods Main results References

Whey protein Citrus pectin Closed-cavity rheometer, 80– HPSEC, fluorescence A novel Maillard reaction method Koch et al.
isolate (WPI) 140  C, 0.75–10 min measurements was applied to highly concentrated (2017)
WPI-highly methylated citrus pectin
blends at elevated temperatures.
Soy protein Dextran Dry heating, 60  C, RH 79%, SDS-PAGE, OPA, FTIR Oil droplet size was decreased, and Boostani
isolate 8 days emulsion stability was improved. et al.
(2017)
Casein hydro- Acacia seyal Dry heating, 60  C, RH 75%, TNBS, HPSEC Conjugates prepared from A. seyal Hou et al.
phobic peptide polysaccharide 3 days polysaccharide and CHP with a (2017)
(CHP) degree of hydrolysis of 1.5%
presented the best emulsifying
properties.
Soy protein Soy hull hemi- Dry heating, 60  C, RH 75%, Color, FTIR, free amino acids Optimized SHH-SPI conjugates Wang
isolate (SPI) cellulose 7 days exhibited the substantially improved et al.
(SHH) emulsification capacity. (2017)
WPI Maltodextrin Dry heating, 80  C, RH 79%, OPA Emulsifying and colloid stabilizing Ding et al.
3h properties were improved. (2017)
Canola protein Gum arabic Wet heating, pH 7.0, 90  C, SDS-PAGE, HPSEC Conjugate-stabilized emulsions Pirestani
isolate 15 min showed smaller droplet size, lower et al.
creaming index, and higher resis- (2017)
tance to pH and heat.
β-lactoglobulin Gum Acacia Dry heating, 60  C, RH 79%, TNBS, SDS-PAGE Emulsion stabilities against storage, Bi et al.
Seyal 0–48 h pH, temperature, and salt were (2017)
improved.
β-lactoglobulin Chitosan Wet heating, pH 6.0, 40  C, 0– SDS-PAGE, fluorescence Emulsifying properties were Mengíbar
7 days measurements improved. et al.
(2017)
H. Zhang and L. Deng
Soy Dextran Macromolecular crowding, pH SDS-PAGE, TNBS Degree of conjugation grafting car- Weng
β-conglycinin 7.0, 95  C, 6 h; Dry heating, ried out in macromolecular crowding et al.
65  C, RH 75%, 96 h environment was generally higher (2016)
than that produced by dry heating
method.
Buckwheat Dextran Ultrasound-assisted wet FTIR, CD, fluorescence Emulsifying properties of the conju- Xue et al.
protein isolate heating, 70  C, 80 min; Wet measurements gates obtained by the ultrasound (2017)
heating 70  C, 40 h treatment were improved, compared
5 Emulsifying Properties

to those obtained by classical

heating.
Peanut protein Maltodextrin Ultrasound-assisted wet OPA, SDS-PAGE High-intensity ultrasound promoted Chen et al.
isolate (PPI) heating, pH 7.0, 70  C, 10– the production of glycated PPI. (2016b)
100 min
197
198 H. Zhang and L. Deng

Fig. 5.9 Schematic representation of Maillard reaction between proteins and polysaccharides. The
formed conjugates are used to improve stabilization of emulsions

fraction onto the oil–water interface accounted for the improved emulsification. As
for okra hydrocolloid mucilage, it has both proteins and characteristic slimy poly-
saccharides. Heating at 100  C for 6 h resulted in the conjugates that showed better
emulsifying property than the ones formed between okra polysaccharides and bovine
serum albumin (Temenouga et al. 2016). Lately, brea gum, which is an exudate from
Cercidium praecox tree, was purified and subjected to thermal treatment at 110  C
for 24, 48, 72, and 96 h, respectively. The thermally treated brea gum presented
changes in the molecular mass of its protein fractions and in its color parameters as a
result of non-enzymatic browning reactions. The modifications in the gum structure
produced an increase in its surface activity and improved its emulsifying/stabilizing
capacity, leading to corn oil-in-water emulsions stable for several months (Castel
et al. 2018).

9 Applications in Food Industry

The hydrocolloid stabilized emulsions have showed a number of applications in the

food industry, such as dairy products, meat products, beverages, etc. (Fig. 5.10).
Beverage emulsions are a unique class of emulsions, which are consumed in a highly
dilute form. The emulsions in both the concentrate and diluted forms must be stable
at least for 6 months. Gum arabic is the most widely used emulsifier as well as a
stabilizer in beverage emulsions. The most accepted alternative to gum arabic for
beverage emulsions is modified starches, whose lipophilic and hydrophilic groups
can be balanced by modifications (Tan 2004). There are other hydrocolloids with
good emulsifying properties to be used in beverage emulsions, for example, locust
bean gum. In ice cream, hydrocolloids have been applied for various functions,
especially as a stabilizer, fat replacer, and cryoprotectant (Javidi and Razavi 2019).
Structurally, it consists of a complex matrix of fat globules, ice crystals, air bubbles,
and a continuous phase of unfrozen water with dissolved sugars, proteins, and salts
(Daw and Hartel 2015). Gelatin almost exclusively serves as a stabilizer in the ice
5 Emulsifying Properties 199

Fig. 5.10 Major applications for hydrocolloid emulsifiers in food industry

cream industry, but has gradually been replaced by the polysaccharides of plant
origin due to their increased effectiveness and reduced cost. Blends containing locust
bean gum, guar, and carrageenan are excellent stabilizing systems for ice cream.
Sausages are made of a fine homogenate by adding water and salt into chopping
meat, in which the dispersion and emulsification of pork fat is further achieved.
Solubilization of muscle protein is associated with the interactions with salt solutions
during blending of meat. Then, a stable emulsion is formed due to the formation of a
surrounding layer of the salt soluble proteins at the surface of the released fat. In
sausage products, more myofibrillar proteins are extracted, thus heat treatment can
contribute to the formation of a stronger gel. Sodium caseinate, soy protein isolate,
guar gum, xanthan, gellan gum are commonly used in meat products as emulsifiers.
Due to the high calorific value and/or the fat-related health problems caused by
high-fat diets, the demand for reduced-fat counterparts in food products has been
developed in recent years (Azeredo et al. 2019). Some studies of fat replacement
have been performed using different hydrocolloids in various products, such as ice
cream, mayonnaise (Golchoobi et al. 2016), sausages (Pintado et al. 2018). Several
structuring approaches have been reported to mimic the fat texture. Emulsion gels
have been fabricated by casein, whey protein isolate, soy protein isolate as emulsi-
fiers (Freire et al. 2018). Double emulsions (W/O/W) have been fabricated to
maintain the contact area between fat and the oral surface in the mouth. Pickering
emulsions have been successfully stabilized by zein, starch granules, cellulose
crystals, whey protein microgels, crosslinked casein gel particles as potential fat
replacers (Javidi et al. 2019; Wang et al. 2018b). Nanoemulsions are more stable
against creaming, sedimentation, coalescence, and flocculation, and have higher
surface area and more free energy, so they are suitable carrier systems. Recent
researches have shown that the nanoemulsions stabilized with proteins were more
stable than those stabilized by low molecular weight surfactants due to the formation
200 H. Zhang and L. Deng

of an elastic film by proteins, which can inhibit coalescence of droplets in the

dispersed phase. In addition, the bioactive encapsulated nanoemulsions have been
prepared by stabilization of hydrocolloids, such as β-lactoglobulin (Ali et al. 2016),
pectin/whey protein complexes (Esfanjani et al. 2015; Gharehbeglou et al. 2019).

10 Future Prospects

Over the last decade, a growing number of studies have focused on the characteri-
zation of the emulsifying properties of food hydrocolloids, leading to a fundamental
understanding of the mechanisms of their interfacial functionalities. Apart from the
known hydrocolloid emulsifiers, more and more naturally extracted biopolymers
will be explored as a novel candidate for applications in foods, inspired by the
consumer preference for clean label ingredients. Many studies have also shown that
the protein-polysaccharide conjugates induced by Maillard reaction may present the
improved emulsifying property compared to the single components, indicating the
potentials to be used in beverages and dairy products. However, the main challenge
for developing such novel emulsifiers is to select a conjugated fraction with high
functionality in emulsions under a certain environmental condition for the desirable
formulated products. Therefore, in future more studies are needed to understand the
relationship between the conjugate structures and functional properties before use in
practical applications.

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Chapter 6
Liquid Foaming Properties

Yongguang Guan

Abstract Liquid foams are thermodynamically unstable colloidal systems with a

vapor phase incorporated in a continuous phase, which can be considered as a
composition of gas, foaming agents, and liquid. In this chapter, the foam formation
and microstructure were firstly introduced to understand foaming processes and
basic structural components in a foam system. Then, the foam instability mecha-
nisms comprising drainage, coalescence, and disproportionation were introduced
with an insight into the factors of floatation, coalescence, and Ostwald ripening for
foam destabilization. Afterwards, the foam colloidal and functional properties
including foam ability, foam stability, and stabilization strategies focusing on
improving foam overrun and mean life by employing various foaming agents such
as proteins, polysaccharides, and Pickering fine particles were discussed. Finally,
potential applications of stable liquid foams in food industry manufactures of ice
cream, beer, bread, food-grade packages, and foam-mats were illustrated. We expect
this chapter may provide fundamental knowledge for readers to better understand
food-grade liquid foams.

Keywords Liquid foam · Interface · Thin film · Foaming agents · Foam stability

1 Formation and Microstructure

Liquid foam is a multiscale system composed of gas, foaming agents, and liquid. Its
formation is governed by transportation, penetration, and reorganization of the
molecules at the gas–liquid interface, depending on surfactant size, surface visco-
elasticity, and conformational flexibility (Wilde and Clark 1993). In foaming

Y. Guan (*)
Department of Food Science and Engineering, School of Agriculture and Biology, Shanghai
Jiao Tong University, Shanghai, China
e-mail: yguan@sjtu.edu.cn

© Springer Nature Singapore Pte Ltd. 2021 207

Y. Fang et al. (eds.), Food Hydrocolloids,
https://doi.org/10.1007/978-981-16-0320-4_6
208 Y. Guan

Fig. 6.1 High-energy emulsifiers and homogenizers used for foam preparation. Gray circles and
ovals denote gas in bubbles. This figure is plotted referring the work from Urban and co-workers
(Urban et al. 2006)

processes such as employing high pressure, membrane, ultrasound, rotor-stator

homogenizer, and disc whipper (introduced by Urban and co-workers (Urban et al.
2006) and shown in Fig. 6.1), energy is supplied from mixer into liquid to incorpo-
rate gas and, therefore, forms a cohesive gas–liquid interfacial layer (Djelveh and
Gros 1995), which tears large bubbles to smaller ones primarily through simple shear
flow and elongational flow (Fig. 6.2). After these foaming processes, two major
foams, i.e., bubbly and polyhedral foams are fabricated (Fig. 6.3). The liquid bubbly
foam is determined to be stable only in a high viscosity system such as syrup at high
concentrations. Meanwhile, the polyhedral foam is considered to be generated
through the bubble floatation originated from the initial bubbly foam (Fig. 6.4). A
whole polyhedral foam structure can be denoted in Fig. 6.5, the separated gas–liquid
system by surfactants supports foam with disperse phase of gas in bubbles and
continuous phase of liquid in thin film. Moreover, the two adjacent interfacial layers
and intermediate thin film (or thin liquid film) constitute a lamella (Wasan and
Nikolov 2008). Although most of the surfactants, i.e., the surface active agents
such as proteins and polysaccharides, are always adsorbed at the gas–liquid interface
to decrease the interfacial tension, a portion of them can be presented in thin film
(Wasan and Nikolov 2008), which is likely to form gel-like network. Thus, from a
colloidal viewpoint, the stable liquid foam can be seen as a complex soft condensed
matter rather than a simple solid, liquid, or gas. In this case, the interaction forces
among bubbles are primary normal stresses without tangential forces (Labiausse
6 Liquid Foaming Properties 209

Fig. 6.2 Two types of foam breakup through simple shear flow and elongational flow. Circles and
ovals denote gas in bubbles. Particles with different shapes and colors were shown at the interfaces
of bubbles

Fig. 6.3 Two foam types, i.e., bubbly and polyhedral foams. Circles and polygons denote gas in
bubbles

et al. 2007). The whole structure of foam follows the Plateau’s equilibrium law, that
is, four bubbles form a basic unit with one intersection, and each three bubbles
constitute a Plateau border (Weaire and Fortes 1994). The curvature radius of a
Plateau border is 10 nm to 1 mm depending on the synergetic interaction of liquid
fraction, surface tension, and interfacial forces. The length of a Plateau border is
approximately one-third of bubble diameter. The thin liquid film exists in two
adjacent bubbles with the thickness of 0.01 nm to 1000 nm, which is the minimum
distance among bubbles. The dihedral angles of two adjacent lamellae and Plateau
210 Y. Guan

Fig. 6.4 The formation pathway of polyhedral foam from the initial stage of bubbly foam
generating an intermediate foam after floatation, coalescence, and even the Ostwald ripening
(will be discussed below), and finally generating polyhedral foam. Circles and polygons denote
gas in bubbles

borders are 120
and 109.47
, respectively (Stamenović 1991), which was elabo-
rated by Weaire and Phelan (Weaire and Phelan 1996).
The morphologies of bubble interface membrane are formed attributing to the
Laplace pressure primarily including compression driven force, phase separation,
and buckling evolution, which generate various microstructures such as polygonal,
bean, snowflake dendritic, and mesh structures. Electron microscopies can be used to
observe bubble interfacial layer microstructures. For instance, a polygonal micro-
structure of the liquid foam membrane surface generated by gas in 1,2-distearoyl-sn-
glycero-3-phosphatidylcholine monolayer was observed by the transmission elec-
tron microscopy (TEM). The evolutionary process of this polygonal microstructure
was demonstrated by the transfer from a smooth microstructure in the initial stage of
bubble formation to a wrinkling one during bubble storage attributing to the Laplace
pressure (Kim et al. 2003). A regular hexagonal microstructure of elastic liquid foam
membrane with equivalent diameter of 50–100 nm was also observed in a sucrose
stearate-syrup homogeneous system. The formation of this hexagonal foam mem-
brane microstructure is driven by the Laplace compression because of the
mismatched curvature between the bubble core and the self-assembled surfactant
membrane (Dressaire et al. 2008). Additionally, in a multi-agent foam system, phase
separation is a main factor to generate diverse bubble interface membrane micro-
structures. The phase separation is mainly caused by the changes in temperature,
shear rate, and liquid film viscosity (Borden et al. 2006, 2004), that is, the bubble
interface diversity is attributed to the classical interface compression mechanism. For
example, the L-α-1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) can be iso-
lated from the liquid foam system with multiple mixed surfactants during membrane
melting and cooling, generating bean microstructure after the compression of the
Langmuir film (Nandi and Vollhardt 2003; McConlogue and Vanderlick 1997). In
addition, unstable growth of crystal nucleus with limited diffusion behavior also
appears in the process of viscous film quenching, forming snowflake dendritic
microstructure of the bubble interface membrane (McKiernan et al. 2000). Repeated
compression and expansion of the foam monolayer can promote the formation of
reticular microstructure at the liquid foam membrane interface. For instance, with a
6 Liquid Foaming Properties 211

Fig. 6.5 The simulated microstructure of bubble in liquid foam system is plotted referring the work
of Weaire and Phelan (Weaire and Phelan 1996). Polygons denote gas in bubbles

rapid cooling under the ultrasonic treatment, the phospholipid cluster can interact
with water molecules through hydrogen bonds, which further increases the foam
membrane interface branch and therefore generates net membrane microstructure
(Borden et al. 2004).
212 Y. Guan

2 Instability of Foams

Liquid foams are thermodynamically unstable colloidal systems through floatation,

coalescence, and Ostwald ripening to destabilize foams (Fig. 6.6) (Green et al.
2013), which is also denoted as three fundamental mechanisms, i.e., drainage,
coalescence, and disproportionation (Horozov 2008).

2.1 Drainage

The drainage is a process that the lamella thickness becomes thin and consequently
ruptures, attributing to the separated fluid between the thin film and bubble under the
gravity due to the lighter density of bubble than liquid (Dickinson 1992). Once the
liquid in the lamellae enters the Plateau borders, it will flow rapidly and be separated
from thin liquid film caused by the drainage (Verbist et al. 1996; Rio et al. 2014). In a
complete foam system, the drainage continues until the static pressure gradient in the
gravity direction balances the gravitational acceleration. At this moment, the static
pressure gradient is maintained by the disjoining pressure because of a very thin
liquid film and therefore cannot further reduce the interfacial potential energy. The
thickness of the thin liquid film is decreased following the height increase of the
foam system (Verbist et al. 1996). The primary factor affecting discharge is thin
liquid film characteristics, suggesting substances in the thin film with strong liquid
holding capacity can slow down the drainage and increase the liquid foam stability
(Verbist et al. 1996). A simulated diagram of drainage is plotted referring to
published works (Weaire and Phelan 1996; Saint-Jalmes 2006) and shown in
Fig. 6.7.

Fig. 6.6 Schematic diagrams of unstable factors of floatation, coalescence, and Ostwald ripening
for destabilizing foams. Circles denote gas in bubbles
6 Liquid Foaming Properties 213

Fig. 6.7 The simulated diagram of drainage is plotted referring published work (Weaire and Phelan
1996; Saint-Jalmes 2006), showing decreased lamella thickness, increased size, and losing spherical
structure of bubbles with the increase in foam height, caused by the downward flow of liquid under
the gravity, which suggests an accelerated rate of drainage when the liquid from the thin film of
lamellae enters the Plateau borders

2.2 Coalescence

The coalescence is a process of bubble merging that a lamella between adjacent

bubbles ruptures, causing the two bubbles combination, subsequent collapse, and
loss of desirable structure and texture (Rio et al. 2014; Murray and Ettelaie 2004;
Disalvo 1988; Marrucci 1969). A simulated bubble coalescence process is shown in
Fig. 6.8. The coalescence is determined by the balance between capillary and
disjoining pressures. That is, the disjoining pressure arises from forces between
the two interfaces of the liquid lamella (Damodaran 2005; Oboroceanu et al. 2014),
while the capillary pressure occurs because of the force between the gas and liquid
phase, which is calculated by the Young–Laplace equation (Eq. 6.1, 6.2, and 6.3,
Eq. 6.1, 6.2, and 6.3) (Lian et al. 1998; Thitakamol and Veawab 2009).
 
1 1
ΔP ¼ γ þ ð6:1Þ
R R0
2πxγ sinθ ¼ VΔρg þ πx2 ΔP ð6:2Þ
2γ
ΔP ¼ ð6:3Þ
R

where ΔP is the pressure difference between the inside and outside of a bubble at any
point on the interface, γ is the surface tension, R and R0 are the radii of curvature at
the bubble apex, x is the Cartesian coordinate at any point of the bubble profile, θ is
the angle of the tangent to the bubble profile, V is the volume below a point plane, Δρ
is the difference of density between the two phases, and g is the gravitational
214 Y. Guan

Fig. 6.8 The simulated process of bubble coalescence suggests the thin film between two adjacent
bubbles is dying out along with the loss of the liquid, a subsequent combination of interfacial layers,
and a final rupture of the combined layer. This bubble coalescence process finally results in the
incorporation of small bubbles to form big bubbles and collapses the foam structure. The dark blue
ovals denote liquid. The gray circles and ovals denote gas in bubbles

acceleration. When the surface is a sphere, R is equal to R0, then Eq. 6.1 can be
transferred to Eq. 6.3 (Thitakamol and Veawab 2009; Benjamins et al. 1996).
Furthermore, the main factors of the generated imbalance between capillary and
disjoining pressures are the weaker surfactant molecular attraction against repulsive
force and the lack of liquid fluidity in thin liquid film since the Marangoni effect
(Marrucci 1969). An intact surfactant layer is capable of reducing the gas–liquid
phase interfacial tension. However, when the intermolecular attraction among sur-
factants is weaker than the repulsive force, the originally compact surfactant layer is
ruptured, leading to bubble coalescence. Additionally, as the lamellae of the adjacent
bubbles are disturbed by external factors (e.g., temperature) to produce locally
thinner liquid film, the flow of liquid in lamellae produced by the Marangoni effect
will form the Marangoni flow under the surface tension gradient, restoring the
thinner liquid film timely (Christenson and Yaminsky 1995). That is, the liquid
rapidly moves from a lower bubble surface tension film along the tension gradient to
6 Liquid Foaming Properties 215

Fig. 6.9 The simulated Marangoni effect denotes a protective effect of bubbles against coalescence
through generating the Marangoni flow under the surface tension gradient and restoring the thinner
liquid film timely. The light blue background denotes liquid, and gray circles denote gas in bubbles.
The surfactants are denoted and composed of hydrophilic heads with dark blue and green ovals and
hydrophobic chains with orange needles

a higher one (Fig. 6.9). However, when the fluid mobility of the liquid in thin film is
decreased, the efficiency of restoring the thinner liquid film is therefore reduced,
leading to the rupture of lamellae and bubble coalescence (Marrucci 1969).

2.3 Disproportionation

The disproportionation, driven by the difference of Laplace pressure between

smaller and larger bubbles, is another important destabilization mechanism causing
foam destabilization (Ettelaie et al. 2003). In foam disproportionation, gas molecules
in high-energy smaller bubbles have higher solubility in liquid, and thereby
are released in a single molecular form from original smaller bubbles (Fig. 6.10).
The released gas molecules coalesce into larger bubbles until the foam reaches the
minimum specific surface area and energy (Rio et al. 2014; Murray and Ettelaie
2004; Disalvo 1988). In an unstable foam system, the disproportionation becomes
drastic as the dilatational modulus is larger than one-half of the surface tension (Rio
et al. 2014; Murray and Ettelaie 2004). In addition, smaller bubbles have shorter
curvature radius compared to bigger ones, which means the inner Laplace pressure
of a smaller bubble is greater than that of a bigger one, although the Laplace pressure
within bubbles also depends on the interfacial elasticity of the surfactant layers
around the bubbles and their shrinkage extents. The Laplace pressure difference
between smaller and bigger bubbles facilitates gas molecule diffusion from smaller
to bigger bubbles driven by layer surface tension and finally reduces the specific
surface area of gas–liquid interface. The disproportionation in foams is analogous to
the Oswald ripening in emulsions but more dramatically caused by the smaller size
of gas molecules with better liquid film permeability (Murray and Ettelaie 2004;
Disalvo 1988).
216 Y. Guan

Fig. 6.10 The simulated disproportionation suggests gas molecules can diffuse from bubble
through the gap between surfactants of interfacial layer, leading to a decrease in original bubbles
and therefore foam destabilization. The light blue background denotes liquid. The gray circles and
curves denote gas in bubbles and gas molecules, respectively. The surfactants are denoted and
composed of hydrophilic heads with dark blue and green ovals and hydrophobic chains with orange
needles

Generally, the liquid foam drainage caused by gravity can be prevented when the
static pressure gradient in the gravity direction is equal to the gravitational acceler-
ation. Increasing the liquid holding capacity in thin film can significantly slow down
the drainage (Bisperink et al. 1992; Narsimhan and Xiang 2018). The bubble
coalescence caused by lamella rupture can be effectively inhibited by choosing
appropriate interfacial stabilizer with stronger intermolecular attractions and surface
activity (Lian et al. 1998; Christenson and Yaminsky 1995; Bisperink et al. 1992).
The disproportionation is a very slow process and can be inhibited by building dense
interfacial layer and stable thin liquid film (Ettelaie et al. 2003; Bisperink et al.
1992). However, a liquid foam system is always in thermodynamic non-equilibrium
state, resulting in an unfulfillable complete prevention of gas molecule dispropor-
tionation (Kornev et al. 1999). That means the disproportionation is very difficult to
6 Liquid Foaming Properties 217

be prevented since even the well-packed interfacial layers at gas–liquid interface can
only provide limited barriers against the gas diffusion through the transient gaps
among surfactant molecules. The drainage, coalescence, and disproportionation
synergistically disturb foam stability, showing time-dependent non-equilibrium
characteristics. When drainage occurs, gas molecule diffusion from bubbles is
facilitated by fluid flow in thin liquid film. While the disproportionation facilitates
the coarsening of the bubble size, accelerating the drainage and coalescence.

3 Improvement of Foam Ability and Stability

3.1 Physicochemical Characteristics

The foam ability and stability are two important parameters to evaluate the foaming
properties. The foam ability is denoted as the percentage of overrun based on the
volumes of fresh foam and totally whipped liquid, which can be expressed by Eq. 4
(Mitchell 1986).

V0  V1
Overrunð%Þ ¼  100 ð6:4Þ
V1

where V0 is the foam volume right after whipping, and Vl is the volume of liquid used
for foaming. Higher overrun indicates a higher foam ability of the whipped
materials.
The foam stability is calculated by an exponential decay law that the foam height
is expressed in Eq. 6.5 and can be further expressed as an index of foam stability by
the “mean life” τ value that is used to evaluate the foam stand time and shown in
Eq. 6 (Leike 2002).

H t ¼ H 0  eλt ð6:5Þ
t
τ¼ ð6:6Þ
ln ðH t Þ  ln ðH 0 Þ

where Ht is the foam height after a storage time t, and H0 is the initial height of foam.
The λ here is the foam decay constant. A larger τ value means a more stable foam
system.
The thermodynamics of foaming agent adsorbed layers at the gas–liquid interface
can be expressed by the surface pressure isotherm, which is a relationship between
the surface pressure and the foaming agent concentration (Patino et al. 2008). When
using low molecular weight foaming agents, different supramolecular structures
with covered gas are formed at a higher surfactant concentration than the critical
micelle concentration (CMC). These bubbles built by low molecular weight agents
show little repercussions on the surface pressure, and therefore, their surface
218 Y. Guan

pressure isotherms are sensitive to temperature and pH (Patino et al. 2008; Niño and
Patino 1998). The dependency between surface pressure and adsorption isotherms of
foams prepared by biopolymers suggests a sigmoidal behavior, implying an increase
in the surface pressure with biopolymer concentrations until tending to a plateau
where the surface pressure reaches its maximum value over the range of biopolymer
concentrations, which forms an irreversibly adsorbed monolayer or multilayer
beneath the primary monolayer at the gas–liquid interface (Patino et al. 2008).
These biopolymer-built foams are generally more stable compared to those using
low molecular weight surfactants. The biopolymer adsorption at the gas–liquid
interface is a complex process including (1) diffusion from the solution to the
subsurface, (2) adsorption and unfolding at the interface, and (3) reorganization or
rearrangement of segments previously adsorbed at the interface (Patino et al. 2008),
which critically depends on the chemical constitutions of biopolymers and foaming
systems.
The interfacial dilatational capacity, reflected by the interfacial dilatational mod-
ulus, provides the fundamental insight into the foam ability and stability from the
interfacial perspective. The interfacial dilatational modulus (ɛd) can be expressed as a
linear response of film to sinusoidal deformation at a certain frequency and
suggested by Eq. 6.7 (Maldonado-Valderrama et al. 2008).

ɛd ¼ ɛ þ iωη ð6:7Þ

where ɛ is the interfacial elasticity modulus (i.e., storage modulus) reflecting the
ability of the layer restoring its interfacial tension after stress, which measures the
resistance to deformation of the layer. While η is the interfacial viscosity, measuring
the speed of the relaxation processes that restore the equilibrium after the distur-
bance. And the ωη is the viscosity modulus (i.e., loss modulus), reflecting the energy
loss of the layer after a stress induced deformation and, therefore, measures the
capacity of the layer adapting a deformation (Maldonado-Valderrama et al. 2008).
The interfacial viscosity of the protein adsorbed layer appears to be a key factor in
the formation of foams (i.e., foam ability), whereas the interfacial elasticity is more
concerned with the foam stability.
Based on the above mechanism, the increase in foam ability is expected to utilize
low molecular weight surfactants with higher interfacial viscosity and surface
activity. Nevertheless, the improvement in foam stability is expected to use high
molecular weight biopolymers to prevent or delay gas–liquid phase separation and to
reduce the gas–liquid interfacial energy. That is to prevent drainage in thin liquid
film, coalescence of bubbles, and disproportionation of gas molecules. Smaller
amphipathic surfactants have a higher interfacial viscosity and surface activity
with faster diffusion rate to form interface layers to pack gas when the foam are
formed in liquid. Larger biopolymers and their aggregates with different supramo-
lecular configurations such as fibrils, micelles, and crystals have lower interfacial
viscosity and cannot quickly diffuse to the gas–liquid interface because of larger size
compared to the smaller ones, but they are capable of irreversible adsorption at the
interfacial layer and/or dispersing in lamellae and Plateau borders, which can
6 Liquid Foaming Properties 219

effectively increase the interfacial elasticity and thin film viscosity in lamellae and
Plateau borders for improving foam stability.
Wettability and spreading of particles at the interface can stabilize or destabilize
foams, depending on their properties (Narsimhan and Xiang 2018). Similar to the
Pickering emulsion that was first reported in the early 1900s by Pickering (Pickering
1907), the particle-stabilized strategy is important in foam stabilization. At present,
these particles constituted by larger biopolymers or their aggregates can prevent
bubble coalescence or rupture via steric hindrance in lamellae and inhibition of
drainage in both lamellae and Plateau borders. In addition, these particulate-
stabilized foam systems with an appropriate particle size require a higher energy to
remove particles off from the interface for the destabilization when particles have a
three phase contact angle close to 90
(Tzoumaki et al. 2015) and sizes ranging from
10 nm to 30μm (Hunter et al. 2008). In this case, the interfacial area occupied by the
particles allows the adsorption energy per particle to reach several thousands of kT
and therefore is capable of preventing the symmetrical shrinkage of bubbles. The
kinetic shrinkage of a single bubble suggests the existence of a critical bubble size,
above which the bubble surface is covered rapidly by particles for the inhibition of
bubble dissolution, while, below this critical size, the bubbles shrank too fast to be
stabilized. The critical bubble size, commonly with the diameter of 60 to 80μm
(Dickinson et al. 2004), depends on the size of the particles or their aggregate forms
in dispersions (Dickinson et al. 2004; Aveyard et al. 1994). Oversize dimension of
particle, probably higher than 30μm, reduces the diffusion coefficient and therefore
foaming properties or form a possible sublayer assisting to stabilize lamella struc-
ture. While, too smaller particles with size less than 10 nm may lose their mechanical
properties and further destabilize the foam stability (Lavoine and Bergström 2017;
Lam et al. 2014). Moreover, particles with higher aspect ratio can better stabi-
lize foams compared to the particles with spherical shape, which is speculated that
the anisotropic particles promote greater capillary deformations at the interface
(Campbell et al. 2009), generating stronger capillary attractive forces between
adjacent particles at the interface and therefore leading to higher interfacial packing
and higher mechanical rigid (Madivala et al. 2009). When such particles with
irregular shape and appropriately large size are adsorbed at the interface, they can
create a very close-packed structure at the gas–liquid interface to generate a colloidal
barrier. This compactly covered interfacial layer can prevent or even completely
inhibit the foam destabilization caused by coalescence and disproportionation
(Tzoumaki et al. 2015; Dickinson 2010). Therefore, the surface free energy for
particle adsorption at the interface can be calculated by Eq. 6.8.

G ¼ γ 1S A1S þ γ 2S A2S þ γ 12 A12 ð6:8Þ

where G is the surface free energy for a particle adsorption at interface, the γ 1S, γ 2S,
and γ 12 are the interfacial tensions between phase 1 and the particle, phase 2 and the
particle, and immiscible phases 1 and 2, respectively. The A1S, A2S, and A12 are the
contact area of phase 1 with adsorbed particle, phase 2 with the adsorbed particle,
220 Y. Guan

and the contact area between the immiscible phases 1 and 2 eliminated by the
adsorbed particle at the interface (Lam et al. 2014; Kralchevsky et al. 2005, 1992).
Equation 6.8 can be transferred to Eq. 6.9 when the particle is a spherical shape.

G ¼ ΔE ¼ πr 2 γ 12 ð1  j cos θ12 jÞ2 ð6:9Þ

where r is the particle radius, γ 12 the interfacial tension of the immiscible phases, and
θ12 the three phase contact angle (Lam et al. 2014).
Based on Eqs. 6.8 and 6.9, the particle cannot be removed from the interface until
the extrinsic energy is higher than the surface free energy (G). When spherical
particles adsorbed at the gas–liquid interface have larger interfacial tension and
contact angle closer to 90
, more energetic adsorption of these particles at the
biphasic interface is determined. That means such particles cannot be easily
displaced from the interface by random fluctuations. However, if the particles are
too hydrophobic with contact angle observably higher than 90
, they can bridge the
surfaces of more than one bubbles resulting in bubble coalescence (Aveyard et al.
1994), which is similar to the antifoaming mechanism of surfactants with very low
hydrophile–lipophile balance (HLB) values (always lower than 3.0). Besides the
above mechanism focusing on the improvement of foam stabilization based on the
adsorption of particles at the interface, the non-adsorbed particles still play an
important role in improving foam stabilization through building stable thin film. In
general, the non-adsorbed particles can be firmly trapped in lamellae and Plateau
border when these particles have reasonable size, shape, as well as liquid holding
capacity, which can provide additional repulsive interaction force based on steric
hindrance of these non-adsorbed particles between gas–liquid interfacial layers
(Murray and Ettelaie 2004). The greater repulsive interaction force can be deter-
mined when the non-adsorbed particles are better in-film packed with highly ordered
structures. Additionally, these non-adsorbed particles can block Plateau border and
therefore sufficiently slow down or completely inhibit drainage (Murray and Ettelaie
2004). All these adsorbed or non-adsorbed particles with the function to build
interface and thin film (in lamellae and Plateau border) stabilization have been
investigated in multiple literature researches. Here, focusing on the food-grade
particles, they are classified as proteins, polysaccharides, and inorganic particles
with the anisotropic sharps of fibers, rods, and sheets, and are comprehensively
discussed in this chapter with a simulated diagram shown in Fig. 6.11.

3.2 Foaming Agents

3.2.1 Protein

Most protein molecules have hydrophilic–hydrophobic amphipathic chemical struc-

ture, considered as surface active macromolecules to stabilize liquid foams (Szilvay
et al. 2007). Generally, the hydrophobic groups of proteins are capable of bonding
6 Liquid Foaming Properties 221

Fig. 6.11 The foam stabilization by using biopolymers at interface or in liquid continuous phase
with different mechanisms suggests through the electrostatic repulsion of interface (a), the low
molecular weight polymers such as hydrolysates adsorbed at the gas–liquid interface (b), the large
molecular weight polymers or aggregates attached with the interfacial layer (low molecular weight
polymers) as a sublayer (c), the co-adsorption of low and large molecular weight polymers at the
interface (d), the covalent linking (e.g., Maillard-type conjugation) of hydrophilic–hydrophobic
biopolymers adsorbed at the interface (e), the biopolymer particle such as clay, fiber, rod, and
amorphous nanoparticles adsorption at the interface based on the Pickering mechanism of stabili-
zation (f), the formation of gel-like network in lamellae and Plateau border with a strong liquid
holding capacity and increased steric hindrance among bubbles (g), and the particle adsorption at
the interface and in lamellae and Plateau border to increase the mechanical strength (h). The light
blue background denotes liquid, and gray circles denote gas in bubbles

with gas molecules by hydrophobic forces, while their hydrophilic groups with water
molecules by hydrogen bonds. The amphipathic protein generates adsorbed layer at
interface or sublayer that attaching interface to improve the interfacial rheological
viscoelasticity and to form cohesive three-dimensional gels for packing gas (Szilvay
et al. 2007). Higher interfacial activity and thicker interfacial layer are expected for
foam stability, which strongly depends on the conformational aggregation and
flexibility of protein. In principle, a reasonable protein layer can provide an energy
barrier to decrease the rate and extent of bubble shrinkage that results from higher
bubble-interfacial elasticity and viscosity, which suggests to be crucial for stabilizing
foam by slowing down the gas disproportionation according to the interfacial elastic
mechanism (Murray and Ettelaie 2004). In addition, macromolecular proteins are
always composed of covalently bonded amino acid residues, meaning an infrequent
thermodynamical rupture of bubble membrane compared to that by small molecular
surfactants. Also, electrostatic repulsion between protein hydrophilic groups at
adjacent bubble layers can enhance the foam stability by preventing bubble coales-
cence. Meanwhile, dispersed protein molecules in liquid film lamellae are also likely
to provide dramatical steric hindrance against dynamic collision of adjacent bubbles,
222 Y. Guan

which is much more possible to prevent bubble coalescence. Another important

foam stability mechanism is that protein aggregates in thin film, i.e., continuous
phase, can undergo a percolation process due to confinement, leading to a gel-like
network formation, which is possibly to slow down the drainage in thin film. In brief,
proteins influence both bulk foaming properties (e.g., overrun and stability) and
interfacial properties (e.g., adsorption rates and interfacial rheology), and therefore
are desirable foaming candidates. The commonly used food proteins to form stron-
ger protein films for stabilizing foams are ovomucin, soy protein, whey protein,
casein, and their aggregates or hydrolysates.

Ovomucin

The ovomucin, accounting for 3.5% of total egg white protein, being responsible
for the thick gel characteristics of liquid egg white, has a long linear molecular chain
with many coiled regions at its side chains, appears a randomly coiled structure, and
hence shows a highly polymerized steric configuration (Omana et al. 2010). Early
studies suggested that the ovomucin was an excellent foaming agent with a higher
foam overrun and τ value (Hammershoj et al. 2008; Hammershoj and Qvist 2001),
because of its larger molecular size and intrinsic viscosity (Kato et al. 1985).
Hydrolyzing ovomucin at a hydrolysis degree of 15 to 40% was found to further
improve foaming ability (i.e., overrun), but little enhanced effect on foam stability
(Hammershoj et al. 2008). Recent studies proposed that the increased foam ability
mainly depended on the ovomucin network degradation to form small-sized
fragments and protein unfolding, suggesting a break of S-S bonds by extra energy
input to generate -SH groups, which is associated with the decrease in fibril size.
Further inputting energy led to the re-aggregation of the new-generated fragments
with the transformation of -SH groups to SS bonds, associating with partial recovery
of ovomucin network (Fig. 6.12a) (Brand and Kulozik 2016). Therefore, this
degradation of ovomucin by inputting energy such as under high hydrostatic pres-
sure or high intensity ultrasound was promising to generate small particles with
higher solubility and flexibility, resulting in a rapid adsorption at the gas–liquid
interfacial layer to encapsulate gas (Gharbi and Labbafi 2019). For example, ultra-
sonically assisted foaming ovomucin was found to generate 4.9-fold higher foam
overrun with clear foam microstructures observed by SEM (Fig. 6.12b) compared to
that without the ultrasound radiation (Fig. 6.12c), which is caused by ovomucin
molecular rearrangement to improve protein solubility and therefore to decrease
gas–liquid interface tension (Gharbi and Labbafi 2018). A very interesting finding
suggested that aggregates and fragments of ovomucin degradation products could be
formed under a lower ultraviolet irradiation, which adsorbed at the interfacial layer
and increased the interfacial viscoelasticity to improve the foam overrun. Neverthe-
less, larger aggregates of ovomucin hydrolysate were produced under ultraviolet
irradiation at higher dose. These larger aggregates could not be adsorbed at the
interfacial layer but effectively block the Plateau borders and slow down the
drainage of liquid in lamellae to improve the foam stability (Gharbi and Labbafi
6 Liquid Foaming Properties 223

Fig. 6.12 The schematic diagram proposing the degradation of the ovomucin network to form
small-sized fragments and protein unfolding with the break of S-S bonds to form -SH groups at the
initial stage of the extra energy input and a subsequent re-aggregation of the new-generated
224 Y. Guan

2019). All these findings demonstrated that the well-known foam ability and stability
of ovomucin make it an ideal candidate for foaming agent.

Soy Protein

The soy protein and its hydrolysates are commonly used as food-grade foaming
agents. Native soy protein is mainly composed of 7S and 11S globulins and shows
high solubility (>90%) at alkaline but lower solubility as the pH is decreased and
close to the isoelectric point (pI ¼ 4.5) (Wolf 1970). The relatively weaker hydration
of soy protein is always deemed as limited foaming ability because of its compact
tertiary structure at environmental pH values lower than 7. Utilizing modified soy
proteins with improved conformational flexibility as foaming agents is an inchoate
means to improve foam ability and stability. Early researchers fabricated cross-
linked soy protein-glutaraldehyde biopolymers with a significant decrease in surface
hydrophobicity compared to that by native soy protein, which allowed greater tensile
strength with both improved foaming ability and stability (Park et al. 2000). A recent
study reported that the hydrolyzing soy protein at a low hydrolysis degree of 0.4%
was capable of increasing the surface activity with improved foam overrun. Further
increasing soy protein hydrolysis degree contributed little in improving foam ability
compared to the native soy protein, which was likely due to the formation of a stable
surface film by the hydrophobic hydrolysates. In addition, the foam stability was also
improved by the hydrolyzed soy protein at a lower hydrolysis degree of 0.4%
because of the maintained considerable viscoelasticity of the surface films and
water-holding capacity that could prevent liquid drainage in thin film. However,
further increasing soy protein hydrolysis degree led to a decreased viscoelasticity of

Fig. 6.12 (continued) fragments through the transformation of -SH groups to SS bonds at a further
extra energy input with partial recovery of ovomucin network (a) (Gharbi and Labbafi 2019), the
SEM images of ovomucin foams prepared with (b) and without (c) a 360-W ultrasound treatment
(Gharbi and Labbafi 2018), the AFM images of the whey protein fibers for foaming after 20 h of
heating at 80
C and concentration of 2% w/w at pH 2 before (d) and after whipping (e)
(Oboroceanu et al. 2014), the confocal images of whey protein fluid gels made at pH 5 (f) and
8 (g) with a green fluorescent probe suggesting the aggregated fluid gels at the gas-liquid interface at
pH 5, and partially dispersed in liquid continuous phase with an increase in bulk and interfacial
viscoelasticity (Lazidis et al. 2016), and a proposed interfacial structure of the SC/TA/OSA-starch
complex at pH 6.0, suggesting a resistant capability of mechanical compression and extension to
improve foam stability (h) (Zhan et al. 2019). The light blue background denotes liquid, and dark
blue irregular shapes denote particles. The schematic diagrams of (a) and (h) were plotted by
referring the corresponding literature. The images of (b)–(g) were reproduced from the
corresponding references (Lian et al. 1998; Wolf 1970; Chen et al. 2018) with permission
6 Liquid Foaming Properties 225

foam liquid films, causing the promoted drainage with possible foam collapse
(Martínez et al. 2009). Recently, applying physical means to rearrange soy pro-
tein molecular configuration to improve the foam ability and stability aroused an
interest of researchers. Rearranging soy glycinin hydrolysates to form long semi-
flexible 11S fibrils and peptides was found to provide much better foam stability at
pH of 5 to 7 even at a very low concentration of 0.1% w/w. The mechanism was
suggested that the adsorption kinetics of soy protein at gas–liquid interface was
dramatically enhanced by the hydrolytic aggregates of fibril clusters and peptides
and the formation of highly elastic surface layer by the peptides. The improved gas–
liquid interface activity provided a potential to decrease the bubble layer rupture,
which was capable of preventing coalescence (Wan et al. 2016). A novel and
fairytale study reported that soy protein treated by subcritical water at 120
C
generated larger soluble aggregates with a lower aggregation degree and flexible
conformation because of the intermolecular hydrophobic interactions, resulting in a
higher surface activity at the gas–liquid interface and a significantly improved foam
ability and stability (Wang et al. 2019). Therefore, it can be seen that the soy protein
and its hydrolysates are promising foaming agents in food industry.

Whey Protein

Dairy proteins are always used to fabricate foam with distinguished foam ability and
stability. The whey protein is one of the main dairy proteins constituted by
β-lactoglobulin, α-lactalbumin, bovine serum albumin (BSA), and the immunoglob-
ulins (Bryant and McClements 1998). Environmental factors such as pH and tem-
perature play important roles in whey protein conformational aggregation and
flexibility, managing whey protein surface activity, rheological viscoelasticity, and
surfactant layer morphologies, which dramatically affects its foam ability and
stability. Early study reported that heating 5-% w/w whey protein in aqueous at
70
C for 1 min promoted whey protein conformational change and partial molecular
aggregation, forming a mix system of monomers and soluble aggregates. Such
soluble whey protein aggregates after heating at this condition were primarily
formed by physical intermolecular attractions rather than covalent bond (e.g.,
sulfhydryl-disulfide interchange), which generates an increased interfacial area and
foaming activity index with flexible conformation for desirable interfacial layer
materials. In this case, denser foam with smaller average bubble size was measured
with the maximum foam ability at the monomer to polymer mass ratio of 6:4 and the
maximum foam stability at 4:6 (Zhu and Damodaran 1994). Whey protein long
fibrils prepared in long-time of thermal treatment (80
C and 20 hrs) at pH 2 showed
much more significant improvement in foam ability and stability than native
non-fibrillar whey protein, which was caused by the formation of thick layer with
increased viscoelasticity adsorbed onto the gas–liquid interface and the generation of
gel-like network in continuous phase. In addition, long fibrils of whey protein
can adsorb at gas–liquid interface to increase the thickness of the liquid film
and therefore to form polymeric structures, which was observed by atomic force
226 Y. Guan

microscope (AFM) and shown in Fig. 6.12d and e (Oboroceanu et al. 2014). These
stabilizing preconditions are capable of preventing liquid drainage or bubble coa-
lescence. Another interesting finding applying whey protein fluid gels fabricated at
heat induced gelation within the turbulent flow field at pH of 5 and 8 was demon-
strated to produce very stable foams (Fig. 6.12f and g). The proposed mechanism to
stabilize foams suggested that the smaller and free proteins (monomers or soluble
oligomeric whey proteins) could diffuse quickly to the gas–liquid interface and
reduce interfacial tension to promote foam generation. Subsequently, the larger
protein aggregates (whey protein fluid gels) filled the free space by both adsorbing
at the interfacial layers and existing as a network in continuous phase, thereby
increased the flexibility of the interfacial layer and the local bulk viscosity, which
improved foam stability primarily through inhibiting drainage and secondary
through increasing interfacial elasticity. Therefore, employing whey protein fluid
gels can prominently increase foam stability without significantly changing foam
ability compared to using native whey protein (Lazidis et al. 2016). It is noteworthy
that besides the thickness, viscosity, and flexibility of the interfacial layer, the lateral
electrostatic repulsive interaction is still an important factor affecting bubble inter-
facial layer stability. A recent study reported that although transglutaminase
repolymerized whey protein isolate thermolysin hydrolysates (TR-WPITHs) repre-
sent a larger size than 100 nm comparing to transglutaminase repolymerized whey
protein isolate (TR-WPI) with size less than 100 nm, the strong electrostatic repul-
sive interactions between adsorbed TR-WPITHs highly ruptured and destabilized
the interfacial layer, which led to a faster bubble coalescence (Chen et al. 2018).
Thus, it can be seen that not only the size or surface activities (viscosity and
flexibility), but also the electrostatic properties play a role in foam stability (Bhat
and Karim 2011). Another research reported that binding polyphenols with whey
protein improved the foam ability but decreased the foam stability. The mechanism
of improving foam ability was generating a more effective protein crosslinking with
polyphenols to form soluble composites adsorbed at interface, which increased the
interfacial viscosity (Cao et al. 2018). However, phenolic compounds suppressed the
dilatational elasticity of protein film via weakening protein interactions at the
interface, which led to weakening foam stability.

Casein

Casein is an alternative important protein in dairy, composed of four major fractions,

i.e., αs1-, αs2-, β-, and κ-casein in the mass proportion of 3:0.8:3:1 (Aoki et al. 1996),
differing in many respects including charge (αs1- > αs2- > β- > κ-casein), sensitivity
to ion-induced precipitation (αs2- > αs1- > β- > κ-casein), and foaming and
emulsifying properties (Kruif et al. 2002). The αs1-, αs2-, and β-casein fractions
6 Liquid Foaming Properties 227

show a stronger gas–liquid interfacial adsorption than κ-casein, and among them,
β-casein shows the highest surface activity (Fang and Dalgleish 1993). Sodium
caseinate (SC), a primary sodium salt of casein, is prepared by reacting casein
with sodium hydroxide and subsequently neutralizing the system pH to neutral.
Casein and SC are distinguished by their good foaming properties such as forming
strong gas–liquid interfacial layers, which is extremely able to diminish external
disturbances that can prevent liquid film rupture, reflected by the interfacial dilata-
tional modulus shown in Eq. 6.9 (Langevin 2000; Wilde 2000). Based on the
principle of foam properties with the interfacial dilatational modulus, small and
flexible β-casein has higher foaming ability due to its more rapid diffusion onto the
surface. The interfacial viscosity of interface layers produced by individual β-casein
showed a positive correlation with interfacial thickness (Martinez-Pedrero et al.
2018), suggesting that the individual β-casein at pH of 8.7 showed the highest
foam ability, attributed to the dramatically amphipathic structure at this pH condition
and the fastest diffusion rate for the adsorption onto gas–liquid interface (Barackov
et al. 2012). In addition, the foam stability strongly depended on casein aggregate
size. Casein micelle aggregates prepared at 4
C (average particle size of ~500 nm)
suggested a remarkably stable property compared to that prepared at 20
C without
aggregate formation (average particle size of ~200 nm) or SC (Chen et al. 2017). The
mechanisms based on the interfacial elasticity and thin film properties were eluci-
dated that the peptides, individual caseins, and small micelles could be quickly
adsorbed onto gas–liquid interface to form a primary interfacial layer. Subsequently,
the larger sized casein micelle aggregates were randomly incorporated and attached
as sublayers with a simultaneously reversible adsorption of peptides and individual
caseins pushing out of the interface and therefore form a heterogeneous thin film.
This interfacial layer consisting of patches of peptides, individual caseins, and casein
micelles and attached sublayers consisting of casein micelle aggregates showed a
significantly increased elasticity modulus at a frequency of 0.005 Hz than that
without casein micelle aggregates, which increased the interfacial dilatational mod-
ulus and was demonstrated an improved stability against film rupture (Chen et al.
2017). However, many researchers found that the increase in thin film viscosity was
a more dominant factor prompting foam stability compared to increasing interfacial
elasticity (Georgieva et al. 2009; Wierenga and Gruppen 2010). The thin film
constituted by casein micelle aggregates was more heterogeneous with large aggre-
gates stuck in the lamellae and Plateau borders, which dramatically increased the
thin film viscosity and effectively slowed down or even stopped the liquid drainage
(Chen et al. 2017). Further homogenizing large casein micelle aggregates to form
smaller-sized micelles or individual caseins however could not maintain the film
viscosity, which led to an instability of foam system (Chen et al. 2017). Same
viewpoint suggested that aggregates did not inset into the gas–liquid interface but
might be attached to interface as sublayers and dispersed in the thin film to stabilize
liquid foam (Fameau and Salonen 2014). In recent studies, the complexes consti-
tuted by SC (1.0% w/w), tannin acid (TA, 0.3% w/w), and octenyl succinate starch
(OSA-starch, 1.0% w/w) showed a significantly improved foam stability compared
to that prepared by SC only at an equal concentration and pH 6.0. This investigation
228 Y. Guan

suggested that although the polyphenols (such as TA) decreased the surface pressure
leading to an inhibition of foam ability, the interfacial elasticity was significantly
increased, generating a higher dilatational modulus of gas–liquid interface especially
at a higher polyphenol concentration, which improved the foam stability. Further-
more, the addition of octenyl succinate starch increased the strain softening in
expansion and the strain hardening in compression, which generated a possible
segregated interfacial structure of SC/TA patches and OSA-starch patches
(Fig. 6.12h) with a resistant capability of mechanical compression and extension
(Zhan et al. 2019, 2018).

Gelatin

Gelatin is a combination of denatured fibrous proteins mainly produced from porcine

and bovine skin and bone and is composed of heterogeneous protein/polypeptide
mixture of α-chain (single chain), β-chain (two covalently cross-linked α-chains),
and γ-chain (three covalently cross-linked α-chains) (Benjakul and
Kittiphattanabawon 2019; Ramos et al. 2016). Gelatin can be classified into type
A and type B gelatins prepared through acid and alkaline hydrolysis of collagen,
respectively (Benjakul and Kittiphattanabawon 2019). The alkaline hydrolysis of
collagen generally transforms glutamine and asparagine to glutamic acid and
aspartic acid and therefore results in higher contents of aspartic acid and glutamic
acid proportions in type B gelatin than those in type A (Duconseille et al. 2015). The
most abundant sources of gelatin are pig skin (46%), bovine hides (29.4%), pig and
cattle bones (23.1%), and fish (1.5%) (Duconseille et al. 2015), having molecular
weight ranging from 15 to 400 kDa (Benjakul and Kittiphattanabawon 2019). The
difference in amino acid compositions, molecular weight distributions, and sources
significantly affects the foaming property of gelatin. Amino acid compositions of
gelatin primarily depended on the sources of animal species and processing means.
Generally, gelatin has a number of glycine (33%), proline (12%), alanine
(11%), and hydroxyproline (10%) residues but contains limited amino acid resi-
dues of histidine, methionine, and tyrosine (Karayannakidis and Zotos 2016).
Among them, the proline and hydroxyproline residues, denoted as imino acid
residues, play a critical role in rheological properties such as foaming property
because of their hydrophobic structures. Gelatins with higher imino acid residue
content are mainly from the sources of tropical animals with 194–225 imino acid
residues per 1000 amino acid residues compared to those from temperate animals
with 150–173 imino acid residues per 1000 residues (Benjakul and
Kittiphattanabawon 2019; Abedinia et al. 2017; Kittiphattanabawon et al. 2016).
Furthermore, the molecular weight of gelatin primarily depends on the temperature
of collagen hydrolysis. Higher hydrolysis temperature facilitates rupture of intra- and
intermolecular bonds of collagen protein chains, resulting in a more adequate
degradation of collagen and generating lower molecular weight gelatin (Benjakul
and Kittiphattanabawon 2019; Gomez-Guillen et al. 2002; Alfaro et al. 2015). Thus,
it can be seen that gelatin obtained at a higher temperature with shorter chain length
6 Liquid Foaming Properties 229

and/or α-component (α-chain) is more likely to be employed for improving foam

ability, while gelatin prepared at a milder process with longer chain length and more
β- and γ-chains is appropriate for improving foam stability.
A conventional application of gelatin is development of marshmallow, a typical
foam colloid, because of its acceptable viscosity, hardness, bloom strength, and
foaming stability (Tan and Lim 2008). Broad application of gelatin as foaming agent
was introduced in recent literature (Poursamar et al. 2015; Huang et al. 2017;
Phawaphuthanon et al. 2019). Poursamar and co-workers applied gelatin as a base
material to crosslink each other via glutaraldehyde and to fabricate porous scaffolds
by a gas foaming method, which is promising to manufacture artificial tissue
engineering scaffolds (Poursamar et al. 2015). However, a lower mechanical
strength of the gelatin-built scaffolds, as a weakness, may be a concern in practical
application. Gelatin–sodium alginate composites at a mass ratio of 5:1 and pH of 3.5
dramatically improved foam stability compared to gelatin only because of the
electrostatic attraction induced intermolecular aggregation between gelatin and
sodium alginate to generate charge-neutralized complexes, although a decreased
foam ability was determined caused by the liquid viscosity (Phawaphuthanon et al.
2019). Huang and co-workers found that the addition of fish gelatin into egg white
protein in subcritical water was capable of improving both foam ability and stability
(Huang et al. 2017). The increased foam ability by the addition of fish gelatin was
caused by a reduction in the gas–liquid interfacial tension, while the improved foam
stability was demonstrated via building an interfacial viscoelastic network at gas–
liquid interface with the increased surface dilational rheological behavior, inhibiting
drainage in lamellae and bubble coalescence. These investigations above proved that
the addition of gelatin in foaming agents is potential to improve foam properties
especially stability.
Based on the discussions above, we can speculate that the ovomucin aggregates,
soy protein fibrils, whey protein long fibrils, and casein micelle aggregates with
larger sizes are very suitable to improve foam stability. The protein hydrolysates or
individual protein with lower molecular weights such as ovomucin hydrolysates at a
hydrolysis degree of 15–40%, soy protein hydrolysates at hydrolysis degree of 0.4%,
and individual casein or casein hydrolysis are in favor of improving foam ability.
Therefore, referring recent literature, proteins at various forms such as aggregates,
fibrils, and hydrolysates are promising foaming agents in food and relevant fields.

3.2.2 Polysaccharides

The irregularly shaped polysaccharides with poly-dispersion in size and morphology

are known as effective stabilizers against foam destabilization. The stabilization
mechanism of these polysaccharides as foaming candidates is same as proteins. That
is, increasing the elasticity of interfacial layers and sublayers and improving the
viscosity and steric hindrance of lamellae and Plateau border. Three primary poly-
saccharides, i.e., cellulose, chitosan, and starch have been demonstrated as excellent
foam agents (Dickinson 2017) and will be discussed in the following parts.
230 Y. Guan

Cellulose

Cellulose is a kind of linear polysaccharide consisting of β-(1,4)-linked

glucopyranose units, which can be extracted from natural biomass such as the
walls of plant cells, and obtained after further isolation as macroscopic fibers,
microfibrillated cellulose, and nanofibrillated cellulose (Tingaut et al. 2012), which
constitutes the largest portion (~50%) of the total biomass in nature (Mathur and
Narang 1990). Cellulose has a semi-crystalline structure with various crystallinity
degrees in nature, caused by its constitution of both crystalline and amorphous
substructures (Tingaut et al. 2012). The amorphous portion of cellulose is capable
of partial degradation via the acid treatment to form smaller-sized microcrystalline
and nanocrystalline celluloses that contain abundant hydroxyl groups at the surface.
These fiber-like native, microfibrillated, nanofibrillated celluloses, and their hydro-
lysates always have higher aspect ratio and liquid holding capacity, which is a
potential to enhance the elasticity of interfacial layers or sublayers and the viscosity
of liquid film and therefore makes them as ideal candidates for foaming agents. The
larger dimensional cellulose fibers, with an average length of 2.3 mm and a
spherical-shaped diameter of 35 to 40μm, from wood pulp were reported to generate
stable foams after mixing with sodium dodecyl sulfate. Fundamental investigation
on the viscoelasticity of film demonstrated that a mass of cellulose fibers was
dispersed in the continuous phase resulting in the increase in thin film viscosity
and the formation of gel-like network in both lamellae and Plateau borders, which
could slow down the drainage. However, scarcely bound cellulose fibers at the
interface were determined, speculating that the diffusion rate of larger sized cellulose
fibers to the interface of gas–liquid interface was enormously slower than that of
sodium dodecyl sulfate when the bubbles were formed during whipping (Al-Qararah
et al. 2013). In addition, foams prepared by the mixture of cellulose fibers and
sodium dodecyl sulfate were found to have a reduced bubble size compared to those
prepared by the sodium dodecyl sulfate only. The magnitude of the size reduction
was greater along with the increase in the cellulose fiber percentage. The decrease in
bubble size along with the addition of cellulose fibers was likely due to the locally
enhanced shear forces because of the increase in the breaking tendency of the larger
bubbles by cellulose fiber inertia. The decrease in bubble size was always observed
in stable region of foam instead of in an unstable one (Al-Qararah et al. 2013). The
addition of smaller dimensional nanofibrillated cellulose fibers in the presence of
octylamine was also reported to improve the foam stability, which was attributed to
the reduction of the surface tension at gas–liquid interface as well as the charge
density of nanofibrillated cellulose fibers and more hydrophobic properties (Cervin
et al. 2013). In this case, smaller nanofibrillated cellulose fibers were able to diffuse
to the interfacial layer accompanied by the octylamine during the foam formation
(Cervin et al. 2013), enhancing the elasticity of interfacial layer and further the foam
stability. Comparing to both researches above, it is not difficult to find that the
factors for foam stabilization utilizing different cellulose fiber types are primarily the
size and surface hydrophobicity. Smaller cellulose nanofibrils (approximately 4 nm
in width and 500–1000 nm in length, mixing with octylamine and filtrated through a
6 Liquid Foaming Properties 231

200 nm filter) show faster diffusion rate and stronger hydrophobic forces with
surfactants than larger cellulose fibers (an average length of 2.3 mm and diameter
of 35 to 40μm), while the larger cellulose fibers cannot be adsorbed at the interface
and more likely to attach to the interface as sublayers or to disperse in thin film of
lamellae and in Plateau border. Similar theory can explain that casein micelle
aggregates with average particle size of ~500 nm form a sublayer attaching to the
gas–liquid interface rather than forming only a simple gas–liquid interface discussed
above in this chapter. Furthermore, hydrophobic-modified celluloses were also
utilized to improve the foam stability (Wege et al. 2008; Jin et al. 2012; Wang
et al. 2017; Hu et al. 2016). Zhong and Velev reported an effective means using
physically modified hydrophobic cellulose derivative, i.e., hypromellose phthalate to
develop super stable liquid foam. The hypromellose phthalate particles were deter-
mined to be more easily adsorbed onto the gas–liquid interface created during shear
blending, resulting in highly stable foams for insisting 1 month when the concen-
tration of hypromellose phthalate in stock solution was ca. 1% w/w and was shown
in Fig. 6.13a with a schematics of the forming mechanism in Fig. 6.13b (Wege et al.
2008). Another study demonstrated that the ethylcellulose particles were capable of
adsorption at the gas surface to stabilize foam based on the Pickering mechanism
observed through a cryo-SEM investigation and shown in Fig. 6.13c (Jin et al.
2012). A recent study found that cellulose nanocrystal particles twined by methyl-
cellulose coils was liable to prepare thermostable foams. The prepared liquid foams
showed the lowest density when the concentrations of cellulose nanocrystal particles
and methylcellulose were 2.5% w/w and 0.5% w/w, respectively, implying the
highest gas loading capacity. In this case, these foams were determined to suffi-
ciently robust against defoaming at temperature of 70
C for 6 hrs, which was likely
due to that the reinforced methylcellulose fibrillar gel by cellulose nanocrystal
particles could increase the gas–liquid interfacial elasticity and film viscosity
(Hu et al. 2016). So far, sufficient evidences based on scientific researches have
demonstrated that the foam stability could be remarkably strengthened ascribing the
enhanced viscosity of thin film by the appropriate large dimension of celluloses with
various types and also the increased elasticity of interfacial layer or sublayer,
although the addition of celluloses decreases the diameter of bubble size (Wege
et al. 2008; Hu et al. 2016). Thus, it follows that celluloses with various structures are
promising to be desired foam stabilizers.

Chitin/Chitosan

The chitin is another linear polysaccharide as a foam stabilizer and mainly obtained
from the shells of crustaceans, insects, and microorganisms, which is the second
most common portion of biomass in nature (Mathur and Narang 1990). The chem-
ical structure of chitin contains β-(1,4)-2-acetamido-2-deoxy-D-glucopyranose
units, which can be transferred to chitosan by the deacetylation using hot concen-
trated alkali, although the commercial chitosan still contains 20–25% of N-acetyl
groups (Mathur and Narang 1990). The chitosan is similar to cellulose in chemical
232 Y. Guan

Fig. 6.13 The foam system prepared by shearing 10% w/v hydrophobic cellulose particles in
1 mol/L acetate buffer at pH 4.2 and the images taken by a camera and an optical microscope (a)
(Wege et al. 2008), the schematics of the forming mechanism of this hydrophobic cellulose particle
foam (b) (Wege et al. 2008), the cryo-SEM image of the ethyl cellulose particle-stabilized foam in a
0.25% w/w xanthan solution at pH 3.0 shown with a cross-sectional view of the bubble surface (c)
(Jin et al. 2012), the typical crossed-polarized optical micrograph of the chitin nanocrystal stabilized
foam at chitin nanocrystal concentration of 1% w/w and pH of 7.0 (d), the microstructure observed
by a SEM with low- and high-resolution microscopic images (e) (Tzoumaki et al. 2015), and the
optical microscope images of foam film prepared by stirring a mixture of 4% w/w sodium dodecyl
sulfate and 0.5% w/w KYPAM-2 (a kind of polyacrylamide) without (f) and with (g) 20% w/w
starch particles in aqueous (Zhang et al. 2015). The images were reproduced from the corresponding
references (Hunter et al. 2008; Jin et al. 2012; Wang et al. 2017; Asghari et al. 2016) with
permission
6 Liquid Foaming Properties 233

construction and contains hydroxyl groups along its backbone with a molecular
weight ranging from 50,000 to 2,000,000 Daltons (Poole 1989). However, because
of the introduction of amine groups in side chains, the variation of the zeta-potential
at different pH is opposite against celluloses, exhibiting a pH-dependent water-
solubility at pH lower than 6 (Lam et al. 2014), causing by protonation with positive
charges at the zeta-layer. The chitosan polymers, however, form uncharged self-
aggregated particles with different types when increasing the pH higher than pKa in
solution, resulting from the deprotonation of the -NH3+ groups (Liu et al. 2012). In
general, chitosan alone represents weak foaming properties such as producing very
lower foaming overrun (Poole 1989). After covalently or non-covalently binding
with other polymers, chitin and chitosan showed enhanced foaming properties via
improving the rheological characteristics of interface and thin film. For instance,
carbon nanotubes-chitosan composites were utilized to develop foams with ordered
lamellar structure and enhanced mechanical strength, which presented excellent
thermal stability (Yan et al. 2016). Crosslinking chitosan with cellulose or hemicel-
lulose as foaming agents is another desirable method that is capable of improving the
foam stability (Guibal et al. 2013; Salam et al. 2011). The composite of cellulose
fibers with chitosan allowed changing the mechanical and microstructural properties
of foams, suggesting a restriction of bubble size reduction during drying, resulting
from a reinforcing structure against the collapse of the chitosan network and a
possible prevention of chitosan chain aggregates (Guibal et al. 2013). Chemically
modified chitosan by covalently linking hemicellulose citrate as foaming agent was
found to form elastic, highly porous, and stable foams, which was likely due to the
increase in viscoelasticity of interfacial layer ascribing the covalent hemicellulose
citrate–chitosan complexes (Salam et al. 2011). The use of chitin nanocrystals, as
colloidal rod-like particles with average dimensions around 240 nm in length and
18 nm in diameter (Tzoumaki et al. 2010), to stabilize aqueous foams has been
reported in a recent study (Tzoumaki et al. 2015). The chitin nanocrystal stabilized
foams without the presence of surfactants were prepared by acid hydrolysis of the
original chitin in hydrochloric acid at temperature of 95
C and pH of 3.0 for 90 min
and a subsequent neutralization to a final pH of 7.0 by using sonication as a high-
energy input technique that can produce better dispersed chitin nanocrystals with a
higher surface coverage on bubbles and shown as a crossed-polarized optical
micrograph in Fig. 6.13d. These chitin nanocrystal stabilized foams presented a
stable volume life time of several days when the concentration of chitin nanocrystals
was 1% w/w, which was much longer than those prepared by proteins or low
molecular weight surfactants with generally stable time of several minutes
(Tzoumaki et al. 2015). As shown in Fig. 6.13e, chitin nanocrystals are uniformly
covered at a whole bubble surface and filled in thin film to build a stable foam
structure. The Pickering mechanism on foam stability by such chitin nanocrystals
here was elaborated as that no existing of larger electrostatic repulsion between the
chitin particles at pH 7.0 resulted in an irreversibly packing of the chitin nanocrystals
at the interface, especially with a greater amount of particles to form a colloidal
armor at a higher nanocrystal concentration and thereby provided additional inter-
facial stability against coalescence and disproportionation (Tzoumaki et al. 2015).
234 Y. Guan

Besides the interfacial stabilization, another primary factor improving the foam
stability by chitin nanocrystals was the network formation in thin film between
bubble interfacial layers. The anisotropic rod-like particles were liable to form
gel-like network because they could more easily entangle, overlap, or form orien-
tated domains than spherical-shaped particles and therefore provided an extra barrier
against drainage or collapse of the foams. This gel-like network could even form
solid-like lamellae along with the increase in the chitin nanocrystal concentration,
which further prevented drainage and collapse for improving foam stabilization
(Tzoumaki et al. 2015).

Starch

Starch, obtained from grains, is the most abundant carbohydrate in the human diet.
Similar to cellulose, starch is a semi-crystalline polymer that can produce nanosized
crystalline or amorphous particles after hydrolysis, regeneration, and mechanical
treatment (BelHaaj et al. 2013; Corre et al. 2010). Acidic and enzymatic hydrolysis
of starch can fabricate starch nanocrystals with the average sizes of 15–40 nm and
500 nm, respectively (Corre et al. 2010; Buléon et al. 1998). While regenerating
starch with subsequent co-crystallization and crosslinking forms crystalline
nanocrystals with average size of 28–51 nm and amorphous nanoparticles of
50–100 nm, respectively (Angellier et al. 2004; Bondeson et al. 2006). Mechanical
treatment of starch such as microfluidizer is a high-energy means that can form
smaller-sized amorphous nanocolloids with average size of only 5–20 nm (Siqueira
et al. 2008). These crystalline and amorphous particles are promising to reinforce
and improve the barrier properties of thin film (Corre et al. 2010) such as in foam
systems based on the Pickering mechanism. For instance, the addition of starch
particles in a mixture of sodium dodecyl sulfate (4% w/w) and KYPAM-2 (a kind of
polyacrylamide) (0.5% w/w) for having an insight into the foam stabilization based
on the film dilational rheological behavior was revealed in recent literature,
suggesting that the dilational viscoelasticity modulus, dilational elasticity modulus,
dilational viscosity modulus, and foam strength were enhanced by the addition of
starch particles. A thicker film could be clearly observed in the presence of starch
particles with concentration of 20% w/w than a thinner film without starch particles
and shown in Fig. 6.13f and g. These particles effectively gathered in Plateau border
and slowed down the drainage leading to the formation of dilatant fluid with an
increased viscosity of lamellae (Zhang et al. 2015). Further in-depth investigation
suggested that the addition of hydrophobic ocentyl succinic anhydride modified rice
starch particles with an average particle size of 150 nm and a contact angle with
water drop of ~90
enhanced foam stability 12 folds without compromising overrun
compared to that by native starch particles. The improvement of foam stability
attributed to the interaction of the ocentyl succinic anhydride modified starch
particles and protein was first through the increase in the interfacial dilatation
elasticity and viscosity, which resulted in a dramatic inhibition of initial drainage
rate. Secondly, the ocentyl succinic anhydride modified starch particles provided a
6 Liquid Foaming Properties 235

robust energy barrier by forming a more concentrated film at gas–liquid interface to

prevent the disproportionation. That is the crowding of modified starch particles at
the interface increased the effective interfacial concentration of protein and thus
improved the foam stability (Asghari et al. 2016). Less hydrophobic and smaller
ocentyl succinic anhydride modified rice starch particles with an average size of
90 nm and a contact angle with water drop of ~65
was also employed to stabilize
foams, although the degree of stability was significantly lower than that utilizing
hydrophobic and bigger ocentyl succinic anhydride modified starch particles
(enhancement of 2 folds) (Asghari et al. 2016). This is caused by the thermody-
namic exclusion of starch particles from the gas–liquid interface because a smaller
particle needs less surface free energy for desorption than a larger particle. The
excluded particles, however, can be dispersed in thin film with a relatively stronger
liquid holding capacity due to smaller contact angle and thus increase foam stability
through retardation of drainage rate. Thermal-treated rice starch particles with a
much larger particle size of 10.8μm and a small contact angle with water drop of
~38
also showed effective synergy with protein to improve foam stability. The
dominant stabilization mechanism was very different compared to that using ocentyl
succinic anhydride modified starch particles, suggesting no enhancement of the
interfacial dilational moduli because of their larger size and more hydrophilic nature.
However, the thermal-treated starch particles trapped by the hydrodynamic pressure
increased the viscosity of thin film, which formed a barrier to drainage and finally
contributed to the overall structural stability of foam (Asghari et al. 2016). Therefore,
it can be seen that these starch particles with different sizes and rheological proper-
ties can synergistically maintain the foam structure with proteins, obeying the
mechanisms of increasing interface viscoelasticity and thin film viscosity.
Other polysaccharides that are also important in foam formulation and stabiliza-
tion include but not limit alginate, carrageenan, and xanthan gum. These polysac-
charides are expected to increase foam stability via improving rheological properties
of the interfacial layers, lamellae, and Plateau border and still need further investi-
gation as a potentially interesting research topic on foaming properties.

3.2.3 Inorganic Particles

The inorganic particles such as carbon particles, silicates, and clays have micro- or
nano-dimensional irregular structures and large surface area so that they can be
employed to stabilize colloidal systems, e.g., emulsions and foams (Lam et al. 2014;
Gonzenbach et al. 2006). Incorporation of inorganic particles into biopolymers to
form composites can improve foam stability. Three fundamental mechanisms elab-
orating the foam stability in detail by the inorganic particles are summarized here
(Murray and Ettelaie 2004; Dickinson 2010). Firstly, the inorganic particles are
dispersed in the thin film, which increases the viscosity and liquid holding capacity
of the continuous phase via forming gel-like network and therefore slows down the
drainage in lamellae. From the perspective of the microstructure of bubbles, the
inorganic particles are also gathered in the Plateau borders, causing the increase of
236 Y. Guan

regional area and curvature radius and a corresponding decrease of the Laplace
pressure. While enhanced surface pressure generated by the enrichment of inorganic
particles in the Plateau border dramatically strengthens the Gibbs–Marangoni effect,
finally improves the foam stability. Secondly, the addition of inorganic particles is
capable of increasing mechanical strength of lamellae and elasticity of interfacial
layer or sublayer. The inorganic particles dispersed in thin film and adsorbed at
interfacial layer or sublayer can resist external thermal and mechanical interference
and therefore prevent the liquid film rupture and further bubble coalescence. At last,
the inorganic particles always have highly ordered structure forming compact gas–
liquid interface layer with a higher density than conventional liquid, and hence, can
reduce the gas molecule diffusion from bubbles and prevent the disproportionation.
In brief, the addition of inorganic particles in foams is a promising strategy to
improve the mechanical features of liquid film, which can be used to prepare super
stable foams.

4 Application in Food Industry

Stable foam system plays a critical role in food structural design, contributing to
preparing ice cream (Pei and Schmidt 2010), beer (Iimure et al. 2012), bread (Glenn
et al. 2001; Shogren et al. 2002), food package (Andersen et al. 1999), foam-mat
drying (Karim and Wai 1999), etc. Ice cream is a multiphase foam-type colloid
matrix generally consisting of air, fat, proteins, polysaccharides, sugars, water, etc.
The development of stable bubbles in ice cream has been suggested as the process
that the emulsion with partially coalesced fat globules is first formed. During the
aging step, the fat crystals are formed with a simultaneous rearrangement of the fat
globule membrane leading to the lowest free energy state. In subsequent whipping
and dynamic freezing, the fat globules with proteins adsorbed onto the air surface,
together with ice crystals are dispersed by the shear forces in continuous phase. In a
whole ice cream system, the air bubbles usually in the range of 20 to 50μm are
partially coated by fat globules with and without fat globule membrane protein and
further coated by whey protein and casein. That is, the fat globules with membrane
protein, whey protein, and casein consist of the air–water interfacial layer. Addi-
tionally, sugars and polysaccharides are primarily dispersed in thin film forming a
freeze-concentrated aqueous solution. Therefore, the ice cream can be seen as a
constitution of discontinuous bubbles, a network of biopolymers (including fat,
proteins, and polysaccharides) surrounding the air bubbles, ice crystals, and a
continuous unfrozen aqueous solution (Goff 1997; Eisner et al. 2005).
Beer is one of the most widely consumed alcoholic drinks and the third most
popular drink after water and tea in the world. An important evaluation standard of
beer trait is its foam quality, which is characterized by stability, quantity, lacing,
whiteness, creaminess, density, viscosity, and strength (Iimure et al. 2012; Bamforth
1985). Among these characteristics, foam stability is the most important, which is
promising to be improved by the addition of food-grade biopolymers such as
6 Liquid Foaming Properties 237

proteins and non-starch polysaccharides (Evans et al. 1999). The high molecular
weight protein Z (35,000 to 40,000 Da) and low molecular weight lipid transfer
protein (5000 to 15,000 Da) obtained from malts are positively correlated with foam
stability (Evans and Sheehan 2002), mainly attributed to the interfacial hydropho-
bicity and viscometric activity. While the β-glucan and arabinoxylan, i.e., non-starch
polysaccharides, play a part role in increasing the viscosity of the continuous phase
(Evans et al. 1999). Additionally, filling beer with nitrogen has been applied to
improve foam stability in beer industry because of the decreased gas (nitrogen)
solubility in liquid and therefore to inhibit the disproportionation.
Bread is prepared from dough consisting of flour and water and is a prominent
staple food in large parts of the world. An important factor affecting the texture and
sensory of bread is embedded bubbles comprising up to 70% of the whole bread
volume. Therefore, controlling the bubble size and number, especially retaining their
stability within the bread baking process, becomes necessary to improve the bread
quality with overall desirable structure and texture. Generally, the bubbles undergo
instability processes primarily including drainage, coalescence, and disproportion-
ation. However, much higher viscoelastic modulus of the dough prevents the floating
of bubbles against gravity, and therefore reduces bread foam drainage. Whilst, dis-
proportionation and coalescence are likely to be the main factors leading to bread
foam instability (Mills et al. 2003; Gan et al. 1995). The starch-gluten matrix
involving in building the structure of foams suggests that the low molecular weight
lipids and surface active gluten proteins are the primary components adsorbed at the
liquid films lining the bubbles. The soluble arabinoxylan in continuous phase
ulteriorly increases the viscosity of thin liquid film and thereby further improves
the stability of foam structure (Mills et al. 2003).
The application of food-grade foams in packaging is a possible alternative in food
industry. Important properties of the food packaging materials include safety, bio-
degradability (Brant et al. 2018), resistance to heat transfer (Ahmadzadeh et al.
2015), low moisture susceptibility (Ahmadzadeh et al. 2015), high mechanical
strength (Ahmadzadeh et al. 2015), and low cost of production (Singh et al. 2008).
The cellulose-modified montmorillonite nanocomposite is one of such materials
with potential in developing food-grade foams (Ahmadzadeh et al. 2015). The
nanofoams prepared by high speed shearing the cellulose matrix incorporated with
exfoliated surface-modified montmorillonite clay platelets suggest excellent
mechanical and barrier properties and thermal insulation performance even though
at a low concentration of exfoliated surface-modified montmorillonite clay platelets
(Ahmadzadeh et al. 2015). This cellulose-modified montmorillonite nanocomposite
constructed foam agent is promising to be used for chilled and cooked chains in food
transportation. Moreover, starch and protein are also two available materials that are
potential in foam development because of their thermoplasticity and biodegradabil-
ity (Mensitieri et al. 2011).
Foam-mat drying is a promising food drying technology to obtain desirable food
structure and inactivate microorganism. Polysaccharides and proteins and their
composites including cellulose (Rajkumar et al. 2007; Hardy and Jideani 2017),
ovalbumin (Rajkumar et al. 2007; Hardy and Jideani 2017; Franco et al. 2015),
238 Y. Guan

maltodextrin (Hardy and Jideani 2017; Febrianto et al. 2012), and gum Arabic
(Hardy and Jideani 2017; Febrianto et al. 2012) are commonly utilized additives
for the foam-mat drying process. During a typical foam-mat drying process, the
liquid foods are whipped into stable foams and then dried in air or heating condi-
tions. At this moment, the foams are required to be stable and retain an open
structure, which is capable of being dried rapidly. In theory, the degree of drying
in the foam-mat drying process is very high resulting from the rapid massive increase
in the gas–liquid interface, which occurs in more than one constant rate periods due
to the periodic bursting of bubble successive layer and therefore exposing new
surface for thermohydrodynamic mass transfer. This foam-mat drying process is
suitable for heat sensitive, viscous, and sticky food materials that is unable to be
dried through common drying means such as spray drying (Hardy and Jideani 2017).

5 Conclusions and Future Prospects

In conclusion, foam formation, microstructure, stabilization mechanisms, common

food foaming agents, and their applications are introduced and discussed in this
chapter. Drainage, coalescence, and disproportionation are the primary mechanisms
destabilizing liquid foams, which can be prevented through three potential strategies
including the enhancement of liquid holding capacity in both lamellae and Plateau
borders, increase of space steric hindrance against the contact between adjacent gas–
liquid interfaces, and improvement of the interface elasticity to resist interfacial
tension. Pickering stabilization employing amphiphilic biopolymers is critical for the
development of stable foams and therefore is worth in-depth study. Improving foam
stability is likely to be achieved at the expense of foaming ability when utilizing a
strategy to increase the interfacial elasticity. How to improve both foam stability and
ability by using a simple and routine operation needs a deep thinking. For instance,
the development of foams based on a viscoelastic variable medium such as syrup
mixed medium having phase transition capacity between liquid and solid. In brief,
although foam as one of the essential elements in modern food systems has been
applied with a long history, there is much work still needs to be done to further
improve their properties especially stability. We expect this chapter may provide
fundamental knowledge for readers to better understand food-grade liquid foams.

Acknowledgements Yongguang Guan sincerely thanks Prof. Qixin Zhong at the University of
Tennessee Knoxville for his kindly guidance in professional knowledge of liquid foams during
post-doctoral training.
6 Liquid Foaming Properties 239

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Chapter 7
Tribological and Sensory Properties

Sandip Panda and Jianshe Chen

Abstract Eating, functionalized by mouth physiology, is performed through a

series of processes which collectively helps in food ingestion, preparing the food
for swallowing, ab-initio digestion, and food sensory perceptions. Sensory proper-
ties of food are typically defined by its texture, flavor, and color. Unlike flavor and
color, characterizing texture perceptions remain a daunting task because of varie-
gated in-mouth breakdown mechanisms of food depending on several influencing
factors. Therefore, it always remains a persisting challenge to correlate instrumental
outputs with texture perception. Over the recent decade, principles of tribology—the
subject of friction, wear, and lubrication––have been recognized in food sensory
research in order to adopt novel instrumental approaches for texture perceptions.
This idea of incorporating tribological principles stems from the availability of
friction that arises while the tongue manipulates food over the palate during oral
processing. Eventually, the terminology such as oral tribology has been introduced,
and the subject is rapidly gaining maturity for food sensory applications especially to
demonstrate some highly specific sensory descriptions and to define a quantifiable
metric for those sensory descriptions. This chapter will revisit the various principles
and applications of tribology in pertinence to texture characteristics of food in
general and edible hydrocolloids in particular while attempting to identify potential
research gaps and future research scopes.

Keywords Food oral processing · Soft tribology · Oral lubrication · Sensory

perception · Saliva

S. Panda · J. Chen (*)

Laboratory of Food Oral Processing, School of Food Science and Biotechnology, Zhejiang
Gongshang University, Hangzhou, Zhejiang, China
e-mail: jschen@zjgsu.edu.cn

© Springer Nature Singapore Pte Ltd. 2021 245

Y. Fang et al. (eds.), Food Hydrocolloids,
https://doi.org/10.1007/978-981-16-0320-4_7
246 S. Panda and J. Chen

1 Introduction

While our grandmothers, mothers, and great chefs work hard to formulate great
recipes of all times to fill our plate with delectable dishes, scientists and researchers
are also relentless to find the mystery behind senses of eating. How do we sense food
while eating? This is a valid question worthy of scientific interrogations. Food oral
processing involves complex and dynamical physico-chemical processes which
occur over a shorter time scale (from few seconds to a few minutes at most) inside
mouth; and our sensory perceptions during the oral processing depend on a number
of factors. The evolution and synchronization of these oral processes and various
influencing factors are critical to the success behind the perception and pleasure of
eating. Food components vary widely in terms of its structure, texture, and chemis-
try. However, our perceptions may vary based on the oral physiology and health,
condition of saliva, and psychophysical factors such as culture, the geographical
location, and of course the availability of food resources, and many hitherto
unknown factors. All these factors lead this subject of food oral processing and
sensory perception towards many folds of complexity. Influence of many of these
factors on food texture perception and mouthfeel are still not well known.
Majority of food sensory research until a decade back was limited to bulk
mechanics and rheological experiments and expert panels’ assessment despite the
realization on the importance of tribology by as early as 1980 (Chen 2009). How-
ever, in recent time, there has been a strong inclination towards understanding and
enabling methods and principles of tribology in food oral processing. Tribology,
being primarily an engineering subject, covers the topics of contact mechanics,
friction, wear, and lubrication studies. In the early stage of developments, the subject
was growing around mechanical, industrial, and orthopedic applications. However,
bringing this subject in to food oral perception research is relatively nascent and
opening a new era in food sensory research especially in the direction to adopt more
of an instrumental approach in food texture characterization. Laguna and Sarkar
(2017) reported sub-quadratic growth in research publication data based on the
search with the key word, “oral tribology,” over the preceding decade up to
May, 2017.
Concerning the thematic limitations, this chapter will primarily focus on appli-
cations of tribological principles in food oral processing emphasizing the case
studies on food hydrocolloids. In general, food hydrocolloids refer to wide spectrum
of edible components; nevertheless, a handful of model hydrocolloids will be
referred here in pertaining to various case studies. In the following sections, begin-
ning with an introduction to engineering tribology, this chapter will briefly introduce
various concepts and methods of tribology being applied in food oral processing
research. Inside oral cavity, tongue and palate constitute a soft tribological system;
so, discussions on soft tribology section will be given little more elaboration in this
context. Following on, various experimental and analytical techniques will be briefly
covered. Few case studies on tribological assessment of food hydrocolloids are also
discussed. Finally, the positive hindsight on the prospects of quantitative framework
7 Tribological and Sensory Properties 247

of food sensory perception based on tribological assessment along with associated

challenges and future research scopes will be discussed before the chapter concludes.

2 Tribology Basics

Tribology is the subject to study the mechanics and chemistry of interfaces between
two surfaces which are moving relative to each other. Beyond the applications in
engineering and design of industrial machinery, tribology in recent years has enabled
us to understand and explain seemingly diverse phenomenon ranging from macro to
cellular scale events such as movement of tectonic plates and glacial ice blocks,
animal locomotion and physiology (Stachowiak 2017), and even in cancer growth
(Pitenis et al. 2017). Therefore, concepts of tribology, at this stage, draw attention
from many disciplines of science and engineering. It is nevertheless important to
discuss some of the founding concepts of this subject as part of the present chapter.
Figure 7.1 depicts a phenomenological schema of various independent and
interdependent events which are likely to occur when two relatively moving surfaces
come in contact to form a sliding interface. In a usual sliding process that involves
two or three bodies in relative motion, the sliding interface experiences a series of
physico-chemical interactions and phenomenological consequences such as friction,
wear, and corrosion.
Phenomenological complexity and multi-physical interactions at the interface
throw enormous challenges to engineers attempting to design and optimize machine
elements such as bearings, gear teeth, piston ring-liner, artificial orthopedic joints,

Mechanical loading Environmental loading: temperature, pressure,

humidity, electro-magnetism, radiation, chemistry

lubrication
friction wear corrosion

adhesion third particle

wear debris/
lubricant retaining crack

Fig. 7.1 Schema of contact phenomenology of two interacting surfaces

248 S. Panda and J. Chen

Fig. 7.2 Contact between

two spherical bodies (the
deformation, δ, shown here
is the cumulative
deformation of the surface/
point of contact and the
down ward direction is
assumed for representation)

and more. In the context of food oral processing, tribology of soft oral surfaces,
saliva, and food ingredients in pertinence to assess food sensory properties has
offered novel set of complex problems in the field and has gathered inter-disciplinary
experts to collaborate in this new knowledge development process.
Contact mechanics, macroscopically, in between the nominal boundaries of the
physical dimensions of the contacting surfaces, or, microscopically, in between two
individual asperities often dominantly impact on the magnitude of friction. Based on
classical continuum mechanics, Hertz has derived the first predictive theory for the
force-displacement relationship between two spherical bodies building up a circular
contact under an ideal hemi-spherical pressure distribution (Timoshenko and
Goodier 1951).
Based on the Hertz theory, the mathematical relationships such as contact load-
deformation and contact area-deformation can be derived in consistent with Fig. 7.2,
where W is the load, a is the radius of circular contact area, and δ is the deformation
and the units of the quantities are as per SI system.

Contact load : W ¼ KR1=2 δ3=2 ð1aÞ

Contact radius : a ¼ ðRδÞ1=2 ð1bÞ

Maximum pressure at the geometric center of contact area,

pmax ¼ 1:5W=πa2 ð1cÞ

 2 
1
P 
where K ¼ Hertzian modulus, K ¼ ð4=3Þ 1  v2i =E i and R ¼
2 1 i¼1
P
1=2Ri , (ν ¼ Poisson’s ratio)
i¼1
7 Tribological and Sensory Properties 249

Nominal boundary of
contact, Anominal = π a 2

Actual contact spots, Ai

Areal = ∑ Ai
Areal < Anominal

Fig. 7.3 Contact area hypothesis: circular boundary is representing the nominal contact area and
black dots are representing actual contact spots over asperities introduced by randomness, hierar-
chy, and scale of surface roughness

However, the Hertz theory encountered limitations in some practical instances.

For example, contact between highly smooth and clean elastic solids under small
external load unlikely to follow Hertz theory. In such cases, surface energy associ-
ated with the contacting surfaces actively influences the local contact condition, and
hence the concept of adhesive contact theory was developed at later stage (Johnson
et al. 1971; Fuller and Tabor 1975). Furthermore, every solid surface has small scale
geometric features which are called asperities. Distribution of these asperities of
varying shapes and sizes over the surface space forms the random and hierarchical
micro-geometric structure on the surface popularly known as surface roughness
(Panda et al. 2017). The shape, size, and distribution of asperities are naturally built
for biological surfaces and inherited through the controlled production processes for
engineering surfaces. Surface roughness is albeit another important consideration in
tribology studies. This inherently introduces the randomness, hierarchy, and the
scales at which the surfaces come in to contact. Some of these effects in asperity
interactions are largely off-limits to observations. In short, the surface roughness
introduces the difference between the nominal contact and the real contact area,
where nominal contact area is defined by macro-geometric boundary and the real
contact area is the summed-up area of all tiny contact spots (Fig. 7.3).
Surface roughness or asperities have been found to have much greater impact on
the contact condition and the resulting friction and wear (Greenwood and
Williamson 1966; Whitehouse and Archard 1970). It is eventually understood that
the contact pressure experienced at the tiny contact spots of individual asperities is
much higher than the nominal pressure over macro-contact area since Areal < Anominal.
Pressure over asperities often exceeds the strength of materials at interfacial junc-
tions and results in microscopic material failures known as wear.
250 S. Panda and J. Chen

Fig. 7.4 Dry and lubricated contact scenarios

Wear of materials at the sliding/rolling interfaces resulting from the micro-

mechanical failures, adhesion, and/or chemical actions are almost inevitable. A
preventive action is therefore crucially important to avert excessive wear as well
as to alleviate friction at the interface. Thereby, the idea of enabling a sustainable and
protective film at the tribo-interface has been developed. Functions of such films are
usually engineered to prevent the asperities and surfaces from coming to direct
contact during sliding operation, and thereby preventing wear and reducing friction.
The mechanism of interfacial film formation and its functions are commonly termed
as lubrication. The subject of lubrication in industrial context has been well devel-
oped and optimized over the entire latter half of the previous century (Stachowiak
2017). However, in the last few decades, the knowledge of lubrication has been
extended to several fields such as bio-medical, personal and beauty care product
development, food oral processing, and in many other fields. In all these contexts,
the use of a lubricating media such as a fluid is vital. In natural systems, the lubricant
is naturally present such as synovial fluid in orthopedic joints and saliva in tongue–
palate system; whereas in engineering systems, the lubricant is synthetically devel-
oped and applied depending on applications. In food oral processing context, saliva
and some food compounds such as fat act as lubricating media. It is important to
reiterate here that the usual understanding of lubrication is to enable easy in sliding
by reducing friction. Figure 7.4 schematically demonstrates the difference between
dry and lubricated contacts along with the description of Amonton–Da Vinci’s laws
of friction (Hutchings 2016).
Engineering insights of lubrication is usually manifested by the Stribeck frame
work. This frame work describes the variation of coefficient of friction with respect
to the product of sliding velocity, lubricants’ viscosity, and inverse of normal load.
Figure 7.5 shows a typical schematic of the Stribeck curve. This is often referred to
distinguish between different regimes of lubrications termed as boundary, mixed,
elasto-hydrodynamic, and purely hydrodynamic lubrication. For simplicity, the
product representing the abscissa of Stribeck curve is termed as bearing parameter.
7 Tribological and Sensory Properties 251

Fig. 7.5 Schematic of the Stribeck framework (η ¼ dynamic viscosity, U ¼ sliding velocity;
W ¼ normal load)

From this graph, it can be apparently understood that with the increase in the
bearing parameter, ηU/W, the coefficient of friction drops and reaches to minima
before starts rising slowly again. This gradual rise in COF occurs in full film
condition at high speed and attributes to the fluid viscous friction and turbulence.
Understanding and utilization of Stribeck curve has been increasingly important
in oral lubrication context. Firstly, because it is hard to generalize the governing
regime of lubrication concerning one typical kind of food–saliva mixture, so
presenting the friction response over a range of parametric inputs might give better
clarification on friction–sensory relationship. Secondly, it is also naive to claim an
absolute value of friction coefficient for a system or material; so, it is more invig-
orating to produce a map of friction coefficient as a function of parametric inputs. In
some attempts to simplify, friction coefficient is often shown as function of speed
instead of the product, ηU/W, in the Stribeck curve. Nevertheless, use of only sliding
speed in the abscissa of Stribeck curve is actually a compromise since the precise
load variation in between the oral surfaces and the real viscosity of the
non-Newtonian food–saliva mixture is yet to be known.
Applications of tribology for sensory studies are rapidly emerging. While the
model of tongue–food/saliva–palate sliding system can be easily recognized, the
challenges persist in establishing an appropriate tribological set-up to replicate this
tribo-system. This is critically important to note here that the performance of
lubrication is collectively dependent on the whole system and surroundings. There-
fore, lubrication and/or friction are not intrinsic properties of any specific material
such as food articles in the present context. Some of the most influencing parameters
are load, speed, lubricant’s viscosity, temperature, interfacial chemistry, surface
252 S. Panda and J. Chen

roughness, and materials’ properties. Moreover, oral surface materials are soft
biological tissues which exhibit typical visco-elastic behavior. Therefore, mechanics
and tribology of soft materials ought to be understood in some details.

3 Soft Tribology in Oral Processing

Soft tribology basically deals with the studies on governing principles behind
tribological performances of soft materials such as elastomers, biological tissues,
and bio-polymers (Pitenis et al. 2017). From an engineering point of view, solid or
semi-solid material systems below elastic moduli of 100 MPa are usually considered
as soft materials; however, in biological structures, materials below 100 kPa are
commonly found and possess an even ultra-soft characteristic. The low elastic
modulus governs the load-deformation behavior of these material systems, the
system encounters large deformations even under extremely low load, which
makes these systems vulnerable to inaccurate measurements and linear theories of
mechanics often become inadequate to describe some of these behaviors. It is likely
that soft material behaviors are somewhere in between the solid and fluid constitutive
characteristics, so a combined constitutive behavior termed as visco-elasticity needs
to be properly evaluated for the soft material systems. From the tribological per-
spectives of soft materials, large deformation leads to a larger area of contact under
small load which drastically reduces the contact pressure, and thereby, exhibits a
unique frictional response which is different from most engineering materials such as
metals, alloys, and hard polymers.
Any tribological pair can be categorized as hard–hard, hard–soft, and soft–soft
systems based on the contacting materials’ constitutive behaviors in relative to each
other. Measurements and theories have been optimized over the years to bring in our
present day understanding on the behaviors of the engineering systems to deal with
hard–hard and, to some extent, hard–soft contacts. These developments for conven-
tional systems are nevertheless limited to capture the behavior of soft–soft systems;
where each material has non-linear, time dependent, visco-elastic characteristics
which often limit the use of linear theories of mechanics. Both in nature and
engineering applications, numerous examples of soft–soft systems can be found.
Understanding and capturing the behavior of soft–soft tribo-systems therefore have
burgeoning research scopes to bring in novel applications and to optimize the
existing applications for societal needs. In particulate to oral systems, both tongue
and palate are made up of biological tissues and constitutively soft on their surfaces
as well as bulk. Noteworthy, the palate is comparatively harder than the tongue.
Thereby, tongue–palate system is an excellent example of soft tribology applications
in nature which is crucial to food oral processing and sensory perceptions. Figure 7.6
shows a schematic depiction of tongue–palate system.
In the process of eating, at certain stage the tongue manipulates food by sweeping
it on the surface of the upper palate. At this stage the friction that arises at the
interfaces between food and tongue contributes to certain amount of sensory
7 Tribological and Sensory Properties 253

Fig. 7.6 Schematic of tongue–palate tribo-system

perceptions such as smoothness, creaminess, and slipperiness (Kokini et al. 1977;

Kokini 1987).
Saliva keeps the oral surfaces protected from bacterial colonization and irritated
rubbing. One can perform simple voluntary experiments to check the saliva starved
situation on oral surfaces: for example, dry up the tongue and palate by wiping the
surfaces with a piece of cotton and then allow tongue to slide on the palate; the
irritation can be easily felt. This kind of oral irritation results from the rise in friction
by the absence of saliva. In tongue-palate systems, presently it has been well
recognized that salivary film works as a lubricant. Friction coefficients of mechan-
ically stimulated saliva roughly fall in between 0.02 and 0.45 when tested in a PDMS
(poly-di-methyl-siloxane) ball-on-disc tribo-system; and interestingly, friction coef-
ficient of saliva remained always lower while compared to fresh water under various
tribological testing conditions (Bongaerts et al. 2007b). Typically, salvia consists of
nearly 99% water and around 1% of other components which include mostly pro-
teins, enzymes, and some inorganic elements. Therefore, the low friction coefficient
of saliva as compared to water may be attributed to certain other major components
such as mucin proteins. Overall, the viscosity, the coating ability, and the lubricating
behavior of saliva are governed by the intertwining actions of various mucins. More
detailed information about saliva and the functions of mucin can be drawn from a
recent review on age-saliva relationships (Xu et al. 2019), a special issue on food–
saliva interactions (Mosca et al. 2019), and a model demonstrating the anchoring of
MUC5B mucin on the oral epithelial cells (Ployon et al. 2016).
In pertinence to the lubricating characteristics of saliva, tribological experimen-
tation can be an important instrumental approach for assessing typical sensory
attributes such as astringency. Astringency is thought out to be due to high friction
out of saliva starved situation or saliva breakdown during oral processing of a variety
of foods and beverages such as fruits, tea, and wine (Upadhyay et al. 2016; Laguna
and Sarkar 2017).
It has been practically important to analyze the soft tongue-palate tribo-system in
presence of salivary fluid and food article in order to establish friction-sensory
relationships. This is a complex natural system; nevertheless, the theory of soft
254 S. Panda and J. Chen

elasto-hydrodynamic (soft-EHL) lubrication received much appreciation in this

context (de Vicente et al. 2006; Bongaerts et al. 2007a). In usual engineering
lubrication studies, Stribeck curve accommodates elasto-hydrodynamic lubrication
regime as a threshold frictional response before the full film hydrodynamic lubrica-
tion. This regime is special because of its dependency on two apparently important
factors: (1) the elastic response of contacting materials; and (2) lubricant’s piezo-
viscous characteristics. Several empirical correlations manifest dramatic increase in
the viscosity of lubricants with respect to pressure (Sargent 1983); and this phenom-
enon is known as piezo-viscous. Thereby, under piezo-viscous situation, an increase
in contact pressure results in thickening of the lubricating fluid.
In much opposed to the hypothesis of classical EHL theory, in soft contacts––
where elastic moduli are in the order of few MPa or few kPa––an increase in contact
load is easily accommodated by more deformation of the soft materials. More
contact deformation in turn alleviates contact pressure. This can be simply checked
by deploying Hertz contact equations (Eqs. 1a, b, and c) for equal W and R, and
varying K for a hard–hard, hard–soft, and soft–soft contacts. The dramatic reduction
in contact pressure for soft–soft contacts results in trivial piezo-viscous influences.
Also, more deformation at the contact allows lubricant to easily spread out and might
result in further alleviation of the piezo-viscous impact. Recently, Masjedi and
Khonsari (2017) estimated trivial differences (<0.5% error for central film thick-
ness) between piezo-viscous and iso-viscous solutions for mixed-EHL contacts of
soft materials having elastic moduli of 100 MPa. It is therefore fair to consider an
iso-viscous condition for the soft-EHL contacts. Overall, the visco-elastic behavior
of soft materials and lubricating characteristic of salivary fluid jointly define the
frictional response of tongue–food/saliva–palate tribo-system.
In the theory of lubrication, Reynolds’ equation governs the flow and pressure
development in the mixed, EHL, and hydrodynamic regimes. The equation is
fundamentally a reduced form of the well-known Navier-Stokes’ equation which
governs the fluid mechanics. A detailed discussion on the derivation and solution of
the Reynolds’ equation is beyond the scope of the present chapter. However, it is
important to include some contextual solutions of the Reynolds’ equation: de
Vicente et al. (2006) solved the Reynolds’ equation for soft-EHL problems
concerning food colloids (e.g. xanthan gum, guar gum, etc.) as lubricating media
and estimated an empirical formulation of friction coefficient in EHL regime as
given below:

0:75ðSRRÞðηU Þ0:34
μEHL ¼ ð2Þ
R0:09 W 0:12 K 0:22

In the above expression, SRR is the slide to roll ratio. SRR can be defined for any
given tribo-pair mechanisms (e.g. gear teeth, ball/roller bearings, ball-on-disc, etc.).
Mathematically, it is the ratio of relative sliding velocity to the mean sliding velocity
at the center of contact. For instance, in case of a ball-on-disc tribometer, if
Uball 6¼ Udisc, then:
7 Tribological and Sensory Properties 255

Table 7.1 Tribological case studies on food hydrocolloids

Coefficients and indices
to fit “master” Stribeck
Reference Experimental details curve
(Bongaerts • Ball-on-disc (PDMS-PDMS) h ¼ 4.75; k ¼ 0.11;
et al. 2007a) • Ball dia. ¼ 19 mm l ¼ 0.07; m ¼ 2.7;
• Composite RMS roughness ~ 27.4 nm n ¼ 0.5; B ¼ 3.8e-5
• SRR ¼ 0.5; W ¼ 1.3 N; U ¼ 1–2400 mm/s (107 < ηU < 2)
• Samples: Water; Corn syrup (95%)
(Krop et al. • Ball-on-disc (PDMS-PDMS) h ¼ 11; k ¼ 0.0065;
2019) • Ball dia. ¼ 19 mm l ¼ 0.075; m ¼ 1;
• Composite cla roughness ~50 nm n ¼ 0.55;
• SRR ¼ 0.5; W ¼ 2 N; U ¼ 1–1000 mm/s B ¼ 3.3e-5
• Samples: hydrogels (κCarrageenan; κC + locust (106 < ηU < 10)
bean gum; κC+ calcium/sodium alginate)
RMS Root mean square, CLA Center line average, SRR Slide to roll ratio

SRR ¼ jU ball  U disc j=U, where U ¼ ðU ball þ U disc Þ=2 ð3Þ

Physically, the value of SRR determines whether the contact is sliding or rolling
motion dominated. SRR can be 0 for a pure rolling condition and 2 for a pure sliding
condition. Moreover, a value of SRR below 1 means that the contact is mostly rolling
and above 1 means it is mostly sliding. Notably, SRR of the tongue–palate system is
hitherto unknown; nevertheless, a value of 0.5 is usually taken in oral tribology
experiments. This is albeit counter intuitive. In consistent with the expression of
SRR, if either disc or ball is static, then SRR ¼ 2. This means, if one element in the
tribo-pair is fixed or quasi-static, then the contact predominantly slides. In tongue-
palate system, the palate is almost quasi-static with respect to the tongue. Therefore,
an SRR of more than 1 seems more appropriate choice for oral tribology
experiments.
Bongaerts et al. (2007a) attempted to fit a “master” Stribeck curve by covering
entire regimes of lubrication and proposed an empirical expression of friction
coefficient assuming power law characteristics:
 
νBoundary  μEHL 
μ ¼ μEHL þ where, μEHL
1 þ ðηU=BÞm
¼ kðηU Þn and μboundary ¼ hðηU Þl ð4Þ

Further, the coefficients h, k, and indices l, m, and n can be estimated by fitting

experimental data with the above equation. Moreover, the value of B is the upper
limit of ηU for boundary lubrication regime for any given case. It is important to note
here that not all but many experimental data may be fitted with the above equation to
plot a “master” Stribeck curve. Table 7.1 shows data from two cases on tribological
testing of food hydrocolloids, where the above equation has been used to obtain the
“master” Stribeck curve.
256 S. Panda and J. Chen

Significant differences in these two experimental cases are in food samples, tribo–
pair roughness, load, and range of speeds. Substantial changes in the fitting param-
eters are in the values of h and k; notably, h and k are power law coefficients in
boundary and EHL lubrication regimes (Eq. 4), respectively. In logarithmic scales,
these coefficients determine the intercept on friction coefficient axis. Physically, an
increase in h means the boundary friction value rises towards the lower limit of ηU,
and a drop in the value of k means, the limiting friction for starting-up EHL regime is
reduced. This means, in the case of (Krop et al. 2019), the boundary and mixed
regime are elongated as compared to the EHL regime. Therefore, two different
studies on different hydrocolloid samples produced two different Stribeck curves.
This is a caveat; and the idea of producing generic “master” Stribeck curve for
hydrocolloid samples need more data for further optimizations. In fact, some impor-
tant effects such as surface roughness, hydrophobicity, and presence of surface
active elements are hitherto not included. These effects significantly influence the
performance of biological surfaces such as the tongue.
Tongue surface is biologically textured with two main types of papillae (Sarkar
et al. 2019) covering nearly 70% of the frontal surface area: (1) filly form, without
any taste buds and with hair like appearance on top; (2) fungi form, containing taste
buds, and has mushroom like appearance. The filly form hairs high around 250 μm
are most protruding and taking part in active sliding friction while tongue swipes
over the palate. These altogether constitute an intricate micro-geometric structure on
the surface of tongue. Saliva introduces further complexity. Mucin in saliva forms a
salivary pellicle of thickness up to 100 nm by getting adsorbed on the base surface of
tongue, and this salivary pellicle holds the fluidic structure of the saliva. The salivary
pellicle thickness varies and at some point may be nearly vanishing during oral
processing. This leads to a saliva starved situation. Sarkar et al. (2019) postulated
three types of adsorbed film formation: (1) saliva-rich/deficient film; (2) saliva–food
mixture dominated film; and (3) food dominated film. One or more of these
adsorption films implicate the food–saliva chemistry which in turn impact on the
friction and mechano-sensation during oral processing and generates a series of
sensory perceptions such as astringency, creaminess, smoothness, etc.
Furthermore, the soft and protruded papillae textures constitute a spongy structure
on the tongue surface. In the presence of salivary papillae and other surface active
agents, the microscopic spongy maze on the tongue surface may store certain
amount of salivary fluid and mechanically squeeze it out under pressure. This
mechanism may possibly develop a salivary fluid film whenever the tongue applies
pressure on food/palate. This may resemble the tongue surface structure as
poroelastic material system. Poroelasticity is usually exhibited by a bi-phasic mate-
rial system, where a spongy solid structure retains a fluid; and the load-deformation
behavior is governed by the solid-fluid interaction. Mammalian cartilage is a striking
example of poroelastic structure made up of collagen, water, and synovial fluid
(Neville et al. 2007). In fact, tongue surface as a poroelastic structure is still a
conjecture; and it is clearly naive at present to accept mechanistic behavior of tongue
as closely similar to that of cartilage. Future research on the mechanistic aspects of
tongue–food–palate contact is likely to bring in more insights in these aspects.
7 Tribological and Sensory Properties 257

4 Experimental Techniques in Oral Tribology

Characterization

In pertaining to tribology–sensory studies, an appropriate in vitro experimental

methodology is vital in order to ascertain the situations inside oral cavity as closely
as possible. Rheology and bulk mechanical experiments have dominated the food
oral processing and sensory relationships over many years. The seminal work of
Kokini et al. (1977) has produced ab-initio empirical models to establish the role of
*friction* in addition to *flow* to provide texture perceptions such as smoothness,
slipperiness, and creaminess:

creaminess / ðthicknessÞ0:54  ðsmoothnessÞ0:84 , where smoothness

/ 1=friction

It can be naively understood from the above correlation that certain texture
perceptions depend more on the tribological behavior of the food articles during
oral processing.
After a decade, Hutchings and Lillford (1988) have drawn philosophical perspec-
tives on how eating and sensation, both of which are dynamical in nature, could best
be correlated with the instrumental methods. The observations hypothesized a three-
dimensional “mouth process model” to demonstrate the criterion of swallowing of
food following a “breakdown path” which is unique to food, individual eater, and
eating occasions. In their model, one typical criterion plane defines the “degree of
lubrication,” where the other two planes are on the basis of “degree of structure,”
and “time.” The model emphasized the importance of “degree of lubrication” and
appended difficulties to define it. According to this model, the normal trajectory of
food breakdown with respect to time in most cases follows a downward direction in
the degree of structure and in the increasing direction in the degree of lubrication;
nevertheless, in some exceptional cases such as for peanut butter or sesame paste, the
trajectory may move opposite to the normal path at the initial stage of oral process by
quickly absorbing saliva before taking the normal direction (i.e. decreasing the
degree of structure and increasing the degree of lubrication) (Nishinari et al. 2019).
Overall, it is understood that a single instrumental approach can be very inade-
quate to bring in comprehensive correlation between instrumental findings and
sensory perceptions, and possibly, a combination of instrumental methods ought to
be designed. Surprisingly, this model did not correspond to the earlier findings of
Kokini et al. (1977) in regard to the in-mouth lubrication and friction–sensory
relationships. Further, despite its comprehensiveness, the Hutchings and
Lillford model remained almost unrecognizable until the end of previous decade
due mainly to dearth of sophisticated instrumental techniques (Chen 2009).
The importance of in-mouth lubrication during food oral processing for both
swallowing and sensory perception has been eventually realized. These develop-
ments led towards a paradigm shift in the food sensory research which is turning
towards the regimes of tribology and rheology instead of mere rheology and bulk
258 S. Panda and J. Chen

mechanics. Chen and Stokes (2012) have illustrated the changes in the governing
mechanisms of eating as a function of oral processing time. They highlighted the
change in length scale of food articles which undergo changes from centimeter
during ingestion to micron/sub-micron scale during swallowing. This change in
length scale is governed initially by mechanical breaking, fracture, and bulk defor-
mation and gradually by saliva mixing, moistening, and shearing. Overall, it was
understood that eating or food oral processing is a complex dynamical process and
so is the sensory perception. Therefore, the sensory attributes also evolve, which
means the mouthfeel at an early stage of oral processing may be different than at later
stage for the same food component, where the early stage mouthfeel depends largely
on bulk mechanical properties and rheology and the later stage, feelings are more
linked to thin film shearing resulting in friction and lubrication.
Tribological experiments on food articles have since been recognized as much
important as rheology measurements and quality descriptive sensory statistics. Also,
in the intermediate stage of oral processing, rheology and tribology jointly contrib-
utes to mouthfeel factors in an implicit manner. This framework is particularly useful
to classify the growing vocabulary of sensory descriptions based on driving mech-
anisms of oral processing: mechanics; rheology; tribology; and rheo-tribology.
These typical mechanisms can be adopted in instrumental techniques, and further
the instrumental outputs can be linked to typical sensory descriptions. Overall, three
mechanical instruments, namely, texture analyzer, rheometer, and tribometer, have
been adopted and being continuously optimized for analyzing foods and colloids. A
comprehensive assessment of food articles for sensory attributes can be largely
possible by one or more of these instruments. The instrumental outputs can eventu-
ally be calibrated to a metric for instrumental sensory descriptions. Table 7.2 is
furnished with some details about these experimental techniques and attached
sensory descriptions.
It must be noted here that tribology measurements of food articles depend heavily
on the systems and surroundings and thereby tribological parameters (friction/
lubrication) are not intrinsic properties of food compounds. With these caveats, it
is vital to have better understanding of the systems being used for in vitro tribolog-
ical assessment which include: the instrument, model tribological pairs, model food
items, application of saliva, and system operating variables and surrounding
environments.
In early stages of oral tribology studies, varieties of tribo-contact configurations
and contacting materials were tested. Pradal and Stokes (2016) have reviewed
different types of tribo-configurations. Due to available variations on the choice of
instruments, material pairs, and model food systems, it is hard to argue over,
advantage of one specific system over others. However, there should be clear
understanding of the system and surroundings being used. Amongst all, a specialized
commercial tribometer, namely, mini traction machine (MTM) has been most
frequently used and eventually popularized. The machine is a modified ball-on-
disc system; where a combined sliding-rolling motion of the contact is given by
rotating both ball and disc and maintaining a constant slide to roll ratio. A schematic
of this system can be seen in the last row of Table 7.2, the geometric figure is a
7 Tribological and Sensory Properties 259

Table 7.2 Instrumental methods in food oral processing in correspondence to sensory studies
Representative test Textural
Test type Geometric configuration outputs attributes
Texture property Hardness,
analysis (Mechanical Springiness,

Force
compression) Crispness

Displacement
(To characterize bulk
mechanical strengths)
Rheology Viscosity (=tangent of the curve) Thickness,
(Flow, squeezing, and Pasty,

Shear stress
bulk shearing) Smoothness,
Slipperiness,
Creaminess
Shear strain rate
(To characterize constitu-
α tive behavior and
viscosity)
Tribology Astringency,
Coefficient of friction

(Thin film shearing, Smoothness,

sliding, and rolling) Slipperiness,
Creaminess

viscosity*speed/load

(To characterize friction at

different lubricating
regimes)
Note: Textural attributes as noted in the last column are not the direct outcome of the test outputs
obtained from any of the tests. In fact, the relationships between test results and sensory attributes are
often complex and depend on additional parameters. For example, to use the force-displacement
curve as an assessment of hardness, one needs to know the size and shape of the sample

representative tribometer; however, in the actual MTM machine the ball holder is
usually tilted with respect to the disc plane in order to avoid the spinning of the ball
with respect to holder axis.
Recent advances in instrumentation and material science have greatly augmented
the applications of tribological experiments in food sensory research. Currently,
PDMS rubber (E~ 1–100 MPa) is the most popular material being used as model
tribo-pairs. Soft and visco-elastic behavior, tunable mechanical properties, and
excellent formability, which enables advanced manufacturing technology such as
3D printing to process PDMS in desired shapes, sizes and properties, are the reasons
behind the popularity of PDMS. However, properties of PDMS are still more than
ten times higher than the maximum pressure experienced in oral conditions (Sarkar
et al. 2019). This means the effect of contact deformation on lubrication in PDMS–
PDMS contact cannot be extrapolated to draw the similar effects in biological
contacts. In fact, the challenge persists to address two main aspects here: first, the
260 S. Panda and J. Chen

elastic response of the model components which should be equivalent to biological

components; and second, the model surface microstructure and chemistry should
mimic the biological surfaces.
Other important aspects of oral tribological experiments are proper and adequate
application of saliva to model food samples and to the system. In fact, it is
challenging to address the complex salivating process in in vitro experiments.
Currently, either simulated saliva or artificial saliva is being used to apply on the
model surfaces and samples before or during experiments. Presence of saliva on
tribo-surfaces is critical to the frictional response of the system; and maintaining the
situation in a tribometer set-up needs challenging arrangement such as submerging
the PDMS ball/disc in to saliva and/or establishing a supply system to keep applying
saliva while the tribometer is running.

5 Case Studies on Tribological Evaluation of Food

Hydrocolloids

In particulate to food hydrocolloids, Chojnicka-Paszun et al. (2014) examined the

tribology–sensory relationships for model solutions of polysaccharides with protein
particle dispersions. Three selected polysaccharide stocks are locust bean gum,
pectin, and xanthan; spherical protein particles were mainly abstract from whey
protein isolate/locust bean gum gel. Tribological evaluations were correlated with
quantitative descriptive analysis (QDA). An attempt has been made in this chapter to
summarize some of these findings in Table 7.3 below.
Looking in to these case studies on polysaccharides, it appears that tribological
evaluation correlates weekly with the sensory attributes as characterized by QDA
scores. In the absence of particle, lubricating ability of xanthan solution is most
superior followed by pectin and LBG, respectively. This order is likely to change
with the add-on protein particles. Except for pectin, the presence of particles caused
dramatic changes in the lubricating ability of xanthan. In the presence of larger
particle size, friction coefficient of xanthan solution increases. On the other hand, the
change in “powdery” sensation is more prominent in case of xanthan. In fact, LBG
without particle has poorest lubrication and paltry powdery sensation; whereas
xanthan with large protein particles has superior powdery sensation despite
diminishing lubricating ability. These observations are highly counterintuitive,
and, therefore, the “powdery” attribute could not be directly linked to friction.
Thereby, this attribute can possibly be linked to other mechanical characteristics
such as hardness or elasticity of the tiny protein spheres.
It is intriguingly intuitive to link “slippery” and “stickiness” with respect to
lubricating ability of the samples. Nevertheless, “slippery” and “stickiness” loosely
relates to tribological evaluations of the polysaccharide solutions. These weak
correlations are likely to be influenced by other factors, and may vary sample to
sample as well as individual to individual. Overall, the poor reflection of tribological
Table 7.3 Relationships between lubricating ability, protein particle size, and sensory attributes for polysaccharide solutions (data from Chojnicka-Paszun et al.
2014)
Parameters Ordering of the samples
Without any particles With small protein beads of size 15 μm With large protein beads of size 56 μm
7 Tribological and Sensory Properties

Lubricating abilitya Xanthan>Pectin>LBG Pectin>Xanthan>LBG Pectin~LBG>Xanthan

(W ¼ 2N;
U ¼ 5–100 mm/s)
Sensory attributesb Powdery Pectin>Xanthan>LBG Xanthan>LBG>Pectin Pectin>LBG>Xanthan
Slippery Xanthan>LBG>Pectin
Stickiness Pectin>LBG>Xanthan
Filminess
Sliminess
a
The ordering of lubricating ability of samples, xanthan (2%), pectin (2.25%), and LBG (1%) are based on significant differences in the values of friction
coefficient plotted in the Stribeck curve
b
The ordering of the sensory attributes of samples is based on marginal differences in the respective QDA scores
261
262 S. Panda and J. Chen

evaluation on sensory attributes indicates that despite tribology has become a vital
instrumental approach, nevertheless, additional measurements like fracture proper-
ties, hardness, etc. should supplement the tribological evaluation in order to achieve
more conclusive correlations.
Polysaccharides are quite common as thickeners in food articles; thereby,
tribology–sensory relationships for these compounds are of specific interest.
Zinoviadou et al. (2008) studied the role of saliva in tribology, rheology, and
spreadability of cross-linked starch and LBG. The addition of saliva dramatically
reduces the apparent viscosity, moderately increases friction and slightly enhances
spreadability (lower contact angle) for starch samples. Overall, these comparisons
attempted to capture the significance of saliva–polysaccharide interactions in oral
processing and sensory implications. Furthermore, starch microstructure is more like
“spherical” granules, and these shapes can be responsible for low friction; however,
after being exposed to saliva, these granules breakdown. Therefore, sustenance of
low friction for starch–saliva mixture as compared with pure starch sample was
identified as a function of the rate at which the starch granules get affected by saliva
induced digestion. The impact of oral processing time on shape and size breakdown
of food compounds is therefore critical to its tribological properties and subsequent
sensory perceptions. In fact, this study was not conclusive on sensory properties
linked to the findings.
In another interesting study, Nguyen et al. (2017) made an appreciable attempt to
mix some of the hydrocolloids (gelatin, xanthan, Carrageenan, and modified starch)
with low fat skimmed yogurt (<0.1% fat) in order to arrive at a full fat yogurt
experience. The study employs texture, rheology, tribology, and QDA assessment
with the selected samples and total of eight different sensory attributes: thickness,
smoothness, creaminess, powdery, stickiness, lumpiness, oily coating, and residue
coating. From QDA statistics, the gelatin was found to be most influencing hydro-
colloids to push the sensory attributes of skim yogurt for enhanced thickness,
smoothness, and creaminess. The QDA assessment was in agreement with the
instrumental assessments. This study implicates the utilization of hydrocolloids as
fat replacements and the establishment of tribology–sensory relationship for their
successful characterization.
Recently, Krop et al. (2019) have presented an extensive study on the relation-
ships between tribology, rheology, and sensory attributes for κ-Carrageenan and
some inhomogeneous gels prepared by mixing κ-Carrageenan with locust bean gum,
sodium/calcium alginate, etc. In particular to “slippery,” the Pearson’s correlation
coefficients for QDA scores of “slippery” with respect to fracture stress, fracture
strain, and COF at 50 mm/s appear to be 0.80, 0.80, and 0.82, respectively. These
correlations are strikingly consistent and good indicators that slippery is linked to
COF and fracture properties; nevertheless, the authors pointed out that “slippery” is
indeed a difficult perception and panelists ought to be properly trained to score this
attribute. Additionally, on the contrary with the empirical model of Kokini (1987),
the “smoothness” perception was not found to be correlated with any of the
instrumental outputs; and this situation was attributed to the composite nature of
the samples used in the study. Based on the assessments of comprehensive
7 Tribological and Sensory Properties 263

experimental observations (fracture properties; viscosity; and tribology), descriptive

sensory analysis, and statistical correlations among the various quantities, the study
established certain relationships between mechanical (fracture) and flow properties
(viscosity), and texture attributes (smoothness, slippery, pasty, etc.) of hydrocolloids
especially at early stages of oral processing.
At the later stages of oral processing, surface properties become more dominant to
produce thin film on the oral surfaces, so lubrication/friction characteristics are vital
to establish tribology–sensory or rheo–tribology–sensory relationships (Chen and
Stokes 2012). In fact, at the early stages of oral processing of hydrocolloids,
simultaneous actions of fracture, flow, and friction are crucial to determine sensory
perception. It is important to mention here that food hydrocolloids structures may
vary widely and ample number of inhomogeneous gel structure may be produced,
there by the behavior of food structures under oral processing might vary accord-
ingly. Follow-up studies on hydrocolloids are therefore needed to further consolidate
the fracture–flow–friction–sensory relationships.

6 Challenges and Future Prospects

Challenges associated with tribology-sensory research basically originate from the

fact that tribological parameters are system dependent and not intrinsic properties of
the food itself. The friction/lubrication parameters are highly sensitive to the tribo-
pair material system, model food colloids, operating variables, and environmental
influences. Thereby, it requires a number of variables to be controlled in order to
mimic the system as nearly as possible to the actual oral processing system. Further,
replication of actual oral surfaces is yet to be achieved. Hierarchical surface texture
and bulk properties of biological tongue enhances this complexity by manifolds.
There have been recent advancements towards a better understanding of tongue
surface texture and the mechanics (Funami 2016). Human variation of tongue
topography in relation to oral tribology has also been recently investigated in some
details (Wang et al. 2019). It can be easily realized that the friction/lubrication
characteristics in actual oral processing has strong dependence on the “filly” and
“fungi” form structures of the tongue surface. At this stage, it is important to
incorporate more of the tongue surface features, material property variations, and
surface characteristics such as hydrophobic effects. The motion and dynamics of oral
processing are important especially when friction is to be evaluated. Thereby,
implementation of actual oral motion (the ratio of rolling/sliding and impact, etc.)
and the degrees of freedom that the tongue enjoys can be taken up in follow-up
researches.
Saliva–food interaction is another important influencing factor to the sensory
perceptions and eating experience as it was aptly put in words, “what is perceived
in-mouth is a food–saliva mixture rather than the food on the plate” (Mosca and
Chen 2017). Thereby, the food–saliva interactions in many cases may result in new
compounds as well as very different microstructures, which may eventually impact
264 S. Panda and J. Chen

on the friction and lubrication scenarios. These important effects in oral tribology
experiments are yet to be captured.
Sensory perceptions are dynamical functions of food breakdown length scale and
oral processing time. It is therefore vital to estimate the duration that a model food
samples needs to be exposed in the tribometer in order to synchronize with the
“breakdown path.” These factors must be duly incorporated and optimized to
corroborate the friction–sensory relationships.

7 Summary

The idea of enabling tribological principles in food sensory property assessment

basically stem from three basic understanding: (1) saliva is a lubricant; (2) tongue–
food/saliva–palate is a natural sliding system; (3) the friction that arises in this
natural sliding system relates to a handful of sensory perceptions. The fundamental
aim behind this idea is to use friction coefficient in order to arrive at some standard
quantitative metric that will determine a particular sensory attribute attached to a
particular food component. The impact that these ideas can bring are multifaceted:
for example, to devise fat replacements having equal pleasure of fat in order to
challenge obesity, to reduce the cost of employing expert sensory panel, to design
food for orally impaired patients, and more.
However, tribological experiments depend largely on the systems and surround-
ings and the system output, usually, the friction coefficient can be variable for same
food articles being tested at different set-ups in two different laboratories. Therefore,
research community in the field may agree on some standards or protocols for
tribological testing with food samples which can avoid redundancies and anomalies.
The “master” Stribeck curve is another interesting idea; after proper optimization, a
standard “master” curve representing a specific group of fundamental and integrated
food articles (e.g. hydrocolloids, dairy colloids, etc.) can be very useful in the field.
Further, the tribological characterization is bringing in more comprehensiveness in
sensory analysis; and correlation between instrumental outputs with typical sensory
descriptions with the help of quantitative metrics can be a striking breakthrough in
food sensory studies.

Acknowledgements Authors acknowledge financial support for this work by the Natural Science
Funding Council of China (grant number 31871885).

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Chapter 8
Coating and Film-Forming Properties

Qian Xiao

Abstract Hydrocolloid-based coatings and films, produced from polysaccharides,

proteins, and their blends, have emerged as alternatives to synthetic polymers for
food and packaging applications, because they are edible, versatile, renewable, and
biodegradable. Coatings are formed as a thin layer directly onto food surfaces by
dipping, spraying, brushing, fluidized bed, or panning method. By contrast, films are
standalone pre-formed materials either placed between food components or sealed
into pouches, and they are manufactured by wet- or dry-casting method. Overall,
hydrocolloid-based coatings and films possess excellent barrier properties to CO2,
O2, and oil under certain conditions, but moderate water vapor barrier properties.
Their formation mechanisms are closely correlated with conformation of biopoly-
mers, their aggregation and crystalline state, as well as their interactions with
additives and water. This chapter discusses the existing and potential applications
of coatings and films, focusing on the developments and trends of hydrocolloid-
based coatings and films for the food industry.

Keywords Hydrocolloids · Coatings · Films · Physicochemical properties ·

Applications

1 Introduction

Petrochemical-based plastics, such as polyethylene (PE), poly(ethylene terephthal-

ate) (PET), polypropylene (PP), polyvinylchloride (PVC), have dominated the food
packaging market for their functionality, lightweight, ease of processing, and low
cost (Siracusa et al. 2008). Despite these advantages, increased use of plastic
packaging materials has led to serious ecological problems, since they are neither
fully recyclable nor biodegradable. While the materials can be incinerated to reclaim

Q. Xiao (*)
School of Food Science and Technology, Hunan Agricultural University, Changsha, Hunan,
China
e-mail: qianxiao@hunau.edu.cn

© Springer Nature Singapore Pte Ltd. 2021 267

Y. Fang et al. (eds.), Food Hydrocolloids,
https://doi.org/10.1007/978-981-16-0320-4_8
268 Q. Xiao

Fig. 8.1 Summary of characteristics and advantages of hydrocolloid-based coatings and films

the energy, this end-of-life approach can produce toxic compounds, including furans
and dioxins, such as those produced from burning PVC (Marsh and Bugusu 2007).
To address these issues, edible coatings and films have emerged as an alternative to
synthetic petroleum-based polymers for food packaging because they are versatile,
renewable, and biodegradable (Siew et al. 1999). They have the potential to delay the
deterioration of food products and to prolong their shelf life due to their selective
barrier properties against oxygen, carbon dioxide, water vapor, and flavor com-
pounds (Giancone et al. 2008). Global edible packaging market is expected to reach
USD 1097 million by 2023, from USD 697 million in 2016, growing at a compound
average growth rate (CAGR) of 6.81% (Edible packaging-global market outlook
from 2017 to 2023 2017).
Polysaccharides, proteins, lipids, and composites derived from these materials,
can be used as base materials to prepare edible coatings and films (Gennadios et al.
1996). Hydrocolloids based on polysaccharides and proteins are used extensively for
the formation of coatings and films for food preservation, because of their desirable
mechanical and gas barrier properties. Besides providing protective function, coat-
ings and films can act as nutritious food ingredients due to the unique nutritional and
functional properties of hydrocolloids (Viebke et al. 2014). A scheme illustrating the
main characteristics of hydrocolloid-based coatings and films is shown in Fig. 8.1.
Generally, there are no fundamental differences in material composition between
coatings and films, other than their method of manufacture. Coatings are formed as a
thin layer directly onto food surfaces by dipping, spraying, brushing, fluidized bed,
8 Coating and Film-Forming Properties 269

or panning method (Andrade et al. 2012). By contrast, films are standalone

pre-formed material either placed between food components or sealed into pouches,
and they are manufactured by wet- or dry-casting method (Janjarasskul and Krochta
2010). The performance and functionality of hydrocolloid-based coatings and films
are evaluated by their mechanical properties, barrier effects against oxygen (O2),
carbon oxygen (CO2) and water vapor, and thermal stability. These characteristics
are strongly correlated with material compositions, manufacture methods, and the
end-use conditions (e.g., relative humidity, temperature, and pH) (Rojas-Graü et al.
2009).
This chapter provides an overview on different categories of hydrocolloids for
coating and film formation. Methods of preparation, forming mechanisms, and the
physicochemical properties for coatings and films are also discussed. Finally, recent
developments and trends for packaging applications involving hydrocolloids are
summarized.

2 Components of Coatings and Films

Hydrocolloid-based coatings and films are produced from polysaccharides, proteins,

their blends, and/or food-grade additives. Their functional, organoleptic, nutritional,
and mechanical properties are modified by addition of food-grade additives, includ-
ing plasticizers, antimicrobials, antioxidants, anti-browning and crosslinking agents,
nanofillers, colorants, and flavors (Otoni et al. 2017). The main components for
formulation of hydrocolloid-based coatings and films are summarized in Fig. 8.2.

2.1 Polysaccharides

Polysaccharides are nontoxic and naturally occurring biopolymers. The polysaccha-

ride film-forming materials include starch and starch derivatives, cellulose deriva-
tives, alginate, carrageenan, chitosan, various plant gums (pectin, konjac, locust
bean gum, and guar gum) and microbial gums (pullulan, xanthan) (Cazón et al.
2017). Although polysaccharide-based coatings and films have superlative barrier
properties to CO2, O2, and oil under certain conditions, and high strength and
structural integrity, they tend to present a poor barrier to water vapor due to their
hydrophilic properties (Yang and Paulson 2000).

2.1.1 Starch and Starch Derivatives

Starch, an agricultural biopolymer found in a variety of plants, is a mixture of

amylose and amylopectin whose content varies depending on its botanic origin
(LeCorre et al. 2011). Amylose, a nearly linear biopolymer of α-1,4 anhydroglucose
270 Q. Xiao

Fig. 8.2 Main components for formulation of hydrocolloid-based coatings and films

units, is known to form a coherent and relatively strong films (Campos et al. 2011).
In contrast, amylopectin is a highly branched biopolymer of short α-1,4 chains
linked by α-1,6 glucosidic branching points occurring every 25–30 glucose units
(Durrani and Donald 1995). Its branched structure leads to form brittle and
non-continuous films (De Azeredo et al. 2014). In comparison with native starch,
modified starches, like acetylated starch, hydroxypropyl starch, and oxidized-starch,
have been reported to form stronger and more flexible films (López et al. 2008, 2010;
Hu et al. 2009).

2.1.2 Cellulose Derivatives

Cellulose, the main component of plant fibers, is essentially a linear high-molecular

weight biopolymer of D-glucose units linked through β-1,4 glycosidic bonds. The
close packing of cellulose chains makes it highly crystalline, fibrous, and insoluble
in water (Wang et al. 2016). Water-soluble cellulose derivatives, such as methylcel-
lulose (MC), hydroxypropyl methylcellulose (HPMC), carboxymethylcellulose
8 Coating and Film-Forming Properties 271

(CMC), and hydroxypropyl cellulose (HPC), possess good film-forming character-

istics (Dhall 2013). Among them, MC films show the lowest hydrophilic properties,
whereas the water vapor permeability of HPMC and CMC films is relatively high
(Sánchez-González et al. 2009; Kester and Fennema 1989). In addition, the substi-
tution type and degree in cellulose derivatives are critical factors determining the
performance of cellulose-based films (Espinoza-Herrera et al. 2011).

2.1.3 Chitosan

Chitosan is a functional biopolymer derived from chitin by deacetylation in alkaline

media. It consists of randomly distributed β-(1,4)-2-acetamido-D-glucose and
β-(1,4)-2-amino-D-glucose units, with the latter usually exceeding 60% (Kim et al.
2006). Chitosan has a wide spectrum of activity and high killing rate against Gram-
positive and Gram-negative bacteria (Chung and Chen 2008). The antimicrobial
activity and film-forming ability of chitosan are correlated to its degree of acetylation
or deacetylation, and molecular weight (Hosseinnejad and Jafari 2016). Owing to its
outstanding characteristics, chitosan could be potentially utilized as the antimicro-
bial packaging materials to improve food quality and shelf life.

2.1.4 Polysaccharides Extracted from Seaweed

Alginate, a linear polysaccharide extracted from brown seaweed, is composed of

variable proportions of β-D-mannuronic acid (M block) and α-L-guluronic acid
(G block) linked by 1,4 glycosidic bonds. The block copolymer consists of homo-
polymeric regions of M- and G-blocks, separated by regions that contain M and G
units (Fu et al. 2011). The proportion and distribution of these blocks determine the
physicochemical properties of the biopolymer (Lacroix and Le Tien 2005). Alginate
dissolves readily in water to form homogeneous film-forming solutions, which upon
drying can yield coherent, and transparent films that have a wide range of food
applications (Xiao et al. 2012).
Carrageenan is an anionic linear polysaccharide, extracted from edible red sea-
weeds of Rhodophycea class. It is formed by alternate units of D-galactose and
3,6-anhydrogalactose linked by α-1,3 and β-1,4 glycosidic linkage (Cosenza et al.
2014). There are three types (κ, ι, and λ) of carrageenan with varying number and
position of sulfate groups on the galactose dimer (Liu et al. 2015). In comparison
with ι-carrageenan films, κ-carrageenan films showed the higher moisture barrier and
mechanical properties, except for its flexibility (Paula et al. 2015).

2.1.5 Pectin

Pectin consists of linear homo-galacturonan (α-1,4-galacturonic acids) chains inter-

spersed with branched rhamnogalacturonan (α-1,4-galacturonic acid to
272 Q. Xiao

α-1,2-rhamnose) chains (Jolie et al. 2010). According to its degree of esterification

(DE), pectin can be classified as high-methoxyl pectin (HMP, DE > 50%) and
low-methoxyl pectin (LMP, DE < 50%) (Espitia et al. 2014). The mechanical, water
barrier properties and thermal stability of HMP films are better than that of LMP
films (Lorevice et al. 2016).

2.1.6 Pullulan

Pullulan is an extracellular and water-soluble microbial polysaccharide produced by

Aureobasidium pullulans. The linear polymer mainly consists of maltotriose units
interconnected to each other by α-(1,6) glycosidic bonds, which are responsible for
the flexible conformation and the ensued amorphous character of this polysaccharide
in the solid state (Sutherland 1998). This unique linkage pattern endows pullulan
with distinctive physical properties to form film that is strong, transparent, and with
low permeability to oil and oxygen (Xiao et al. 2012, 2015).

2.2 Proteins

Proteins used for film-forming materials can be categorized into two groups based on
their origin of sources: plant-derived proteins, such as corn zein, soy protein, and
wheat gluten, or animal-derived proteins like casein, whey protein, gelatin, and
collagen proteins (Han 2014). Depending on amino acid composition and sequence,
the structure of protein can be random coil, fibrous, or globular. For globular proteins
(i.e., soy protein, wheat gluten), they must be denatured by heat, acid, and/or solvent
to shape extra extended structures that are required for film formation (Dhall 2013).
Overall, protein-based coatings and films display considerably lower O2 and CO2
permeability and CO2/O2 permeability ratio, and moderate mechanical and water
vapor barrier properties (Song and Zheng 2014).

2.2.1 Corn Zein

Corn zein, a prolamin protein, has a molecular weight ranging from 18 to 45 kDa. As
a relatively hydrophobic protein, the hydrophobicity of zein is related to its high
content of non-polar amino acids residues including leucine, alanine, and proline
(Shukla and Cheryan 2001). Corn zein dissolves in aqueous ethanol solution to form
the glossy, greaseproof, and brittle films through the hydrophobic, hydrogen, and
limited disulfide (SS) bonds between zein chains (Ghanbarzadeh et al. 2007).
8 Coating and Film-Forming Properties 273

2.2.2 Wheat Gluten

Wheat gluten, an ethanol-soluble protein in wheat flour, is composed of gliadin and

glutenin. Gliadin is monomeric protein with molecular weight of 28–55 kDa, while
glutenin is aggregated protein linked by interchain SS bonds with molecular weight
of about 500 to 10,000 kDa (Wieser 2007). Glutenin films presented higher mechan-
ical strength and lower water vapor permeability than gliadin films (Hernández-
Muñoz et al. 2003). Moreover, the purity of wheat gluten has positive effect on the
appearance and mechanical attributes of wheat gluten films (Gennadios et al. 1993).

2.2.3 Soy Protein

Soy protein is comprised of two major components, 7S (β-conglycinin) and 11S

(glycinin), representing 37% and 31% of soy protein, respectively. 7S is rich in
asparagine, glutamine, leucine, and arginine residues with a molecular weight of
180 kDa. 11S has a molecular weight of 320–360 kDa and contains 20 intramolec-
ular SS bonds (Kumar et al. 2002). Films made from 11S fraction are smooth and
opaque, whereas 7S films exhibit transparent and creased appearance (Kunte et al.
1997). At low relative humidity (RH), O2 permeability of soy protein isolate (SPI)
films was lower than that of films based on low-density polyethylene (LDPE),
methylcellulose, starch and pectin, respectively (Song et al. 2011a).

2.2.4 Casein and Caseinate

Casein mainly consists of five fractions including αs1, αs2, β, κ, and δ-casein, and
their sizes vary from 11.5 to 25 kDa. Among them, β-casein is the most interesting
one, as it produces films with lower permeability to water vapor than other milk
protein (Mauer et al. 2000). Caseinate is a mixture of casein monomers and small
aggregates formed after removing of colloidal calcium phosphate from casein
micelles. Compared to casein, caseinate, particularly for sodium caseinate, is more
soluble and has better film-forming capacity. Films produced from sodium caseinate
possess excellent barriers to O2, CO2, and aromas, and thermal resistance (Khwaldia
et al. 2004a).

2.2.5 Whey Protein

Whey protein includes β-lactoglobulin, α-lactalbumin, bovine serum albumin,

immunoglobulins, lactoferrin, and proteose-peptones (Mulvihill and Ennis 2003).
Films prepared from whey protein isolates (WPI) exhibited promising mechanical
features, as well as moderate moisture permeability and good oxygen barrier prop-
erties, compared to the synthetic polymer films, e.g., low-density polyethylene
274 Q. Xiao

(LDPE), high density polyethylene (HDPE), PVDC, cellophane, and polyester

(Khwaldia et al. 2004b).

2.2.6 Gelatin

Gelatin is an animal protein obtained by hydrolysis of collagen. It is a combination

of many fractions varying in size, including the whole α-chain of tropocollagen
molecule (a trimer of around 330 kDa that aggregates to form the larger collagen
structures) and hydrolytic fragments of parts of the α-chains (Boran and Regenstein
2010). Gelatin films display effective barriers against O2 and aromas at low or
intermediate RH, but weak water resistance due to its hydrophilic nature. Further-
more, their mechanical properties are closely related to the renaturation level of
gelatin (Bigi et al. 2004).

2.3 Food-Grade Additives

2.3.1 Plasticizers

Plasticizers are low molecular weight compounds with non-volatile compounds.

Their primary role is to enhance the flexibility and processability of hydrocolloid-
based coatings and films. However, their barrier properties are impaired as result of
the increased free volume and molecular mobility after plasticizers addition
(Sothornvit and Krochta 2005; Vieira et al. 2011). Food-grade plasticizers mainly
include glycerol, sorbitol, polyethylene glycol, sucrose, glucose, fructose, mannitol,
xylitol, fatty acids, and monoglycerides (Vieira et al. 2011).

2.3.2 Polysaccharide Nanofillers

Nanofillers (at least one dimension smaller than 100 nm) provide reinforcement
effects due to their high aspect ratio and surface-to-volume ratios (Crosby and Lee
2007). Considering the application and safety for hydrocolloid-based coatings and
films in food packaging, the polysaccharide nanofillers, e.g., cellulose nanoparticles,
cellulose nanocrystals, starch nanoparticles, starch nanocrystals, chitin
nanowhiskers, and chitin nanofibers, have been used as excellent candidates for
improvement of their mechanical, barrier, and thermal properties (Otoni et al. 2017).

2.3.3 Antimicrobial Additives

Incorporation of antimicrobial compounds into packaging materials provides inhib-

itory effects against spoilage and pathogenic bacteria by maintaining active
8 Coating and Film-Forming Properties 275

compounds on food surface (Gennadios et al. 1997). There are several categories of
antimicrobial compounds that have been employed in hydrocolloid-based coatings
and films, including organic acids (sorbic and its potassium salt, acetic acid, and
malic acid), polypeptides and bacteriocins (lysozyme and nisin), plant essential oils
(cinnamon, oregano, rosemary, and lemongrass), and polyphenols (flavonoids and
phenolic derivatives) (Franssen and Krochta 2003).

3 Preparation Methods

3.1 Preparation of Hydrocolloid-Based Coatings

3.1.1 Spray Coating

Spray coating is a commonly used technique for food coatings, especially for fruits
and vegetables. In this process, food products are placed on a rotating platform, then
the coating-forming solution forms droplets and distributes them over the food
surface by means of a set of spraying nozzles (Debeaufort and Voilley 2009). The
main advantages of this technique offer uniform coating, thickness control, and the
possibility of multilayer applications, such as using alternating sodium alginate and
chitosan solutions (Ustunol 2009).

3.1.2 Dip Coating

Dip coating involves submerging food products into a vat containing coating
solution. After dipping the products and draining away excess coating, it is dried
either at room temperature or with the aid of a dryer (Andrade et al. 2012). The
advantage of this method is to obtain good uniformity around the irregularly-shaped
and rough food surface. Several problems may occur by using this method, such as
coating dilution, build-up of trash or dirt, and microorganism growth in the dipping
tank (Andrade et al. 2012).

3.1.3 Fluidized-Bed Coating

Fluidized beds are categorized by three different configurations: top spray, bottom
spray, and rotating-fluidized bed. The conventional top-spray method has a greater
possibility of success in the food industry compared to other methods (Andrade et al.
2012). As presented in Fig. 8.3, the coating solution is sprayed through a set of
nozzles onto the surface of fluidized particles to form a shell-type structure. Its
application focuses on the functional ingredients and food additives, i.e., leavening
agents, enzymes, vitamins, minerals, and spices (Chen et al. 2009).
276 Q. Xiao

Fig. 8.3 Schematic of

top-spray fluidized-bed
coating process, adapted
from (Dewettinck and
Huyghebaert 1999) with
permission

Fig. 8.4 Schematic of pan

coating process, adapted
from (Agrawal and Pandey
2015) with permission

3.1.4 Pan Coating

The schematic of pan coating process is displayed in Fig. 8.4. As shown, the coating
solution is sprayed into a rotating bowl (referred to as pan), and the food particles are
tumbled within the pan to distribute the coating solution over their surface. Forced
air, either ambient or elevated temperature, is utilized to dry the coating (Agrawal
and Pandey 2015). Pan coating is mainly used for the confectionery and chocolate
industries or particularly small food items like nuts and raisins (Andrade et al. 2012).
8 Coating and Film-Forming Properties 277

3.2 Preparation of Hydrocolloid-Based Films

3.2.1 Wet Method

The wet method, also known as solvent casting, can be sub-classified to bench
casting and continuous casting, respectively. The bench casting is commonly uti-
lized to fabricate films at laboratory scale as it is simple and cost effective. In this
method, the film-forming solution is deposited over a rimmed plate, and then
followed by drying to produce a cohesive and free-standing film.
Continuous casting is more suitable for industrial applications, because it requires
less space and labor. As shown in Fig. 8.5, film-forming solution is uniformly spread
on a continuous steel belt that passes through a drying chamber. The dried film is
then stripped from the steel belt and wound into film roller. The advantage of this
method is optimizing uniformity, heat transfer, and drying efficiency, while avoiding
expense of a separate substrate (Rossman 2009).

3.2.2 Dry Method

Dry method, i.e., compression molding and extrusion processing, is based on the
thermoplastic properties of polysaccharides and proteins. In the presence of plasti-
cizers, at low moisture levels and high temperatures and with pressure, biopolymers
acquire a viscoelastic behavior that allows them to be shaped for the production of
films (Gómez-Estaca et al. 2016). In general, compression molding is studied at
laboratory scale as a precursor to extrusion with the aim of determining the suitable
processing conditions (Hernandez-Izquierdo and Krochta 2008).
Extrusion processing is a highly efficient manufacturing method with commercial
potential for large-scale production of biopolymer films (Fishman et al. 2000). The

Fig. 8.5 Schematic of continuous casting technique to prepare hydrocolloid-based films, adapted
from (Borges et al. 2015) with permission
278 Q. Xiao

Fig. 8.6 The configuration of one-screw extruder, adapted from (Borges et al. 2015) with
permission

configuration of one-screw extruder is presented in Fig. 8.6. The extruder basically

consists of an endless screw inside a barrel with a double casing that permits control
of temperature. The biopolymer is fed from a hopper and pushed by the screw
towards a die (Nur Hanani et al. 2012). To date, dry method has been successfully
used in preparation of starch, alginate, wheat gluten, soy protein, and whey protein
films (Mendes et al. 2016; Hernandez-Izquierdo and Krochta 2008; Azevedo et al.
2017; Ciannamea et al. 2014).

4 Microstructural and Physicochemical Characterization

The microstructural characteristics (such as chemical, crystalline structure, and

morphology) of hydrocolloid-based coatings and films are closely correlated with
their packaging performance (e.g., mechanical, barrier, and thermal properties).

4.1 Structural Analysis

Microscopy and spectroscopic techniques have been utilized to study the architec-
ture and structure of hydrocolloid-based films at micro and nanometric scales.
Ultrastructural and internal structure in films have been characterized by confocal
laser scanning microscopy (CSLM), while scanning electron microscopy (SEM) and
atomic force microscopy (AFM) are more used to study their surface and cross-
section morphology (Arzate-Vázquez et al. 2012; Andreuccetti et al. 2009). Fourier
transform infrared spectroscopy (FTIR) analyzes the possible functional chemical
groups, conformational transitions, and molecular interactions (Yadav et al. 2014).
Nuclear magnetic resonance (NMR) spectroscopy provides information about the
chemical and physical properties of atoms or their related molecules, as well as
reaction state, dynamics, structure and chemical environment (Karbowiak et al.
2008). For instance, for hsian-tsao gum (HG)-casein films, the hydrogen bonding
interactions and Maillard reactions between HG and casein were revealed by FTIR
8 Coating and Film-Forming Properties 279

data. Meanwhile, NMR analysis indicated that HG addition significantly changed

the mobility of water molecule in casein films (Yang et al. 2015). Other comple-
mentary techniques are also utilized for structural analysis of hydrocolloid-based
coatings and films, such as X-ray diffraction (XRD) to identify the information about
crystalline/amorphous structures, and small-angle X-ray scattering (SAXS) to mon-
itor crystalline and aggregate structures of membrane materials (Bodnár et al. 2007).

4.2 Mechanical Properties

Favorable mechanical properties are essential for packaging materials to perform

their protective functions efficiently. Mechanical properties of selected
hydrocolloid-based films are listed in Table 8.1. A standard method, ASTM-
D882–91, originally developed to evaluate mechanical properties such as tensile
strength (TS), elongation at break (EAB), elastic modulus (EM), and toughness of
commercial plastic, is also applied to hydrocolloid-based films (ASTM-D882-91
1991). As shown in Fig. 8.7, the mechanical parameters are calculated by determin-
ing the relationship between stress and strain, when film is stretched at a set rate
(distance/time). EM, a measure of intrinsic film stiffness, is the slope of the linear
range of the stress–strain curve (Mauer et al. 2000). Toughness refers to the ability of
a material to absorb energy during deformation up to fracture, determined as the area
under the stress–strain curves (Fig. 8.7b). TS is the maximum strength measuring the
resistance of the film, whereas the percentage of EAB is a measure of the stretching
capacity of flexibility of the film prior to breaking. They are calculated by using
Eqs. 8.1 and 8.2:

TS ¼ F=A ð8:1Þ

where TS is the tensile strength (MPa), F is the force (N) at maximum load, and A is
the initial cross-sectional area (m2) of the film specimen.

EAB ¼ 100  ðl  l1 Þ=l1 ð8:2Þ

where EAB is the elongation at break (%), l1 is the initial length, and l is the length of
the film at breaking point.

4.3 Barrier Properties

The basic function of packaging materials is to control mass transfer between food
and the ambient atmosphere. Water vapor in environment transferring to packaged
food results in problematic microbial growth, and undesirable textural changes.
Oxygen can cause deterioration of food due to oxidation of lipids and other
 280

Table 8.1 Mechanical properties and water vapor, oxygen, and carbon dioxide permeability values of selected hydrocolloid-based films
Mechanical properties WVP Gas permeability
TS EAB EM Test (g.m.Pa1. Test O2P (cm3.mm. CO2P (cm3.mm.
Compositesa (MPa) (%) (MPa) conditions h1.m2) conditions m2.d1.Pa1) m2.d1.Pa1) Reference
HMw chitosan 61.82 4.59 – 38  C, 1.34  1010 23  C, 6.65 – (Leceta et al.
90% RH 50% RH 2013)
LMw chitosan 55.83 4.58 – 38  C, 1.53  1010 23  C, 7.70 – (Leceta et al.
90% RH 50% RH 2013)
Pullulan 73.89 1.67 4938 23  C, 7.05  106 23  C, 0.48 – (Xiao et al.
100% RH 23% RH 2012, 2015)
Alginate 74.04 4.60 3047 23  C, 25.75  106 23  C, 8.94 – (Xiao et al.
100% RH 23% RH 2012, 2015)
Sweet potato starch- 33.8 2.02 – 25  C, 7.09  106 23  C, 4.09 – (Shen et al.
chitosan 75% RH 50% RH 2010)
WPC-GLY (40% w/w) 0.72 50 78 20  C, 4.50  106 20  C, 0.2 1.02 (Ramos et al.
50% RH 50% RH 2013)
WPI-GLY (40% w/w) 0.9 55 95 20  C, 3.43  106 20  C, 0.41 1.58 (Ramos et al.
50% RH 50% RH 2013)
Vetch seed protein 1.59 32.08 78.14 – – 25  C, 20.16 24.84 (Porta et al.
55% RH 2015)
Papaya puree-gelatin 6.7 18.09 – 38  C, 6.62  105 23  C, 8.24 10.25 (Tulamandi
(8:1) 90% RH 50% RH et al. 2016)
Papaya puree-defatted 5.13 28.11 – 38  C, 7.52  105 23  C, 8.92 10.89 (Tulamandi
soy protein (8:4) 90% RH 50% RH et al. 2016)
WPI-okra polysaccha- 2.3 31.4 68.2 23  C, 3.23  105 20  C, 5.67 – (Prommakool
ride (1:1) 100% RH 50% RH et al. 2011)
WPI-LBG (0.025% 2.66 11.4 – 20  C, 4.97  107 20  C, 0.78 4.71 (Silva et al.
w/w) 100% RH 50% RH 2016)
a
HMw high molecular weight, LMw low molecular weight, WPC whey protein concentrate, GLY glycerol, LBG locust bean gum
Q. Xiao
8 Coating and Film-Forming Properties 281

Fig. 8.7 (a) Schematic of tension test setup, adapted from (Pham et al. 2008) with permission, (b)
mechanical properties determined from the typical stress-strain curve

oxygen-sensitive components. Thus, water vapor and gas permeability is a vital

property for selecting or tailoring the hydrocolloid-based films.

4.3.1 Water Vapor Permeability (WVP)

Table 8.1 shows WVP values of selected hydrocolloid-based films. These data are
obtained gravimetrically following the ASTM Standard Test Method E96, known as
the “cup method” (ASTM-E96-92 1990). According to this method, a cup with an
open mouth is filled with distilled water or desiccant. The film is sealed on the open
mouth of the cup, the assembly is weighed, and placed under controlled temperature
and RH conditions (Cazón et al. 2017). WVP is calculated according to the com-
bined Fick–Henry laws for gas diffusion through films (Eq. 8.3).

Δw L
WVP ¼  ð8:3Þ
Δt  A Δp

where Δw/Δt is the rate of water gain (g/h), A is the exposed area of the film (m2),
L is the mean thickness of film specimens (m), and Δp is the difference in partial
water vapor pressure between the two sides of film specimens.

4.3.2 Gas Permeability

Oxygen permeability (O2P) and carbon dioxide permeability (CO2P) are evaluated
on the basis of the ASTM D 3985–02 method (ASTM-D3985-02 2002). The films
are sealed between two chambers with each having two channels to the exterior. In
282 Q. Xiao

the lower chamber, O2 or CO2 is supplied at a controlled flow rate to maintain the
pressure constant in that compartment. The other chamber is purged by a stream of
nitrogen, also at a controlled flow. In the case of O2P measurement, the nitrogen flow
leaving this chamber is connected to an O2 sensor installed on-line which measures
the O2 concentration. For CO2P measurement, the nitrogen flow leaving this cham-
ber is collected in a syringe for CO2 quantification by a gas chromatograph
(Cerqueira et al. 2009). The O2P and CO2P of selected hydrocolloid-based films
are listed in Table 8.1.

4.4 Thermal Properties

One key factor that influences the processing and operating temperatures of
hydrocolloid-based coatings and films is their thermal properties. The properties are
investigated by differential scanning calorimetry (DSC), thermogravimetric analysis
(TGA), and dynamic mechanical analysis (DMTA). DSC technique is used to
determine the glass transition temperature (Tg), melting temperature (Tm), crystalli-
zation temperature, heat capacity difference at Tg of hydrocolloid-based coatings and
films (Cheng 2002). TGA is widely employed to examine their decomposition
temperature, weight loss, and activation energy of decomposition (Cheng 2002).
Furthermore, the structural and viscoelastic properties of films are investigated by
DMTA. Dynamic modulus, dynamic loss modulus, temperature of main chain
relaxation, and temperature of local mode relaxation are measured as functions of
temperature and frequency by forced oscillation method (Brown and Gallagher
2011).

5 Film-Forming Mechanism

Understanding the film-forming mechanism is important to predict material proper-

ties of hydrocolloid-based films, which is essential for the optimization of drying and
processing condition. As previously mentioned, both processing methods (wet and
dry) have been widely used to prepare the films. The wet method requires solubi-
lizing the hydrocolloids in a solvent, spreading the solution onto a flat surface, and
then followed by drying to produce a film. The film-forming mechanism involves
conformational change of the biopolymer, as well as solvent-biopolymer and
biopolymer-biopolymer interactions that continue to evolve as the solvent evapo-
rates under different drying conditions (Watanabe et al. 2006; Xiao et al. 2014b).
However, a number of polysaccharides and proteins have capacity to form gel during
film-forming process, and their film-forming mechanism is related to the gelation
mechanism. Although a few researchers proposed that the transition from wet gel
(biopolymer-in-water) to dry film (water-in-biopolymer) is a critical stage during
8 Coating and Film-Forming Properties 283

film-forming process, the complete transition mechanism after gelation have not yet
been fully explained (Szabó et al. 2012).
By contrast, dry method involves heating and mixing biopolymers and plasti-
cizers by extrusion and/or compression molding techniques. Over the course of
extrusion, biopolymer chains denature, dissociate, unravel and align, and then
recombine, crosslink, and aggregate via specific linkages with heat and pressure,
which result in film formation through complete restructuring of biopolymer mole-
cules. Thus, the film-forming mechanism is correlated with conformation changes of
biopolymers, their aggregation and crystalline state, as well as the interactions
among biopolymer, plasticizer, and water.

5.1 Polysaccharide-Based Films

Polysaccharides (with the exception of glycogen, etc.) are long-chain biopolymers

formed from mono- or disaccharide repeating units joined together by glycosidic
bonds. Owing to the presence of a large number of hydroxyl and other polar groups
in their structure, hydrogen bonds and/or electrostatic interactions have a crucial
function in film formation (Han 2014). Polysaccharide films are fabricated by
disrupting interactions among polysaccharide segments and forming new
intermolecular hydrophilic interactions and hydrogen bonding (Rhim and Ng 2007).

5.1.1 Formation Mechanism of Solvent Casting Films

For starch films, their formation mechanism depends on the starch concentration and
amylose content. At relatively high concentration, aggregation and packing of
swollen granules dominated the film formation, whereas both coil-to-helix transition
and aggregation of double helices were operative during the film formation from
dilute starch solutions (Liu 2005). Xiao et al. (2014a, b) elaborated the formation
mechanism of pullulan and alginate films by monitoring the conformational change
of polysaccharides, water-polysaccharide, and polysaccharide-polysaccharide inter-
actions during drying. As pullulan drying process progressed, the oxygen atoms at
the C5 and C6 carbons of the D-glucopyranose ring might preferentially form
hydrogen bond with water or pullulan molecules, resulting in more-ordered structure
with increased interchain interactions in pullulan films. Moreover, the less-ordered
structure domain of the pullulan was first affected during drying, followed by
pullulan skeleton segments. Finally, conformational changes in pullulan chains
occurred as the drying process completion (Xiao et al. 2014b). In the course of the
formation of alginate film, the oxygen atoms at the C2 and C3 carbons of the
pyranose ring preferentially formed hydrogen bond with water or alginate mole-
cules, while the skeletal vibrations of pyranose ring (e.g., C-C and C-O-C groups)
were less perturbed than the stretching vibrations of COO group and O-H bending
vibration of alginate with drying (Xiao et al. 2014a). The film-forming mechanism of
284 Q. Xiao

Flammulina velutipes polysaccharide might be associated with the intermolecular

and intramolecular hydrogen bonds between polysaccharide chains and the forma-
tion of β-glycosidic bonds upon drying (Du et al. 2016). Li et al. (2019) proved that
the electrostatic interactions and hydrogen bonds are crucial in fabricating the
multilayer films based on chitosan and alginate by layer-by-layer (LbL) technique.
Strong intermolecular interactions occurred among the amino, carboxyl, and
hydroxyl groups of the chitosan and alginate.

5.1.2 Formation Mechanism of Extruded and Compression-Molded

Films

Pushpadass et al. (2009) reported that glycerol and/or water destroyed the crystal-
linity of native starch, then the starch fragmentation converted into thermoplastic
starch with heat and shear. During extrusion process, the inter- and intra-hydrogen
bonds of starch would be unraveled when the glycerol was added into starch, and
the new hydrogen bonds between starch and glycerol were formed simultaneously
(Pushpadass et al. 2009). Afterwards, the starch recrystallization induction process
among the helical amylose molecule occurred during cooling, which led to the
Vh-type crystalline arrangement (Azevedo et al. 2017). According to Gao et al.
(2017), neat alginate granules were largely de-structured by glycerol and water,
and glycerol increased the mobility of alginate chains while promoting the crys-
tallization of alginate chains with structural reorganization during compression
molding.

5.2 Protein-Based Films

The main formation mechanism of protein films involves denaturation of the protein
initiated by heat, solvent, or change in pH, followed by association of extended
peptide chains through new intermolecular interactions, such as covalent (SS bond
or crosslinking) and electrostatic, hydrophobic, or ionic interactions between protein
chains (Janjarasskul and Krochta 2010).

5.2.1 Formation Mechanism of Solvent Casting Films

The formation of intact and water-insoluble WPI films was realized by heat dena-
turation of aqueous protein solution (Pérez-Gago and Krochta 2002). Heat denatur-
ation unfolded whey protein and promoted the exposure of SH and hydrophobic
groups. The unfolded protein might then undergo intermolecular interactions
(hydrogen bonds, hydrophobic, covalent and electrostatic interactions). It is note-
worthy that the cohesion of WPI films relied principally on the intermolecular SS
bonds via sulphydryl/disulphide (SH/SS) exchange reactions (Guckian et al. 2006).
8 Coating and Film-Forming Properties 285

On the other hand, WPI had the ability to form water-soluble films without heat
denaturation. Since most of the hydrophobic and SH groups are buried in the interior
of WPI molecule, their film-forming mechanism involves the intermolecular hydro-
gen bonding between protein molecules, rather than the hydrophobic and covalent
interactions (Guckian et al. 2006; Pérez-Gago et al. 1999). Ciannamea et al. (2014)
also proved that hydrogen bonds and hydrophobic interactions played a more
important role in the formation of soy protein films. During the film formation of
11S, along with disappearance of its α-helices and disordered structures, the
intermolecular hydrogen bonds between β-sheet segments predominated the aggre-
gation of 11S (Robert et al. 2001; Subirade et al. 1998). Similar to the 11S films, the
high density of intermolecular hydrogen-bonded β-sheets were conducive to the
formation of gliadin network during drying (Mangavel et al. 2001). According to
Pankaj et al. (2014), the film-forming mechanism of caseinate was attributed to their
random coil structure which allowed them to form extensive intermolecular hydro-
gen, electrostatic, and hydrophobic bonds, resulting in increased interchain
cohesion.
In comparison, the formation mechanism of gelatin films is related to the tem-
perature during drying due to thermo-reversible gelation behavior of gelatin. When
the gelatin films were prepared below the helix-coil transition temperature, partial
renaturation of collagen in gelatin took place, which resulted in the formation of a
collagen-like triple-helix structure. Moreover, the partial renaturation only took
place during the advanced stage of drying (Ghoshal et al. 2014). On the contrary,
a helix structure was rarely formed in gelatin films when they were dried above the
helix-coil transition temperature.

5.2.2 Formation Mechanism of Extruded and Compression-Molded

Films

For compression-molded soybean protein films, the high temperature promoted the
crosslinking between soybean proteins through intermolecular SS bonds, either from
free sulfhydryl (SH) groups or through SH/SS exchange reactions, which
predominated the formation of film matrix (Ciannamea et al. 2014). During extru-
sion process, the aggregation and reorganization of wheat gluten molecules were
principally related to the formation of intermolecular SS crosslinking bonds via
oxidation of SH groups and SH/SS exchange reactions between glutenin and gliadin
(Lagrain et al. 2010). The formation schematic of intermolecular SS bonds between
glutenin and gliadin during heat processing is illustrated in Fig. 8.8.
286 Q. Xiao

Fig. 8.8 Schematic of the formation of intermolecular SS crosslinking bonds between glutenin and
gliadin during heat processing. Adopted and modified from (Lagrain et al. 2010) with permission

6 Applications and Recent Developments

Oxidation, microbial spoilage, and metabolism are the main causes of deterioration
of food products. Thus, the primary function of packaging materials based on
hydrocolloids is to maintain the quality and safety of food products during storage
and conveyance. Normally, fruits and vegetables have short shelf life due to its
perishable nature. Hydrocolloid-based coatings and films may act as a semiperme-
able barrier to selectively control the exchange of CO2, O2, and ethylene, resulting in
the reduction in ethylene levels, ripening, respiration rate, and water loss on fruits
and vegetables (Valencia-Chamorro et al. 2011). Several studies shown in Table 8.2.
have demonstrated the ability of hydrocolloid-based coatings and films carrying
bioactive compounds to retard browning reactions and microbial growth in fruits and
vegetables, especially the minimally processed (MP) fruits and vegetables. Ramos-
García et al. (2012) reported that lime essential oil incorporated into chitosan-
beeswax blend coatings on tomato showed strong inhibitory effect against Rhizopus
stolonifer and Escherichia coli DH5α during storage at 12 and 23  C. Sarengaowa
et al. (2018) coated the fresh-cut “Red Fuji” apples with alginate coatings containing
thyme oil, cinnamon oil, and/or oregano oil, and observed that reduction of total
coliform, yeast and mold counts in comparison with control and alginate-coated
samples. Meanwhile, the respiration rate, weight loss, firmness, and browning
reactions in fresh-cut apples stored at 4  C were significantly decreased.
Recently, the development of multilayer and nanomultilayer coatings based on
hydrocolloids, formed by LbL deposition technique, gained much attention for the
preservation of fruits and vegetables. For instance, the multilayer coatings based on
gelatin and chitosan predominantly enhanced physiological quality and reduced the
8 Coating and Film-Forming Properties 287

Table 8.2 Recent applications of hydrocolloid-based coatings and films in fruits and vegetables
Coating/film
composites Form Food products Main benefits References
Fruits and
vegetables
Pectin-pullulan- Coatings Strawberry Reduced weight loss, (Treviño-
chitosan with (Fragaria fruit softening and Garza et al.
sodium benzoate ananassa) microbial growth (total 2015)
and potassium aerobic counts, molds,
sorbate and yeasts), delayed
alteration of color and
total soluble solids
content
Alginate with Coatings Strawberry Inhibited the (Peretto et al.
carvacrol and Escherichia coli O157: 2014)
methyl H7 and Botrytis
cinnamate cinereal
Alginate-pectin Coatings Blueberry Improved the firmness, (Mannozzi
significantly reduced et al. 2017)
growth kinetics of
yeasts and mesophilic
aerobic bacteria
Gum arabic- Coatings Avocado Reduced the anthrac- (Bill et al.
Aloe vera- (Persea ameri- nose incidence during 2014)
chitosan com- cana Mill.) the postharvest supply
bined with chain, inhibited myce-
thyme oil lial growth of
Colletotrichum
gloeosporioides
Chitosan- Coatings Longan Reduced weight loss, (Lin et al.
carrageenan (Dimocarpus respiration rate and 2018)
longan) color changes
Chitosan-cas- Coatings Guavas Reduced total aerobic (de Aquino
sava starch with (Psidium mesophilic bacteria, et al. 2015)
essential oil guajava L.) mold and yeast counts,
extract from exhibition lower titrat-
Lippia gracilis able acidity value
Pea starch-guar Coatings ‘Valencia’ Reduced fruit respira- (Saberi et al.
gum with shellac oranges tion rate, ethylene pro- 2018)
and oleic acid duction, weight and
firmness loss, and peel
pitting
CMC-chitosan Bilayer coatings Citrus fruit Increased fruit firm- (Arnon et al.
ness, and enhanced 2014)
fruit gloss
Gum arabic with Coatings Banana and Delayed ripening, (Maqbool
cinnamon oil papaya weight loss, fruit firm- et al. 2011)
ness, and titratable
acidity, fungicidal
effects against
(continued)
288 Q. Xiao

Table 8.2 (continued)

Coating/film
composites Form Food products Main benefits References
Colletotrichum musae
and Colletotrichum
gloeosporioides
HPMC-beeswax Coatings Plums Reduced water loss, (Navarro-
(Cv. Angeleno) flesh softening and Tarazaga
internal breakdown et al. 2011)
HPMC with Coatings ‘Formosa’ Reduced the respiration (Choi et al.
oregano essen- plum rate, ethylene produc- 2016)
tial oil tion, total weight loss
Chitosan with Coatings Cherry Decreased Escherichia (Cui et al.
Artemisia annua tomato coli O157:H7 2017)
oil
Chitosan-bees- Coatings Tomato No growth of Rhizopus (Ramos-
wax with lime stolonifer and García et al.
essential oil Escherichia coli DH5α 2012)
Pectin-chitosan Nanomultilayer ‘Tommy Presented a lower mass (Medeiros
coatings Atkins’ loss, lower total soluble et al.
mangoes solids and higher titrat- 2012a, b)
able acidity
Starch-gelatin Coatings Refrigerated Enhanced appearance (Fakhouri
red crimson and decreased weight et al. 2015)
grapes loss
Aloe vera-gum Coatings Button Reduced weight loss, (Mohebbi
tragacanth mushroom color changes and et al. 2012)
softening
Chitosan-banana Films Asparagus Inhibited growth of (Pitak and
flour and corn Staphylococcus aureus Rakshit
2011)
MP fruits and
vegetables
Cassava starch- Coatings Minimally Reduced respiration (Garcia et al.
potassium processed rate, increased water 2010)
sorbate strawberry vapor resistance
Gellan with Coatings Fresh-cut Significantly reduced (Tomadoni
geraniol strawberry microbial counts et al. 2018)
Alginate- Nanomultilayer Fresh-cut Lower values of mass (Souza et al.
chitosan coatings mangoes loss, pH, 2015)
malondialdehyde con-
tent, browning rate,
soluble solids
κ-Carrageenan- Nanomultilayer Whole and Presented lower mass (Medeiros
lysozyme coatings fresh-cut loss, and total soluble et al.
‘Rocha’ pear solids and higher titrat- 2012a, b)
able acidity
SPI with ferulic Coatings Fresh-cut Effective in controlling (Alves et al.
acid apples their weight loss and 2017)
firmness
(continued)
8 Coating and Film-Forming Properties 289

Table 8.2 (continued)

Coating/film
composites Form Food products Main benefits References
Alginate with Coatings Fresh-cut Significantly inhibited (Sarengaowa
thyme/ cinna- ‘Red Fuji’ the microbial growth, et al. 2018)
mon/oregano oil apples respiration, weight loss,
firmness and browning
Gelatin-chitosan Multilayer Fresh-cut Effective inhibition of (Poverenov
coatings melon the total microbial et al. 2014)
growth
Sodium alginate Nanoemulsion Fresh-cut A greater inactivation (Salvia-
with lemongrass coatings Fuji apples of Escherichia coli Trujillo et al.
essential oil during storage time 2015)
Whey protein- Crosslinked Fresh-cut Reduced the weight (Rossi
pectin coatings apples loss, prevented micro- Marquez
bial growth et al. 2017)
Chitosan Coatings Fresh-cut Decreased in total (Moreira
broccoli mesophilic and et al. 2011b)
psychrotrophic bacteria
counts, inhibited open-
ing florets
Starch with Coatings Minimally Decreased counts of (Santos et al.
carvacrol processed Escherichia coli O157: 2016)
pumpkin H7, and Staphylococ-
cus aureus
Whey protein- Crosslinked Fresh-cut Reduced the weight (Rossi
pectin coatings potatoes and loss, prevented micro- Marquez
carrots bial growth et al. 2017)

bacteria, yeast, and fungi counts of fresh-cut melons (Poverenov et al. 2014). Souza
et al. (2015) reported that the nanomultilayer coatings, made of alginate and
chitosan, considerably inhibited putrefaction of fresh-cut mangoes during 14 days
at 8  C. At the end of the storage period, the lower values of mass loss, pH,
malondialdehyde content, and browning rate were observed in the coated mangoes.
Furthermore, nanoemulsion-based sodium alginate coatings with lemongrass essen-
tial oil at 0.5% or 1% (v/v) were created to completely inhibit the natural microflora
of fresh-cut Fuji apples during 2 weeks at 23  C. The application of this coating on
fresh-cut apples exhibited a faster and greater inactivation of Escherichia coli during
storage time compared with conventional emulsions (Salvia-Trujillo et al. 2015).
Rossi Marquez et al. (2017) reported that transglutaminase crosslinked coatings
prepared from whey protein and pectin were able to totally prevent the weight loss
of fresh-cut potato and carrot at least until the sixth day of storage, which also
maintained the phenolic and carotenoid content of fresh-cut carrot during storage.
Meat, poultry, and seafood products are common sources of proteins, yet sus-
ceptible to the spoilage microorganisms and food-borne pathogens. Thus, the
hydrocolloid-based coatings and films with antimicrobial and/or antioxidant
290 Q. Xiao

Table 8.3 Recent applications of hydrocolloid-based coatings and films in meat, poultry, and
seafood
Coating/film
composites Form Food products Main benefits References
Chitosan with Nanoencapsulation Lamb meat Retention of the (Pabast
Satureja plant coatings good quality char- et al. 2018)
essential oil acteristics,
improvement of
microbiological
safety, and exten-
sion of shelf life
Chitosan– Nanocomposite Ground meat Decreased lactic (Dehnad
nanocelullose films acid bacteria et al. 2014)
population
Sodium caseinate Films Ground beef More pronounced (Emam-
with pomegranate against gram- Djomeh
peel extract positive bacteria et al. 2015)
compared with
gram-negative
bacteria
Distiller dried Films Pork meat Decreased lipid (Yang et al.
grains-soluble oxidation 2016)
protein with tea
extract
Perilla seed meal Films Pork sausages Reduced the micro- (Song et al.
protein with clove bial growth, and 2015)
oil decreased peroxide
value and
thiobarbituric acid
value
Chitosan with Films Cooked cured Decreased the (Ruiz-
thymus moroderi ham counts of aerobic Navajas
and piperella mesophilic bacteria et al. 2015)
essential oil and lactic acid bac-
teria, and lipid
oxidation
WPI with oreg- Coatings Chicken breast Decreased counts (Fernández-
ano/clove essen- fillets of total mesophilic Pan et al.
tial oils aerobic, 2014)
enterobacteriaceae,
Pseudomonas spp.,
and lactic acid
bacteria
Sodium caseinate Nanoemulsion Chicken breast Significantly (Noori et al.
with ginger essen- coatings fillets decreased the total 2018)
tial oil aerobic psychro-
philic bacteria,
maintained food
color
(continued)
8 Coating and Film-Forming Properties 291

Table 8.3 (continued)

Coating/film
composites Form Food products Main benefits References
Chitosan-cyclo- Films Chicken breast A bactericidal (Higueras
dextrin with fillet effect against et al. 2014)
carvacrol Staphylococcus
aureus and
Escherichia coli
O157:H7
Skate skin gelatin Films Chicken Inhibited the (Lee et al.
with thyme essen- tenderloin growth of Listeria 2016b)
tial oil monocytogenes and
Escherichia coli
O157:H7
Chitosan with Coatings Ready-to-eat Reduced the (Guo et al.
lauric alginate deli Turkey growth of Listeria 2014)
ester meat innocua
Sunflower seed Films Smoked duck Decreased popula- (Song et al.
protein-red algae meat tion of Listeria 2013)
with grapefruit monocytogenes
seed extract
Chitosan with Coatings Japanese sea Reduced the (Qiu et al.
citric acid/licorice bass TVB-N levels, 2014)
extract (Lateolabrax showed antioxidant
japonicas) and antimicrobial
effects
Alginate with Vc/ Coatings Refrigerated Efficiently (Song et al.
tea polyphenols bream inhibited the 2011b)
(Megalobrama growth of total via-
amblycephala) ble counts, chemi-
cal spoilage, and
water loss
Chitosan with Coatings Refrigerated Maintained lower (Li et al.
grape seed extract red drum pH values, 2013)
and tea (Sciaenops inhibited the degra-
polyphenols ocellatus) dation of ATP and
fillets lipid oxidation
Chitosan Coatings Frozen Atlantic Maintained the (Soares
salmon color, controlled et al. 2015)
microbial activity
Chicken feather Films Smoked Decreased the (Song et al.
protein-gelatin salmon populations of 2014)
with clove oil Escherichia coli
O157:H7 and
Listeria
monocytogenes,
decreased peroxide
and thiobarbituric
acid value
(continued)
292 Q. Xiao

Table 8.3 (continued)

Coating/film
composites Form Food products Main benefits References
Gelatin with lem- Films Sea bass slices Retarded growth of (Ahmad
ongrass essential lactic acid bacteria, et al. 2012)
oil psychrophilic bac-
teria and spoilage
microorganisms,
lowered changes of
color, K value, and
total volatile base
nitrogen
Quince seed Films Rainbow trout Decreased peroxi- (Jouki et al.
mucilage with fillets dation values, 2014)
thyme/oregano reduced the
essential oil changes of color,
texture, and lipid
oxidation
Alginate-chitosan Multilayer coatings Shrimp Reduced the bacte- (Kim et al.
with grapefruit (Litopenaeus rial count and the 2018b)
seed extract vannamei) off-flavor
Chitosan/gelatin Films Fresh shrimp Decreased counts (Shahbazi
with Ziziphora of bacterial, and 2018)
clinopodioides population of
essential oil and Listeria
pomegranate peel monocytogenes
extract
Starfish gelatin Films Crab sticks Inhibited the (Lee et al.
with vanillin populations of 2016a)
Listeria
monocytogenes

compounds are produced to prolong their shelf life (Table 8.3). The incorporation of
grape seed extract and tea polyphenols into chitosan coatings predominantly delayed
the degradation of ATP and lipid oxidation of red drum during refrigerated storage
(Li et al. 2013). Song et al. (2011b) reported the efficacy of alginate coatings
enriched with Vc and tea polyphenols in inhibiting the growth of total viable counts,
reducing chemical spoilage, and improving sensory quality of refrigerated bream
compared to uncoated samples. According to Kim et al. (2018b), the multilayer
coatings, based on alginate, chitosan, and grapefruit seed extract, were fabricated to
reduce the bacterial counts and off-flavor of shrimp stored at 4  C.
As shown in Table 8.3, the hydrocolloid-based coatings and films with
nanoemulsion, nanoencapsulation, and nanocellulose have been created to extend
shelf life of meat and seafood products. Dehnad et al. (2014) proved that the
application of nanocomposite films based on chitosan and nanocellulose on ground
meat decreased lactic acid bacteria population up to 3.1 logarithmic cycles (com-
pared with nylon packaged sample) at 25  C during 6 days of storage. Noori et al.
(2018) showed that the addition of ginger essential oil nanoemulsion into sodium
8 Coating and Film-Forming Properties 293

caseinate coatings caused significant decrease of total aerobic psychrophilic bacteria

of refrigerated chicken fillets during 12 days. The chitosan coatings included with
nanoencapsulated Satureja plant essential oil were developed by Pabast et al. (2018)
to improve the microbiological safety and prolong shelf life of lamb meat during
chilled storage. Additionally, new plant extracts, as well as hydrocolloids based on
non-conventional sources have been developed as potential ingredients of coatings
and films (Shahbazi 2018; Jouki et al. 2014; Lee et al. 2016a; Ruiz-Navajas et al.
2015).
Cheese is nutritious food derived from milk. The shelf life of cheese is limited due
to the uncontrolled and extensive fungal and bacterial proliferation on its surface.
Table 8.4 shows some recent applications of antimicrobial coatings and films based
on hydrocolloids in cheese. WPI coatings included with thyme and clove essential
oils were produced by Kavas et al. (2015) to prolong the shelf life of semi-hard
kashar cheese. The application of this coating on cheese retarded the growth of
Listeria monocytogenes, Staphylococcus aureus, and Escherichia coli O157:H7
during 60 days of storage. Nanolaminate coatings based on alginate and lysozyme
by LbL technique were fabricated to preserve “Coalho” cheese (Medeiros et al.
2014). After 20 days, coated cheese showed lower values of mass loss, pH, lipidic
peroxidation and higher titratable acidity in comparison with uncoated cheese
(Medeiros et al. 2014). Kim et al. (2018a, b) wrapped the Mozzarella cheese with
chicken bone gelatine films containing cinnamon bark oil (1% w/v) and observed the
reduction in the population of Listeria monocytogenes on mozzarella cheese during
20 days storage. In the current market, the commercialized hydrocolloid-based
coatings, RIOCOBERT and RIOCOBERT PLUS (Becor Barbanza Ltd., A Coruña,
Spain) effectively inhibited the growth of fungi on cheese (Fuciños et al. 2017).
For bakery and nuts products, most applications are hydrocolloid-based coatings
rather than films. The coatings made from potato starch with potassium sorbate and
citric acid were applied to extend shelf life of mini panettone (Ferreira Saraiva et al.
2016). Pinto et al. (2015) coated the cashew nuts with starch-cashew tree gum blend
coatings to reduce moisture absorption, lipid oxidation, and the loss of crisp texture
of nuts. Apart from that, hydrocolloid-based coatings are an additional method to
improve unit operation efficiencies in the food industry. For example, they were
applied in frying pre-treatments to reduce oil content in deep-fat fried products, such
as chicken breasts (Dragich and Krochta 2010), potato chips (Hua et al. 2015), and
fish cake (He et al. 2015). In osmotic dehydration processes of fruits and vegetables,
hydrocolloid-based coatings can prevent large solute uptake without noticeably
affecting water loss (Rodriguez et al. 2016; Azam et al. 2013).

7 Future Perspectives

Although hydrocolloid-based coatings and films have been utilized in food products,
their mechanical and water barrier attributes are still weaker compared to those of
synthetic plastic materials. Several approaches (e.g., bilayer, multilayer,
294 Q. Xiao

Table 8.4 Recent applications of hydrocolloid-based coatings and films in cheese

Coating/film Food
composites Form products Main benefits References
Water chestnut Coatings Mongolian Delayed weight loss and the (Mei et al.
starch-chitosan cheese microbial growth 2013)
containing perilla
oil
WPI-guar Coatings Cheese Decreased water loss, hard- (Ramos et al.
gum-sunflower ness, and color change, 2012)
oil with inhibited pathogenic or
natamycin and contaminant
lactic acid microorganisms
Sodium casein- Coatings Cheese Significantly inhibited the (Moreira
ate-chitosan growth of mesophilic bac- et al. 2011a)
teria, psychrotrophic,
yeasts, and molds
Galactomannan Coatings Ricotta Against Listeria (Martins
with nisin cheese monocytogenes et al. 2010)
WPI with thyme Coatings Semi-hard Significant effect on the (Kavas et al.
and clove essen- kashar antimicrobial activity 2015)
tial oils cheese against Listeria
monocytogenes
WPI with ginger Coatings Kashar Inhibited the growth of (Kavas et al.
essential oil cheese Escherichia coli O157:H7 2016)
and Staphylococcus aureus
Alginate- Nanolaminate ‘Coalho’ Lower values of mass loss, (Medeiros
lysozyme coatings cheese pH, lipidic peroxidation, et al. 2014)
microorganisms’ prolifera-
tion and higher titratable
acidity
Sodium caseinate Films Mini red Against Listeria innocua (Cao-Hoang
with nisin Babybel during storage at refriger- et al. 2010)
cheese ated temperatures
Starch with Films Port Salut Controlled Saccharomyces (Ollé Resa
natamycin and cheese cerevisiae and Listeria et al. 2016)
nisin innocua growth
Zein-carnauba Films Fresh A significant reduction in (Ünalan
wax with Kashar initial Listeria et al. 2013)
lysozyme cheese monocytogenes counts
Puffer fish skin Films Gouda Inhibited the Listeria (Lee et al.
gelatin with cheese monocytogenes growth, 2016c)
Moringa oleifera retarded the lipid oxidation
Lam. leaf extract
Chicken bone Films Mozzarella Displayed antimicrobial and (Kim et al.
gelatine with cin- cheese antioxidant activities, 2018a)
namon bark oil inhibited Listeria
monocytogenes
Red algae with Films Cheese Inhibited the growth of (Shin et al.
grapefruit seed Escherichia coli O157:H7 2012)
extract and Listeria monocytogenes,
decreased peroxide and
thiobarbituric acid values
8 Coating and Film-Forming Properties 295

crosslinking, and bio-nanocomposite films, etc.) are employed to ameliorate prop-

erties of hydrocolloid-based coatings and films. Among them, incorporation of
polysaccharide nanofillers into hydrocolloids to produce bio-nanocomposites has
gained increasing attention in recent years, due to their edibility, remarkable physical
performance, and functional properties (Otoni et al. 2017). Thus, this type of
bio-nanocomposites is expected to be a promising area of research in the future.
On the other hand, the nanodelivery systems, such as nanoencapsulation,
nanoliposomes, nanoemulsion, and nanolaminate, have emerged to enhance the
performance of bioactive agents and improve their effectiveness in preserving food
products. Currently, they are developed as the effective tools to augment the
functionality of hydrocolloid-based coatings and films (Aloui and Khwaldia 2016).
Future research should focus on the development of hydrocolloid coatings and films
based on nanodelivery systems as well as their interactions with food products.
As a bottom-up approach, the structure-properties of hydrocolloid coatings and
films should be studied further. Practically important properties such as WVP, TS,
and EAB must be correlated with molecular structure and mobility in the solid state
to further develop the utilization of polysaccharides. For instance, dextran,
consisting of α-1,6 glycosidic linkages, shows a poor film-forming capacity in
comparison with pullulan or amylose. In addition, dextran shows the largest molec-
ular mobility in the solid state, followed by pullulan and amylose. The physico-
chemical properties and molecular mobility of dextran, pullulan, and amylose in the
solid state are quite different from each other because of the different modes of
glucosidic linkages (Nishinari et al. 1985, 1992). Overall, hydrocolloids as packag-
ing materials still need scientific research to improve their properties, quality and
marketability. Further studies include (1) embracing big data and artificial intelli-
gence (AI) in research and development, e.g., for process simulation, classification,
pattern recognition, and transfer learning; (2) developing new techniques, equip-
ment, machines for large-scale industrial implementation and applications.

Acknowledgemensts The author thanks Prof. Loong-Tak Lim of University of Guelph (Guelph,
Canada), for his helpful suggestions and review of manuscript.

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Chapter 9
Self-assembling Properties

Huiyan Zeng

Abstract A simplistic view of food is that food molecules are assembled into
hierarchical structures. As the two main components in foods, the self-assembling
properties of food proteins and polysaccharides determine the nutritional value,
texture, appearance, taste, odour, and shelf-life of foods. In this chapter efforts are
first made to provide an overview of the concepts, mechanisms, and forces of self-
assembly in a broad context, followed by the specific discussion of the self-assembly
of food proteins and polysaccharides. The advancements of the self-assembled
nanostructures with various morphologies and functionalities are summarized and
discussed to provide a guideline for designing desired food structures and broaden-
ing the applications of food proteins and polysaccharides. These self-assemblies may
also benefit the health of the consumer, when considering their journey in the body,
i.e. the disassembly and reassembly processes. We hope that a better understanding
of the self-assembly rules of food proteins and polysaccharides will spark food
scientists to develop novel functional foods to meet future consumer demands.

Keywords Self-assembly · Proteins · Polysaccharides · Nanostructures · Molecular

forces

1 Introduction

Self-assembly is a ubiquitous process throughout nature and technology (Whitesides

and Boncheva 2002; Whitesides and Grzybowski 2002; Mendes et al. 2013). Nature
uses specific self-assembly of molecules to organize elaborate structures that possess
unique biological functions (Luo et al. 2016). From ordered protein aggregates (e.g.,
viral capsids, collagen and actin filaments, flagella), topologically programmed
nucleic acids (e.g., DNA duplexes, RNA triplexes), to complicated nucleosomes

H. Zeng (*)
University of Strasbourg, CNRS, Strasbourg, France
e-mail: huiyan.zeng@ics-cnrs.unistra.fr

© Springer Nature Singapore Pte Ltd. 2021 307

Y. Fang et al. (eds.), Food Hydrocolloids,
https://doi.org/10.1007/978-981-16-0320-4_9
308 H. Zeng

and ribosomes, these self-assembled structures could perform a number of functions,

such as genome packaging, structural support, force generation, and information
storage and transmission (Goodsell and Olson 2000; Saenger 2008; Luo et al. 2016).
An in-depth understanding of molecular self-assembly is important not only to
reveal the mechanisms of these beneficial biological processes, but also to provide
valuable treatments for human diseases, such as the neurodegenerative diseases that
are caused by abnormal protein self-assembly (Dobson 2003; Chiti and Dobson
2006). Self-assembly is also in the forefront of biotechnology and nanotechnology,
as it provides an excellent tool to build a broad of complex architectures that cannot
be easily achieved by other methods (Lee 2007).
Self-assembly is not a new concept in the food sector, and is omnipresent in both
natural and processed foods. Typical examples include the formation of casein
micelles in milk, oil-bodies in seeds and starch spherulites in plants, and the gelation
of pectins in jelly, micelles in yogurt and soy proteins in tofu (Dickinson and Leser
2007; Ravichandran 2010; Sagalowicz et al. 2017). Two food components, protein
and polysaccharide, are the main self-assembly units in the foodstuffs. The assembly
of food proteins and polysaccharides has attracted much attention over the past two
decades, mainly due to the excellent tech-functionalities of the resultant
nanostructures, such as the assembled nanofibrils that form transparent hydrogels
at low concentration and room temperature, aggregates that stabilize emulsions and
foams, and nanoparticles that deliver drugs and nutrients (Veerman et al. 2003;
Kroes-Nijboer et al. 2012; Yao et al. 2015; Hu et al. 2019). In addition, self-
assembly is correlated to food texture, taste, and appearance. For instance, the
beverage appearance is greatly marred by the self-assembly-induced precipitation.
More importantly, as mentioned above, the hierarchical structure of biopolymers is
directly linked to their unique biological functions. As such, the elaborate structures
generated from the self-assembly of proteins and polysaccharides may bring specific
nutritional values or functions to the consumer. All these examples point out that
food scientists should have a comprehensive understanding of the self-assembly of
proteins and polysaccharides. In this chapter, efforts are made to provide the
concepts, mechanisms, and molecular forces for the self-assembly of food proteins
and polysaccharides, to summarize and discuss the assembled nanostructures in the
food sector, as well as to explain how the self-assembly affects food quality and
functions.

2 Physical Aspects of Self-assembly

2.1 Self-assembly

Self-assembly is a special kind of aggregation whereby this process occurs toward

the state of minimum free energy, mainly through non-covalent interactions, such as
electrostatic interactions, van der Waals interactions, hydrogen bonding, hydropho-
bic interactions, metal coordination bonds, and steric and depletion forces (Lindoy
and Atkinson 2000; Lee 2007; Ninham and Nostro 2010; Jiang et al. 2011; Padua
9 Self-assembling Properties 309

Fig. 9.1 Schematic

illustration of Gibbs free
energy landscape of the
different thermodynamic
states in self-assembly
process. Adapted with
permission from Sorrenti
et al. (2017). Copyright
2017 RSC

and Nonthanum 2012; Billon and Borisov 2016; Sundararajan 2016; Wang et al.
2016; Sorrenti et al. 2017). Despite the entropy loss as the ordering of self-assembly
building units, the self-assembly is energetically favourable because the entropic
cost is greatly offset by the enthalpy gained from the non-covalent interactions
(Rajagopal and Schneider 2004; Padua and Nonthanum 2012).
From the thermodynamic point of view, self-assembly is a process that minimizes
Gibbs free energy. In general, it brings the entity closer to a thermodynamic
equilibrium state (#1 in Fig. 9.1) (Whitesides and Boncheva 2002; Roy et al.
2016; Sorrenti et al. 2017). Many involved intermolecular interactions, as mentioned
above, enable the system to explore different configurations (i.e. walk along the
energy landscape), and to find the most stable one (Sorrenti et al. 2017). In some
cases, the self-assembly process may cause metastable or kinetically trapped states
(i.e. non-equilibrium states, #2 and #3 in Fig. 9.1) (Sorrenti et al. 2017). Due to the
low energy barrier, the metastable structures will eventually evolve into the thermo-
dynamic equilibrium state, even without any intervention. In contrast, the system
will be kinetically trapped in state #3 because of the high energy barrier, if there is no
intervention (Wang et al. 2016; Sorrenti et al. 2017). This means that the outcome of
the self-assembly process, e.g. the morphology of assembled nanostructures,
strongly depends on the experimental parameters and protocols (Sorrenti et al.
2017). In other words, the desired pathway can be rationally selected by appropriate
preparation methodologies, resulting in the assemblies with targeted features starting
from the same building blocks. This is crucial to develop materials with optimized
functionalities (Sorrenti et al. 2017).
From the force point of view, self-assembly can be defined as a delicate balance
of the attractive (driving force) and repulsive (opposition force) intermolecular
forces (Whitesides and Boncheva 2002; Lee 2007; Kedracki 2015; Billon and
Borisov 2016; Roy et al. 2016). The driving forces bring the self-assembly units
together, while opposition forces are in balance with the driving forces (Lee 2007;
Kedracki 2015; Billon and Borisov 2016). Besides, many biological and
bio-mimetic systems show a unique directionality during self-assembly processes,
as well as in many food systems, e.g. the formation of protein nanofibrils and
310 H. Zeng

Table 9.1 Common forces in self-assembly

Attractive driving forces Repulsive opposition forces Directional forces
Electrostatic attraction Electrostatic repulsion Electrostatic interaction
Van der Waals interaction Steric repulsion Hydrogen bonding
Hydrophobic interaction Coordination bond
Hydrogen bonding π-π stacking
Depletion force
Coordination bond
π-π stacking

nanotubes, and polysaccharides helices (Graveland-Bikker and de Kruif 2006; Lee

2007; Cao and Mezzenga 2019; Fittolani et al. 2019). The force responsible for these
directional self-assembly processes is known as directional force or functional force
(Lee 2007; Kedracki 2015; Billon and Borisov 2016). Hydrogen bond, coordination
bond, electrostatic interaction, and π-π stacking are the most common directional
forces (Lee 2007; Wang et al. 2016). These forces can be a part of a driving or
opposition force, but sometimes act exclusively as directional force (Lee 2007).
Therefore, self-assembly is an equilibrium of three classes of forces: driving,
opposition, and directional forces, as displayed in Table 9.1 (Lee 2007). The self-
assembly process is quite random when only the first two classes of forces are in
action. Most of the colloidal self-assembly processes belong to this category. When
the third class of forces is involved with the first two classes of forces, the self-
assembly processes are directional and often functional, leading to the formation of
highly ordered or specific functional structures. In engineering, these three classes of
forces can be greatly affected by a number of external parameters, such as pH, ionic
strength and type, temperature, solvent type, mechanical treatments (pressure, shear,
extension, sonication), or chemical treatments (Lee 2007). Therefore, self-assembly
can be triggered, altered, or terminated by controlling these external parameters,
thereby managing the desired assembly and the assembled nanostructures.

2.2 Forces in Self-assembly

In the self-assembly process, whether it occurs at an atomic-, molecular-, colloidal-,

or macro-length scale, the non-covalent forces rather than the chemical forces play
vital roles (Lee 2007). Even though these non-covalent forces are weak individually,
a large number of such forces in the final can be significant. We first briefly illustrate
these forces and then give an example of a combination of two forces—DLVO
(Derjaguin–Landau–Verwey–Overbeek) theory.
9 Self-assembling Properties 311

2.2.1 Electrostatic Interaction

Electrostatic interaction appears universally for charged objects. In nonpolar media

(e.g. vacuum, air, organic nonpolar liquids), the electrostatic interaction is governed
by the Coulomb’s law (Lee 2007; Sundararajan 2016). The interaction potential U(x)
z1 z2 e2
between two charges of Q1 and Q2 is expressed as: U ðxÞ ¼ 4πε Q1 Q2
0ε x
¼ 4πε 0ε x
, where z1
and z2 are the ionic valence of each charge, e is the elementary charge, ε0 is the
dielectric permittivity of vacuum, ε is the relative dielectric permittivity, and x is the
distance between two charges (Lee 2007). Due to U(x)~x1, the electrostatic inter-
actions in ion-free media can extend over long distance. In polar media (e.g. water,
polar organic liquids), free ions in solutions are able to move to oppositely charged
interfaces, resulting in the formation of a kind of molecular condenser, known as
electrical double layer (Tadros 2013). The thickness of the double layer
(i.e. screening length or Debye q length) decreases with the increase in free ion
ffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
1 ε0 εkB T
concentration, written as κ ¼ 2103 N e2 I , here I is the ionic strength (mol/L),
A

kB is the Boltzmann constant, T is the absolute temperature, NA is the Avogadro

number. The range of the electrostatic forces is then typically: κ1 ¼ 10 nm
(at I ¼ 1 mM), 3 nm (at 10 mM), 0.8 nm (at 150 mM), and 0.3 nm (at 1000 mM)
at room temperature (Ninham and Nostro 2010). Different from the long-range
electrostatic forces in nonpolar media, the electrostatic forces between objects in
polar media become short-ranged, or even can be eliminated by increasing salt
content due to the screening effect (Ninham and Nostro 2010; Tadros 2013).
Therefore, the strength of electrostatic interactions is largely dependent on the
solution ionic strength and pH. The flexibility of polymer chain and the charge
distribution in polymer chain are also significant factors (Cao et al. 2016a).

2.2.2 Van der Waals Interaction

Van der Waals force is generated by dipole or induced-dipole interaction at the

atomic and molecular levels, including three contributions: Keesom interaction
(dipole–dipole), Debye interaction (dipole–induced dipole), and London interaction
(instantaneous induced dipole–induced dipole) (Lindoy and Atkinson 2000;
Parsegian 2005; Lee 2007; Ninham and Nostro 2010). All these interactions have
a scaling of U(x)~x6, thus the van der Waals force quickly vanishes at long
distances between interacting atoms (Lee 2007; Ninham and Nostro 2010). The
van der Waals force between two atoms is weak, but its total collective contribution
to molecular interactions can be substantial (Lindoy and Atkinson 2000; Li and
Alessandra 2018). For instance, for two identical interacting colloids with a radius of
R, the van der Waals interaction potential is U ðxÞ ¼  12x AR
, here A is Hamaker
constant (A  3kBT for proteins) (Hamaker 1937; Parsegian 2005; Lee 2007;
Israelachvili 2011). Thus, the interaction between two protein monomers could be
evident (U(x) ¼ kBT ) by considering R ¼ 4 nm and x ¼ 1 nm.
312 H. Zeng

2.2.3 Hydrogen Bonding

Hydrogen bond is an attractive force between a hydrogen atom which is covalently

bound to an electronegative atom (X-H, donor) and another electronegative atom
bearing a lone pair of electrons (Y, acceptor), depicted as X-HY (Lindoy and
Atkinson 2000; Lee 2007; Ninham and Nostro 2010; Sundararajan 2016). The
common hydrogen bond donors include C-H, O-H, N-H, P-H, F-H, Cl-H, Br-H,
I-H, while N, O, P, S, F, Cl, Br, I, alkenes, alkynes, aromatic π-clouds are the
common acceptors (Lindoy and Atkinson 2000). Hydrogen bond is considered to be
a quite strong and directional interaction (Lee 2007). It is generally stronger than the
van der Waals interaction, but weaker than covalent and ionic bonds (Lee 2007). The
directionality of hydrogen bond results from the hydrogen bond–capable molecules
always interacting only through specific sites (Lee 2007). It is a key player in the
assembly of protein and polysaccharide systems as almost all constituent units in
protein and polysaccharide are capable of forming hydrogen bonding.

2.2.4 Hydrophobic Interaction

Hydrophobic interaction describes the energetic preference of nonpolar objects to

interact with other nonpolar objects in the presence of aqueous solution (Motiejunas
and Wade 2006). This short-range attractive interaction is mainly an entropic effect,
but also affected by enthalpy contribution (Motiejunas and Wade 2006; Lee 2007).
When a nonpolar molecule is present in the aqueous solution, a highly ordered
hydrogen bond network around the nonpolar molecule is formed to minimize the
disruption of this nonpolar object, i.e. the formation of a structured water “cage”
(Schaeffer 2008). In the process of nonpolar molecular association, the nonpolar
molecule system obviously loses its entropy, but the water system gains a significant
increase of entropy that overcomes the entropy loss of nonpolar molecules (i.e. total
ΔS is positive) (Lee 2007; Schaeffer 2008). Moreover, the enthalpy is increased as
some of hydrogen bonds that form the “cage” are broken in the association process,
but this effect is limited compared to the entropic effect (Lee 2007; Schaeffer 2008).
Therefore, the Gibbs free energy change is negative (ΔG ¼ ΔH  TΔS, ΔS ¼ large
positive value, ΔH ¼ small positive value), implying that the hydrophobic effect is
spontaneous. Similar to van der Waals forces, hydrophobic interactions are individ-
ually weak, but the total contribution to molecular interactions can be significant
(Schaeffer 2008). The strength of hydrophobic interaction is associated with the
solubility of the nonpolar molecules as well as the quality of the steric match
between the molecules (Schaeffer 2008). Most proteins (possessing nonpolar
amino acids) and some polysaccharide derivatives (such as MC and HPMC) show
a significant hydrophobic character.
9 Self-assembling Properties 313

2.2.5 Steric Repulsion

Steric repulsion is a common force between colloidal particles when water-soluble

polymers are tightly adsorbed or grafted onto the surface of colloidal objects
(Fig. 9.2a) (Lee 2007). It mainly arises from the loss of configuration entropy
when two polymer layers are overlapped (Cooper 2014). Steric repulsion is a
long-range force and can reach up to ~10Rg (Rg is the gyration radius of the polymer
chain) (Lee 2007). For effective steric repulsion, water-soluble polymers in the
diffuse layer must satisfy the following three requirements: firstly, the polymers
should be tightly anchored to the colloidal particles; secondly, part of the polymer
chain should extend into the bulk solution; thirdly, there is no significant exposure of
the colloidal surface (Cooper 2014).

2.2.6 Depletion Attraction

Depletion force is the common force for the colloidal particles considering the
presence of non-adsorbing polymers (Fig. 9.2b) (Lee 2007; Lekkerkerker and

Fig. 9.2 Schematic illustration of steric repulsion and depletion attraction between two colloidal
spheres. (a) Steric repulsion between polymer-adsorbed colloids. (b) Depletion attraction between
two colloidal particles in the presence of non-adsorbing polymers or molecules
314 H. Zeng

Tuinier 2011). A depletion region of polymers is generated when the colloidal

particles are close enough to each other (smaller than the size of polymers, 2Rg),
as the polymers are being squeezed out of this region (Lee 2007). The osmotic
pressure force that is exerted by the polymers in the outside region (outside of
depletion region) exceeds that in the inside region. This, therefore, induces a net
attractive force between the colloidal particles (Lee 2007; Lekkerkerker and Tuinier
2011). The depletion force can determine the stability of colloids when there are no
other significant attractive forces (Lee 2007). It can be strengthened by increasing
polymer concentration and molecular weight (Lee 2007). Colloids with low curva-
ture (e.g. nanorods) experience this attraction more strongly as the depletion attrac-
tion scales with the excluded volume of the colloids.

2.2.7 DLVO Theory: A Case of the Combination of van der Waals

Attraction and Electrostatic Repulsion

The DLVO theory is a useful tool to describe the self-assembly process of charged
colloids, which is the combination of two inter-colloidal forces: van der Waals force
that acts as the attractive force and electric double-layer interaction as the repulsive
force. Their total interaction potential U(x) can be written as (Adair et al. 2001;
Mezzenga and Fischer 2013):

2πσ 2 R κx AR
U ð xÞ  e 
κ 2 ε0 ε 12x

σ and R are the net surface charge density and radius of colloidal particles, respec-
tively, x is the separated distance of a pair of colloids. The plots of this equation at
different ionic strengths are shown in Fig. 9.3, by considering R ¼ 5 nm, σ ¼ 20 mC/
m2, T ¼ 298 K. The electrostatic force gives a positive term and varies with the
solution ionic strength. In contrast, the van der Waals attraction gives a negative term
and is independent from ionic strength (Fig. 9.3a) (Adair et al. 2001). The sum of
these two forces at different ionic strengths is given in Fig. 9.3b. At low ionic
strengths, particles have net repulsion at large and intermediate separations, and the
approaching of colloids requires high kinetic energy due to the high energy barrier.
Thus, in these cases, colloids are separately suspended in the solution. At high ionic
strengths, the energy barrier is lowered and particles can overcome it more easily,
leading to the aggregation or self-assembly (Adair et al. 2001).
9 Self-assembling Properties 315

Fig. 9.3 An example of DLVO interaction potential. (a) Electrostatic repulsion potential versus
particle distance x at different ionic strengths, and van de Waals attraction potential versus particle
distance x. (b) The sum interaction potential of electrostatic and van der Waals forces. R ¼ 5 nm,
σ ¼ 20 mC/m2, T ¼ 298 K are used for the calculation

2.3 Self-assembly of Food Proteins and Polysaccharides

2.3.1 Protein Self-assembly

Understanding the mechanisms and processes of protein self-assembly is essential to

biologists and medical scientists, since it is related to many biological and physio-
logical activities, as mentioned above. The understanding of food protein self-
assembly is equally important to food scientists due to a broad range of food-
related implications and applications. For instance, protein self-assembly can be
harnessed for protein purification through phase separation or crystallization, or can
be problematic during storage (Carpenter and Manning 2002; Flickinger 2013;
McManus et al. 2016).
Most natural food proteins possess globular or fibrillar conformations (Mezzenga
and Fischer 2013; Nicolai 2019). These protein structures result from a combination
of the numerous interactions between amino acids, i.e. the self-assembly of poly-
peptide chain. Depending on the side group, the amino acids in food proteins can be
divided into: nonpolar, polar, and ionic, which mainly contribute to hydrophobic
interactions, hydrogen bonding, and electrostatic interactions, respectively. In glob-
ular conformations, the polypeptide chain is folded into compact spherical shape
with most of the nonpolar amino acids buried in the interior, and the polar and ionic
amino acids predominately located at the surface (Fig. 9.4a) (Mezzenga and Fischer
2013; Jones 2015; McManus et al. 2016; Cho and Jones 2019). The driving forces
for this configuration include the hydrophobic effect as well as other forces, such as
hydrogen bonding that contributes to the formation of protein secondary structure
316 H. Zeng

Fig. 9.4 Schematic illustration of food protein self-assembly and the resultant nanostructures. (a)
The self-assembly of polypeptide chain into globular proteins. (b, c) The self-assembly of natural
proteins into crystals (b) or amorphous aggregates (c). These processes are often reversible since the
protein structure is remained. (d) Protein denaturation leads to the (partial) unfolding and/or
hydrolysis of proteins. (e) The unfolded and hydrolyzed proteins can further assemble into amyloid
fibrils or amorphous aggregates. These processes are often irreversible

(α-helices and β-sheets) (Jones 2015). Indeed, hydrogen bonding is prevalent in

proteins by considering all amino acids containing amine- and carbonyl-groups.
Fibrous proteins often have specific amino acid sequences that favour twisting of the
polypeptide. For example, gelatins are rich in Glycine-X-Y sequence that supports
the twisting of gelatin chains with the formation of triple helixes; X and Y are mostly
proline and hydroxyl-proline (Russell et al. 2007; Hafidz et al. 2011; Cao et al. 2015;
Jones 2015).
9 Self-assembling Properties 317

Since globular proteins are generally viewed as colloids, a number of models for
interpreting colloid self-assembly can be used to understand the self-assembly
behaviour of globular proteins. The simplest model is the DLVO model, as discussed
above, which could interpret many protein behaviours, such as the aggregation of
proteins by adjusting pH or adding salts. However, DLVO cannot explain certain
protein behaviour, such as protein crystallization at high salt contents (Piazza 1999;
Mezzenga and Fischer 2013). Other studies indicate that the protein aggregation may
originate from the presence of depletion forces (Mezzenga and Fischer 2013;
McManus et al. 2016). It is worth noting that many models are employed to
understand protein self-assembly with varying degree of success, yet no model
captures all protein aggregation features (Mezzenga and Fischer 2013). This is
possibly caused by the existence of many other forces (e.g. hydrophobic effect,
hydrogen bonds, specifically ionic bindings) and effects (e.g., surface charge distri-
butions, molecular recognition) that contribute to the complexity of protein self-
assembly (Mezzenga and Fischer 2013). The self-assembly of globular proteins, in
native state, can lead to the formation of crystals or amorphous aggregates (Fig. 9.4b
and c), and this process is often reversible.
Protein self-assembly could also start from the denatured, unfolded, or hydro-
lyzed proteins, which is actually more frequently observed in the food systems
(Fig. 9.4d). Loss of native structures leads to changes in the capacity of those
proteins to interact with each other, and further determine their ability to form
supramolecular assemblies (Jones 2015). For instance, the exposure of nonpolar
amino acids by unfolding the globular proteins supports the formation of
intermolecular forces (Jones 2015; Li et al. 2018). Knowledge of protein character-
istics in the chosen environment is essential to the desired assembly, and there are
mainly two routes for the protein self-assembly at the denaturation condition:
fibrillization and random aggregation, leading to the formation of amyloid fibrils
and amorphous aggregates (Fig. 9.4e and f). The most studied condition for trigger-
ing this type of self-assembly is heating proteins at various pHs and ionic strengths
(van der Linden and Venema 2007; Nicolai and Durand 2013; Jones 2015; Schmitt
et al. 2016; Cao and Mezzenga 2019). At pH in the neighbourhood of protein
isoelectric point (IEP) or at high ionic strengths, the effective charge of the protein
is remarkably suppressed so that amorphous aggregates (i.e. large fractal dimension)
generate during thermal treatment, arising from the loss of opposition electrostatic
repulsion (right side of Fig. 9.5). As the solution pH leaves from the protein IEP, the
effective charge of the protein increases and aggregates become relatively less
amorphous (middle of Fig. 9.5). Instead, protein aggregation produces fibrous
structures when the pH is significantly far from IEP, since the highly effective
charge on the protein surfaces makes them favourable for interactions only among
discrete regions on the protein surface (left side of Fig. 9.5). A notable example can
be found in β-lactoglobulin protein system. The morphology of protein aggregates
remarkably depends on the solution pH: amyloid fibrils and fibrous strands formed at
pH 2 and pH 7, respectively (far from β-lactoglobulin IEP); particulates formed at
pH 5.2 ( IEP); microgels formed at pH 4.7 and 5.9 (near IEP) (Schmitt et al. 2016).
318 H. Zeng

Fig. 9.5 Schematic representation of the effect of pH and ionic strength on protein self-assembly
during heat treatment. Adapted with permission from van der Linden and Venema (2007). Copy-
right 2007 Elsevier

2.3.2 Polysaccharide Self-assembly

Based on the charge nature, polysaccharides can be classified into cationic

(chitosan), anionic (alginate, pectin, carrageenan, gellan gum, hyaluronic acid),
and neutral (agarose, pullulan, dextran) (Kontogiorgos 2015; Stephen et al. 2016).
Despite that most food polysaccharides only consist of 1–3 constituent units, the
type of linkages, isomeric forms, esterification, the branching and periodicity of
constituent units, and the wide range of molecular weight contribute to their great
diversity in structure and property (Kontogiorgos 2015; Stephen et al. 2016). A
notable example is that of cellulose and amylose. They have the same repeating unit
(glucose), but the different linkage (β-D-(1!4) in cellulose and α-D-(1!4) in
amylose), which leads to their extremely different digestion and assembly behav-
iours. Repulsive interactions in polysaccharide self-assembly often are the steric
repulsion and sometimes electrostatic repulsion for charged ones. Attractive forces
are the van der Waals interaction, hydrogen bonding, and sometimes the hydropho-
bic interaction and ionic binding for certain polysaccharides. Hydrogen bonding and
ionic binding are the important directional forces in polysaccharide self-assembly.
Similar to food proteins, temperature, pH, and salt are the most common triggers
for the self-assembly of food polysaccharides. Either increasing or lowering tem-
perature can induce the self-assembly (Nishinari and Zhang 2004; Stephen et al.
2016). Agaroses, carrageenans, or gellan gums experience the transitions of coil-to-
helix and helix-to-super helix in the cooling process, driven by the formation of
hydrogen bonding and ionic binding. The helical structures in polysaccharides are
still controversial, could be single, double, or triple helixes (Fig. 9.6b) (Schefer et al.
2014; Cao et al. 2016b; Stephen et al. 2016; Fittolani et al. 2019). In contrast,
methylcellulose and hydroxypropyl methylcellulose are assembled in the heating
process, mainly driven by the hydrophobic force. During heating, the solvated cage-
like structures (formed through hydrogen bonds between water and cellulose deriv-
atives) are destroyed and thereby hydrophobic regions are exposed, resulting in the
9 Self-assembling Properties 319

Fig. 9.6 Typical polysaccharide structures at molecular levels. (a) Left: “egg-box” model of
alginate or pectin in the presence of divalent ions, e.g. Ca2+; Right: coordination of Ca2+ in a cavity
created by a pair of guluronate sequences. The open circles represent Ca2+ ions and the black dots
represent the oxygen atoms possibly involved in the coordination with Ca2+. Adapted with
permission from Fang et al. (2007). Copyright 2007 ACS. (b) Single, double, or triple helical
structures in polysaccharides, such as carrageenan (single helix), agarose (double helix), curdlan
(triple helix). (c) Hydrophobic association in cellulose derivatives, (e.g. methylcellulose,
hydroxypropyl methylcellulose). At relatively high temperatures, the solvated cage-like structures
(formed through hydrogen bonds between water and cellulose derivative chains) are destroyed and
thereby hydrophobic regions are exposed, causing the formation of hydrophobic junction zones.
Adapted with permission from Li et al. (2001). Copyright 2001 ACS

formation of hydrophobic junction zones (Fig. 9.6c) (Li et al. 2001, 2002; Shen et al.
2016).
For ionic polysaccharides, pH and salt not only affect the electrostatic force but
also have other important effects. The effect of pH is correlated to the dissociation
constant (pKa). The charge magnitude of ionic polysaccharides depends on the
solution pH relative to the pKa. For instance, alginate (pKa  3.8) is slightly negative
(or near  at pH 2 and strongly negative at pH 7, by referring pH  pK a ¼
 neutral)
log 10 CO 2 =½CO2 H ). The protonation of COO at pH 2 not only weakens the
electrostatic repulsion but also enhances the hydrogen bonding due to COOH with
high hydrogen bond forming ability, resulting in the self-assembly (or gelation) of
320 H. Zeng

alginates at low pH (Draget 2009; Draget et al. 2016). The mechanism for the
assembly of chitosan (pKa  6.3) through increasing pH is considered to be similar,
i.e. transition of NH3+ to NH2 (Yi et al. 2005; Pillai et al. 2009; Zargar et al. 2015;
Shen et al. 2016). Salt plays a vital role in the electrostatic force via altering the
Debye screening length, as discussed above. More importantly, some specifically
ionic bindings make polysaccharides with a complex self-assembly behaviour. For
example, multivalent cations, e.g. Ca2+, specifically bind to alginate or pectin,
causing the formation of an ordered “egg-box” structure (Fig. 9.6a) (Fang et al.
2007). It should be noted that, different from protein systems, the assembly induced
by temperature, pH, or salt is often reversible for polysaccharide systems.

3 Self-assembled Nanostructures

Nanostructured materials are the forefront of many fields due to their unique and
outstanding properties, and self-assembly is broadly considered as a promising
approach to produce these nanostructures. In principle, the nanostructures generated
from food proteins and polysaccharides could further enrich the versatility of
nanostructured materials in terms of category and function due to their nutritional
value, biodegradability, biocompatibility, safety, etc. Here we summarize the food
protein and polysaccharide nanostructures with different morphologies, and their
formation conditions and potential applications.

3.1 Protein Self-assembled Nanostructures

Under certain conditions, proteins can self-assemble into a variety of structures with
different sizes and morphologies, including crystals, nanofilaments, nanotubes, and
amorphous aggregates. In this part, we will first introduce a naturally assembled
protein nanostructure and then discuss the nanostructures produced by the
processing.

3.1.1 Natural Self-assembled Nanostructure—Casein Micelles

Milks contain large quantities of protein-based nanostructures, known as casein

micelles. These colloidal particles, typically have an average diameter of ~200 nm,
are naturally assembled from the phosphoproteins—caseins (αs1-casein, αs2-casein,
β-casein, κ-casein) and calcium phosphate, driven by the forces of hydrogen bond-
ing, ionic bridging, hydrophobic interaction, electrostatic interaction, and van der
Waals attraction (Dalgleish 2011; de Kruif et al. 2012; Jones 2015). Within casein
micelles, the balance of the hydrophobic and hydrophilic amino acids not only
allows formation of this micelle nanostructure, but also helps in retaining the
9 Self-assembling Properties 321

individual character of the micelles (i.e. adequately stable as a suspension)

(Kontogiorgos 2015). Although the composition and forces in casein micelles are
well understood, their structure, especially the interior structure, remains a matter of
debate (Dalgleish 2011; Mezzenga and Fischer 2013). Various models have been
proposed, but it is generally agreed that calcium phosphate is responsible for forming
salt bridges between phosphoseryl residues on the β- and α-caseins, and κ-casein is
predominantly distributed on the micelle surface and contributes to stabilizing
micelles (Mezzenga and Fischer 2013; Cho and Jones 2019). Although it is not
possible to duplicate the exact assembly of the casein micelles, casein proteins have
been demonstrated to assemble micelles-like structures by reincorporating calcium
phosphate and citrate ions at milk-relevant contents (Jones 2015). Besides, many
methods are available to disassemble and reassemble natural casein micelles, which
are useful to create novel nanostructures for controlled delivery purposes (Jones
2015; Cho and Jones 2019).

3.1.2 Amorphous Aggregates

Amorphous protein aggregates could be generated from the self-assembly of native

proteins (i.e. in a mild condition without the protein denaturation process). This type
of aggregates is often reversible due to the lack of significant changes in protein
structures and the absence of strong forces. For instance, the clusters formed in high
concentration lysozyme protein solution are reversible; the clusters are disassembled
by lowering protein concentration (Lu et al. 2008). In contrast, most amorphous
aggregates in the food sector are produced by protein unfolding and then assembly,
which are generally irreversible because of the significant changes in protein struc-
tures and the significant aggregation interactions between proteins. The formation of
stable suspensions of amorphous aggregates requires an intermediate surface charge
and low protein concentration, otherwise leading to the formation of precipitates or
bulk gels (Nicolai and Durand 2013; Schmitt et al. 2016). In most cases, it is not
straightforward to form homogeneous nanoaggregates by simply heating globular
protein solution. Particularly, Schmitt et al. (Schmitt et al. 2009) found that stable
suspensions of roughly spherical protein nanoparticles (with a hydrodynamic radius
of ~200 nm) can be formed by heating β-lactoglobulin without added salt in two
narrow pH ranges (around pH 4.6 and around pH 5.8).

3.1.3 Nanofilaments

The formation of filamentous nanostructures needs more specified and stringent

conditions than amorphous aggregates. Typically, two common filamentous struc-
tures could be produced from food proteins: strand-like objects formed when heating
proteins at neutral pH and low salt content; amyloid fibrils formed when heating
proteins at low pH and low salt content. These filamentous structures are the
promising materials owing to their unique properties. For instance, the high aspect
322 H. Zeng

ratio allows them to form gels or significantly increase solution viscosity at very low
protein concentration (Veerman et al. 2003; Kroes-Nijboer et al. 2012).
In the formation of strand-like objects, the intermolecular interactions at certain
sites (e.g. disulphide bonds) bring protein monomers together and lead to directional
growth, where electrostatic repulsion is in balance with these forces to prevent
structure collapse (van der Linden and Venema 2007; Nicolai and Durand
2013; Nicolai et al. 2011). The strands are structurally less ordered than amyloid
fibrils due to the fact that the protein chains are confined (low unfolding and
hydrolysis extent) (Nicolai and Durand 2013). The resultant strands at neutral pH
often have diameters less than 10 nm and lengths between tens and hundreds of
nanometres (Nicolai and Durand 2013).
Amyloid fibrils are characteristic with a cross-β structure in which continuous
hydrogen-bonded β-sheets run along the fibrils (McManus et al. 2016; Chiti and
Dobson 2017; Eisenberg and Sawaya 2017). These nanofibrils formed from different
proteins are similar: unbranched filamentous structures with a few nanometres in
diameter and up to several micrometres in length (McManus et al. 2016; Chiti and
Dobson 2017; Eisenberg and Sawaya 2017). Heating proteins at low pH and low
ionic strength is often used to prepare food protein amyloid fibrils. In this procedure,
the protein monomers are first hydrolyzed and unfolded, and then assembled into
amyloid fibrils. Hydrophobic interactions, hydrogen bonding, and sometimes π-π
stacking are the dominant attractive forces for holding the protein nanofibril struc-
tures. In contrast, electrostatic repulsion is the main opposition force to prevent the
structure collapse. Indeed, the final fibril morphology is largely affected by the
solution ionic strength, since it modulates the strength of electrostatic interactions.
Long semiflexible fibrils are generated at low ionic strength, whereas short wormlike
fibrils prevail at higher ionic strength (Loveday et al. 2010, 2017; Cao and Mezzenga
2019). This arises from the fact that the strong electrostatic repulsion at low ionic
strength leads to the attachment of building blocks to the growing fibrils in a well-
ordered arrangement, whereas at higher ionic strength the growth of fibrils is more
haphazard and chaotic (Loveday et al. 2017; Cao and Mezzenga 2019). Other
factors, such as pH, temperature, protein concentration, stirring speed, co-solvent,
and some chemicals could also greatly affect the protein fibrillization process and the
final nanofibril morphology (Cao and Mezzenga 2019).

3.1.4 Nanotubes

Nanotube is one of the most promising materials from the last century. Carbon
nanotubes could be used to build probes and sensors, to store energy and hydrogen
gas, and to serve as field emission displays and radiation sources, etc. (Baughman
et al. 2002; de Volder et al. 2013). Peptide nanotubes are also of immense interest
due to their diverse bio-functionalities which lead to numerous potential applications
in nanotechnology as well as in biomedicine (Scanlon and Aggeli 2008; Hamley
2014). Indeed, it has been proved that the nanotube structure provides superior drug
9 Self-assembling Properties 323

a b
enzyme Ca2+

α-lactalbumin hydrolysed nanotube

molecules 50 nm

Fig. 9.7 The formation of α-lactalbumin nanotubes. (a) Schematic illustration of the self-assembly
of partially hydrolyzed α-lactalbumin into nanotubes in the presence of Ca2+. (b) TEM image of
negatively stained nanotubes. The dark line in the middle corresponds to the hollow of the
nanotube. Reproduced with permission from Graveland-Bikker and de Kruif (2006). Copyright
2006 Elsevier

loading and uptake, and improved release profiles of therapeutics, compared to the
spherical counterparts (Geng et al. 2007; Tiwari et al. 2017). Nanotubes, generated
from food proteins, could even have new possibilities in food, pharmaceutical, and
cosmetic fields, due to their nutritional value, biodegradability, and biocompatibility.
Unfortunately, food protein-derived nanotubes are relatively less studied and less of
concern. To the best of author’s knowledge, food protein nanotubes have so far only
been reported for α-lactalbumin, lysozyme, and albumin (Graveland-Bikker and de
Kruif 2006; Lara et al. 2013; Tiwari et al. 2017).
The formation of α-lactalbumin nanotubes includes two steps: first, the proteins
are partially hydrolyzed by enzymes, second, the hydrolyzed proteins are self-
assembled into nanotubes in the presence of suitable multivalent ions (Fig. 9.7)
(Graveland-Bikker and de Kruif 2006). These assembled nanotubes typically have a
diameter of ~20 nm and few micrometres in length. The prerequisites to form these
nanotube structures are at an intermediate protein concentration and in the presence
of appropriate cations. Various di- and tri-valent cations could trigger this self-
assembly, including Ca2+, Mn2+, Zn2+, Cu2+, and Al3+, except Ba2+ and Mg2+.
Lysozyme nanotubes are generated by heating proteins at pH 2 and 90  C for 30 h
(Lara et al. 2013). Under this condition, lysozyme proteins are first hydrolyzed and
then assembled into amyloid fibrils with multi-stranded helical ribbon morphology.
In the final stage, the helical ribbons progressively closed into nanotubes. Hence,
these lysozyme nanotubes can also be recognized as a state of amyloid fibrils. The
nanotube diameter is dominated by the initial helical ribbons width and the folding
angle, which ranges from ~40 to 150 nm. It should be noted that many protein
amyloid fibrils possess multi-stranded helical ribbon morphology (Lara et al. 2011),
and thereby could be the source to produce nanotube structures.
Albumin nanotubes are formed by heating proteins (80–85  C) in the presence of
glutathione and paclitaxel, which respectively function as the accelerator of protein
unfolding and the nucleation core of self-assembly (Tiwari et al. 2017). By exposing
buried nonpolar residues, glutathione greatly boosts the interaction of albumin and
hydrophobic paclitaxel. Afterwards, the crystallization of the paclitaxels that are
324 H. Zeng

located in the core contributes to the growth of nanotubes. In this case, nanotubes
have a diameter of 70–120 nm and length of up to few micrometres.

3.2 Polysaccharide Self-assembled Nanostructures

The most common function of food polysaccharides is food structuring, which

requires the creation of structures up to millimetres or centimetres with specific
mechanical properties, that is, the formation of three-dimensional macrostructures
(such as bulk gels) (Kontogiorgos 2015; Stephen et al. 2016; Foegeding et al. 2017).
Therefore, in food structuring applications, the assembly of polysaccharides
involves several length scales, ranging from atomic, molecular, microscopic to
macroscopic scales (Kontogiorgos 2015). Current functions related to polysaccha-
rides are not only limited to food structuring, but also include nanoplatforms for
targeted delivery and biomedical imaging (Saravanakumar et al. 2012; Debele et al.
2016; Swierczewska et al. 2016). For example, due to the specific cellular recogni-
tion of some polysaccharide colloidal nanoparticles, the drug, gene, or nutrient
delivery systems derived from these nanoparticles show superior performances
(Saravanakumar et al. 2012; Salatin and Yari Khosroushahi 2017). In these appli-
cations, individual nanoparticles should be retained and their aggregation must be
avoided (Kontogiorgos 2015). Some assembling approaches, e.g., gelation triggered
by salt, pH, temperature alteration, electrostatic complexation of opposite charged
polysaccharides, have been employed to prepare polysaccharide nanoparticles, but
often in a mechanical intervention and/or a low polymer concentration to prevent the
bulk gelation. Controlling polysaccharide self-assembly in nanometre length scale is
not as easy as that in protein system, because dispersions of polysaccharides in
aqueous solutions exhibit very low interfacial tension (Kontogiorgos 2015). There-
fore, most of self-assembly-derived polysaccharide nanoparticles are formed by the
aid of other methods or chemical modification.
A common approach to produce polysaccharide nanoparticles with controllable
size or shape is to first establish the liquid droplets and then self-assemble poly-
saccharides in these confined droplets (Burey et al. 2008; Shewan and Stokes 2013;
Joye and McClements 2014). Extrusion and emulsification always are used to
produce these droplets, and the size and shape of nanoparticles are controllable by
altering the applied experimental conditions. It should be noted that a “switching”
effect can be built into these polysaccharide nanostructures that respond to stimula-
tion in vitro or in vivo, due to the reversible feature of polysaccharide assembly
process (Myrick et al. 2014). Another prominent approach to produce polysaccha-
ride nanoparticles, especially when designing delivery nanoplatforms, is through the
assembly of hydrophobic segment-grafted hydrophilic polysaccharides, i.e. the
assembly of amphiphilic copolymers, as discussed below (Myrick et al. 2014;
Debele et al. 2016; Swierczewska et al. 2016). Such copolymer assembled
nanostructures is known as promising drug carriers, and even could lower drug
toxicity because the hydrophilic polysaccharide parts are often less absorbed by
9 Self-assembling Properties 325

Fig. 9.8 Schematic illustration of cellulose nanofibrils and nanocrystals produced from fibre cell
walls by mechanical and chemical treatments, respectively. Reproduced with permission from Salas
et al. (2014). Copyright 2014 Elsevier

normal tissues but can accumulate in cancerous tissues through the enhanced
permeability and retention effect (EPR effect) (Myrick et al. 2014; Keservani and
Sharma 2018).
Besides the above-mentioned methods, the assembled nanostructures can also be
separated from many natural materials since they are already existent in nature but
are highly structured. For example, cellulose nanofibrils and cellulose nanocrystals
can be dissociated from fibre cell walls by mechanical, chemical, enzymatic treat-
ment, or a combination thereof (Fig. 9.8) (Xu et al. 2013; Salas et al. 2014; Patel
2018). The abundance of OH groups in cellulose chains facilitates the formation of
hydrogen bonding, resulting in the assembly into highly ordered structures
(i.e. crystalline regions) that alternate with disordered structures (i.e. amorphous
regions) (Salas et al. 2014; Patel 2018). Cellulose nanocrystals are usually produced
through ultrasonic acid hydrolysis, in which most of the amorphous regions are
degraded and the crystalline parts are remained (Salas et al. 2014; Patel 2018). The
yielded cellulose nanocrystals often possess a diameter of 10–50 nm and a length of
several hundred nm (Habibi et al. 2010; Xu et al. 2013). In contrast, cellulose
nanofibrils contain both amorphous and crystalline regions and have a larger aspect
ratio than cellulose nanocrystals (Salas et al. 2014; Patel 2018). These cellulose
nanostructures have gained great attention in the scientific community, including in
food science, due to a wide spectrum of unique properties such as high aspect ratio,
excellent mechanical strength and inherent abundance. They can be used to stabilize
326 H. Zeng

emulsions and foams, to prepare hydrogels and aerogels, and to fabricate food-grade
packing materials, etc. (Salas et al. 2014; Coffey et al. 2016; Ullah et al. 2016; Patel
2018).

3.3 Protein-co-polysaccharide Self-assembled

Nanostructures

Surfactants (e.g. mono- or diglycerides) have the unique property of self-assembling

into a broad range of nanostructures, from micelles and vesicles to membranes and
cubic phases, as these molecules possess discrete hydrophobic and hydrophilic
moieties (Smart et al. 2008; Jones 2015). Despite that proteins and polysaccharides
could self-assemble into several nanostructures as discussed above, these structures
are not as diverse and controllable as the specific structures assembled from surface-
active agents. This is due to the lack of a significant anisotropic distribution of
hydrophobic and hydrophilic moieties in food polymers (Jones 2015). One method
to increase this anisotropy is to attach a second component, forming copolymers. A
number of chemical techniques could be used to generate these copolymers, but
many of them are unfavourable for food formulations (Jones 2015).
Maillard reaction, as one of most common food chemical reactions, is widely
used in the food sector to improve food tastes and appearances. It is also an ideal
approach to produce food-grade copolymers, typically protein-co-polysaccharide.
During Maillard reaction, a reducing end of a polysaccharide and a free amine of a
protein are conjugated, with the formation of covalently bonded protein-co-polysac-
charide (Kato 2002; Oliver et al. 2006; Jones 2015; de Oliveira et al. 2016). In this
type of copolymers, the protein part is often the relatively hydrophobic component
that forms the internal phase of micelle-like structures (Fig. 9.9) (Smart et al. 2008;
Jones 2015). These food-grade copolymers are the ideal platforms to encapsulate
and deliver bioactive compounds. For instance, casein-co-maltodextrin assembled
colloids have very high stability, and could reduce the oxidization of encapsulated
vitamin D (Markman and Livney 2012; Jones 2015).

4 Tech-functionalities

Proteins and polysaccharides are widely used in the food sector as thickeners, gelling
agents, emulsifiers, foam stabilizers, fat replacers, and so on (Phillips and Williams
2009). Self-assembly could happen at different length scales, produce diverse
structures, continuum at different time scales (Whitesides and Grzybowski 2002).
The appropriate control of self-assembly can produce novel foods and enable new
applications. For example, the self-assembly-induced phase separation could pro-
duce diverse structures and thus lead to different texture properties. Moreover,
9 Self-assembling Properties 327

Fig. 9.9 Different geometries could be produced from protein-co-polysaccharide in selective

conditions. Adapted with permission from Smart et al. (2008). Copyright 2008 Elsevier

the assembled nanostructures generally possess better tech-functionalities than the

individual proteins and polysaccharides, owing to their specific morphologies and
structural alterations (e.g. high aspect ratio and heat resistance for protein
nanofibrils). Many examples are mentioned above, such as forming cold-set gels,
stabilizing emulsions and foams, and delivering drugs and nutrients. It is worth to
note that some polysaccharides are capable of binding with a particular group of
receptors at the cell surfaces, thereby can be engineered to prepare desired platforms
to enhance the bioavailability of the loaded biomolecules, including food nutrients
and bioactive compounds (Schmitt et al. 2016; Swierczewska et al. 2016; Salatin and
Yari Khosroushahi 2017).

5 Disassembly and Reassembly in the Gastrointestinal

Tract

As discussed above, the self-assembly is extremely vital to both natural and

processed foods as it determines food appearance, texture, shelf-life, etc. However,
the final functions of foods mainly depend on their biological fates in the gastroin-
testinal tract. As is well known, the macromolecular assembly plays a key role in
biological phenomena; analogously, food polymer assembly process in the body
certainly affect food functions. Assembly-related processes, such as self-assembly,
disassembly, and reassembly, are present in the human digestion and absorption
processes. During eating, foods are first broken down in the mouth and then entered
into stomach and intestine. Afterwards, foods are degraded into small molecules that
328 H. Zeng

are absorbed by intestinal walls and eventually enter into the bloodstream. The
optimal control of the disassembly of food protein and polysaccharide
nanostructures can bring not only the basic nutritional value but also other additional
functions, e.g., by delivering incorporated bioactive compounds to the targeted sites
(Mcclements et al. 2009; Joye and McClements 2014; McClements 2014; Yao et al.
2015). In contrast, inappropriate disassembly could cause some safety issues. For
example, both high-speed (leading to high local concentration) and wrong-site
release (such as in the colon altering the gut microbiota) of vitamins will harm the
consumer health (McClements and Xiao 2017). Moreover, due to the condition
change in the gastrointestinal tract, e.g. pH, the assembled nanostructures could
further self-assemble into advanced nanostructures that alter the biological fates of
food proteins and polysaccharides by controlling their digestion and absorption
abilities. Besides, some degraded molecules can reassemble into nanostructures in
the gastrointestinal tract, such as the amyloid fibrils formation in the stimulated
gastric condition (Bateman et al. 2011). Understanding the self-assembly, disassem-
bly, and reassembly of food protein and polysaccharide nanostructures during
digestion and absorption processes is vital to maximize food nutritional value and
improve food quality and even safety.

6 Summary, Challenges, and Future Scope

Self-assembly is commonly seen in both natural and processed foods, which is of

vital importance to control food quality, functions, and even safety. It can be
employed to innovate functional foods, whereas uncontrollable self-assembly may
lower food quality and even cause safety issues. Toward the state of minimum free
energy and the equilibrium of three classes of forces—driving, opposition, and
directional forces, is the basic principle of the self-assembly of proteins and poly-
saccharides. The specific forces and mechanisms in certain food systems are not well
understood, which arises from the complexity of food systems, and requires a deeper
investigation in the future by learning from synthetic polymer systems. A variety of
protein and polysaccharides nanostructures with unique properties and functionali-
ties, such as micelles, nanofibrils, and nanotubes, could be produced by the self-
assembly approach. Yet, some nanostructures are only reported to be generated from
a few food sources under very specific experimental conditions. Future research
needs to understand the generic feature of these nanostructures, and thereby to
extend their sources as well as categories. It is worthy of noticing that many
nanostructures, such as amyloid fibrils and amorphous aggregates, are recently
considered to be a generic feature of proteins. The self-assembly behaviour of
copolymers has been significantly investigated in polymer science but is far from
thorough understanding in food science, which calls for more efforts in the future.
Milliard reaction is not the only chemical reaction in the field of food chemistry. In
addition, an increasing number of food-grade cross-linkers were found in recent
years; this may also open the doors to produce protein-co-polysaccharides. In order
9 Self-assembling Properties 329

to successfully exploit self-assembly in practical applications and to ensure efficient

scale-up, a high level of control is also required.
Although some progress has been made in understanding how proteins and
polysaccharides assemble into nanostructures and how external factors determine
nanostructures properties, studies on how self-, dis-, and re-assembly in gastrointes-
tinal tract control the specific functions of foods are still in the infant stage. They are
crucial for human health as these assembly processes can modulate food-body
interactions and the biological fate of food components. Know-how on them could
provide the guidance to produce foods and generate new functions in food products
to benefit the consumer. In summary, future research on the assembly of food
polymers and the resultant nanostructures is extremely indispensable.

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Chapter 10
Flavour Delivery

Matthias Schultz

Abstract Flavourings are usually complex blends of solid or liquid compounds that
need to be turned into a format useful to the food or beverage manufacturer. They
often require protection during processing, transport, and storage to make sure that
the consumer has a delightful and authentic flavour experience during food and
beverage consumption. Flavour delivery systems provide flavourings in liquid
format as emulsions or in dry format embedded in a solid matrix or core-shell
capsule.
Food hydrocolloids play an indispensable role in the development and production
of flavour delivery systems due to their emulsifying, viscosifying, glass forming,
film forming, and other functionalities. The use of food hydrocolloids in flavour
delivery systems is reviewed in this chapter mainly considering the scientific liter-
ature of the last two decades. Emphasis is put on the role of food hydrocolloids for
the different technologies currently used to produce flavour delivery systems and on
the structural concepts which their performance is based on. Interaction between
hydrocolloids and flavouring compounds often influences flavour partitioning and
release, and thorough knowledge is needed to perfectly balance the composition of
the flavouring.
Current trends such as consumption of healthier food, changing life style, and
labelling transparency are impacting the way flavour delivery systems are being
designed.

Keywords Flavourings · Flavour delivery · Dry flavours · Beverage emulsions ·

Flavour release

M. Schultz (*)
Givaudan International SA, Kemptthal, Switzerland
e-mail: matthias.schultz@givaudan.com

© Springer Nature Singapore Pte Ltd. 2021 335

Y. Fang et al. (eds.), Food Hydrocolloids,
https://doi.org/10.1007/978-981-16-0320-4_10
336 M. Schultz

1 Introduction

1.1 Flavour and Flavourings

During consumption of food and beverages substances causing aroma or taste

sensations impart the perception of flavour. Aroma perception is commonly trig-
gered by the contact of compounds released from food and beverages with receptors
in the nose. These compounds can enter the nose either directly from the headspace
over the food or beverage (orthonasal perception) or through the oral cavity
(retronasal perception). Taste molecules interact with receptors on the tongue or in
the oral cavity and cause sensations like sweet, sour, salty, bitter, and umami.
Despite being mostly related to the senses of smell and taste, flavour perception
also involves textural, visual, and motorial aspects, all processed in a variety of brain
systems (Fig. 10.1) to make flavour perception one of the most complex human
behaviours (Shepherd 2006).
Substances imparting flavour perception by the senses are called flavourings.
Flavourings can be liquid or dry substances. They can be single chemical com-
pounds (like vanillin) as well as complex blends of materials of natural (e.g.,
essential oils, plant extracts, juice concentrates) and/or synthetic origin. Their chem-
ical diversity is vast (Uhlemann and Reiß 2010); often they are complex blends of
compounds displaying a huge variety of physicochemical properties such as melting,
boiling and flash points, solubility, volatility, partitioning coefficients, and chemical
reactivity (Table 10.1). Being given a range of just 16 flavouring compounds, a
flavourist would be able to compose different fruit flavourings by changing the ratio
of these compounds (Table 10.1, Grab 1998). The flavouring concentrate represents
the complete flavour profile in the most concentrated form, but is often diluted with a
carrier (ethanol in the case of the examples in Table 10.1). In practice, however,
flavourings are often not created by using only individual chemical substances, but
contain essential oils, botanical extracts, and the like (Wright 2010).

1.2 Flavour Delivery and Flavour Delivery Systems

Only a perfectly balanced composition at the point of flavour release immediately

before or during consumption will deliver an authentic flavour profile. The process
between preparation of a flavouring by a flavour company and contact with the
consumer’s receptors, however, can take from a few weeks to several years
(Fig. 10.2).
Any changes in composition caused by untimely release or the reaction of
flavouring constituents during blending, packaging, transport, storage, application
into food or beverage products and cooking, baking, frying or other food preparation
steps will change the composition of the flavouring and affect flavour perception.
Turning a flavouring into the right format for application into food or beverages
 10
Flavour Delivery

Fig. 10.1 The dual olfactory system, a Brain systems involved in aroma perception during orthonasal olfaction (sniffing in), b Brain systems involved in aroma
perception during retronasal perception (breathing out), with food in the oral cavity. Air flows indicated by dashed and dotted lines; dotted lines indicate air
carrying odour molecules. ACC, accumbens; AM, amygdala; AVI, anterior ventral insular cortex; DI, dorsal insular cortex; LH, lateral hypothalamus; LOFC,
lateral orbitofrontal cortex; MOFC, medial orbitofrontal cortex; NST, nucleus of the solitary tract; OB, olfactory bulb; OC, olfactory cortex; OE, olfactory
epithelium; PPC, posterior parietal cortex; SOM, somatosensory cortex; V, VII, IX, X, cranial nerves; VC, primary visual cortex; VPM, ventral posteromedial
337

thalamic nucleus (Reprinted with permission from Shepherd 2006, Copyright Springer Nature)
338 M. Schultz

Table 10.1 Examples of simple flavouring compositions, data from Grab (1998); physical data:
log octanol-water partitioning coefficients (logPow) calculated by ChemDraw® Professional (Perkin
Elmer 2019), boiling temperatures (Tb) from Reineccius (1994)
Apple Banana Pear Pineapple Tb


Flavouring compound parts by weight logPow C (at mm Hg)
Alcohols
1-Butanol 30 5 30 1 0.97 117–118
2-Methyl butanol 30 5 50 5 1.37
1-Hexanol 30 5 40 1.80 156–157
Esters
Amyl acetate 50 10 20 5 1.62 149
Isoamyl acetate 5 150 5 5 1.53 143
Ethyl butyrate 5 40 10 10 1.37 120
Amyl butyrate 5 30 20 20 2.69 185–186
Heptyl acetate 5 5 100 5 2.45 192–193
Ethyl 2-methylbutyrate 5 10 5 20 1.93 132–133
Allyl caproate 5 5 5 120 2.55 180–188
Citronellyl acetate 5 5 40 1 3.05 119–121 (15 mm)
Aldehydes
Hexanal 100 1 5 1 1.33 130–131
(E)-2-Hexenal 100 10 30 5 1.31 47–48 (17 mm)
Benzaldehyde 0.1 0.2 0.2 0.1 1.78 179
Others
Vanillin 1 30 1 30 1.27 170 (15 mm)
Eugenol 0.1 2 0.2 0.1 2.57 254
Solvent added to make up to 1000 parts by weight
Ethanol 693.8 686.8 638.6 770.8 0.07 64

(e.g., an instantly soluble powder or sufficiently stable emulsion), protecting it from

adverse changes and releasing it at the right point in time is accomplished by
capturing it in a flavour delivery system. For better protection of the flavouring,
especially against oxidation, a physical barrier might be erected between the
flavouring and the environment. For example, liquid flavouring droplets are embed-
ded in a solid polysaccharide matrix or in a core-shell capsule. Such systems are
referred to as flavour encapsulation systems. Hence, flavour delivery systems com-
monly bridge the gap between the flavouring and food and beverages, enabling food
manufacturers to more easily apply flavourings in their products with optimum
protection and performance, and providing the desired consumer experience
(Schultz and Ringgenberg 2008; Bouquerand et al. 2012; Ubbink 2013).
10 Flavour Delivery 339

Fig. 10.2 The journey of a flavouring from the flavour house to the consumer

Table 10.2 Flavour encapsulation technology overview

Particle size Flavour loada
Min Max Min Max
Technology μm μm wt% wt% Release mechanism
Extrusion 200 2000 8–10 15 Dissolution
Spray drying 20 100 20 25 Dissolution
Multi-stage spray drying 50 150 20 25 Dissolution
Fluid bed granulation 200 2000 10–15 20 Dissolution
Coacervation 20 800 40 70 Breakage, diffusion
Submerged nozzle 800 5000 70 90 Breakage, dissolution
a
Flavour load ¼ wt% of flavourings in the delivery system

1.3 Flavour Delivery Technologies

Table 10.2 presents an overview of the most important technologies to manufacture

solid flavour delivery systems together with their main characteristics. A useful
comparison of technologies and principles for product design and process engineer-
ing has also been published by Uhlemann et al. (2002), Doorn and Campanile
(2006), Madene et al. (2006), Uhlemann and Reiß (2010), Bouquerand et al.
(2012), and Ubbink (2013).
For the preparation of flavour powders the flavouring is usually dissolved or
emulsified in a carrier material melt (for extrusion) or solution (e.g., for spray
340 M. Schultz

drying). Almost all technologies require a subsequent drying step to remove excess
water. In order to homogeneously distribute the particles with other powder food or
powder beverage constituents, subsequent powder blending is often required. Food
hydrocolloids play an important role as emulsifiers and wall or matrix materials
capturing and protecting the flavouring (Sect. 2).
Water soluble liquid flavourings for beverages and liquid foods can often be
applied in highly concentrated form and do not need a flavour delivery system. In
some cases, solvents are added to mediate the generation of a homogeneous solution
of individual constituents and facilitate application. In case the flavour is no longer
sufficiently soluble in the food or beverage application, a flavour emulsion is
produced that can be easily diluted, e.g. in a beverage. Again, food hydrocolloids
are an integral part of such emulsions and act as emulsifiers and stabilizers (Sect. 3).
The desired mechanism by which the flavouring substances should be released
(e.g., dissolution, diffusion, mechanical breakage, instant or delayed release) during
food and beverage consumption (Sect. 4) plays a role when selecting the most
suitable technology for a specific application.

2 Functionality of Hydrocolloids in Dry Flavour Delivery

Systems
2.1 General Concepts of Matrix Encapsulation

A matrix surrounding flavouring droplets in powder particles needs to fulfil certain

criteria to be efficient. It needs to be stable against environmental conditions, in
particular against humidity, have good mechanical resistance and be dense enough to
prevent even the smallest flavouring molecules from permeating out and reactants
like oxygen molecules from permeating into the particle (Quellet et al. 2001). Dry
flavour delivery systems usually need to dissolve in water immediately, since typical
applications they are blended in—like instant beverages, soups and sauces—all need
to dissolve rapidly.
Food hydrocolloids fulfil certain tasks to contribute to the functionality of dry
flavour delivery systems (Bouquerand et al. 2012). First, a certain reduction of
interfacial tension is often needed to achieve homogeneous distribution of the
flavouring in the matrix carrier solution as finely dispersed droplets. Hydrocolloids
like octenyl succinate modified starches or acacia gum are commonly employed, but
other emulsifiers are also possible. Second, to provide the mechanical and protective
carrier properties of the matrix material, carbohydrates are used and processed in a
way to solidify them in their amorphous state during drying. Traditionally, starch
hydrolysis products are widely used to form a dense, glassy matrix. Being of
comparatively high molecular weight carbohydrate macromolecules leave voids in
the matrix structure that can facilitate permeation. Hence, third, and for oxidation
sensitive flavourings in particular, lower molecular weight molecules usually not
10 Flavour Delivery 341

being hydrocolloids are added to further densify the matrix. We will have a more
thorough look at hydrocolloid emulsifiers and matrix materials in the following
sections.

2.1.1 Emulsifiers

Emulsifiers need to facilitate formation of flavouring droplets during emulsification

and stabilize them for the time between spray feed preparation and the drying step.
This could cover a period of time between several minutes and a few days. Hence,
the stability requirements for feed emulsions are lower than for beverage emulsions
(see Sect. 3.2.1), which require several months of stability. Therefore, for feed
emulsions, it is easier to compromise between emulsifier performance and cost.
Traditionally, acacia gum, also known as gum arabic, is used. In particular, gum
derived from trees of the species Acacia seyal shows sufficient performance for dried
products. In some cases, acacia gum is used as both emulsifier and wall material
(Bertolini et al. 2001; Fang et al. 2005).
Alternatively, several types of starches modified by reaction with octenyl succinic
anhydride (OSA) are used as a very efficient emulsifier. The introduction of the
hydrophobic octenyl chain to the hydrophilic starch molecule provides the
amphiphilicity required for interfacial activity. Furthermore, the efficiency of octenyl
succinate modified starches is very much affected by other structural parameters like
the degree of branching (DB), degree of octenyl succinate substitution (DS), and the
distribution of octenyl chains in the macromolecule (Fig. 10.3, Sweedman et al.
2013; Tizzotti et al. 2013; Zasypkin and Porzio 2004).
Proteins have been proposed and tested as emulsifiers for the production of
flavour powders; however, a few disadvantages have prevented proteins from
broad application. They often have limited solubility in the pH range used to prepare
feed emulsions, and tend to react with some flavouring constituents, especially
aldehydes. They can, however, be useful for the encapsulation of other food ingre-
dients (Reineccius 2019).
Other polysaccharides such as sugar beet pectin (Paramita et al. 2010), cellulose
derivatives (Zhou et al. 2018), and modified guar gum (Sarkar et al. 2013) have been
tested as emulsifiers for feed emulsions for dry flavours. Often, their interfacial
activity is not sufficient due to their hydrophilic character unless they are chemically
modified or have a fraction of protein bound to polysaccharide chains providing
amphiphilic functionality such as in sugar beet pectin (Williams et al. 2005).

2.1.2 Matrix Materials

Starch hydrolysis products such as dextrins, maltodextrins, and glucose syrups are
commonly categorized by a parameter called dextrose equivalent (DE) which is a
measure of the reducing power of a material relative to that of glucose (being
assigned a reducing power of 100). That means the larger the DE the shorter the
 342

Fig. 10.3 Schematic representations of OSA-modified starches used by Tizzotti et al. (2013), ● ¼ grafted OSA groups, DS ¼ degree of substitution,
DB ¼ degree of branching, Rh ¼ hydrodynamic radius of gyration, Mw ¼ weight average molecular weight. Wi-x, Wm-x, Ri-x and Gm-x: W, R and G stand for
Waxy Maize, Rice or Gelose 80, respectively. Subscripts i and m indicate the alcohol used during the acid hydrolysis reaction (isopropanol or methanol,
M. Schultz

respectively) (Reprinted with permission from Tizzotti et al. 2013, Copyright Elsevier B.V.)
10 Flavour Delivery 343

polymer chain and the lower the molecular weight. Starch hydrolysates with DE
values below 20 are referred to as maltodextrins, while those with DE greater than
20 are typically referred to as glucose syrups, glucose solids, or corn syrup solids.
Being derived from natural products, they all have a comparatively broad molecular
weight distribution, often with a certain amount of oligomers present (White Jr. et al.
2003).
Starch hydrolysis products play an outstanding role as matrix materials as they
unify a number of advantageous properties including good water solubility, com-
paratively low viscosity and high glass transition temperature. The former two are
important to dissolve them at rather high solid content, reducing the amount of water
that needs to be removed during the drying process.
The concept of glass transition temperature, Tg (Liu et al. 2006), is especially
important to produce powders with low hygroscopicity, which is, in turn, related to
desirable transport, storage stability and powder flow properties. When being dried,
starch hydrolysis products solidify in an amorphous state, also referred to as glassy
state. Glass formers undergo a second order phase transition at Tg where they turn
from a brittle and hard glass into a somewhat rubbery, viscoelastic material. The rate
of mass transport within the rubbery state is much greater than in the glassy state.
Hence, at and above Tg, integrity of the powder is no longer maintained. Tg is a
function of DE as well as of the water activity, aw, in a powder. Careful control of Tg
and aw is of huge practical importance, as low DE materials have a beneficially high
Tg. However, due to their high molecular weight they form a more permeable matrix,
causing lower retention of volatile chemicals and limited oxidative protection of the
flavouring compared to matrices with a mixture of low and high molecular weight
polymers.
Research on the relationship between maltodextrin molecular weight and glass
transition temperature has been reported by Avaltroni et al. (2004). An increase in aw
results in a reduction of Tg as the water has a plasticizing effect on the carbohydrate
matrix (Fig. 10.4). This may have devastating consequences if the Tg is reduced
down to the transport or storage temperature of a powder, leading to the formation of
lumps or agglomeration of the powder into a solid block. On the other hand, drying a
powder down to very low aw may considerably increase drying time and manufactur-
ing cost. Normand et al. (2019) studied water diffusion in the 18 DE maltodextrin/
water system and the drying kinetics by collecting the evaporated water through a
condenser.
The concept of free volume has been introduced to better explain the occurrence
of voids in the structure of glassy carbohydrates (Roussenova et al. 2013). Positron
Annihilation Lifetime Spectroscopy (PALS) was found useful to measure free
volume in maltooligomer matrices (Roussenova and Alam 2013). The plasticizing
properties of water and substances like glycerol have also been determined
(Roussenova et al. 2014).
A common approach to densify carbohydrate matrices is the addition of small
molecular weight molecules to fill the subnanometer-size voids in the glassy matrix.
Typical materials are sucrose, maltose, and trehalose, which work well to substan-
tially improve oxidative stability of flavourings, but again reduce Tg (Sillick and
344 M. Schultz

Fig. 10.4 Tg measured for maltodextrins as a function of the water content of the sample. Open
scatters: squares DE ¼ 2, circles DE ¼ 10, triangles DE ¼ 19, stars mixture of DE ¼ 2 and DE ¼ 19.
Solid squares: used from the literature (Benczédi et al. 1998) for extruded starch (Reprinted with
permission from Avaltroni et al. 2004, Copyright Elsevier B.V.)

Gregson 2010). Therefore, thorough knowledge of the material science of carbohy-

drate glasses is a prerequisite for the successful development of dry flavour delivery
systems (Fig. 10.5).
More recently, in studies of hydrophobically-modified starch-sucrose blends
using wide-angle x-ray scattering and PALS, partial amorphous-amorphous phase
separation in the powders was detected (Tedeschi et al. 2016) and confirmed by
Dynamic Scanning Calorimetry (DSC) and 1H solid state NMR (Hughes et al.
2018). The temperature dependence of the local free volume as a function of blend
composition and water content results in complex phase separation behaviour
(Hughes et al. 2016).
Other matrix materials have been tested, but have not found broad application due
to performance, cost or labelling issues. However, as in recent years the use of
materials like maltodextrins has become less preferred due to increasing sensitivity
of consumers regarding food ingredients that they are less familiar with, it is
expected that other materials will gain importance as matrix materials.
10 Flavour Delivery 345

Fig. 10.5 Trends of Tg vs. aw for selected sucrose (S)/18 DE maltodextrin blends (Reprinted with
permission from Sillick and Gregson 2010. Copyright Elsevier B.V.)

2.2 Use of Food Hydrocolloids to Produce Dry Flavour

Delivery Systems

2.2.1 Extrusion

Extrusion is a general term for processes which include the transport of a compar-
atively viscous material under pressure through a die. The die configuration defines
the shape and size of the extrudate. Typical extrusion technologies for flavour
encapsulation are melt injection, also known as ram extrusion, and melt extrusion,
a process carried out in a screw or twin-screw extruder (Porzio 2008; Tackenberg
and Kleinebudde 2015; Lazou and Krokida 2017).

Ram Extrusion

This is the older process where a sugar or corn syrup solids melt is blended with the
flavour, the system is pressurized by a gas and injected through a die into a bath of
cold solvent (usually isopropanol) where the melt immediately solidifies and is
broken into particles by rapid agitation (Porzio 2008). Thus, particles with a shape
described as rods, threads or strands are obtained (Fig. 10.6a). Hydrocolloids do not
play a major role in this process, although addition of hydrocolloids is discussed in
the literature to modify certain properties and produce matrices with a specific
functionality. For example, the use of agar has been described in a matrix further
346 M. Schultz

Fig. 10.6 Scanning Electron Microscopy (SEM) images of extruded particles prepared by a ram
and b twin-screw extrusion, scale bars 1 mm

containing maltodextrins or starch hydrolysates and flavouring compounds such as

cinnamic aldehyde or menthol to produce soft-gelled particles when dispersed in
water with good flavour retention (McIver et al. 2002).

Screw or Twin-screw Extrusion

In this process, a carbohydrate polymer matrix is molten or plasticized in an extruder

and conveyed by a screw or co-rotating twin-screws to the die. A liquid flavouring
can be added at the inlet feed or at any stage during the extrusion process, and, if
needed, be emulsified in the presence of an emulsifier. Single-screw extruders are
rarely described, whereas twin-screw extruders with co-rotating screws have become
common to produce flavour delivery systems (Porzio 2008; Tackenberg and
Kleinebudde 2015). A broader variety of shapes is possible due to different die
configurations (Fig. 10.6b). Carbohydrate polymer matrices play an important role
here and are used as such or in combination with low molecular weight sugars or
sugar alcohols (Tackenberg et al. 2014). Recent reviews have been published by
Tackenberg and Kleinebudde (2015) and Castro et al. (2016a).
In a series of articles, Tackenberg et al. (2015a, b) describe the encapsulation of
orange terpenes in maltodextrin matrices. They studied the variation of sucrose,
water, and orange terpene content in the extrusion matrix to assess the influence of
feed composition and processing parameters on crystalline content, Tg, orange
terpene retention and limonene content in the orange terpenes. Good encapsulation
efficiency and storage stability are confirmed under optimum conditions. Castro
et al. (2016b) varied the DE of maltodextrins in a melt extrusion process using
different plasticisers (water, glycerol, sorbitol) and found no linear dependence
between maltodextrin DE and glass transition temperature in extrudates.
Compared with spray dried systems, a main disadvantage of extrudates is their
comparatively low flavouring load. Snyder and Zhang (2017) aimed to increase the
load of cold pressed orange oil in a matrix containing maltodextrin and octenyl
10 Flavour Delivery 347

succinate modified starch by varying the DE of the maltodextrins. The lower the DE
the higher was the retained orange oil after extrusion. The maximum retained oil
value for an optimum composition and screw configuration was in the range of
12–15 wt%.
Native starches have been studied as extrusion matrices as well. Emin and
Schuchmann (2013) simulated the flow of plasticized maize starch during extrusion
processing by Computational Flow Dynamics. The results suggest that a strong
increase in zero shear viscosity has no influence on the flow, whereas a slight change
in viscosity at the shear thinning region leads to significant increase in pressure drop
along the mixing zone of the extruder. Simulation results were validated quantita-
tively by experimental data.

2.2.2 Spray Drying

For many years, spray drying has been the work horse technology to encapsulate
flavourings in a glassy carbohydrate matrix (Porzio 2007). Typically, a feed solution
is prepared by dissolving or emulsifying a flavouring into a carrier solution and using
an atomizer disk or pressure nozzle to spray from the top into the chamber of a spray
drier. Hot air is blown from the bottom of the dryer that evaporates the water in the
spray droplets within a very short period of time, and a glassy matrix is formed
wherein the flavouring molecules are dissolved or finely dispersed as droplets.
Particle diameters of the dry powder are small (Fig. 10.7a, b), typically in the
range between 20 and 100 μm (Fang and Bhandari 2012; Ré 1998).
In order to increase the flowability and wettability of the powder, spray drying
can be combined with an agglomeration step. In a so-called multi-stage drier, the
finest particles are returned into the drying chamber and more feed emulsion is
sprayed onto their surface. In addition, an internal or external fluid bed can be
applied for further agglomeration, and particle diameters of 50–150 μm are com-
monly obtained (Fig. 10.7c, d).
As for delivery systems prepared by extrusion (Sect. 2.2.1), there is a range of
requirements for well performing spray dried flavour powders. These include
mechanical (physical) stability, long-term flavour (chemical) stability through ade-
quate protection in a sufficiently dense matrix, consistent powder integrity during
transport and storage at varying temperatures and limited hygroscopicity. These
properties are related to physical and structural parameters such as water activity,
bulk density, and glass transition temperature that need to be carefully controlled.
Besides thorough understanding of the spray drying process parameters, the com-
position of the powder matrices surrounding the flavouring molecules or droplets is
key, and hydrocolloids play a major role. Cost and energy consumption consider-
ations are becoming more and more important as well.
Within the last decades a multitude of materials including many hydrocolloids
has been tested for their capability to form matrix materials and some reviews are
available (Soottitantawat et al. 2015; Reineccius and Yan 2016; Jafari et al. 2008;
348 M. Schultz

Fig. 10.7 Schematic and SEM images of spray and multistage spray dried particles, a spray dried
particle: flavour droplets (orange) embedded in a glassy carbohydrate matrix (yellow), b SEM
image of spray dried particles, scale bar 20 μm, c multistage spray dried particle (colours same as in
a), d SEM image of multistage dried particles, scale bar 10 μm

Gharsallaoui et al. 2007). The mechanisms of permeation of flavouring and oxygen

molecules through the matrix are still under investigation.
Upon ageing a glassy system moves toward equilibrium below Tg, a process
called structural relaxation occurs, resulting in a decrease of energy and free volume
and an increase of structural order. Enthalpy relaxation times of spray dried carbo-
hydrate matrices containing volatile flavour compounds were determined by differ-
ential and isothermal calorimetric methods and correlated with flavour retention and
formation of oxidation products (Sahni et al. 2015). Greater enthalpy relaxation time
appears to be well correlated with better stability and may become a useful predictor
of stability for both loss of volatile flavouring compounds and oxidation.
High solid content in the feed solution or emulsion is important to keep drying
times short (less water to be evaporated), to limit exposure of volatile or thermolabile
flavouring compounds to heat (reducing flavouring losses) and for cost reasons
(reduced manufacturing cost at high throughput). However, an increase in the
content of dissolved materials in the feed is often limited by their solubility or
viscosity. Consequently, hydrocolloids that have excellent solubility while keeping
suspension viscosity low are most preferred. Not surprisingly, maltodextrins with a
DE of 5–20 work well and are comparatively well studied. Great care was taken in
the characterization of the maltodextrin-water system using Tg as a measure to avoid
stickiness of spray dried powders (Roos and Karel 1991; Normand et al. 2013). The
10 Flavour Delivery 349

effect of DE on the properties of lime oil spray dried particles has been studied by
Campelo et al. (2017). High DE was found to be better for oil retention and
encapsulation efficiency. In a study to develop a model for shelf life prediction of
orange oil, the oxidation stability was found to increase with increasing DE of the
encapsulation matrix, but led to decreased humidity resistance (Subramaniam et al.
2013). By adding high DE carbohydrates, mono- or disaccharides, a compromise has
to be found to produce a shelf-stable product.
Acacia gum is often combined with maltodextrins mainly for its emulsifying
properties. In some cases, it is used as the sole encapsulation material (Bertolini et al.
2001). When compared with other wall materials, it has been found to display better
protecting properties (Krishnan et al. 2005), but the results often depend on the
quality of the gum, the specific flavouring and the stability testing conditions.
Other hydrocolloids are more rarely used. Depolymerized guar gum has been
reported as partial replacement for acacia gum in the encapsulation of mint oil
(Sarkar et al. 2012). Blends of acacia gum with xanthan have also been used
(Outuki et al. 2016).

2.2.3 Freeze Drying and Vacuum Drying

Despite its huge importance for the production of instant coffee, freeze drying has
not found wide application for the encapsulation of flavourings mainly due to the
immense energy input needed and the related processing costs. A solution of matrix
material is first frozen followed by direct sublimation of water under reduced
pressure (Fang and Bhandari 2012).
Vacuum drying is mainly used in the production of the so-called process or
reaction flavours (Kerler et al. 2010), but in some instances for flavour encapsulation
as well. A high solid content dispersion is spread as a thin film in trays, occasionally
foamed to accelerate drying under vacuum. The resulting cake is milled to obtain a
free flowing powder (Ubbink 2013).

2.2.4 Fluidized-Bed Technologies

For flavour encapsulation in a fluidized bed a batch of solid particles is fluidized in a

chamber by blowing air through them. In some equipment configurations, moving
part of the equipment contributes to efficient fluidization. As a result, the particles
behave and move like a fluid. A layer of encapsulated flavouring can be generated on
the surface of the particles by spraying flavour dissolved or emulsified in a carrier
solution into the fluid bed and evaporating the remaining solvent in the hot air
stream. There are different fluid bed geometries available in either batch or contin-
uous mode (Meiners 2012; Guignon et al. 2002; Teunou and Poncelet 2002). Batch
granulators can, e.g., be equipped with rotor, Wurster or top spray inserts. Additional
layers around the particles can be added in a subsequent coating step to further
modify the functionality of the particles (Fig. 10.8).
350 M. Schultz

Fig. 10.8 Cross section of a particle produced by fluidized-bed granulation, a schematic sketch,
b X-ray microtomography image, image width 1.5 mm

The size of the resulting granules depends mainly on the size of the core material,
typical particle diameters are between 500 μm and several millimetres. They are
useful in applications where larger particles are required such as tea bags or visual, in
part coloured cues are desirable such as in confectionery, hard boiled, and chewy
candy products.
Any food grade material that can be fluidized may serve as core material. Typical
core materials used by the flavour industry are amorphous or crystalline mono-, di-,
and oligosaccharides, food acids, plant leaves, fibres, and fruit pieces. Hydrocolloids
are rarely used as core materials, but play a role as matrix material in the flavour layer
protecting the flavouring by forming a glassy matrix as described for the technolo-
gies above.
Materials for the outer coating around the flavour layer include fats and waxes,
but hydrocolloids are rather useful as well. Cellulose derivatives have found broad
application. Nienaltowska et al. (2009) studied the coating quality of three water
soluble cellulose derivatives—methyl cellulose, carboxymethyl cellulose, and
hydroxypropyl cellulose, using glass beads as a core material. The concentration
of the coating solutions and the related viscosity had a profound effect on coating
quality.
Spray dried particles can be further agglomerated and coated in fluidized-bed
equipment. In one study (Turchiuli and Dumoulin 2013), the flavour/carrier emul-
sion was spray dried. Then the fine powders (<30 μm) were agglomerated in a
fluidized bed with hot air by top spraying water or water/flavour/carrier emulsion.
Then agglomerates were coated with a dry thin layer of emulsion.

2.2.5 Coacervation

Coacervation is a phase separation phenomenon in polymer solutions triggered by

changes of temperature and/or pH. In simple coacervation, a polymer phase
10 Flavour Delivery 351

Fig. 10.9 Coacervate beads, a schematic sketch, b microscope image, image width about 500 μm

separates from a pure solvent phase. Complex coacervation is more commonly used
for flavour encapsulation (Thies 2007; Xiao et al. 2014). Typically, an ionic macro-
molecule with interfacial activity is used to emulsify a hydrophobic flavouring in an
aqueous medium. An oppositely charged polymer is present or added which after
adjusting temperature and pH accordingly combines with the first polymer at the
interface of the oil droplets forming a coacervate phase around the droplet. The shell
can be turned insoluble by adding a cross-linking agent generating water insoluble
core-shell capsules with the lemon shape typical for products made by coacervation
(Fig. 10.9).
Hydrocolloids are useful polymers for complex coacervation. There are certain
requirements for optimum performance in forming coacervate phases. Both coacer-
vation partners should have about approximately equal but opposite charge at a pH
where each of them alone is completely soluble (Schmitt and Turgeon 2011).
Chitosan is among the few positively charged polysaccharides at acid pH and has
hence been studied for coacervation processes (Liu et al. 2013; Han et al. 2013;
Butstraen and Salaün 2014), although it is considered food grade in only some
regional markets. Chitosan has, however, a few disadvantages related to solubility
and the mechanical properties of the resulting capsules. Instead usually proteins are
being used, gelatin in particular, as the positively charged coacervation partner
(Schmitt and Turgeon 2011). There is a broader choice of anionic hydrocolloids
for complex coacervation, among them acacia gum (Leclercq et al. 2009a; Yeo et al.
2005), pectin (Silva et al. 2012), agar (Singh et al. 2007), carboxymethyl cellulose
(Lv et al. 2012), and alginate (Devi and Kakati 2013). Many combinations of
proteins and coacervates are possible. Detailed overviews were published by Schmitt
et al. (2009) and Xiao et al. (2014).
Dardelle and Erni (2014) have studied the interplay of wetting phenomena and
fluid viscoelasticity in coacervate/oil/water systems. Coacervate shells containing
gelatin can also be easily cross-linked using aldehydes such as glutaraldehyde or
enzymes like transglutaminase (Dardelle et al. 2011).
Direct encapsulation of complex blends of flavourings in coacervate capsules is
difficult to achieve since some constituents of the blends partition between the oil
core of the later coacervate capsule and the aqueous phase in which coacervation
352 M. Schultz

occurs (Reineccius 2019). An approach to overcome this problem is to produce dry

coacervate capsules with a neutral oil core, and then load them with flavour by
spontaneous partitioning of the flavour into the oil cores through the coacervate shell
(Wieland and Soper 2008; Leclercq et al. 2009b).

2.2.6 Other Technologies to Produce Insoluble Capsules

There are other technologies to prepare insoluble core shell or matrix capsules for the
encapsulation of flavourings. The possibility to cross-link alginate or pectin capsules
by dripping or jetting them into a bath containing calcium ions is used in a variety of
processes. Alginates are derived from seaweed mainly as a sodium salt. They are
structurally characterized by their molecular mass and the ratio of guluronic and
mannuronic acid subunits (the G/M ratio) in the polymer. A high G/M ratio provides
the basis for efficient cross-linking by calcium ions in a so-called egg-box type
structure (Draget 2008). Pectins also form cross-linked gels with bivalent cations,
and are characterized by their molecular mass and their content of methoxy or amide
substituents in the glucopyranose rings (Endreß and Christensen 2009). Low
methoxy content or high amide group content is necessary for them to be efficiently
cross-linked by bivalent cations. Combinations of alginates and pectins can be used
for formation of matrix or core-shell capsules.
In the simplest approach, a flavouring is emulsified into an alginate or pectin
solution, and the emulsion is just dripped through a syringe into a calcium chloride
bath (Petzold et al. 2014). More sophisticated nozzles such as vibrated two-fluid
nozzles allow for the preparation of core-shell capsules with oil in the core and a
polysaccharide as the shell. There are methods for making narrow size-distribution
capsule cores using vibrating nozzles (Brandenberger and Widmer 1998), piezo
driven drop formation in submerged nozzles (Böhmer et al. 2006), electrostatic
extrusion (Lević et al. 2016) or microfluidic channels (Amici et al. 2008). All
these methods have in common that they find applications in the pharmaceutical
and cosmetic industries where small batches are produced and rather expensive
ingredients are encapsulated. The days of the production of flavour delivery systems
by such techniques at a sufficiently large scale are, however, still to come.
Yeast cells were used as thermo-stable flavour delivery systems (Normand et al.
2005; Dardelle et al. 2007). Their cell walls consist of external β-glucan and
mannoprotein layers and were found permeable to both small polar and apolar
molecules in aqueous solution. The reticulated β-glucan network plays the role of
a skeleton, whereas the external protein layer acts like a sieve with a mesh-size
evolving with the water content.
10 Flavour Delivery 353

3 Functionality of Food Hydrocolloids in Beverage

Emulsions

3.1 General Concepts in the Development of Beverage

Emulsions

Beverage emulsions are oil-in-water emulsions delivering flavour, in most cases

cloudiness, and in some cases additional ingredients like colours, juices, juice
concentrates or vitamins in the final beverage. For the convenience of the beverage
manufacturer, the emulsion is supplied as a beverage concentrate that needs to be
only diluted with water, sweetener, and food acid to provide the final soft drink,
energy drink, flavoured water, squash or syrup, depending on the dilution ratio.
Flavourings which are highly water soluble do not need to be emulsified and are
often supplied as such or diluted in a solvent. Citrus flavourings—from orange,
lemon, lime, mandarin, tangerine, and similar fruits—containing essential oils are
not sufficiently soluble in the ready to drink beverage and hence need to be
emulsified, unless the least soluble ingredients are removed, which often has adverse
consequences for an authentic flavour profile.
Industry requirements for food hydrocolloids in beverage emulsions have been
reviewed (Schultz 2010). Dickinson (2009) lists some important performance related
requirements such as fast reduction of interfacial tension at the oil–water interface,
strong adherence to the interface once they are adsorbed, and protection of newly
formed droplets against flocculation and coalescence. There are additional needs
arising from quality assurance, economical, regulatory, and consumer perception
considerations. Those include constant quality and performance, regulatory accep-
tance, reasonable and constant price, and consumer acceptance.
Beverage emulsions are oil-in-water emulsions with a typical oil content of
5–25%. The oil phase consists of the flavouring and, in emulsions for cloudy
beverages, of the so-called weighting agents. Weighting agents such as glycerol
esters of different types of tree rosin or sucrose acetate isobutyrate increase the
density of the oil phase to bring it closer to that of the water phase in order to delay
creaming of oil droplets and ring formation in the emulsion and beverage. In case an
oil soluble colorant, e.g. a carotenoid, is included, it also needs to be dissolved in the
oil phase. Optionally, antioxidants like tocopherols or butylated hydroxyanisole may
be part of the oil phase. The water phase contains emulsifiers and, in part, stabilizers,
water soluble colorants and typically a preservative system. Traditionally, sodium
benzoate or potassium sorbate are added as preservatives in combination with citric
acid to convert benzoate and sorbate into the corresponding acids that imply the
preserving functionality.
The emulsions are in general produced by separately preparing the water and oil
phases to completely dissolving all the ingredients in each. During a
pre-homogenisation step the oil phase is slowly added to the water phase under
high shear, followed by a subsequent high-pressure homogenisation step that can
apply one or more passes through a high-pressure homogenizer. For emulsions for
354 M. Schultz

Fig. 10.10 Typical droplet size distributions of emulsions after high-shear (pre)homogenisation
(dashed curve) and high-pressure homogenisation (solid curve)

cloudy beverages the emulsion droplet diameter is reduced from below about 5 μm
after pre-homogenisation down to 300–800 nm (Fig. 10.10). Droplets in this size
range identical with the wavelength range of visible light provide optimum cloud-
iness in the beverage by effective light scattering.
Recent overviews of beverage emulsion materials science and technology are
available (Given Jr. 2009; Schultz 2010; Piorkowski and McClements 2014; Dick-
inson 2018).

3.2 Hydrocolloids Used in Beverage Emulsions

3.2.1 Hydrocolloids as Emulsifiers

Dickinson (2018) has summarized why and which hydrocolloid emulsifiers work
well in food emulsions. Besides having surface activity resulting in the reduction of
interfacial tension and interfacial adsorption at a timescale relevant for emulsion
preparation, they should not be prone to strong aggregation and gelation. Only a
small number of hydrocolloids satisfy these requirements in a way that makes them
useful as primary emulsifiers, whereas many hydrocolloids are good stabilizers due
to their functionality as thickeners and viscosifiers.
10 Flavour Delivery 355

Acacia gum

Acacia gum, for the application as emulsifier and stabilizer in beverage emulsions, is
harvested from Acacia senegal trees as the exudate produced by the tree to close
wounds of natural origin or from tapping. It is a complex polysaccharide whose
exact structure is still under debate (Al-Assaf et al. 2005; Renard et al. 2006; Sanchez
et al. 2008; Mahendran et al. 2008; Williams and Phillips 2009; Atgié et al. 2019).
When fractionated by Size Exclusion Chromatography, a high molecular weight
arabinogalactan-protein fraction constituting about 10 wt% of the gum was identified
and found to be mainly responsible for its emulsifying properties. The largest
fraction, 85–90 wt% of the gum, despite containing some protein as well, is mainly
arabinogalactan and contributes to the stabilizing properties of the gum. The smallest
fraction, mainly peptides, makes another 1–2 wt% of the whole gum. Much effort
has been spent to assure consistent gum quality for beverage emulsions—a chal-
lenging task due to the dependence of the exact composition of the gum on climate,
soil, age of trees, and other factors (Al-Assaf et al. 2003). The content of the
emulsifying arabinogalactan-protein fraction is an important but not the only
parameter.
In an attempt to bind more protein to polysaccharides to increase the content of
the arabinogalactan-protein fraction, acacia gum was subjected to a physical treat-
ment in dry form at elevated temperature. The resulting gum showed improved
emulsifying efficiency and has since then been commercialized (Al-Assaf et al.
2007).
Another more recent development is chemically modified A. seyal gum to get its
performance closer to that of gum from A. senegal (Bi et al. 2017). As for starch, a
hydrophobic octenyl chain can be introduced to the polysaccharide chains of the
gum structure (Shi et al. 2017). Indeed, a strong improvement of emulsifying power
can be achieved, although the treated gum loses its natural reputation due to the
chemical modification step.

Ghatti gum

Another tree exudate with excellent emulsifying properties is gum ghatti collected
from Anogeissus latifolia trees growing in India (Al-Assaf et al. 2009). Compared
with acacia gum the protein content is higher (Ido et al. 2008) and more evenly
distributed throughout the gum’s molecular structure. The component of gum ghatti
adsorbed at the oil–water interface has higher surface coverage and the amount of
gum ghatti adsorbed is more than three times that of acacia gum (Katayama et al.
2018). Consequently, gum ghatti can be used at lower concentrations than acacia
gum (Ido et al. 2008; Al-Assaf et al. 2008) and would be expected to play an
increasingly important role if regulatory admittance in major markets could be
achieved.
356 M. Schultz

Octenyl Succinate Modified Starch

Like for flavour powders, octenyl succinate modified starch plays an important role
as emulsifier in beverage emulsions (Trubiano 1995; Viswanathan 1999). However,
different starch properties are desirable than for spray dry emulsions. While modified
starches for spray dry feed emulsions should provide low viscosity to maximize the
solid content in the feed, it is generally desirable for modified starches for beverage
emulsions to provide higher viscosities to the water phase, contributing to long-term
stabilization of the emulsions. Hydrolysed starches from waxy maize or corn are
good substrates for chemical modification due to their high content of amylopectin
and their good and almost complete solubility (Zhao et al. 2018b). Starches of
different origin are also being studied (Jain et al. 2019). Compared with acacia
gum, the interfacial tension is more efficiently decreased and smaller emulsion oil
droplet diameters can be obtained at comparable pressures during homogenisation
(Qian et al. 2011). However, the formed interfacial layers have lower shear elasticity
than those formed by acacia gum (Erni et al. 2007).

Sugar Beet Pectin

Pectins are polysaccharides that display little interfacial activity unless they contain
substantial amounts of protein (Funami et al. 2006; Williams et al. 2005). One such
pectin is isolated from sugar beet and was studied as emulsifier in beverage emul-
sions (Bai et al. 2017). The emulsifying efficiency was found to be good, however,
only small amounts could be applied due to the very high molecular weight and high
viscosity.

Other Food Hydrocolloid Emulsifiers

Many other hydrocolloids have been proposed as emulsifiers for beverage emul-
sions, among them corn fibre gum (Yadav et al. 2009; Bai et al. 2017), mesquite gum
(Román-Guerrero et al. 2009), fenugreek gum (Huang et al. 2001; Kaltsa et al.
2016), and spruce galactoglucomannan (Mikkonen et al. 2009). None of them has,
however, yet found broad industrial application for the emulsification of flavours.

3.2.2 Hydrocolloids as Emulsion Stabilizers

Guar gum and carboxymethyl cellulose have been tested as emulsion stabilizers
alone and in combination by Izadi and Emam Djomeh (2015). A combination of
0.5% guar gum and 0.2 wt% carboxymethyl cellulose was found to work best.
However, long-term stability tests have not been reported and stability would be
expected to be low due to comparatively large particle size. Nevertheless, a range of
10 Flavour Delivery 357

hydrocolloids, due to their thickening and viscosifying properties, can considerably

stabilize beverage emulsions.
Xanthan gum has been used in combination with acacia gum in orange oil
emulsions (Mirhosseini et al. 2008a). Xanthan as well as tragacanth gums can
stabilize emulsions due to the formation of a phase separated microstructure
containing oil rich compartments (Moschakis et al. 2005; Drakos and Kiosseoglou
2006). Furthermore, the shear thinning properties of xanthan gum contribute to
stabilization; the structure is maintained as long as the emulsion is not subjected to
high shear or heavy movement.
Given that many flavours in beverage emulsions contain essential citrus oils the
use of citrus pectin as a stabilizer deserves attention from a labelling perspective. The
emulsifying capacity of citrus pectins of different origin has been studied (Akhtar
et al. 2002; Leroux et al. 2003; Schmidt et al. 2015, 2017; Zhao et al. 2018a),
however, its combination with conventional emulsifiers appears more promising
(Verkempinck et al. 2018).

4 Flavour Release

While flavourings need to be captured and protected during processing, transport and
storage, efficient release is required at the point of consumption. For foods and
beverages containing dry delivery systems fast and complete dissolution is required
in the food or beverage product immediately before consumption or in the oral
cavity. Aroma perception depends on the availability of flavouring molecules in the
gas phase; hence, the partitioning of flavouring molecules from the food matrix plays
an important role and is influenced by the interaction with food components such as
hydrocolloids and fat. Taste is perceived in the oral cavity only if the taste molecules
reach the receptors on the tongue.

4.1 Flavour Release from Delivery Systems

While instantly soluble flavour powders release the flavour upon dilution into the
water phase and from there into the gas phase, water insoluble capsules display
release by diffusion in an aqueous environment or by mechanical disruption,
e.g. when chewing. The effects of texture and structure on the retention of aroma
compounds during processing and storage and on the aroma release and perception
during consumption were reviewed by de Roos (2003). The release of cherry and
peppermint flavouring compounds encapsulated by melt extrusion was measured by
Gunning et al. (1999). The effects of water content and temperature variation on the
release into the headspace over the extruded powders were studied. The largest
amounts of release occurred when the matrix was above its glass transition
temperature.
358 M. Schultz

Release from gelatin/acacia gum coacervate capsules was measured by monitor-

ing the headspace of a vessel containing the capsules to proton transfer reaction mass
spectrometry (PTR-MS). No effects of cross-linking or wall/core ratio on volatile
release in hot water could be found for any of the volatiles studied. When comparing
real-time release of the prepared coacervates to a spray dried equivalent, there was
no difference in the release from hot water but the release was slower when
coacervates were added to ambient-temperature water. Volatile release was primar-
ily determined by compound partition coefficients (oil/water and water/air) and
temperature (Leclercq et al. 2009b).
In some specific applications prolonged or delayed release is desirable, e.g. in
chewing gum. This can be achieved by hydrocolloid and other coatings added to
granulates (see Sect. 2.2.4) that take time to first dissolve the coating before the
flavour is released. Sequential flavour release from foods like chewing gums,
confectionery products or bars are possible by combining fast with delayed release
systems.
Flavourings included in emulsions and pastes are released into the headspace
upon consumption, but the process is far from being simple due to partitioning of
flavouring compounds in the water and oil or fat phase (de Roos 2000; Taylor and
Linforth 2001) or interaction with food components, e.g. fat and protein phases in
dairy products (Mao et al. 2017).
Very few publications deal with flavour release from typical beverage emulsions
with hydrocolloid emulsifiers and stabilizers whereas there is ample literature on
release from emulsions stabilized with small molecular weight emulsifiers and
surfactants. Mirhosseini et al. (2008b) study the influence of main emulsion com-
ponents (acacia gum, xanthan gum, and orange oil) on the release of target volatile
flavour compounds released from a model orange beverage (diluted orange beverage
emulsion) into the headspace.

4.2 Flavour Release from Food

Flavour release from food products and their sensory perception has been widely
studied (van Ruth and Roozen 2010). This section only deals with food systems
where food hydrocolloids play a role. Polysaccharides affect retention, release, and
perception of flavourings, e.g. the viscosity and swelling ability of a hydrocolloid
influences the release of flavour molecules from the matrix. In a study of the release
from polysaccharide films of cellulose derivatives, fast dissolving polysaccharides
were found to result in a quick burst of flavour at high intensity that tapered more
quickly whereas slowly dissolving films gave a slower onset and a more consistent
release over time (Cook et al. 2018).
Release of 20 flavour compounds from xanthan thickened food model systems
with different viscosities were studied (Bylaite et al. 2005). Limonene and some of
the esters and aldehydes exhibited decreased air-liquid partitioning coefficients in
the presence of xanthan, indicating that the release of these volatile aroma
10 Flavour Delivery 359

compounds was reduced due to interaction with the xanthan matrix. The release of
43 flavour compounds was reported from viscous solutions containing
λ-carrageenan and sucrose (Bylaite et al. 2004). No overall effect of λ-carrageenan
was found, except with the most hydrophobic compounds. Analysis of flavour
release under non-equilibrium conditions, however, revealed a suppressing effect
of λ-carrageenan on the release rates of aroma compounds, and the extent of decrease
in release rates was dependent on the physicochemical characteristics of the aroma
compounds, with the largest effect for the most volatile compounds. ι- and
κ-carrageenans did not appear to cause a significant difference in odour intensity
for flavour molecules of different chemical classes, except for ethyl hexanoate,
which was found to be retained by ι-carrageenan. Nevertheless, in-mouth aroma
perception was significantly different between the two systems (Juteau et al. 2004).
The influence of food composition and temperature on the release of a strawberry
flavouring in a system simulating yoghurt with a fruit preparation syrup has also
been described (Nongonierma et al. 2006). Agar gels were used as encapsulating
medium for limonene (Piazza and Benedetti 2010). The flavour binding capacity of
amylose, a major component of starches is also well known (Conde-Petit et al. 2006;
Arvisenet et al. 2002; Yeo et al. 2016). Flavour-protein interactions and their
influence on flavour release and flavour perception are described by Wang and
Arntfield (2017).
Control of flavour release is important when fat is reduced in certain foods. In
part, hydrocolloids are added as thickeners to products to compensate for the loss of
thickness and texture. Arancibia et al. (2015) demonstrated that thickener type and
concentration and fat content significantly affect in vivo aroma release from a fat
reduced dairy dessert. The delivery to the nasal cavity of the most lipophilic
compound (linalool) mainly depended on fat content while thickener type and
concentration mainly affected the release of the least lipophilic compound (cis-3-
hexen-1-ol).
Such interactions between flavourings and other food ingredients constitute a
challenge for the formulation of the flavouring. Flavourists as well as flavour
delivery experts need to be aware to balance the ratio of individual compounds. In
an exemplary article, Yang et al. (2011) describe reformulating a commercial
strawberry flavouring that delivered a highly acceptable flavour in a pectin-sucrose
gel, but did not perform as well in a chewy candy containing sugar, protein and fat.

5 Summary and Outlook

Development and innovation in the flavour industry will continue to be driven by the
demand of consumers for fresh and authentic eating and drinking experiences.
Flavour delivery systems seen as the essential bridge between the flavourings and
food and beverages will play a key role. Delivery systems allow capturing,
protecting, and releasing flavourings, and food hydrocolloids play an indispensable
role for all these functionalities.
360 M. Schultz

In a recent article (Givaudan Science and Technology Team 2017), three key
areas have been identified to be important to be successful in the highly competitive
area of designing state of the art flavour delivery systems. First, the growing need to
replace chemically modified ingredients with natural substances and those that are
more label-friendly. Second, the industry must develop a foundational understanding
of how these materials behave in flavour applications; and finally, it must also
improve the efficiency and sustainability of processing technologies:
When it comes to the material basis of delivery systems, customers are always keen to make
more appealing declarations on the end product labels. As an example, modified starches
have historically been used as encapsulation matrices in spray drying thanks to the solubility
in water and the ability to form a robust encapsulating matrix around the flavour. The
challenge is now to either develop natural starch-based materials, which can provide the
same performance, or research alternative materials from different sources.
A scientific approach to the understanding of how materials behave is also essential. While
much is already known, there is work to be done to close the gaps in the existing knowledge,
for example to pinpoint how specific flavours might be best protected in an encapsulation
matrix.
There is also room to improve the cost effectiveness and sustainability of processing
technologies, such as using less water and working at lower temperatures in the drying
processes, which would save energy and reduce the CO2 footprint.

The introduction of new functional hydrocolloids is often restricted by legislative

hurdles and lack of consumer acceptance. Materials only processed by physical
methods with a natural and healthy reputation will be preferred and most successful
in the market. Natural materials (McClements et al. 2017), dietary fibres (Gidley and
Yakubov 2019), and plant proteins (Wang and Arntfield 2017) have the potential to
replace traditional emulsifiers and encapsulation materials in case there are labelling,
performance and cost benefits. Sustainability concerns will play an increasing role;
sourcing and supply including the use of food processing side streams will also
become more important.

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Chapter 11
Encapsulation and Targeted Release

Bin Liu, Lulu Jiao, Jingjing Chai, Cheng Bao, Ping Jiang, and Yuan Li

Abstract This chapter described the definition of encapsulation and food delivery
systems/food carriers, benefits for encapsulation, and various delivery systems
classified by structure and function. Firstly, the classification of food bioactive
compounds and the reason for encapsulation were discussed. Later, the structure
classified delivery systems of core–shell carriers, Pickering emulsions, complex
coacervates, gels, and self-assemblies of cross-linked biopolymer were described.
Due to the delivery of bioactive compounds via the oral route is restricted by various
physiological barriers including harsh gastrointestinal (GI) pH conditions, enzymes
in GI, the mucus layer, and the epithelium, intelligent delivery systems are promising
strategies to protect bioactive molecules from degradation and improve their bio-
availability. Then the novel intelligent carriers that are responsive to the oral route,
pH, enzymes, and cell receptors were discussed. Next, the cellular uptake mecha-
nisms of food carriers were summarized and analyzed. In addition, the gastrointes-
tinal in vitro and in vivo models to evaluate the function of the carriers were
compared, and the applications of carriers in different types of food were reviewed
and analyzed. Finally, the future design and development trends and practical
applications of the carriers are prospected. This comprehensive multidisciplinary
chapter provides useful guidelines and inspirations for the design of intelligent
carrier systems, which were exploited for the encapsulation of bioactive com-
pounds and improving their low solubility, poor stability, and low bioavailability
during the processing, storage, and orally uptake.

Keywords Intelligent delivery systems · Food colloid-based carriers ·

Encapsulation · Food bioactive compounds · Bioavailability

B. Liu · L. Jiao · J. Chai · C. Bao · P. Jiang · Y. Li (*)

College of Food Science and Nutritional Engineering, China Agricultural University, Beijing,
People’s Republic of China
e-mail: yuanli@cau.edu.cn

© Springer Nature Singapore Pte Ltd. 2021 369

Y. Fang et al. (eds.), Food Hydrocolloids,
https://doi.org/10.1007/978-981-16-0320-4_11
370 B. Liu et al.

1 Introduction

Great attention has been paid to bioactive compounds derived from foods over the
past few decades since they may be beneficial to the health by decreasing the risks of
numerous disorders, including obesity, cancer, diabetes, and cardiovascular diseases
(CVDs) (Oh 2016). Such beneficial effects can be attributed to their effects against
inflammation, cancer, oxidation, hypertension, and hyperlipidemia, apart from the
fundamental nutritional effects. Such food-originated bioactive compounds cover
phytochemicals (Velderrain-Rodríguez et al. 2014), proteins (Dhaval et al. 2016),
bioactive peptides (Dhaval et al. 2016), vitamins (Katouzian and Jafari 2016),
bioactive polysaccharides (PLS, Jiang et al. 2016), and fatty acids (FAs, Kruk
et al. 2016). Such compounds show high sensitivity to diverse environmental
stimuli, including heat, light, oxygen, and low pH in the processing and storage
processes (Gleeson et al. 2016). Besides, more efforts should be made to manage the
problems occurring when bioactive ingredients are administered orally. For instance,
some consumers discover the unpleasant flavor of certain bioactive ingredients when
they are administered orally. Besides, hydrophobic compounds are poorly soluble,
which greatly prevents them from being dissolved into the water medium and
absorbed in the gastrointestinal tract of human body. However, such compounds
are unstable in the processing and storage conditions (like heat, oxygen, and light
exposure), which decreases their functional efficiency. Generally, many of the
bioactive ingredients display low absorption rates via small intestinal epithelium.
Foods will be digested and degraded in acidic environment by a variety of enzymes
(pepsin, amylase, pancreatin) and by the mucus barriers that cover the GI tract
luminal epithelium (Chen et al. 2011). Besides, only limited food ingredients can
diffuse through the intestinal mucus or permeate across the intestinal epithelium,
which has seriously impeded their bioavailability. On the other hand, food ingredi-
ents can hardly be absorbed by cells even though they permeate the intestinal mucus.
Only limited polyphenols can be absorbed in cells based on the carrier-regulated
transport, which is related to the mechanism of “multiple-drug resistance”. There are
multiple glycoprotein efflux pumps on epithelial cells, which function to pump the
polyphenols absorbed back to intestinal lumen (Chan et al. 2004).
For overcoming the above-mentioned problems, a number of delivery systems
based on food colloids are synthesized, which can mask the unpleasant compound
taste, dissolve the hydrophobic compounds in water through combining them in the
food hydrocolloid milk protein microcapsules, protect against the harmful environ-
ment, and improve the permeability across the epithelial and intestinal mucus
(Livney 2010). Numerous micro/nanocapsules delivery systems constructed based
on food colloids are able to resist the decomposition of bioactive ingredients in GI
tract. The above-mentioned systems have different structures and sizes and are
utilized to change the cargo release kinetics as well as target delivery to typical
sites. Certain carriers that can penetrate the intestinal mucus are able to rapidly
transport bioactive ingredients through intestinal mucus, as a result, they are mark-
edly absorbed to the systemic circulation when they are administered orally
11 Encapsulation and Targeted Release 371

(Lundquist and Artursson 2016). Nonetheless, certain carriers prepared by

mucoadhesive biomaterials will increase the compound retention time in intestines
and elevate the corresponding local concentrations near epithelial cell surface
(Saura-Calixto et al. 2007). Recently, delivery systems synthesized based on food
colloids become more and more functionalized. But to obtain the more intelligent
carriers, factors including oral cavity responsiveness, enzymes, intestinal pH, over-
coming of intestinal mucus barriers, and physiological biorecognition must be taken
into account in the design of a sophisticated delivery system. The compound food
hydrogels coacervation system has been synthesized to deliver flavor compounds
orally, so as to extend the food retention time and to elevate the sensory perception
(Zhang et al. 2015). Layer-by-layer carriers based on polysaccharide hydrogel can
respond to intestinal pH, which prevent the decomposition of encapsulated com-
pounds and release these compounds into small intestine (Shi et al. 2017). Polysac-
charide/polyaminoacid hydrogel for colon-specific delivery systems assists in the
release of encapsulated compounds by means of the colonic dextranase-mediated
degradation (Casadei et al. 2008). For carriers responsive to receptor that target the
intestinal cells, they markedly enhance the absorption of encapsulated compounds
by cells (Tian et al. 2016). Besides, the food hydrocolloid soy/milk liposomes that
can penetrate the intestinal mucus efficiently promote cargo absorption because they
can rapidly penetrate the mucus barriers to reach the epithelial cells (Li et al. 2017a).
In comparison, the mucoadhesive carriers constructed based on food colloid can
interact with mucus, thus realizing the extended release of the encapsulated com-
pounds (Sheng et al. 2015). As a result, the intelligent delivery systems synthesized
based on food colloids become more and more important, which offer the efficient
platforms to apply these bioactive ingredients in food industry and to promote their
effects and bioavailability.

2 The Encapsulation Technology Developed for Sensitive

Bioactive Compounds

2.1 Factors Influencing the Stability and Bioavailability

of Bioactive Compounds

The most important types of bioactive compounds were summarized in Fig. 11.1
(Taiz et al. 2015; Bao et al. 2019). As shown in Fig. 11.1, the bioactive ingredients
displayed include bioactive peptides (casein, carnosine, α-lactalbumin, angiotensin-
converting enzyme (ACE)), polyunsaturated fatty acids (PUFAs, such as EPA,
ARA, DHA), bioactive polysaccharides (grifolan, lentinan), polyphenols (resvera-
trol, curcumin, tea polyphenols, anthocyanin, quercetin), minerals (Ca, Fe, Zn),
vitamins (VB6, VC, VD, VE), and others including isoflavone, carotenoids,
isothiocyanates, sitosterol, alkaloids, terpenes, and lignin (not presented in
Fig. 11.1) (Taiz et al. 2015). Such bioactive ingredients are suggested to be
372 B. Liu et al.

Fig. 11.1 Overview of food-sourced bioactive compounds. Important types of bioactive ingredi-
ents were summarized, including polyphenols (e.g., curcumin, resveratrol, epigallocatechin gallate
(EGCG), quercetin, anthocyanin), polyunsaturated fatty acid (e.g., docosahexaenoic acid (DHA),
eicosapentaenoic acid (EPA), arachidonic acid (ARA)), vitamins (e.g., vitamin C (VC), vitamin A
(VA), vitamin E (VE)), bioactive polysaccharides (e.g., lentinan, grifolan), bioactive peptides (e.g.,
α-casein, L carnosine, α-lactalbumin) and mineral elements (e.g., Zn, Fe, Mg, Ca, Na, P) (Bao et al.
2019). Reproduction with permission from (Bao et al. 2019), Copyright 2020 ELSEVIER. Order
Number: 4958630740831

beneficial to human health (Table 11.1). For instance, flavonoids including apigenin
and quercetin show potent antitumor activities (Batra and Sharma 2013); omega-3
PUFAs can be adopted to reduce the risk of cardiovascular diseases (CVDs)
(Richard et al. 2009); carotenoids like astaxanthin and β-carotene can scavenge the
free radicals (Nagao 2009); while bioactive polypeptides including peptidyl dipep-
tidase A have antihypertensive effect (Corvol et al. 1995); and the plants- and algae-
derived polysaccharides have excellent anti-inflammatory activity (Liu et al. 2015a).
Food nutraceuticals can hardly be adsorbed by cells, which has severely affected
their bioavailability in vivo because their absorption in the human body is only
achieved when they penetrate the GI barriers. In addition, numerous environmental
stimuli have affected the stability of bioactive ingredients. This section analyzed the
vital factors that affected bioactive ingredient absorption, as displayed in Table 11.1.
11 Encapsulation and Targeted Release 373

Table 11.1 The summary of physiological function, absorption mechanisms, and environmental
influencing factors for bioactive compounds including vitamins, polyphenols, polypeptides, poly-
saccharides, and unsaturated fatty acids. Reproduction with permission from (Bao et al. 2019),
Copyright 2020 ELSEVIER. Order Number: 4958630740831
Absorption
Bioactive Influence mechanisms in small
compounds Function Factors intestine Ref.
β-Carotene Antioxidant, antican- Light, heat By passive diffusion (Wang
cer, immune enhance- and/or transporter- 1994)
ment and anti-obesity mediated processes
activities, prevention
of cardiovascular
diseases
Omega-3 poly- Essential for humans Heat, light Carrier-mediated (Yang
unsaturated fatty and animals to treat transport et al.
acid and prevent athero- 2017)
sclerosis and cardio-
vascular diseases
Quercetin Prevention of oxida- Heat, alkali Passive diffusion (Day et al.
tive stress-related and/or transporter- 2003)
chronic diseases mediated processes
Curcumin Antioxidant, anti- Heat, light, Passive diffusion, (Wang
inflammatory, antican- Fe P-glycoprotein pump- et al.
cer, antiviral, and mediated 2017)
antibacterial activities transportation
Resveratrol Anticancer, antioxi- Heat, light Passive diffusion (Gambini
dant, antiaging, anti- and/or transporter- et al.
frailty, anti-inflamma- mediated processes 2015)
tory, anti-allergenic
activities
Vitamin D Facilitating calcium Light, acid Passive diffusion (Reboul
absorption to build-up and/or receptor- et al.
bone preventing can- mediated transport 2011)
cer and cardiovascular
diseases
Vitamin C Action as coenzyme or Heat, light, Na+-dependent (Bianchi
prosthetic group which humidity transporters-mediated et al.
is important to meta- transportation 1986)
bolic activity of human
body
Vitamin A Essential to the forma- Light, acid, As the form of retinol (Harrison
tion of visual chromo- heat, via the chylomicron/ 2012)
phore and hormone humidity lymphatic route or
which maintain the flowed into the portal
vision and promote vein circulation by the
body growth; lipid transporter
strengthen immunity
and scavenge free
radicals
(continued)
374 B. Liu et al.

Table 11.1 (continued)

Absorption
Bioactive Influence mechanisms in small
compounds Function Factors intestine Ref.
Vitamin E Antioxidant, antican- Heat, light, Passive diffusion, (Reboul
cer, antiaging activity humidity receptor-mediated 2017)
transport
Casein peptides Lowering the blood High enzy- Vesicle-mediated (Shen and
pressure by matic degra- transcellular transport Matsui
angiotensin-converting dation, short 2017)
enzyme (ACE)- half-life
inhibitory peptides
Soybean Anti- Easy oxida- Carrier-mediated (Chen
β-Conglycinin hypocholesterolemic, tion, easy transport, receptor- et al.
anti-hypoglycemic, hydrolysis mediated transport 1995)
antioxidant, and prebi-
otic effects
Astragalus Triggering Stickiness, Carrier-mediated (Cheng
polysaccharides non-specific reaction low transport, passive et al.
of the immune system, solubility diffusion 2011)
and have antimicro-
bial, anti-
hypocholesterolemic,
anti-hypoglycemic
antioxidant, and prebi-
otic effects
Fungal Immunomodulating, Heat, light, Clathrin-mediated (Giavasis
polysaccharides anticancer, antimicro- humidity endocytosis 2014)
bial,
hypocholesterolemia,
hypoglycemic, and
health-promoting
properties

First of all, a variety of environmental stimuli can affect the bioactive ingredient
stability, like light, strong acid, oxygen, and heat. For instance, carotenoids have
antioxidant ability, which is achieved by the active conjugated double bonds that can
be oxidized and broken easily (Wang 1994). Moreover, the interactions between
carotenoids and other food ingredients, like fats and proteins, can decrease their
antioxidant ability (Ranhotra et al. 1995). Curcumin shows high sensitivity to heat,
light, and acid, since they are three tightly associated lipophilic moieties constituted
by certain double bonds and phenolic groups (Heger et al. 2014). On the other hand,
the harsh GI tract environment is also a major factor that affects bioavailability. For
instance, the degradation rates of hydroxycinnamoyl acid and flavonoids are 80%
and 84% in the in vitro GI tract, while that of vitamin C is 91% under intestinal
digestion conditions (Vallejo et al. 2004). Further, digestive enzymes, like trypsin in
intestine and pepsin in stomach, are also able to decompose peptides and proteins,
resulting in the decreased activity (Layer and Gröger 1993). Particularly, the
11 Encapsulation and Targeted Release 375

bioactive ingredients are digested or broken before they can exert their functions.
Therefore, digestion in GI tract also represents a physiological barrier that should be
managed to improve ingredient uptake. Mucus layer is the last barrier of absorption,
and it covers the surface of the whole GI tract. Mucus is mainly made up of lipids,
glycoproteins, along with the sloughed cellular materials (Abdulkarim et al. 2015). It
may serve as an intelligent semipermeable membrane, which allows for numerous
nutritional substances exchange in the meantime of preventing pathogen or bacte-
rium permeation onto the surfaces of epithelial cells (Cone 2009). Nonetheless, only
little amount of food ingredients can penetrate the mucus, which possibly suppresses
the availability and absorption of these compounds in enterocytes. Typically, hydro-
phobic bioactive ingredients have been reported to interact with mucin proteins,
thereby slowing down their penetration across the mucus (Liu et al. 2015b).
In this regard, it is of great necessity to design a measure for managing the above
problems (Li et al. 2015b). Encapsulation technique has been developed rapidly, and
used widely applied in many areas such as pharmaceutical and food industries. In
food industries, encapsulation refers to the process where core material microparti-
cles are packed in the wall material for the formation of capsules. Encapsulation
means to isolate the active ingredients in food products with the building blocks of
food-grade biopolymers, and it is identified as an efficient approach for masking the
unpleasant taste and improving stability and solubility of orally administered func-
tional ingredients. In the field of food industry, encapsulation systems have been
widely used for solving the formulation issues induced by the low active ingredient
stability (incompatibility of the active ingredients with the food matrix), or for
controlling active ingredient release or improving the nutrient bioavailability
(Ubbink and Krüger 2006). The encapsulation approach is used for protecting
bioactive compounds (micronutrients, nutraceuticals, polyphenols, antioxidants,
and enzymes), protecting these compounds from the harsh environment during the
finished applications, and controlling cargo release at specific sites. The encapsula-
tion technology is adopted in food industry for maintaining excellent properties and
quality, including taste, texture, water solubility, stability, and color strength in the
processing and storage processes. For bioactive compounds, their absorption rate is
tightly associated with the water solubility, but most of them are hydrophobic, giving
rise to the poor absorption in GI tract. At present, the Pickering emulsions, complex
coacervates, cross-linked polymer gels, core–shell-structured microcapsules,
together with the self-assembled structures, have been extensively utilized for
enhancing the stability, bioavailability, and water solubility of the encapsulated
compounds (Fig. 11.2). The important food delivery systems based on encapsulation
techniques classified by structure are presented in the section below.

2.2 The Food Delivery Systems Classified by Structure

The delivery systems for food bioactive compounds played a vital part in the food
sector. Natural biopolymers, including lipids, proteins, and polysaccharides, are
376 B. Liu et al.

Fig. 11.2 The delivery systems classified by their building structures including (a) complex
coacervates capsules, (b) Pickering emulsion delivery systems, (c) core–shell capsules made by
layer-by-layer techniques, (d) cross-linked polymer gels, and (e) self-assembled nanocapsules (Bao
et al. 2019). Reproduction with permission from (Bao et al. 2019), Copyright 2020 ELSEVIER.
Order Number: 4958630740831

often used as wall materials for preparing novel capsules. The food delivery systems
can be built by various structures such as Pickering emulsions, complex coacervates,
cross-linked biopolymer gels, core–shell-structured microcapsules, along with the
self-assembled structures (Fig. 11.2). Those carriers will be discussed in this section.

2.2.1 Complex Coacervates

Two Dutch physicists named Kruyt and Bun Enberg de Jong created the term of
“complex coacervate” for discriminating it from the one-single polymer coacervate.
Complex coacervate stands for the phenomenon where phase separation occurs
when two polymers with opposite charges are added into water (Sing 2017). The
formation of complex coacervates was driven by the discharge of counterions with
low molecular weight (MW) that conjugated with charged groups on large molecules
and entropy gained during this process (Pergushov et al. 2012). Typically, the most
extensively synthesized complex coacervates consist of proteins with positive
charges and polysaccharides with negative charges (De Kruif et al. 2004). Among
the earliest diverse polysaccharides with negative charges, acacia gum (AG) is
involved in the coacervation with β-lactoglobulin (β-lg) with positive charges
(Schmitt et al. 2001).
Complex coacervates can form the insoluble films that may be additionally
synthesized as microcapsules. Many studies have been conducted to investigate
the delivery systems constructed based on the protein–polysaccharide complex
coacervates. The casein hydrolysate was encapsulated in complex coacervation
formed by soybean protein isolate (SPI)/pectin (Mendanha et al. 2009), of which
the solubility and stability were improved, and the bitter taste was masked. It is also
reported that, the whey protein isolates (WPI)-GA complex coacervate can be
utilized for the encapsulation of L. paracasei subsp. paracasei in yogurts without
changing the texture. It is an efficient carrier used to deliver probiotics to the intestine
(Bosnea et al. 2017). According to the above studies, complex coacervate is a
candidate food carrier to improve bioactive components for their stability. The
11 Encapsulation and Targeted Release 377

charge screening effect of the 150 nM physiological salt can result in complex
coacervates capsule dissociation, while cross-linking can manage such problem.

2.2.2 Pickering Emulsion Delivery Systems

In recent years, wide attention has been paid to the particle-stabilized Pickering
emulsions at food grade because of their excellent colloidal stability. To be specific,
Pickering emulsions are stabilized with the solid colloidal particles but not the
biopolymers or small molecular surfactants. Those particles utilized for stabilizing
Pickering emulsions should be partially wetted by the aqueous and oil phases
(Chevalier and Bolzinger 2013). There is a contact angle θ for the colloidal particles
absorbed onto the interface between oil and water for characterizing solid particle
wettability. The contact angle of water-in-oil (W/O) emulsions produced in the
presence of residual hydrophobic particles in oil phase is over 90
; on the contrary,
that of oil-in-water (O/W) emulsions produced in the presence of residual hydro-
philic particles mainly in aqueous phase is <90
(Binks 2002; Williams et al. 2014).
Typically, emulsions stabilized by colloidal particles are extensively investigated to
examine whether they can be utilized as the delivery systems of food ingredients,
and they are usually classified as 3 types: (1) polysaccharides with hydrophobic
modification: the nanospheres constructed based on amphiphilic starch are suggested
to be effective on stabilizing Pickering emulsions (Tan et al. 2012). Besides, both
modified starch (MS) and microcrystalline cellulose (MCC) are utilized for prepar-
ing the highly stable Pickering emulsions. Additionally, compared with MS parti-
cles, the MCC particles can better improve the lipid stability to resist oxidation, since
they can produce a thick layer surrounding oil droplets, thus preventing lipid
oxidization (Kargar et al. 2012). The vitamin E (VE as in L283) stability is suggested
to be markedly improved when it is encapsulated to the modified tapioca starch
nanocapsules. At 60 days after storage under the temperature of 35
C, the vitamin E
retention rate is about 1/2 of the original value (Hategekimana et al. 2015). (2) Pick-
ering particles prepared based on protein: specifically, zein, the plant protein with
water insolubility, is used for stabilizing Pickering emulsions. Feng and colleagues
used sodium caseinate (NaCas) to modify zein nanoparticles (NPs) on the surface,
and discovered that the resultant nanocomplexes performed better in stabilizing the
O/W Pickering emulsions relative to the unmodified ones (Feng and Lee 2016).
(3) Miscellaneous Pickering particles: Colloidal particles can be utilized to stabilize
the colloidosome-based Pickering emulsions by using those self-assembled particles
at the interfaces, and the modified products show superb embedding together with
controlled-release characteristics. Curcumin is encapsulated into the chitosan–
tripolyphosphate NPs-stabilized Pickering emulsions (Shah et al. 2016). As a result,
the Pickering emulsions become the prospective systems used to engineer the
functional interfaces within the food emulsions. They show relative anti-coalescence
stability because the Pickering particles have great desorption energy. However, it
requires a long time for the equilibration of the rigid Pickering particles shell in the
process of production, making it difficult to obtain emulsion with expected droplet
378 B. Liu et al.

size (Berton-Carabin and Schroën 2015). For solving the above problems, soft
particles including whey protein nanogels can be adopted to be the emulsifiers to
make emulsions, which is a novel way to generate the uniform size of emulsion
droplets (Wu et al. 2015).

2.2.3 Core–Shell Microcapsules Delivery System Prepared by

Layer-by-Layer Technique

As the novel colloidal structure in the encapsulation and controlled release of

bioactive ingredients, the multilayer microparticles synthesized by the layer-by-
layer (LbL) assembly technology have received wide attention (Tong et al. 2012).
Notably, the LbL deposition approach forms a specific structure that has tailored
functionality on the basis of weak interaction across the interacting polymers (Costa
and Mano 2014). The LbL assembly approach has been utilized to prepare the three-
layered microcapsules with the use of octenyl succinic anhydride (OSA)-starch, soy
protein isolate, and chitosan as the wall materials. Three-layer microcapsules display
reduced encapsulation rate in comparison with the one- or two-layer counterparts,
but they show the procedural controlled-release behavior (Noshad et al. 2015). The
poly (L-glutamic acid) (PLGA) with negative charges and poly (L-lysine) (PLL)
with positive charges were assembled onto the oxidized starch microgel particle
surfaces through the LbL assembly technology, and the resultant PLL/PLGA-coated
starch microgels are found to efficiently reduce lysozyme release within the 0.05 M
NaCl solution, which also prevent gel particle degradation via the action of
α-amylase (Li et al. 2012). Nonetheless, there may be certain drawbacks, like the
difficulties in regulating the synthesis process usually accompanied with aggrega-
tion, and assembly dissociation because of salt sensitivity, which have restricted
their applications (Guzmán et al. 2017). Besides, the introduction of other inter-layer
covalent cross-links may stabilize the LbL structure (Han et al. 2017).

2.2.4 Cross-Linked Biopolymer Gels

Polymer gels can be produced through the intermolecular or intramolecular biopoly-

mer cross-links connected based on polymer chains. In general condition, both
physical (non-covalent bonds) and chemical (covalent bonds) cross-links can be
used to form biopolymer network. Of them, chemical cross-linking is extensively
utilized in various fields, such as microsphere preparation, double-layer cross-
linking, and food starch modification. The modified food starch using sodium
tripolyphosphate (STPP) combined with sodium trimetaphosphate (STMP) in the
alkaline environment has enhanced activity of α-amylase to restrict swelling and to
resist decomposition (Woo and Seib 2002). Further, the introduction of
transglutaminase at food grade into the gelatin–maltodextrin microspheres leads to
superb activity in the protection of probiotic lactic acid bacteria that are subjected to
the simulated GI tract environment (Nawong et al. 2016).
11 Encapsulation and Targeted Release 379

In potato starch, the TEMPO ((2,2,6,6-Tetramethylpiperidin-1-yl)oxyl)-oxidation

can be applied in the selective oxidation of primary alcohol groups located at the
sixth position in hexose unit to the carboxyl groups. Such method achieves a
controlled oxidation degree of 30–100%, and the resultant toxicity is negligible
(Li et al. 2009; Jiang et al. 2018). For the charged neutral polysaccharides, such an
oxidation process can enhance the controlled-release property and charge density
(Li et al. 2011). For instance, for oxidized starch microgels cross-linked with STMP,
they exhibit a great loading capacity for anthocyanins (Wang et al. 2013) and
proteins with positive charges (Zhao et al. 2015a). Besides, they can regulate
β-carotene release especially under the intestinal conditions (Wang et al.
2015a, b). Oxidized starch microgels cross-linked with glycerol diglycidyl ether
are reported to regulate lysozyme release and uptake (Zhao et al. 2015a). In addition,
the oxidized konjac glucomannan microspheres are synthesized through the cross-
linking of Fe3+ (Chen et al. 2014; Chen et al. 2016), and they may be decomposed
and release anthocyanins in sequence under the intestinal conditions (Lu et al. 2015).
As the non-covalent cross-linker, tannic acid has been utilized for preparing the
hollow zein NPs that have enhanced resistance to the simulated intestinal digestion
compared with the non-crosslinked samples. But, cross-linked polymers are linked
with the disadvantage of absorption of excessive water, while this is undesired in
food processing. In some cases, the cross-linking agents will develop side reactions
with bioactive compounds existing within the hydrogel matrix (Maitra and Shukla
2014). Further, these compounds involved may show the uncontrolled pulsed release
driven by diffusion. Therefore, a more controllable and cleaner process of cross-
linking should be developed in the near future.

2.2.5 Self-Assembled Delivery Systems

Recently, wide attention has been paid to the self-assembly based delivery systems
(Song et al. 2015). Self-assembly occurs spontaneously, which helps to organize the
biopolymers to micro/nanostructures with clear boundary by means of non-covalent
interactions (such as Van der Waals forces, hydrogen bonding, π-π interactions, and
electrostatic forces) in the absence of any other external intervention (Mendes et al.
2013). It should be noted that, this spontaneous process on the basis of the reversible
and weak bonds or interactions can result in the formation of different nanostructures
because of the “self-error-correcting” and “self-checking” processes controlled
thermodynamically (Baek et al. 2015). For instance, α-lactalbumin is partly hydro-
lyzed to amphiphilic peptides, which contributes to the self-assembly to
nanomicelles that are 20 nm in size on average (Li et al. 2017b). The prepared
nanomicelles can be used to load curcumin at the hydrophobic core and deliver
curcumin to tumor sites specifically. Numerous natural biopolymers in foods,
including proteins and polysaccharides, are found to generate the self-assembled
structure by themselves or with additional functional molecules (Song et al. 2015).
In the case of food biopolymers, self-assembly possibly takes place under the
conditions mentioned below. (1) Polysaccharides self-assembly delivery systems:
380 B. Liu et al.

Fig. 11.3 The illustration of micelles loaded with β-carotene. α-Lactalbumin was partially hydro-
lyzed into amphiphilic peptides, which self-assembled into micelles. During this process β-carotene
can be loaded into the micelles core via hydrophobic interaction (Du et al. 2019). Reproduction with
permission from (Du et al. 2019), Copyright 2020 ELSEVIER. Order Number: 4958631460919

the self-assembled starch NPs can be used to efficiently enhance starch

nanocomposite film thermostability on the basis of the short amylose (Jiang et al.
2016). It is reported that, when linoleic acid is conjugated with hydroxyethyl
cellulose, a novel amphiphilic cellulose derivative can be generated, which can
self-assemble to spherical nanomicelles in water. Such resultant micelles show a
great efficiency in encapsulating hydrophobic components, thus enhancing the
β-carotene solubility as well as bioavailability (Yang et al. 2016). (2) Protein self-
assembly delivery systems: the amphiphilic peptides acquired by the partial hydro-
lysis of α-lactalbumin are able to self-assemble into micelles for encapsulating
β-carotene and curcumin at the hydrophobic core (Li et al. 2017b; Du et al. 2019).
The schematic illustration of α-lactalbumin nanomicelles loaded with β-carotene
was shown in Fig. 11.3 (Du et al. 2019). Besides, the negatively charged
α-lactalbumin nanomicelles are able to deliver multiple antioxidants such as antho-
cyanins and curcumin with enhanced synergistic cellular antioxidant activity (Jiang
et al. 2018). When vitamin D is encapsulated into the β-lactoglobulin self-assembled
capsules, its absorption and solubility are improved (Górska et al. 2012). (3) Lipo-
somes: microbubbles loaded with DOX (doxorubicin)-liposome can be generated
spontaneously according to the ultrasound-mediated self-assembly approach, and
the resultant products can target cancer cells. On the other hand, hydrophilic
polyethylene glycol (PEG) modification, ursolic acid (UA) can be successfully
loaded using the soya lecithin-based self-assembled liposomes. Such self-assembled
liposomes modified with PEG efficiently enhance UA stability and reduce its release
(Zhao et al. 2015b). In addition, Lee and colleagues discovered that, lipid anion
reverse self-assembly took place within the diluted oil through integrating the
saturated phospholipid, 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC),
and the divalent or trivalent cations. The above lipid anions are stable for few
weeks, which can be utilized for encapsulating bioactive components (Lee et al.
2013). For instance, the degradation of curcumin within GI tract is prevented
through liposomes encapsulation, which can markedly increase its plasma level,
suggesting the promoted bioavailability and bioactivity (Takahashi et al. 2009).
To sum up, these delivery systems can serve as the candidate measures to
improve the solubility, stability as well as bioavailability of bioactive components.
Table 11.2 summarizes the benefits, mechanisms of formation and assessed models
11 Encapsulation and Targeted Release 381

Table 11.2 Comparisons of the encapsulated compounds, mechanisms of formation, benefits, and
assessed models applied in investigating the absorption and digestion behaviors of a variety of
carrier systems. Reproduction with permission from (Bao et al. 2019), Copyright 2020 ELSEVIER.
Order Number: 4958630740831
Encapsulated Formation Used
Structures compounds mechanisms models Benefits Ref.
Complex β-Carotene Electrostatic Simulated Improved stabil- (Schmitt
coacervation interactions GI fluid ity, and antioxi- et al.
model dant activity 2001)
Pickering Vitamin E Particles absorp- TIM Improved (Williams
emulsion tion at the oil– models stability et al.
water interfaces 2014)
Spray Carotenoids Atomization at an TIM Improve solubil- (Ye et al.
drying elevated models ity and stability 2010)
temperature
Layer-by- Polypeptide Weak interaction Simulated Slow down the (Costa
layer of the interacting GI fluid enzymolysis and Mano
polymers model 2014)
Cross-linked β-Carotene Intra- or Cell cul- High storage sta- (Tian et al.
biopolymer intermolecular ture bility and 2016)
gels physical or models improved cellu-
chemical cross- lar uptake
links of
biopolymers
Self- Curcumin Non-covalent Simulated Improve solubil- (Qiu et al.
assembly interactions GI fluid ity and 2014)
model bioavailability
Self- Insulin Emulsion Cell cul- Improve phar- (Shan
assembly template ture and macological et al.
animal availability 2015)
models

for those above technologies for diverse components. Therefore, the appropriate
encapsulation technology may be screened for some compounds with diverse
properties.

3 The Different Delivery Systems Classified by Function

Based on the above analysis, food carrier systems can be used to enhance bioactive
compound stability and solubility, yet there are numerous challenges to be solved
when food ingredients are delivered orally, like the ionic strength, pH, mucus
barriers, and enzymes within GI tract. Such problems have great influences on
carrier properties and thus the bioactive compound bioavailability. Bioaccessibility
refers to the proportion of components released from the food substrate into GI tract,
and this helps to facilitate the intestinal uptake (Saura-Calixto et al. 2007). Besides,
bioavailability can accurately indicate the effective use of bioactive compounds
382 B. Liu et al.

indeed absorbed via enterocytes and then entering into the blood circulation.
Research on bioaccessibility mainly emphasizes the digestion in vitro, while bio-
availability focuses on the absorption in vivo. For the sake of improving food
ingredient oral bioavailability, more efforts are needed to develop the advanced
carrier systems to deliver bioactive compounds on demand and release them at the
appropriate time and site. To enhance the responsiveness to intestinal pH value,
retention performance, mucus penetration, and cell absorption of bioactive com-
pounds, it is needed to design the more advanced carriers.
In addition, the smart delivery systems for the controlled release of encapsulated
compounds at suitable time and site through targeted site-specific delivery are in
greater and greater demands. At the same time, oral bioavailability is markedly
improved by the use of carriers that have improved mucus-penetrating capacity and
increased epithelial cell absorption efficiency (Ezhilarasi et al. 2013). Based on the
previous descriptions regarding the microenvironments for different parts in GI tract,
the smart carriers with responsive to the GI tract (including mouth, stomach, colon,
and small intestine) environmental changes should be designed (Table 11.3). Typ-
ically, the smart food delivery systems can be classified as different types based on
their speciality, including pH-responsive, oral-responsive, targeted-delivery,
enzyme-responsive, mucoadhesive carriers, and mucus-penetrating (Fig. 11.4)
(Chai et al. 2018). Firstly, the complex coacervation system can be developed for
the controlled release of flavor compounds in the mouth, so as to extend the food
retention time while increasing sensory perception (Zhang et al. 2015). Besides, the
intestinal pH-responsive carriers can be utilized for protecting the encapsulated
compounds transported into the stomach and releasing these compounds into the
small intestine when the carriers are degraded in the intestine (Garinot et al. 2007).
Colon-specific carriers have been synthesized for releasing the encapsulated com-
pounds after the carriers are degraded by the microbial enzymes in the colon
including dextranase. The ligand-conjugated carriers significantly enhance cell
absorption through the receptor-regulated endocytosis (Jiang et al. 2014). Mucus-
penetrating carriers rapidly penetrate into the mucus barriers and reach the epithelial
cells at an increased level, leading to the enhanced encapsulated compound uptake
relative to the free compounds (Li et al. 2017a). Besides, mucoadhesive carriers may
stay stably in the mucus for a long period and allow for the controlled release of
encapsulated compounds (Sheng et al. 2015). The next section will elaborate each
type of carrier.

3.1 Oral-Responsive Delivery Systems

Generally, flavor compounds include the volatile and sensitive molecules like
ketones, aldehydes, or additional phytochemicals (Wu et al. 2016). The delivery
technique is extensively applied in the field of flavor encapsulation for enhancing
some pleasant flavors while masking the unfavorable ones. For food flavor com-
pounds, their excessively high volatility promotes them to retain in foods for an
11 Encapsulation and Targeted Release 383

Table 11.3 The comparison of structure, responsive mechanisms, and effect for various responsive
delivery systems. Reproduction with permission from (Chai et al. 2018), Copyright 2020
ELSEVIER. Order Number: 4958621407796
Responsive
Responsiveness Structure mechanisms Effect References
Oral responsive Pickering Promote the stability Have an on-demand (Doyennette
emulsion of flavor compounds release in mouth. et al. 2014;
Complex during storage. Have Prolong their reten- Zhang et al.
coacervation a burst release in the tion time and provide 2015)
mouth triggered by the desired sensory
amylase, temperature, experience
or pH
pH responsive Complex The structure Increase their stability (Ezhilarasi
coacervation remained contact at at acidic and digestive et al. 2013;
Spray dry- acidic pH but rup- enzymatic conditions. Liu et al.
ing tured at intestinal Prevent the early 2016b)
Layer-by- neutral pH release at stomach.
layer Specifically arrive to
Self- small intestine for
assembly further absorption
Enzyme Pickering The wall materials Release specifically in (Kumar et al.
responsive emulsion such as resistant the colon for therapy. 2017; Situ
Layer-by- starch or konjac Work as the et al. 2014;
layer glucomannan can be prebiotics Zeeb et al.
degraded by colonic 2015)
microorganism
enzymes such as
dextranase
Receptor Cross-linked Coupling a targeting Improve the cellular (Jiang et al.
targeting biopolymer molecule at the sur- uptake as well as the 2014; Yun
gels face of carriers, then absorption of bioac- et al. 2013)
the carriers will have tive compounds
an enhanced uptake
via receptor-mediated
enterocytosis
Mucus- Layer-by- Quickly and deeply Transport across the (Iglesias
Penetrating layer penetrate through the epithelium and arrive et al. 2017;
Complex mucus barriers with to the epithelial cells Lai et al.
coacervation reduced interaction for absorption and 2009)
with mucin network then largely enter the
systemic circulation
Mucoadhesive Layer-by- Increase the interac- Have a prolonged and (Sheng et al.
layer tion between the car- sustained release 2015;
riers and the mucus manner Shrestha
layer, then prolong et al. 2015)
the retention time of
the carriers on mucus
 384

Fig. 11.4 The illustration of various food intelligent systems. (a) Oral-responsive carriers: prolong the flavor retention; (b) pH-responsive carriers: keep stable
at stomach pH but release specifically at intestinal pH 7; (c) Enzyme-responsive carriers: degraded by colonic enzymes and release specifically in colon; (d)
Targeted-delivery carriers: a targeting molecule conjugated with carrier, which increases the biorecognition with the receptor at the cell surface; (e) Mucus
B. Liu et al.

penetrating carriers: penetrate through the mucus barriers and arrive to the epithelial cells and (f) Mucoadhesive carriers: have a sustained release manner (Chai
et al. 2018). Reproduction with permission from (Chai et al. 2018), Copyright 2020 ELSEVIER. Order Number: 4958621407796
11 Encapsulation and Targeted Release 385

extended period. Their flavor contents may decrease because of the early release in
the process of storage. Typically, encapsulation mainly aims to embed the bioactive
components to the delivery systems for reducing their evaporation while enhancing
chemical stability and solubility in the storage process. Meanwhile, carriers may be
used to release the bioactive components at suitable sites, like in the mouth
(McClements 2017). Currently, the carrier systems are put forward to encapsulate
the flavor components in food substrates, which allow for the controlled and
responsive release in the mouth. For example, the aromatic menthol and menthone
are encapsulated to the amylose α-helix by means of the hydrogen bonding, thus
forming the saliva amylase-responsive starch/flavor complexes. In the process of
storage before and after oral absorption, the flavor is well preserved and slowly
released into the oral cavity when it is absorbed. As suggested by studies on the
simulated saliva fluid release, due to the gradual hydrolysis of starch by the saliva
α-amylase, the aroma experiences controlled release from the complexes, which also
retains for a long time in the mouth (Ades et al. 2012). Additionally, a starch-based
Pickering emulsion is synthesized for encapsulating curcumin, which increases the
resistance to oxygen and heat during storage. Moreover, the starch-based Pickering
emulsion shows high susceptibility to α-amylase within the simulated saliva fluid,
resulting in the early encapsulated curcumin release (Wang et al. 2014). The
multilayer emulsions stabilized by whey protein isolate (WPI)-pectin can be utilized
to be the carrier for delivering the volatile flavor compounds into the mouth. In the
mouth, hydrophobic volatiles, including heptanone, pentanone and ethyl butyrate,
will be released. Meanwhile, the WPI and pectin with negative charges repulse each
other at pH 7.0, giving rise to the weak interaction between them (Benjamin et al.
2013). McClements and colleagues used the casein/alginate complex coacervation
hydrogel to encapsulate the lipophilic bioactive components. Typically, delivery
systems at food grade can efficiently protect the lipid droplets in the storage process,
but they will release lipid droplets in the oral cavity because the hydrogel particles
are dissociated at pH 7.0 (Zhang et al. 2015). In addition, hydrogel particles
constituted by caseinate and gelatine can be adopted for encapsulating the polyun-
saturated lipids to resist oxidation in the process of storage, yet they can be released
in the oral cavity through gelatine melting under body temperature (Zhang et al.
2015). The textural properties perceived are determined by particle size, shape, and
hardness, while the palate can detect particles >ca 10 μm in size (Engelen et al.
2005). Therefore, controlling the oral delivery carriers size <10 μm is of great
importance. On the other hand, the association of carriers with tongue mucus layer
must also be considered.
Moreover, oral delivery systems can also be utilized to mask the unpleasant flavor
in foods. For instance, Jaleh and colleagues prepared the lipid nanocapsules covered
by the carbohydrate biopolymers (low methoxypectin, alginate, and chitosan,) that
contained hesperetin. As suggested by sensory analysis, the synthesized
nanocapsules markedly shielded the bitter taste of hesperetin (Fathi and Varshosaz
2013). In addition, carriers loaded with phenobarbital constructed on the basis of
microemulsions (MEs) to deliver components in the mouth can reduce the bitter taste
of phenobarbital (Monteagudo et al. 2014). The SPI/gelatin complexes-based
386 B. Liu et al.

microcapsules prepared by spray drying can be utilized to diminish the bitter taste of
casein hydrolysate (Favaro-Trindade et al. 2010). On the other hand, to reduce the
unpleasant garlic flavor, the β-lactoglobulin microcapsules have been designed for
the encapsulation of thiosulfinate allicin (Wilde et al. 2016). Further, the oral
delivery systems may be utilized to investigate the sensory perception as well as
oral processing. For instance, the starch NPs loaded with vanillin constructed
according to the electrospray approach are utilized for investigating the vanillin
taste perception after vanillin is taken. The findings suggest that, starch NPs can
enhance the vanillin flavor perception in the oral cavity because the saliva amylase is
gradually degraded in the mouth (Ege et al. 2017). Furthermore, the carboxymethyl
cellulose-based delivery systems allow for the taste perception and controlled release
of sodium in the mouth, thus balancing the sodium flavor (Aditya et al. 2017).

3.2 pH-Responsive Delivery Systems

Various probiotics, peptides, proteins, vitamins, and polyphenols can be easily

decomposed under the acidic environment containing numerous digestive enzymes,
including pepsin in GI tract. Such environment will induce great loss of compounds
before they reach the small intestine. After such components are appropriately
encapsulated, their degradation by the enzymes and acids in stomach can be
prevented (Ezhilarasi et al. 2013). Thereafter, those compounds encapsulated will
be maintained in stomach and released into the small intestine, thus increasing the
specific bioaccessibility. Consequently, functional microcapsules must be respon-
sive to the neutral pH in intestine; in addition, the early decomposition or release at
acidic enzymatic conditions in the stomach must be prevented, thus facilitating the
intestine-specific release.
The delivery system responsive to pH is extensively investigated previously.
Typically, complex coacervation using WPI/gum Arabic (GA) is adopted for deliv-
ering the alive probiotics (L. paraplantarum B1 and L. paracasein E6), and this
approach contributes to maximally retaining the probiotic viability in comparison
with free cells within the simulated gastric fluid. The interaction of WPI with GA
changes from attraction to repulsion, as a result, the controlled release of cells can be
tuned at pH 7.0 (Bosnea et al. 2014). Nonetheless, the pH-responsive coacervate
capsules also have certain limitations, since they are instable at great physiological
salt level of about 150 mM. Polymers with opposite charges can dissociate at this salt
level because of the charge screening effect. This problem can be potentially
managed by cross-linking between polymers with opposite charges. Notably, the
oxidized starch microspheres cross-linked with Fe3+ and encapsulated with
β-carotene can preserve the stabilized structure within the simulated gastric fluid
(pH < 2.0) and efficiently enhance bioactive component release within the simulated
intestinal fluid (pH 7.0) (Wang et al. 2015b). The akin pH-responsive effects can also
be detected in the starch hydrogels-encapsulated nano-emulsion loaded with
β-carotene (Wang et al. 2015a). The oxidized Konjac glucomannan (OKGM)
11 Encapsulation and Targeted Release 387

microspheres coated with chitosan (CS), where anthocyanins and β-carotene are
introduced at the same time, are responsive to the pH value in the intestine (Shi et al.
2017). These microspheres are stable in the acidic stomach (pH < 2.0), which will be
destroyed to release the antioxidants loaded at neutral pH in the intestine (pH 7.0),
thus attaining intestine-specific release. Furthermore, the dual bioactive components
are loaded onto the CS-OKGM microspheres; as a result, microspheres have syner-
gistic antioxidant ability and increased thermostability. Typically, the double-
network microspheres constituted by oxidized polysaccharides cross-linked with
Fe3+/CS oligosaccharides cross-linked with glutaraldehyde also display similar
responsiveness (Lu et al. 2015). Nonetheless, the cross-linked polymer hydrogels
also have certain drawbacks, since they frequently absorb excessive water, while this
goes against food processing. Because of the potent hydrophobic interactions and
hydrogen bonds of polyphenols with human serum albumin (HSA), HSA encapsu-
lation markedly improves polyphenol stability under slight alkaline and neutral pH
conditions (Li et al. 2015a). ()-Epigallocatechin-3-gallate (EGCG) accounts for
vital antioxidant isolated based on the tea extracts, yet it has poor solubility and
stability. In this regard, it is of great importance to encapsulate EGCG for delivery
and improving its stability. By adopting the emulsion solvent diffusion approach, the
encapsulated EGCG is stabilized under acidic condition (pH 1.2) in the stomach or
the rat small intestine. Additionally, the encapsulated EGCG can be released at the
pH condition in the intestine and maintained stable at that in the stomach (Onoue
et al. 2011). For probiotics, their survival ability within host GI tract greatly affects
their activity. Tremendous probiotics are stable in the digestive tract. But the large
amounts of bile salts and highly acidic environment in stomach leads to the
decreased bacterial survival rate. A variety of delivery systems synthesized to
encapsulate probiotics can enhance the viability of bacteria. For instance, some
study has successfully synthesized the new intestine-specific carrier constituted by
one—encapsulated Ca2+-alginate core encapsulated with Lactobacillus-casein as
well as one composite shell consisting of Ca2+-alginate/protamine, which effectively
protects the encapsulated Lactobacillus casein under the acidic gastric condition, but
the compound is released into small intestine rapidly because the core–shell structure
is rapidly degraded (Mei et al. 2014). Additionally, the hydrogel microspheres
consisting of the ethylenediaminetetraacetic acid-calcium-alginate (EDTA-Ca2+-
Alg) system have been successfully synthesized and used to be the pH-responsive
carrier to deliver Lactobacillus rhamnosus ATCC 53103 into the oral cavity. The
prepared system can protect probiotics against the acidic environment in the stom-
ach, but it is slowly gastric released into the intestine with collapsed gel structure at
pH 7.0–8.0 (Zheng et al. 2017).

3.3 Enzymatically-Responsive Delivery Systems

A variety of enzymes, including β-mannanase and dextranases, can be produced by

colonic microflora existing in GI tract. As a result, microcapsules responsive to
388 B. Liu et al.

colonic enzymes may be potentially used as the colon-specific delivery systems

(Pérez-Esteve et al. 2015). Typically, in terms of the colon-specific delivery, Konjac
glucomannans (KGM) are the frequently utilized food polysaccharides. For instance,
it is reported that KGM–hydroxypropyl methylcellulose capsules can be used for the
specific delivery of 5-aminosalicylic acid to colon, due to its effect on avoiding
gastric fluid degradation and release in the colon because of colonic β-mannanase
degradation (Zhang et al. 2014). Besides, the KGM-xanthan gum (XG) based carrier
system is degraded via the colonic β-mannanase but not small intestinal one and it
shows the controlled-release feature for the embedded compounds (Alvarez-
Manceñido et al. 2008). The bioactive compounds-loaded dextran hydrogel shows
high stability within GI tract at physiological conditions, which is specifically
released in colon only because of colonic microbial dextranase-mediated degrada-
tion (Hovgaard and Brøndsted 1995). The glycidyl methacrylate dextran (GMD) and
poly acrylic acids (PAA) based hydrogels are also responsive to dextranase. The
embedded 5-aminosalicylic acid is only slightly released at both intestinal and
gastric pH values, but it is rapidly released from PAA/GMD hydrogels because of
colonic dextranase-mediated glycosidic bond hydrolysis (Kim and Oh 2005). On the
other hand, the CS-coated liposomes (chitosomes) have been designed to be respon-
sive to pancreatic lipase in releasing the carotenoids encapsulated. The CS layer
controls lipase permeability into liposomes present within the small intestine, which
thereby shows the controlled-release feature (Tan et al. 2016). It is interesting that,
the lipid digestion kinetics may be adjusted via the carrier systems because digestive
lipase shows controlled approachability to oil encapsulated lipids; consequently,
they may be utilized for designing the low-calorie foods. For instance, Zeeb and
colleagues (Zeeb et al. 2015) discovered that lipid digestion was adjusted through
changing the microcapsule layer number that covered the oil droplets. For the
double-layer O/W emulsions, their colloidal stability is enhanced, which thereby
facilitates lipase to penetrate the layer and change the encapsulated lipids to
monoacylglycerols and free fatty acids. Nonetheless, in the primary single layer
emulsion, coalescence or flocculation takes place when it is in the stomach environ-
ment; therefore, oil is digested at a slow rate within small intestine because the
surface area exposed decreases. The stable and small emulsion droplets may seem to
have a higher digestion rate. It should be noted that, wall materials (restricted to
certain cleavable polysaccharides specific to intestinal amylase or colonic microbial
enzymes) are the key factors limiting carriers responsive to intestinal enzymes. In
addition, protein only is not appropriate in this application because the protein has
been totally hydrolyzed under the gastric conditions.

3.4 Biorecognition by Specific Receptors Delivery System

Coupling targeted molecules (like peptide ligands) on carrier surfaces may contrib-
ute to the interaction between the molecule with surface receptors in targeted tissue.
Such approach can be used to efficiently enhance bioactive compound absorption in
11 Encapsulation and Targeted Release 389

cells, in particular for certain immunomodulatory compounds (Yun et al. 2013). For
instance, as the new M cell-homing peptide ligand, CKSTHPLSC (CKS9) peptide is
cross-linked with CS NPs and the resultant product can efficiently deliver com-
pounds to the follicle-associated epithelium (FAE) in Peyer’s patches. CKS9 peptide
has markedly improved transcytotic property and binding affinity to M cells;
besides, the specific localization to FAE in Peyer’s patches can be realized in vivo
(Yoo et al. 2010). On the other hand, the solid lipid nanoparticles (SLNs) modified
by CSKSSDYQC (CSK) can be used to be the candidate carriers to transport
hydrophobic drugs, like atorvastatin calcium (ATC), and they are synthesized
through cross-linking peptide ligand CSK with the stearic acid. Specifically, the
SLNs modified by CSK peptide display potent ability to penetrate the mucus into
intestinal cell monolayer and it also significantly enhances the bioavailability of
ATC (Tian et al. 2016). RGD (Arg-Gly-Asp), which is also a peptide ligand, can be
covalently bonded onto the PEGylated PLGA-based NPs loaded with antigen to
specifically target the human M cells. Besides, the RGD-conjugated NPs evidently
promoted compounds to penetrate human FAEs (co-cultures), since RGD ligand
interacts with β1 integrin on the co-culture apical surface. Studies in vivo also
suggested that RGD-conjugated NPs are especially enriched into M cells (Garinot
et al. 2007). Additionally, a seven-peptide (namely, Histidine-Alanine-Isoleucine-
Tyrosine-Proline-Arginine-Histidine, HAIYPRH) that specific to transferrin recep-
tor (TfR) is used to cover the poly (ethylene glycol)-block-poly (ε-caprolactone)
(PEG-b-PCL) micelle cores loaded with coumarin 6, so as to prepare the targeted
NPs (Du et al. 2013). As a result, the synthesized NPs markedly elevate the cell
absorption rate and penetrate cells via the specific clathrin-regulated mechanism
associated with the receptor-regulated mechanism. Moreover, FQS peptide
(FQSIYPpIK), which represents a receptor with high level of conservation that is
expressed in intestinal epithelium, exclusively binds to integrin avb3 receptor.
Cross-linking FQS-TMC with the insulin-loaded NPs constituted based on the
poly(lactide-co-glycolide acid)-monomethoxy-poly(polyethylene glycol) (PLGA-
mPEG) improves the cell absorption rate and promotes glucose-lowering effect
(Liu et al. 2016a). Such specific delivery systems have been extensively utilized in
the field of pharmaceuticals, but they are rarely applied in the food industry. The
green and safe methods like enzymatic modification to conjugate the ligands onto
carriers must be taken into consideration in food applications.

3.5 Mucus-Penetrating and Mucoadhesive Delivery Systems

The mucus layer acts as a barrier for the uptake of bioactive compounds and the
transport of carriers. It can trap the foreign particles because of its adhesion and steric
hindrance and remove them in few seconds to few hours at the anatomical site
(Dunnhaupt et al. 2015). The EGCG-loaded liposomes are found to show increased
interactions with the human intestinal mucus compared with the β-carotene-loaded
liposomes, since the hydrophobic bioactive compounds have certain influence on the
390 B. Liu et al.

mucus layer rheological characteristics, which cannot be achieved by hydrophilic

bioactive compounds. The co-culture systems consisting of Caco-2 (mucus-free) as
well as Caco-2/HT29-MTX (mucus secretory cells) were adopted for investigating
the interactions between mucus and liposome (Li et al. 2017a). Recently, the
“mucus-penetrating particle” (MPP) (Lai et al. 2009) systems enable the efficient
penetration of delivery systems into mucus layers and maximally reach the absorp-
tion membrane, since such vehicles rapidly deliver compounds to the systemic
circulation via the epithelium. Using the electrically neutral and hydrophilic PEG
to decorate the surface of NPs that had weak interaction with mucus can be a possible
strategy to cope with this problem. PEG is the extensively utilized hydrophilic
polymer, which has been usually utilized in coating NPs for improving their ability
to penetrate the mucus. For example, Li and colleagues analyzed how the chain
length (500–5000) of PEG-modifying micelles affected the oral bioavailability and
membrane permeability in the delivery of trans-retinoic acid (ATRA), indicating that
ATRA-PEG micelles with the PEG chain length of 1000 contributed to attaining
superior oral bioavailability (Li et al. 2015a). The dextran–protamine (DEX-Prot)
complex-modified nano-sized lipid carrier can offer the charge-neutral surface,
which thus facilitates its diffusion into the intestinal mucus (Lai et al. 2009). In
addition, different polymers, including DEX, cyclodextrin (HPBCD), aminodextran
(ADEX), PEG, and mannose-amine (MA) modified NPs are compared for their
mucus permeability. As a result, the MA-decorated NPs showed the maximal
penetration into mucus layer (Iglesias et al. 2017). Moreover, carriers should be
further designed for overcoming the lysosomal enzymes-mediated intracellular
degradation. Certain proteases existing in endosomes may trigger the degradation
of proteins, thereby resulting in complete carrier degradation as well as bioactive
component tracking inside the lysosomes. It is necessary to design such carriers for
escaping from lysosome, delivering them to cytoplasm and subsequently entering
into the blood circulation.
Notably, the mucoadhesive delivery system represents the oral delivery approach
frequently utilized to deliver components with poor absorption. It mainly functions
to prolong the retention time as well as accumulation on mucus and to release
the components near the absorption site (Dünnhaupt et al. 2011). On the other
hand, the thiolated polymers, including conjugated poly(acrylates), thiol groups,
CS, and the deacetylated gellan gum derivatives, may also interact with the mucus
glycoprotein subdomains with abundant cysteine through the disulphide bonds, thus
improving the mucoadhesion (Leitner et al. 2003). Palazzo and colleagues (Palazzo
et al. 2017) compared the low-molecular-weight (LMW) CS-coated poly(isobutyl
cyanoacrylate) (PIBCA) NPs with the reduced glutathione (GSH)- and glycol
chitosan (GCS)-modified N-acetyl cysteine (NAC) for their mucoadhesive proper-
ties. As a result, the GCS-GSH-coated PIBCA NPs showed the optimal
mucoadhesive properties. Additionally, it is found that the CS polymer-coated
solid lipid NPs markedly enhance the curcumin oral bioavailability (Ramalingam
et al. 2016). Besides, the mucoadhesive CS-coated porous silicon (Si) nanocarriers
have also been synthesized to release two peptides, namely, the glucagon-like
peptide-1 together with the dipeptidyl peptidase-4 (DPP4). The nanocarriers loaded
11 Encapsulation and Targeted Release 391

with two compounds may release the encapsulated peptides and have improved
mucoadhesion because of the CS surface modification (Shrestha et al. 2015). N-
trimethyl chitosan chloride (TMC), a kind of mucoadhesive polymer displaying
biocompatibility, is utilized for nanocarriers modification, thus improving the spe-
cific mucoadhesive performance. For instance, the TMC-coated poly(lactide-co-
glycoside acid) (PLGA) NPs (TMC-PLGA NPs) have been developed and used to
deliver insulin in the mouth. When TMC-PLGA NPs are administered orally, they
prevent the enzyme-mediated degradation of insulin within GI tract and extend the
retention time of insulin at the absorption site, which is ascribed to TMC’s
mucoadhesive performance (Sheng et al. 2015). More attention should be paid to
strategies for preventing the intestine-specific carriers from adhering to the gastric
mucus.

4 The Mechanisms of Cellular Uptake of Carriers

Generally speaking, many bioactive compounds cannot be efficiently absorbed via

the epithelium. For instance, hydrophobic compounds can be usually absorbed by
means of simple passive diffusion. In addition, vitamin A (VA) can be absorbed by
way of bile salt micelle transport, while polyphenols can be absorbed through the
carrier-regulated transport. However, the presence of efflux pump P-gp in epithelial
cells may contribute to pumping back the absorbed polyphenols into the lumen,
which thus largely restrict their absorption by cells. As a result, it is necessary to use
the delivery carriers for managing the problem of poor bioactive compound absorp-
tion because of the restricted cell absorption. In other words, the delivery carriers can
increase the absorption of bioactive compounds. Carriers may be developed to
penetrate deep into the mucus layer in the meantime of improving the transmem-
brane permeability into the epithelium. Besides, carriers also deliver excessive
components in each package and then actively enter into cells. Further, carriers
also cope with the potential impacts of efflux pumps. Carriers show different cell
absorption pattern from that of bioactive compounds; as a result, it is necessary to
understand the different mechanisms between carriers and cell absorption before a
carrier system is designed and applied. The NPs passive transcellular transport takes
place by the non-specific permeability pathways, which is extremely restricted
because NPs have large size (Ramesan and Sharma 2009). In addition, the energy-
dependent mechanisms may also be applied in transporting NPs by means of active
transport, and it is efficient relative to passive transport (Chen et al. 2011). On the
other hand, active transcellular transport can be divided as 4 diverse mechanisms,
including macropinocytosis, phagocytosis, caveolin-regulated endocytosis, and
clathrin-regulated endocytosis (Fig. 11.5). Of them, the phagocytosis and
endocytosis pathways may possibly take place within M cells. Besides, the
clathrin-regulated endocytosis, caveolin-regulated endocytosis, micropinocytosis,
and endocytosis represent the major pathways related to NP absorption by
enterocytes, which are complicated and may take place through several routes; as
392 B. Liu et al.

C D
A
B

Caveosome Macropinosome

Fig. 11.5 The different active transcellular transport mechanisms of cellular internalization for
foreign delivering particles. (a) Caveolae-mediated endocytosis depends on a multidomain GTPase-
dynamin. (b) Clathrin-mediated endocytosis is a widely shared pathway of nanoparticle internali-
zation. (c) Macropinocytosis displays a poor selectivity for engulfing nanoparticles and the extra-
cellular milieu. (d) Phagocytosis is an actin-based pathway taking place primarily in professional
phagocytes. The figure was modified according to reference (Yu et al. 2016) Reproduction with
permission from (Chai et al. 2018), Copyright 2020 ELSEVIER. Order Number: 4958621407796

a result, the new NPs are greatly needed to enhance transcytosis (Yu et al. 2016). For
example, the soybean Bowman–Birk inhibitor NPs loaded with curcumin (Liu et al.
2017) can be detected in the clathrin-regulated endocytosis pathway. Moreover, the
amphiphilic co-dendrimer porous graphitized carbon (PGC) NPs loaded with
honokiol are mostly absorbed through the clathrin- and caveolae-regulated endocy-
tosis (Guo et al. 2017). Generally, enterocytes specifically absorb NPs with the size
of <100 nm, whereas the M cells in Peyer’s patches are more likely to absorb
NPs > 500 nm in size (Chen et al. 2013).
Hydrophilic bioactive components are mostly transported by the paracellular
pathway. But there are intercellular tight junctions (TJs), as a result, there is a
quite limited intercellular space (about 1 nm), thereby significantly restricting the
diffusion of bioactive components. TJs represent the vital barriers limiting the
intercellular permeation, particularly for the high-molecular-weight (HMW) com-
ponents (Ramesan and Sharma 2009). For the sake of speeding up the paracellular
transport rate, it is an efficient approach to open TJs. NPs cross-linked with N-
trimethyl CS chloride (TMC) are suggested to efficiently open TJs in a reversible
way and enhance insulin uptake in the mouth (Sheng et al. 2015).
11 Encapsulation and Targeted Release 393

5 The Evaluation of Carriers by Digestion and Absorption

Models

For screening the appropriate carriers to deliver diverse bioactive compounds,

multiple models in vitro have been utilized for evaluating the delivery performances.
The in vitro simulated GI model has frequently been used to evaluate the uptake and
digestion of bioactive components together with the corresponding delivery systems,
which do not require human or animal experiment. The in vitro GI models are
associated with the advantages of cost-effectiveness, great reproducibility, and time-
saving property; as a result, they are extensively used for research in the pharma-
ceutical and functional food fields. Such models have offered the fundamental
information before the in vivo experiments are performed (Kong and Singh 2010).
At present, the existing in vitro models are the simulated GI fluid model, SHIME
model, INFOGEST models, HGS model, and TIM model. In addition, the in vivo
animal bioavailability model and in vitro Caco-2 monolayer cell model are
constructed based on living systems. Further, we also conclude the representative
digestion and uptake models, and compare the corresponding merits and demerits, as
presented in Table 11.4. Typically, the simulated GI fluid model represents the
multi-stage stimulator that is reported for the first time, which can be utilized for
investigating factors that affect food ingredient digestion, including enzymatic
activity, ionic components, applied mechanical stresses, and digestion time (Boisen
and Eggum 1991). In addition, the SHIME model represents the multi-stage simu-
lator designed for investigating the association of food ingredients with colonic
microbial composition. It has been extensively used to study in the pharmaceutical,
nutrition, micro-ecology, and general safety assessment fields and to examine the
digestion and efficacy behaviors of nutraceuticals as well as their bioaccessibility
(Alander et al. 1999). The TIM model is also extensively used to research in the
pharmaceutical and food industries for studying the uptake and interactions of
nutrients, drugs, and functional components. Additionally, the food digestion site,
the nutrient uptake rate, and the association of inhibiting factors with stimulating
ones may be studied using the TIM model as well (Ribnicky et al. 2014). Moreover,
the HGS model contributes to mimicking the digestion process in human body and
study those changes in gastric contents and food components in the food digestion
process. In addition, it can also be used to study food and nutrient degradation
kinetics affected by physiological conditions including enzyme production, acid,
and mechanical contraction. According to the COST Infogest international network,
the general standardized and practical static digestion model appropriate for foods on
the basis of physiological conditions is put forward, namely, the INFOGEST model,
and it can be used to investigate different endpoints and is modified for accommo-
dating the more specific requirements (Minekus et al. 2014). As the in vitro cell
model, the Caco-2 cell monolayer can be used for investigating the apparent
permeability (Papp) for reflecting nutrient uptake by enterocytes (Sambuy et al.
2005). However, the Caco-2 monolayer model does not represent the actual in vivo
condition, so the nutrition oral bioavailability must be assessed through calculating
394 B. Liu et al.

Table 11.4 The comparison of various GI digestion absorption models. Reproduction with
permission from (Bao et al. 2019), Copyright 2020 ELSEVIER. Order Number: 4958630740831
Type of models Application Advantage Disadvantages Ref.
Simulated GI Simulate the GI Easy to build up The mechanical (Kong and
fluid method fluid environment and cost-saving forces and the Singh 2010)
and digestion fluid mechanics
time, and analyze that food encoun-
the stability of tered cannot be
food during diges- reproduced in the
tion at the stomach stomach
condition
The simulated Study the interac- The kinetics of The interaction (Kontula
human intesti- tion and composi- chyme pass between et al. 1998)
nal microbial tion of microbial through the GI probiotics and the
ecosystem community in tract can be host cannot be
(SHIME) colon microbiota simulated imitated
TNO’s gastro- Monitor the place Comprehensively Absence of feed- (Yoo and
intestinal model where food simulate the pro- back from the Chen 2006)
(TIM) digested, evaluate cess of digestion, central nervous
the availability of and the process is system and spe-
nutrients absorp- controllable cific hormone
tion and the inter- releases for con-
actions of food trolling the gas-
between stimulat- tric, intestinal
ing and inhibiting motility and
factors secretions, muco-
sal cells and
immune system
Human gastric Study the diges- A more accurate Lack of intestinal (Kong and
simulator tion of foods in stomach wall conditions Singh 2010)
(HGS) stomach movement can be
simulated, and the
gastric secretion
and emptying are
controllable
Cell culture Predict the perme- Easy to build and Lack of mucus (Sambuy
models ability and mecha- standardize, barrier allows et al. 2005)
nism of food require small compounds easy
compounds across amount of sample access to the api-
the small intestinal reflect the absorp- cal cell mem-
cell monolayer tion on the cell brane, neglect
level other complex
factors in vivo
Animal models Study the bio- Offer a living sys- Time-consuming, (Boxenbaum
availability of tem imitating the costly, and some- 1982)
nutrients in vivo full dynamic phys- times inhumane
iological and
physiochemical
events during the
absorption, distri-
bution, and
metabolism
11 Encapsulation and Targeted Release 395

the pharmacokinetics of blood nutrient contents at consecutive time intervals when

they are administered to animals (Bursztyka et al. 2008).

6 The Application of Carriers in Different Types of Food

Numerous requirements should be met in the preparation of carriers. Firstly, carriers

must be of stability and will not release those encapsulated compounds ahead of time
in the stomach but release them in the intestine. Secondly, easy diffusion of carriers
across the intestinal mucus layer should be guaranteed, so that they rapidly reach the
surface of enterocytes and will not be removed by mucus renewal. Besides, the
charge-neutral and hydrophilic components can easily penetrate the mucus (Shan
et al. 2015). Particularly, nanocapsules are able to directly enter into small intestinal
epithelium by active transport. As a result, this significantly improves the bioavail-
ability of the encapsulated bioactive components in such delivery systems. Besides,
these nanocapsules may also open TJs between epithelial cells in a reversible
manner, thus enhancing bioactive compound delivery via the paracellular pathway
(Dodane and Vilivalam 1998). When the bioactive compounds are delivered, TJs
will return back to the physiological status (Li et al. 2015a). Moreover, carrier size
also represents a factor that affects the penetration into mucus and the opening of
TJs. Carriers of a smaller size can rapidly penetrate the mucus barriers while
reversibly opening TJs, thereby releasing bioactive compounds to the capillary
blood (Patel and Champavat 2015). Further, the associations of nanocapsules con-
jugated with target ligands with typical receptors on the surface of epithelial cells can
promote bioactive compound absorption (Jiang et al. 2018).
A variety of sensitive food component delivery systems are synthesized to mask
the unpleasant flavors, which thus increase the water solubility of these components,
protect them against the harmful external environment, and improve their penetra-
tion into intestinal mucus as well as epithelial cells (Augustin and Hemar 2009).
Different technologies are designed in recent years to prepare the food delivery
systems. Initially, complex coacervates, which are constituted by polysaccharides
and proteins, have been the widely used conventional technology used to fabricate
microcapsules (de Kruif et al. 2004). Proteins including whey protein (Weinbreck
et al. 2003), zein (Chen and Subirade 2009), casein (Mendanha et al. 2009), and
gelatin (Saravanan and Rao 2010), or carbohydrates including chitosan (Espinosa-
Andrews et al. 2007) and pectin (Saravanan and Rao 2010) are extensively used to
be the constructing materials. Thereafter, the cross-linked polysaccharides- and
proteins-based hydrogel microparticles have aroused increasing attention due to
their water swellability and great loading capacity (McClements 2017). Then, the
Pickering emulsions are also developed to be the new delivery systems to load
hydrophobic components because of the excellent long-time emulsification stability
on the surface of small surfactants (Chevalier and Bolzinger 2013). Additionally, the
food-resourced Pickering particulate stabilizers are adopted for improving the bio-
availability of lipophilic bioactive compounds, which including lipids,
396 B. Liu et al.

nutraceuticals, or the oil-soluble vitamins (McClements 2018). In recent years, great

attention has been paid to self-assembled NPs due to their numerous merits, includ-
ing the ability of penetration into mucus, high cell absorption and controlled release
compared with the conventional delivery systems (Ezhilarasi et al. 2013). Different
bioactive compounds have diverse chemical structures (lipophilic or hydrophilic),
sensitivity, and biological functions. For instance, a majority of polyphenolic com-
pounds possess the unfavorable flavors or tastes, among which, some are lipophilic,
thus limiting their wide application. Typically, the encapsulation technology can be
used to load polyphenols, which efficiently mitigate the above drawbacks, such as
freeze drying, spray, crystallization, coacervation, liposome, complexation, and
emulsion entrapment (Fang and Bhandari 2010). Most vitamins, including
vitamin A, D, E, and K, have low solubility and oxidation sensitivity, thus restricting
their actual application. On the whole, it is of great significance to develop an
approach for overcoming the above problems while enhancing bioactive component
bioavailability. Micro- or nano-emulsion and nano-encapsulation can markedly
enhance their water stability and dispersibility (Gonnet et al. 2010). The polyunsat-
urated fatty acids (PUFAs), like EPA and DHA, can be easily oxidized. The
encapsulation technology may be adopted for inhibiting the degradation of these
compounds while retaining the resultant product quality (Holser 2011). Numerous
bioactive peptides and proteins show high susceptibility to degradation, but they are
poorly absorbed within GI tract; in addition, microparticles and NPs may be utilized
for encapsulating, retaining, protecting, and specifically delivering the bioactive
proteins to the intestine (McClements 2018).
A number of carrier systems are proposed to deliver bioactive compounds
(Ezhilarasi et al. 2013). Meanwhile, most carrier systems can be utilized to promote
the hydrophobic compound solubility; for example, the successful encapsulation of
procyanidin in zein NPs is achieved, thus evidently enhancing the procyanidin
solubility within the water solutions (Zou et al. 2012). The application of molecular
co-assembly or self-assembly technique in encapsulating vitamin D (VD) into
β-lactoglobulin markedly increases the VD solubility. For β-lactoglobulin
nanocapsules encapsulated with linoleic acid, their solubility increases by 33–45%
(Górska et al. 2012); while for albumin NPs encapsulated with CoQ10, their aqueous
solubility evidently increases (Matsushita et al. 2013). Certain bioactive compo-
nents, including USFAs and vitamins, are unstable in the presence of oxygen, light,
or heat. In this case, the encapsulation technique can be utilized in the food industry
for encapsulating, protecting, and releasing many bioactive components. For
instance, investigators succeed in encapsulating vitamin E (VE) to the modified
tapioca starch nanocapsules, which remarkably enhances the VE stability. At
60 days after storage under the temperature of 4–35
C, VE attains a retention rate
of about 50% of the original value, along with good solubility (Hategekimana et al.
2015). Amphiphilic peptides can be acquired by hydrolyzing caseins and they can
co-assemble to the micellar NPs. By means of hydrophobic interactions, it is
possible to incorporate VD to the hydrophobic cores, thus markedly improving the
thermostability (Haham et al. 2012). Vitamin C (VC) represents another unstable
vitamin in the presence of heat, light, and oxygen (Bendich et al. 1986). For VC
11 Encapsulation and Targeted Release 397

encapsulates with the inorganic layer SiO2 shell, its stability is 95%, but that for the
non-encapsulated VC is just lower than 10% at 4 weeks later (Yang et al. 2003).
Besides, forming nanocomplexes may also enhance the USFA stability.
Nanocapsules constructed based on β-lactoglobulin and pectin protect DHA against
degradation; to be specific, jut about 5–10% loss is detected over 100 h under 40
C
in comparison with 80% loss reported for the non-encapsulated DHA (Zimet and
Livney 2009). Bioactive compounds are associated with off-target effect and low
stability at physiological conditions (enzymes, pH, together with intestinal mucus
barrier) within the GI tract, which results in their poor bioavailability and
bioaccessibility. There are many obstacles preventing the nanocarriers loaded with
bioactive compounds from arriving at the target site because of the complex phys-
iological environment in human body.

7 Future Prospects

As a result, it is necessary to develop the intelligent delivery systems for protecting

and transporting bioactive compounds to target sites. The GI condition-responsive
intelligent carriers may also be synthesized for the specific release of compounds in
the intestine under the following conditions: microbial enzymes, neutral pH, epithe-
lial cell mucus barriers, and specific receptors. Typically, it is possible to deliver
bioactive compounds to specific intestinal sites where they can be further absorbed
and utilized. At present, many reviewers have focused on investigating whether such
compounds are stable under the simulated harsh GI conditions but neglect their real
bioavailability and stability in vivo. Nonetheless, further practical models are needed
because of the existence of factors such as cell absorption, mucus barriers, metab-
olism, and transport mechanisms, which facilitates to the accurate evaluation on the
bioavailability and stability of encapsulated bioactive compounds. For the high-load
intelligent designs, compound stability within GI tract, facile diffusion into the
intestinal mucus layers, enhanced cell absorption, and cell junction opening must
be taken into full consideration. The research directions in the future are the
hydrogel- and self-assembled NPs-based multifunctional nanocarriers with simulta-
neous responsiveness to physiological environment (including enzymes, pH value,
as well as intestinal mucus barrier) within GI tract. More efforts are needed to
explore the nano-sized delivery systems that efficiently promote the stability, bio-
availability, and sensory perception. For such delivery systems, their toxicity in vivo
must be studied from the perspectives of biodistribution, metabolism, and cell
viability. The analysis in this section contributes to developing the more sophisti-
cated carrier systems to prevent disease and promote health. As found in this chapter,
intelligent delivery systems can be utilized for the effective improvement of bioac-
tive compound bioavailability and quality within functional foods in the future,
which shed more lights on the design of intelligent carriers.
398 B. Liu et al.

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Chapter 12
Replacement of Fat or Starch

Cuixia Sun and Yapeng Fang

Abstract A growing public demand for low-calorie foods is stimulating the

researchers and food manufacturers to develop reduced-calorie products due to the
recognized adverse effects of high energy diet on human health. Fat and starch are
two condensed sources of energy, and reducing their intake is a major dietary goal
for the consumer. Currently a variety of available technological options have been
applied to decrease the content of fat or starch in foods. The development of food
hydrocolloids-based fat or starch replacers is one of the most important approaches
for fat or starch reduction because their functionalities allow them to mimic the oral
and flow properties of fat or starch. However, the replacement of fat or starch is not
trivial because both of them play important roles in determining the nutritional,
physical, chemical, and sensory characteristics of foods. How to achieve the replace-
ment of fat while matching as close as possible all the characteristics of full-fat foods
remains a major challenge. This chapter describes the main challenges for the
reduction of fat from the viewpoint of flavour perception and texture quality. Two
strategies for fat replacement are involved, including food formulation optimization
and food structure design. Various hydrocolloids-based formulations created for the
purpose of starch replacement are introduced and the associated principles
discussed. The commercially available fat replacers and their applications in differ-
ent food products are presented.

Keywords Fat · Starch · Replacement · Challenges · Strategies

C. Sun · Y. Fang (*)

Department of Food Science and Technology, School of Agriculture and Biology, Shanghai
Jiao Tong University, Shanghai, China
e-mail: ypfang@sjtu.edu.cn

© Springer Nature Singapore Pte Ltd. 2021 409

Y. Fang et al. (eds.), Food Hydrocolloids,
https://doi.org/10.1007/978-981-16-0320-4_12
410 C. Sun and Y. Fang

1 Introduction

High calorie food intake is associated with the increased prevalence of numerous
chronic diseases, including obesity, type 2 diabetes, hypertension, cancers, gall
bladder disease, and coronary heart disease. Fat and starch are two major sources
of calories in many processed food products. High levels of certain types of
triacylglycerols in the blood have been related to coronary heart disease and obesity
(Bray et al. 2004; Han et al. 2016). Rapid digestion of starch causes a sudden rise in
blood glucose level, which has been linked to diabetes and obesity (Lustig et al.
2012; Svihus and Hervik 2016). Based on the growing awareness about the adverse
effects of high caloric products through nutrition and health claims, reducing fat or
starch intake is becoming a major dietary goal for the consumer. In addition, our ever
more sedentary life styles serve as a driving force to decline the consumption of
high-energy foods.
The World Health Organization (2003) recommends a daily intake of total fat no
more than 30% of dietary energy, of which <10% should be saturated fatty acids.
The UK Food Standards Agency (2008) published its Saturated Fat and Energy
Intake Programme and recommends that the saturated fat consumption should be
reduced from 13.3% of dietary energy to 11%. The 2015–2020 Dietary Guidelines
for Americans emphasize the restriction of trans and saturated fats intake and
recommend the consumption of low-fat or fat-free foods (DeSalvo et al. 2016).
Food reformulation to reduce the content of fat was included in nutrition action plans
of many European countries (Belc et al. 2019). The United States Code of Federal
Regulations (2014) defines low-fat food as: ‘The food has a reference amount
customarily consumed >30 g and contains 3 g or less of fat per reference amount
customarily consumed (RACC); or the food has a reference amount customarily
consumed of 30 g or less and contains 3 g or less of fat per RACC and per 50 g of
food’. In the USA, a low-fat cheese must contain 6 g or less of fat per 100 g of
cheese, while a reduced-fat cheese requires at least a 25% reduction in fat level from
the traditional fat level of the referenced variety. Fat-free cheese is defined as that
with <0.5 g fat per 100 g of cheese (Johnson 2011).
In such a context, researchers and the food industry are highly sensitive to
consumer perceptions and demands for low-calorie foods. However, the replacement
of fat or starch is not trivial because both of them play important roles in determining
the nutritional, physical, chemical, and sensory characteristics of foods. For exam-
ple, as shown in Fig. 12.1, fat not only provides a concentrated source of energy, but
also supplies essential fatty acids and fat-soluble vitamins (A, D, E, and K). Fat
reduction would result in loss of nutritional benefits or even an unbalanced diet
(Colmenero 2000). Besides, fat provides structure in baked goods, influences the
storage stability, and affects the observed lightness of food products owing to the
impact of the fat droplets on light scattering (Chung et al. 2013a). A direct removal
or simple replacement of fat without compensation for specific functions would
cause a considerable change in the organoleptic characteristics of foods, leading to a
poor food quality. For instance, inulin addition at levels of 100% in cake
12 Replacement of Fat or Starch 411

Fig. 12.1 Functions of fat in food products

formulations resulted in remarkable loss of different quality attributes, including

higher water activity and baking loss, lower volume, harder texture, darker colour,
and highly asymmetrical shape (Majzoobi et al. 2018). Starch granules are often
used in food products to provide desirable texture attributes, such as ‘thickness’, and
contribute to the physicochemical properties of foods, including the volume, vis-
cosity, gelation, and stability (Ai and Jane 2015). Any ingredients to replace starch
granules should be able to simulate such characteristics normally provided by
conventional starch granules and meanwhile should contribute less calories and
lower glycemic index (McClements et al. 2017).
Consequently, a number of considerations need to be taken into account for the
feasibility of fat- or starch-reduced products in order to maintain the desired phys-
icochemical and sensory attributes of original foods. In this chapter, major chal-
lenges for the development of low-fat or low-starch foods are discussed from the
aspects of scientific problems and technical limitations. A variety of hydrocolloids-
based formulations developed for the purpose of fat or starch replacement are
introduced and the associated principles discussed. The commercially available fat
replacers and their applications in different food products are presented.
412 C. Sun and Y. Fang

2 Challenges for Fat or Starch Replacement

Based on either omission or replacement of fat or starch, a variety of available

technological options have been involved to quantitatively reduce fat or starch
content and qualitatively modify the fatty acid profiles. For example, food
reformulation is one of the most important approaches to remove, reduce, and/or
replace different components in order to develop healthy products. However, food is
a typically complex colloid system with common characteristics of multiphase,
multicomponent, and multiscale. Fat droplets and starch granules vary considerably
in their size, shape, charge, and behaviour and endow foods with many desirable
functions. Therefore, they are difficult to replace.

2.1 Challenges for Fat Replacement

The elimination or reduction of fat in foods evidently modifies its composition and
structure and also the expected interactions among components, giving rise in most
cases to clearly perceptible changes in flavour and/or texture. Moreover, although
most taste compounds do not dissolve in fats, when fat content is reduced, the salty,
sweet, sour, and umami tastes will be weakened and the bitter taste enhanced
(Metcalf and Vickers 2002).

2.1.1 Flavour Concerns

Sensory preference for fat appears to be a universal human trait because fat contrib-
utes to numerous sensory characteristics of fatty products, including appearance,
texture, flavour and aroma, and mouthfeel. Flavour as one of the most-important
attributes determines consumer selection and satisfaction with fat-reduced foods.
Reduction or elimination of fat directly affects the processes of flavour release and
perception, modifies the signals received by the brain on consuming a particular
food, and may partially determine its acceptance or rejection.

Flavour Distribution and Release

The term ‘flavour’ refers to volatile components that are sensed by aroma receptors
in the nose and non-volatile components that are sensed by taste receptors in the
mouth (Taylor and Linforth 1996). The overall perceived flavour of a food product
usually involves the integration of information from mouthfeel, taste, and aroma
during mastication (González-Tomás et al. 2008). As an extremely complicated
process, flavour perception is dependent on a combination of physicochemical,
biological, and psychological phenomenon, which is perhaps the most multisensory
12 Replacement of Fat or Starch 413

of our everyday experiences (Spence 2015). Fat functions as flavour precursors, and
reducing fat content influences both the distribution of the flavour and the kinetics of
flavour release (McClements and Demetriades 1998). The type and concentration of
flavour molecules mainly determine the perceived flavour. For nonpolar flavour,
decreasing fat content would increase in the aqueous phase flavour concentration,
leading to an intense initial taste perception (Roberts et al. 2003). Even if the amount
of flavour compounds is rebalanced in fat-reduced foods to provide the same
maximum aroma intensity as the full-fat products, the low-fat alternatives still fail
to match the same perception. The major reason is that fat has an impact on time-
intensity profile of lipophilic flavour release. The sustained aroma release relies on
the hydrophobicity and fat content of the product, that is, the higher the hydropho-
bicity and fat content, the slow the aroma release into the aqueous phase (de Roos
2006). In full-fat products, a rich flavour sensation is perceived because flavour
compounds with various degrees of hydrophobicity are released at different rates.
On the contrary, in reduced-fat products, lower amount of fat is insufficient to retain
the aroma compounds and thus causes quick flavour disappearance and lack of
richness (van Ruth et al. 2002).

Flavour–Ingredients Interactions

Food matrix ingredients such as proteins, polysaccharides, and lipids can interact
with flavour compounds (Guichard 2002). Modification of the food formulation by
using fat replacers would change such interaction and thus alter the flavour percep-
tion. The development of fat-reduced products with desired flavour will be only
possible if the knowledge of flavour-ingredients interactions has been well under-
stood (Plug and Haring 1994). Flavour-binding behaviour of fat replacers results in
lower volatilities in aqueous systems, which may explain the decreased flavour
intensity in fat-reduced foods (Godshall 1997; Fischer and Widder 1997). For
example, proteins like β-lactoglobulin, casein, gelatin, and egg albumin can interact
with flavour compounds by reversible or irreversible binding, causing lower vola-
tilities in water phase (Maier 1970). More research is required to explore the
mechanisms of flavour–ingredients interactions and the location of the binding
sites, which can provide the necessary information for the selection of suitable fat
replacers in the development of fat-reduced food products.

2.1.2 Texture Concerns

Texture is another important sensory attribute to assess food quality, such as being
hard or soft, cold or warm, oily or juicy, elastic or flaky, heavy, viscous, or smooth. If
the texture attribute fails to meet our expectations, we may reject the food regardless
of the quality of flavour (Engelen and de Wijk 2012). Fat is an important contributor
to texture in different types of foods, such as thickness of liquid foods (Villegas and
Costell 2007), consistency of semisolids (Tárrega and Costell 2006), and firmness of
414 C. Sun and Y. Fang

solids (Kavas et al. 2004). Fat content has obvious impacts on the food texture in
different ways because fat droplets impart many textural characteristics such as the
viscosity, afterfeel, lubrication, and melting or cooling mouthfeel. Fat reduction
usually results in a dramatic decrease in viscosity, leading to the lower perceived
thickness. The common approach is to add biopolymer or biopolymer mixture to the
aqueous phase to enhance its viscosity. However, biopolymer molecules are not able
to mimic all of the textural characteristics. Particularly, fat has distinctive thermal
properties because of its unique melting point, which contributes the creamy mouth-
feel after the melting of fat crystals at room temperature (Weenen 2005). It was
reported that heat transfer between foods and the oral surface may be an important
factor for fat perception. The perceived food temperature is associated with fat
content, for example, high fat products were perceived as warmer than low-fat
products (Weenen et al. 2003). Therefore, the sensory difference of foods with
different fat contents can be detected based on the fact that lips and tongue are
highly sensitive with the temperature changes (Prinz et al. 2007).
Moreover, numerous sensory characteristics of food emulsions are related to their
rheological properties such as elasticity, viscosity, and viscoelasticity. For example,
the fatty, creamy, smooth, and thick texture of full-fat products is related to the bulk
rheology, thin-film rheology, and colloidal interaction between food and oral surface
(Malone et al. 2003). Removal of fat causes textural problems in reduced or low-fat
ice creams, such as coarseness and iciness, crumbly body, shrinkage, and flavour
defects (Akalın et al. 2008). Cakes presented significantly increased hardness,
elasticity, and decreased specific volume as fat replacement increased above 65%,
leading to lower scores on taste and flavour (Psimouli and Oreopoulou 2013). Fat
reduction is likely to reduce the degree of shear-thinning behaviour, which may have
important implications for the mouthfeel and texture of the product. Therefore, a
thorough understanding of the rheological behaviours and colloidal properties of
foods would provide guidelines for the design of products with the replacement of
fat or starch but without quality loss.
Overall, the functional properties of lipids that must be reproduced include
organoleptic properties, the ability to dissolve lipid-soluble flavours, aeration,
aroma, emulsification, flavour, heat stability, and spreadability. Translating idea
into reality is not a simple task. Hydrocolloids-based fat replacers are generally
polar water-soluble compounds, so it is difficult for them to replace some of the
nonpolar functional characteristics of fats, such as lipid-soluble flavour-carrying
capacity.

2.2 Challenges for Starch Replacement

Starch granules are widely used in foods to create desirable textural attributes
because of its preferable characteristics such as gelling, thickening, and aqueous
solubility. The viscosity is a determining factor for the application of starch in food
products to obtain desirable rheological characteristics (Sarkar et al. 2013).
12 Replacement of Fat or Starch 415

Therefore, the challenge for starch replacement is how to develop a food ingredient
with similar functional attributes as starch granules.

2.2.1 Pasting and Gel Texture Properties Control

The replacement of starch with non-starch hydrocolloids influences the properties of

a starch-based paste, gel, or food product. Mixing gelatin (0.5 wt%) with pectin
(0.01 wt%) formed the hydrogel particles with similar dimensions to swollen starch
granules, and these hydrogel microspheres were shown to have similar rheological
attributes as starch pastes (Wu et al. 2014). However, the unsolved technical and
scientific question is that the physicochemical properties of gelatin are highly
susceptible to temperature because it forms a gel at low temperatures. By the
increase of corn starch citrate (CSC) substitution level, the textural parameters of
wheat starch gels were decreased, such as firmness, cohesiveness, springiness,
gumminess, and chewiness (Hedayati and Niakousari 2018). Addition of glutamic
acid or lysine increased gelatinization temperatures of the cross-linked potato
starches and decreased the G0 and G00 moduli of the modified starch gels while
accelerated the retrogradation process (Gałkowska and Juszczak 2019), which may
cause undesirable changes to food and mostly affect the bakery products. The
substitution of kudzu and lotus starches with soybean soluble polysaccharide
(SSPS) reduced the hardness of the starch/SSPS gels, and the mixtures of starch/
SSPS yielded more liquid-like behaviour than the controls did (Liu et al. 2019). The
modified pasting and gel texture properties may change the sensory properties of the
end product. Therefore, how to control the stability and quality of starch-based foods
is the key point for starch replacement. In addition, the amylose content positively
correlated with its cohesiveness and stringiness (Zhang et al. 2019), and amylopectin
molecular size significantly contributes to gel viscoelasticity (Li et al. 2019). Con-
sequently, the pasting and gel texture properties of starch-based foods would be
influenced after starch being replaced by the non-starch hydrocolloids.

2.2.2 Interactions Control

The compositions of starch-based foods are generally complicated, and the interac-
tions of starch with various food components such as proteins and lipids have been
extensively reported. The challenge of starch replacement lies in the understanding
of the interaction mechanisms of starch and non-starch food constituents to achieve
desirable quality of low-starch foods. Starch–protein interactions affect the rheolog-
ical, pasting, gelatinization, textural, and physicochemical properties of food sys-
tems (Villanueva et al. 2015). In general, the inclusion of proteins increased water
absorption capacity, water absorption index, water solubility index, and swelling
power, decreased the viscosity of gels, and increased their stability, with the effect
being more conspicuous for SPI incorporation (Villanueva et al. 2018). The inter-
action between starch and whey protein mainly through hydrogen bonds restricted
416 C. Sun and Y. Fang

the swelling process of starch granule while accelerated recrystallization after cold
storage (Yang et al. 2019). Complexes between amylose and lipids may significantly
modify the properties and functionality of starch. For example, the solubility of
starch in water is reduced, and the gelatinization temperature is increased after the
complexation with lipids (Copeland et al. 2009). Lipids may prevent gelatinization
by inhibiting hydration of amylopectin chains and retard retrogradation, which
affects starch digestibility (Henry 2009). Starch/carrageenan interactions are espe-
cially involved in dairy products where gelling properties are of primary importance
(Huc et al. 2014). The lower carrageenan charge density, the higher the interaction
between starch and carrageenan (Lascombes et al. 2017). Formation of starch gel is
hindered by the presence of cationic polysaccharide and, therefore, the retrograda-
tion of starch at very early stage can be delayed by addition of chitosan. However,
long-term retrogradation was slightly increased (Raguzzoni et al. 2016).
Overall, a universal starch substitute does not exist. All of the macromolecule
replacers contribute to distinct properties suitable for replicating a limited number of
functions in particular food products.

3 Strategies for Fat Replacement

The traditional dietary advice is to replace fat with low-calorie fruits, vegetables, and
grains, which has not been very effective for reducing fat consumption. Direct fat
removal was evolved as the first strategy to comply with nutritional recommenda-
tions in the 1980s, which worked well for milk, some dairy products, certain
processed meat, but not much else. The approach to fat replacement has changed
in the twenty-first century. Formulation optimization emerged as the second strategy
to reduce fat content in foods, which is one of the most important approaches to
replace fat in modern food technology. The food reformulation refers to the devel-
opment of a range of functional ingredients as fat replacers to reduce fat intake.
There are two large groups of fat replacers: fat substitutes and fat mimetics. In
general, lipid-based fat replacers are fat substitutes, and protein- or carbohydrate-
based fat replacers are fat mimetics (Sandrou and Arvanitoyannis 2000). In addition,
processing technology is the third fat replacement strategy by varying processing
conditions (pH, pressure, ionic strength, time and temperature, mixing order, stirring
speed, etc.) to cause interactions in ingredients or to modify functionalities.

3.1 Food Formulation Optimization

Ideally, fat mimetics should be safe, inexpensive, low calorie, suitable for cooking
applications, yet provide the sensory equivalent of fat texture and flavour, and
maintain the eating quality of foods. The majority of fat mimetics belong to the
groups of polysaccharide and protein hydrocolloids because their functionalities
12 Replacement of Fat or Starch 417

allow them to mimic the oral and flow properties of fat. For example, viscosity
enhancement potential is an important feature in the use of hydrocolloids as emul-
sifying, stabilizing, and bodying agents in low-fat foods. Hydrocolloids as fat
mimetics are able to augment the lubrication of aqueous fluids through the three
lubrication regimes via both viscosity modification and by adsorbing to hydrophobic
substrates (Stokes et al. 2011). Hydrocolloids-based fat replacers are usually divided
into protein-based, carbohydrate-based, and lipid-based.

3.1.1 Protein-Based Fat Replacers

Protein-based fat mimetics are typically produced from egg, milk, whey, soy, or
wheat proteins. Using proteinaceous ingredients to replace fat in food emulsions is
mainly because of their emulsifying and stabilizing capacities. The particle size of
hydrocolloids is important in determining both the taste and mouthfeel of fats in
fat-replaced products. Food particles <3 μm in diameter are not detected by the oral
cavity, and instead the substance feels creamy and smooth. The minimum size of
particles above which humans detect the grittiness depends on the hardness and the
shape of particles (Engelen et al. 2005). Therefore, proteins are often
microparticulated by applying a high shearing force during heating of the proteins.
The obtained small spherical (0.1–2.0 μm diameter) protein gel particles are per-
ceived in the mouth and taste buds as similar to fat with a creamy, smooth texture.
Gelatin is commonly used in low-fat yoghurts due to its melting behaviour at body
temperature, as it forms a thermoreversible gel (Alting et al. 2009). The representa-
tive microparticulated whey proteins (MWP) have been produced and applied in
food systems as fat replacers since the 1980s due to its soft lubrication characteris-
tics. MWP increases the lightness and viscosity of products, and almost all of the
MWP-based systems have a creamy white appearance similar to sauces and dress-
ings, indicating their potential application in the manufacture of reduced-fat foods
(Chung and McClements 2014). For example, the improved texture and rheological
properties of low-fat yoghurt were obtained when whey proteins were added as
microparticles rather than conventional whey protein ingredients such as whey
protein isolate (Torres et al. 2018). In addition, the complex of MWP and high-
methoxyl pectin was prepared as a novel fat mimetic in low-fat mayonnaise
(Fig. 12.2), and two possible hypotheses were proposed to explain the interaction
and distribution of MWP, pectin, and droplets (Sun et al. 2018): (1) in low-fat
mayonnaise (upper figure of Fig. 12.2), MWP particles were adsorbed on the surface
of oil droplets by hydrophobic interaction and formed thick and viscous films, and
pectin formed coatings around interfacial proteins to inhibit flocculation of oil
droplets. (2) in non-fat mayonnaise (lower figure of Fig. 12.2), MWP evenly
distributed among the stable three-dimensional network formed by pectin molecules,
and the stable structure of fat mimetic was maintained by hydrogen bonding,
electrostatic force, and van der Waals interactions (Gentès et al. 2010).
It was observed that the friction levels attained with MWP and dairy fat (DF) at
typical speeds involved in oral processing were comparable, hence demonstrating
418 C. Sun and Y. Fang

Fig. 12.2 Schematic representation of interaction among microparticulated whey protein, pectin,
and oil in low-fat and non-fat mayonnaises. Reproduction with permission from (Sun et al. 2018),
Copyright 2018 Elsevier

the capability of MWP in skim milk dispersions to imitate DF in fluid milk-based

systems from a lubrication point of view (Olivares et al. 2019). Compared with
MWP, superfine MWP (sMWP) exhibited more stable liquid behaviours (Sun et al.
2015a) and could maintain creamy mouthfeel better due to high dispersion stability
of sMWP–pectin–xanthan gum gel mixtures (Sun et al. 2016). In addition, a novel
group of fat globular mimetics (FGMs) was prepared by coating calcium carbonate
particles with a layer of casein–maltodextrin conjugates. Such FGMs were stable in
skim milk during 10-day storage at 5  C, and increased the desirable turbidity and
viscosity of skim milk, which can be used to simultaneously reduce fat and increase
calcium contents of food products (Qu and Zhong 2017).
Animal proteins are rich in necessary nutrients, particularly the essential amino
acids needed for human body. However, they may have a strong allergic effect and
are not suitable to produce food requiring heat treatment, because high temperatures
induce irreversible denaturation of protein, altering the structure of the final product
(Jing et al. 2011). Besides, the consumption of animal proteins would cause prob-
lems associated with biodiversity, land use, water use, climate, human health, and
animal welfare (Aiking 2011). Natural plant proteins show similar physicochemical
properties to animal proteins like water binding capacity and can serve as fat sub-
stitutes in low-calorie food. Soy proteins are increasingly important in the human
diet because of reported beneficial effects on nutrition and health, including lowering
plasma cholesterol, prevention of cancer, diabetes, and obesity, and protection
against bowel and kidney disease (Friedman and Brandon 2001). The addition of
soy protein isolate improved the textural properties of chopped low-fat pork batters
and lowered the cooking loss (Gao et al. 2015). Soy protein hydrolysates (SPH) and
their blends with xanthan gum (SPH/XG) is an alternative choice as a fat replacer in
the production of reduced-fat ice cream since 50% fat-substituted ice cream with
12 Replacement of Fat or Starch 419

SPH/XG (96:4) had an appearance, taste, and texture similar to that of 10% full-fat
ice cream (Liu et al. 2018a). The major limitation of proteins as fat replacers is the
occurrence of molecular interactions between proteins and some volatile com-
pounds, giving rise to unbalanced or even unacceptable sensory profiles (Kühn
et al. 2006).

3.1.2 Carbohydrate-Based Fat Replacers

Carbohydrates are typical fat replacers due to their molecular diversity that gives rise
to various structural and physicochemical properties. For example, carbohydrate-
based fat replacers bind water into a gel-type structure, resulting in lubrication and
flow properties similar to those of fat. Cookies prepared from wheat flour by 15%
supplementation of carbohydrate-based fat replacers showed better attributes in
terms of colour and texture, which were judged to be the best by the panellists
(Majeed et al. 2017). Compared with the relatively limited available choices for
protein-based fat replacers, carbohydrate-based fat replacers include a much larger
family of materials, which can be categorized into digestible polysaccharides
(starch) and non-digestible polysaccharides (gums and cellulose).

Digestible Starch

Among the carbohydrate-based fat replacers, starch is one of the most frequently
used ingredients as it is relatively inexpensive and readily available and capable of
replacing numerous attributes (O’Connor and O’Brien 2011). There are two hypoth-
eses for starch-based fat replacer to mimic the mouthfeel and texture characteristics
of fat. One is that starch would form a three-dimensional gel network structure, and
the water trapped in the gel could provide a fat-like sensation (Alexander 1995). The
other is that the starch could form spherulites with similar size to fat globules
(2–10 μm) during heating and cooling treatment, which provide the lubricating
mouthfeel of fat (Singh et al. 2010). Amylose plays an important role in the
simulation mechanism of starch-based fat replacer because amylose contributed to
the gel structure of starch-based fat replacer (Yang and Xu 2007). With the increas-
ing amylose ratio, spherulites of 2–10 μm diameter, similar to fat globules in size,
started to appear with the increasing amylose ratio, and the potato starch with 85%
amylose ratio presented the better creamy texture (Hu et al. 2019). Starch-based fat
replacers have been applied in a wide variety of low-fat food products including
cheeses, sausages, yoghurt, mayonnaise, and frozen desserts (Peng and Yao 2017).
The concentrations of starch frequently used as fat replacer in cheese normally
ranges from 0.5 to 1.5% (Diamantino et al. 2014).
Different types of starches may present different behaviours as fat replacers.
Compared to native starches, the modified starches produced high paste viscosity
values and showed low retrogradation rates, which can be regarded as promising fat
replacers in cheese (Diamantino et al. 2019). Starch granules remain even after
420 C. Sun and Y. Fang

heating the aqueous starch suspension, but heating at temperatures higher than
130  C leads to complete molecular dissociation (Hanselmann et al. 1996). Starch–
water systems can be classified into three states: the intact granular state, the melted
state, and the solution state. When aqueous potato starch suspensions were heated
and then cooled, spreadable particle gels were obtained with a spherulite morphol-
ogy and a cream-like texture, which is currently applied as a fat mimetic (Steeneken
and Woortman 2009). Citric acid treated sweet potato starch showed fat mimetic
properties as its melting temperature (51.44  C) was close to the melting point of fat
(Surendra Babu et al. 2016). The pasting viscosity of the octenyl succinic anhydride
(OSA) modified mung bean starches (OSA-MS) was found to be higher when
compared with native starch, and cakes prepared from 30% OSA-MS were found
to be highly acceptable by their overall quality score including the best texture,
desirable colour, and mouthfeel (Punia et al. 2019). For acidified milk gels (yoghurt)
with pregelatinized (PG), and both pregelatinized and chemically modified starches,
viscosity/texture values were similar to or higher than those found for full-fat milk
gel (Bravo-Núñez et al. 2019). The corn starch nanocrystals (CSNC) are regarded as
a useful fat replacer/stabilizer for an O/W model emulsion because its addition
resulted in a more solid-like behaviour of the emulsions due to the formation of
nanocrystal network in the continuous phase (Javidi et al. 2019). However, one of
the potential disadvantages of starch-based fat replacer is that it contains calories, so
its overconsumption may lead to problems with overweight and diabetes (Lustig
et al. 2012). Some of these problems may be overcome by using resistant starch
(Parada and Aguilera 2011).

Non-digestible Cellulose Derivatives

Cellulose derivatives show different solubility, emulsifying property, and gelation

characteristics, and they can reassociate with each other to form aggregates that can
be used as fat replacers. Typically, 60–70% of the cellulose microcrystals are
<0.2 mm long, which can form an insoluble dispersion in water. Methylcellulose
(MC) and hydroxypropyl methylcellulose (HPMC) have been used to stabilize air
bubbles, provide lubricity and creaminess, and entrap moisture in a variety of foods,
such as salad dressings and biscuits (Laguna et al. 2014). Microcrystalline cellulose
(MCC) is an uncharged biopolymer with a crystalline structure and could form
a filled particle gel network. Therefore, MCC provides a fat-like mouthfeel and a
softer texture (Fig. 12.3a) when it is added to the ground beef without causing a
disturbance of the protein network. Since carboxymethyl cellulose (CMC) at high
concentrations (>0.5 wt%) is thermodynamically incompatible with meat proteins, it
is not a suitable fat replacer because it weakened the connections within the protein
network (Gibis et al. 2015). As shown in Fig. 12.3b, cellulose nanofibrils (CNFs) at
concentrations of 0.15 and 0.3 wt% were incorporated into low-fat (5 wt%) and
standard ice cream formulations (10 wt%), which improved the sensory properties of
low-fat samples, even after heat shocking the specimens (Velásquez-Cock et al.
2019).
12 Replacement of Fat or Starch 421

Fig. 12.3 (a) Suggested mechanisms of interaction of meat proteins with CMC and MCC () and (b)
effect of cellulose nanofibrils (CNFs) at concentrations of 0.15 and 0.3 wt % on the physicochem-
ical properties of low-fat (5 wt%) and standard ice cream formulations (10 wt%). (a) Reproduction
with permission from (Gibis et al. 2015), Copyright 2015 Elsevier. (b) Reproduction with permis-
sion from (Velásquez-Cock et al. 2019), Copyright 2019 Elsevier

Non-digestible Inulin

Inulin, typically derived from chicory root, belongs to a class of carbohydrates

known as fructans (Kaur and Gupta 2002). As a non-digestible dietary fibre, inulin
can remain stable during processing and successive heat treatment. It is widely used
in replacing dietary fat in baked products, providing nearly the same sensory
characters as of full-fat products while giving only 25–35% energy as compared to
422 C. Sun and Y. Fang

digestible carbohydrates (Keenan et al. 2014). Such fat-substituting property is based

on its ability to stabilize the structure of the aqueous phase, which creates an
improved creaminess. A creamy mouthfeel is achieved when inulin is used as a fat
replacer in dairy products due to its interactions with whey protein and caseinate
(Karaca et al. 2009). Long chain inulin microcrystals could aggregate each other,
interact with water, and eventually agglomerate creating a gel network, thus altering
the product texture and providing a fat-like mouthfeel (Bayarri et al. 2011). The
consumer study revealed that 15% fat replacement by inulin provided acceptable
biscuits, but higher replacement decreased the overall acceptability (Laguna et al.
2014). Inulin addition (2–7%) to replace fat in fresh caprine milk cheese provided a
creamier mouthfeel and added a reasonable flavour with softening effect (Salvatore
et al. 2014). Fermented chicken sausages made with inulin as a partial oil replace-
ment persisted stable without any significant loss of physicochemical, microbiolog-
ical, and sensory characteristics during storage at 4  C for 45 days (Menegas et al.
2013). It was found that inulin fortification of low-fat set yoghurt significantly
reduced syneresis by 59% over full-fat control yoghurt (Rudra et al. 2017). What
is more, inulin has promising gut health properties due to its prebiotic nature and
may increase absorption of nutrients such as calcium. Therefore, it is recommended
as a reasonably high-level fat replacer in crackers, cakes, biscuits, and muffins
(Shoaib et al. 2016).

Other Non-digestible Gums

The performance of gums as fat replacers is determined mainly by their distinct

chemical composition and structure (Saha and Bhattacharya 2010). The principles
that are taken into account in applying gum as a fat mimetic include the rheological
properties of the gel it forms, the effects of temperature and shearing forces on the
functional properties of the gum, and its compatibility with other ingredients in the
foods. Xanthan gum and carrageenan had large spheres of hydration, provided
slipperiness and viscosity, and mimicked the continuous phase of mayonnaise
(De Ruiter and Rudolph 1997). Locust bean gum (LBG) formed non-dissolved
microparticles at relatively high concentrations (0.4%), trapping fat droplets within
its hydrogel particles and helping balance the flavour profile of reduced-fat products
(Chung et al. 2013b). Water-extracted okra gum was found to be effective to make
an ice cream comparable with full-fat ice cream and was used to replace the fat in ice
cream at 0, 22, 44, 55, 88, and 100% to produce super premium (18% fat), premium
(14% fat), regular (10% fat), economy (8% fat), low-fat (2% fat), and zero-fat (0%
fat) ice cream (Aziz et al. 2018). The substitution of fat with okra gum increased the
viscous modulus (G”) of the ice cream, and up to 55% replacement of fat was
feasible to achieve satisfactory ice cream properties (Aziz et al. 2018). Addition of
tragacanth gum (A. gossypinus and A. compactus) to sausage formulation effectively
reduced cooking loss and enhanced oxidative stability, and 0.5% tragacanth
(A. gossypinus) showed an acceptable sensory score of the sausage formulation,
suggesting its potential to be a fat replacer in the reduced-fat sausages (Abbasi et al.
12 Replacement of Fat or Starch 423

Fig. 12.4 Schematic representation of KGM addition as a potential replacer in Mozzarella cheese.
Reproduction with permission from (Dai et al. 2018), Copyright 2018 Elsevier

2019). The addition of pectin in ice cream can cause an increase in viscosity,
overrun, and hardness and a decrease in meltdown of the ice cream. When 0.72%
pectin (w/w) was incorporated into ice cream, a prototype product of ice cream with
45% lower fat content compared to the control was prepared (Zhang et al. 2018).
Konjac glucomannan (KGM) as a natural polysaccharide also exhibits functional
properties as a potential fat replacer in dairy products. As shown in Fig. 12.4, KGM
addition in cheese affected the lightness, increased the moisture, lowered firmness,
and increased the stickiness, and such changes were closer to those of full-fat cheese,
suggesting KGM could improve some characteristics of the fat-reduced Mozzarella
cheeses (Dai et al. 2018). Mozzarella cheese with konjac had lower firmness but
higher meltability and less scorching in pizza bake and exhibited a denser casein
matrix with coalesced fat globules (Dai et al. 2019). Sodium alginate was used to
modify the textural and microstructural properties of low-fat Cheddar cheese up to
91% fat reduction (Khanal et al. 2018).

3.1.3 Combination Systems

Individual fat mimetic has limitations in its ability to cover the full functions of fat.
The combination of different fat replacers may show synergistic interactions and
provide better fat-like qualities which are not easy to achieve by individual fat
replacers (Sikora et al. 2008). For example, single carbohydrate-based and protein-
based fat mimetics may suffer from several sensory and functional limitations such
as poor stability and undesirable mouthfeel. The simultaneous addition of protein
and polysaccharides may induce intermolecular interactions that modify or generate
more desirable functional properties (Gulão et al. 2016).
424 C. Sun and Y. Fang

Protein–Polysaccharide Combination

A combination of polydextrose with MWP is the most suitable fat replacers for soft-
type cookies (Zoulias et al. 2000). The MWP in combination with either modified
starch or locust bean gum (LBG), with or without fat droplets (5%), could be used as
fat mimetics to modulate the texture, appearance, and stability of emulsion-based
food products with reduced calorie such as sauces, mayonnaise, dressings, and dips
(Chung et al. 2014). A nonheated whey protein–high methoxyl pectin mixture can be
used as fat replacer in the skim milk formulations, which yielded a yoghurt texture
resembling the full-fat counterpart because the associative interaction of whey pro-
teins with pectin suppressed whey protein aggregation while maintaining the struc-
turing effects of denatured whey protein in yoghurt (Krzeminski et al. 2014).
Biopolymer-based hydrogel particles consisting of a protein-rich core and a
pectin-rich shell were formed by using a segregation–aggregation phase separation
method. Such particles may be suitable as texture modifiers and fat replacers since
they scattered light strongly to give the hydrogel suspensions a milky white appear-
ance and also led to an appreciable increase in viscosity or gel-like characteristics
(Duval et al. 2015). The mixtures of soy protein isolate (SPI) and cellulose nanofibre
(CNF) with a higher CNF proportion showed increased viscosity, storage modulus,
and loss modulus and a higher tendency of gelation. The targeted low fat, low
calorie, anti-melting, and similar textural taste were achieved when SPI–CNF
complex gels with an SPI:CNF ratio of 7:1 were added to ice cream as a fat replacer
(Fig. 12.5), in which 10% fat was replaced (Sun et al. 2015b).

Polysaccharide–Polysaccharide Combination

Polysaccharides such as pectin and alginates can interact with each other to form
more or less permanent junction zones, providing yield stress and gel structure.
Maltodextrin and xanthan gum yielded increased moisture, hardness, and chewiness
in 66% FR (fat replacer) muffins (Khouryieh et al. 2005). The mixtures of guar gum

Fig. 12.5 Soy protein isolate/cellulose nanofibre complex gels as fat substitutes in dairy products.
Reproduction with permission from (Sun et al. 2015b), Copyright 2015 Springer Nature
12 Replacement of Fat or Starch 425

(GG) and xanthan gum or citrus fibre simulated the function of oil emulsions and
made the low-fat mayonnaise score the same as the full-fat counterpart with sensory
panels (Su et al. 2010). Synergistic interaction between xanthan and pregel corn
starch and also between xanthan and GG was identified by data analysis (Rahmati
et al. 2015). The addition of xanthan gum and GG in low-fat cheese softened the
structure by interfering the casein–casein interaction and cellulose particles that
function similar to fat globules (Murtaza et al. 2017). Polydextrose and GG were
successful fat replacers in biscuits at a relatively high level of FR (70%), with an
increase in perceived taste, flavour, and consumer acceptance (Chugh et al. 2013).
The combination of gum arabic at concentrations of <75 ppm with GG in the
concentration range of 75–170 ppm provided the softest texture of low-fat Iranian
white cheese (Lashkari et al. 2014). The blend of GG and basil seed gum yielded
better creaminess in low-fat ice cream than GG alone (Javidi et al. 2016). Four
combination sets of carboxymethyl cellulose, gum arabic, carrageenan, and xanthan
were used as fat replacers in Labneh (semi-solid yoghurt with high solid content
23–25%), suggesting Labneh water holding capacity in the following order:
xanthan>gum arabic>carrageenan>carboxymethyl cellulose (Saleh et al. 2018).
Mixture of κ-carrageenan, locust bean, and xanthan gums has been added to milk
to make cheese. Majeed et al. (2017) explored the combined potential of pectin and
banana powder as carbohydrate-based fat replacers in cookies, suggesting that the fat
content was reduced from 29.82% to 17.07% by using 15% such complex fat
replacers. Upon using different concentrations of hydrocolloids, low-fat cheese
showed a significant increase in the physiochemical characteristics, yield, and
moisture. Furthermore, organoleptic properties obtained were both highly acceptable
and comparable to full-fat cheese (Alnemr et al. 2016). The hybrid hydrogel
prepared from sodium alginate and pectin by combining both physical and chemical
cross-linking methods using citric acid as the cross linker was proved to reduce up to
50 vol% fat content in chocolate with the highest melting resistance (80  C) (Francis
and Ramalingam 2019).

3.1.4 Lipid-Based Fat Replacers

Lipid-based fat replacers are either chemically synthesized or derived from conven-
tional fats and oils by enzymatic modification. They are usually stable at cooking and
frying temperatures.

Emulsions

Wheat gluten-stabilized high internal phase emulsions (HIPEs) could be promising

substitutes for mayonnaise because HIPEs and mayonnaise might have similar
sensory property and perceived texture such as creaminess, smoothness, and slim-
iness (Liu et al. 2018b). Concentrated emulsions prepared by adding a fish gelatin-
gum arabic mixture at pH 5.0 and 3.6 to olive oil at W:O ¼ 30:70 (w/w) were
426 C. Sun and Y. Fang

Fig. 12.6 Emulsion gels prepared with different protein emulsifiers and gelling agents. Reproduc-
tion with permission from (Freire et al. 2018), Copyright 2018 Elsevier

designed as novel fat replacers in reduced-fat (15% fat) and low-fat (6% fat) Cheddar
cheeses (Anvari and Joyner 2018). Micron to nano-sized fat emulsions prepared
from sodium caseinate and anhydrous butter were used as a source of fat in low-fat
Cheddar without affecting much the texture, chemical and bio-chemical properties of
cheese (Khanal et al. 2019). Food-grade emulsion (O/W) gels (Fig. 12.6) formulated
with a lipid phase rich in n-3 fatty acids and different emulsifiers (sodium caseinate,
whey protein isolate, and soy protein isolate) showed solid-like structure and their
overall appearance is good enough to be used as animal fat replacers with a lower fat
content (Freire et al. 2018). A multiple emulsion refers to the coexisting of water-in-
oil and oil-in-water morphologies within the same system (Dickinson 2011). Mul-
tiple emulsions of water-in-oil-in-water (W/O/W) are particularly suitable for reduc-
ing the fat content of products because some of the fat within the droplets is replaced
by water (Lobato-Calleros et al. 2008). However, it is difficult to ensure that the
multiple emulsions have sufficient stability for commercial applications (Chung
et al. 2016). Multiple emulsions (W/O/W) prepared with olive oil and sodium
caseinate (SC) by two-step emulsification procedure resulted in reduced lipid,
increased protein content, and modified fatty acid composition and were noted as
promising constituents for beef fat replacement (Serdaroğlu et al. 2016). Multiple
emulsions (W/O/W) with an average droplet size of 32 μm containing native
beetroot juice as inner water phase, sunflower oil as oil phase, and 0.5% whey
protein isolate as outer water phase were used to replace fat (11%) in meat products
and also to enhance the product colour (Eisinaite et al. 2017). The emulsion gel
(EG) prepared with gelling agents (chia flour and/or soy protein isolate, inulin,
carrageenan, sodium caseinate, and sodium tripolyphosphate) resulted in a solid-
like fat replacer, which was utilized as an animal fat replacer to prepare soft Bologna
sausage (de Souza Paglarini et al. 2019).
12 Replacement of Fat or Starch 427

Structured Oils

Unsaturated vegetable oils are often used to reduce saturated fats content in meat
products, which improves the fatty acid profiles and also helps in product stability
(Siraj et al. 2015). Oil structuring, or oleogelation, is the process in which edible
liquid oil is immobilized in a three-dimensional gel network of gelators, conferring
solid-fat functionality to liquid oils (Co and Marangoni 2012). This technology is
relatively simple since it refers to the transformation of a liquid oil into a ‘gel-like’
structure with visco-elastic properties. The schematic representation is shown in
Fig. 12.7 (Jiang et al. 2018). Unlike polymers used for hydrogels, such oleogels
utilize small, amphiphilic molecules that self-assemble via non-covalent interactions
forming fibrillar or platelet crystals (Patel and Dewettinck 2016). The interactions
are responsible for gelation, including hydrogen bonding, π-π stacking, electrostatic
and van der Waals interactions (Okesola et al. 2015).
Over the past decade, oleogels have made significant strides towards emulating
desired sensory traits while maintaining healthy nutritional profile of the oil. In
recent years, structuring techniques for liquid oils have received considerable atten-
tion in different fields including food science. Oleogel technology has shown strong
potential as a way to replace hard-stock fats in meat products. Sunflower oil oleogels
structured with monoglycerides and phytosterols at 15:5 weight ratio were used to
replace 50% of the pork backfat in frankfurter sausages without significantly
compromising their physicochemical, textural, and sensorial characteristics, at the
same time providing an enriched polyunsaturated fatty acids lipid profile (Kouzounis

Fig. 12.7 Schematic representation of the process where liquid oil is first used to prepare an oil-in-
water emulsion stabilized by regenerated cellulose (RC) and carboxymethyl cellulose (CMC),
followed by freeze-drying to selectively evaporate the water phase, where further shearing of the
dried oil results in the formation of an oleogel. Reproduction with permission from (Jiang et al.
2018), Copyright 2018 Elsevier
428 C. Sun and Y. Fang

et al. 2017). Ethylcellulose (EC) oleogels have been used to replace colloidal fat
crystal networks comprised of saturated fat of frankfurters (Zetzl et al. 2012). A lipid
combination made with olive, linseed, and fish oils stabilized in a konjac gel matrix
was created to reduce pork backfat in pork patties (Salcedo-Sandoval et al. 2015).
Canola oil was structured with foam-structured hydroxypropyl methylcellulose
(HPMC) into solid-like oleogels. Such an HPMC oleogel was used as an animal
fat replacer for saturated fat-reduced meat patty, and the highest sensory acceptabil-
ity was obtained at a 50% replacement level (Oh et al. 2019). Oleogels appeared to
be the most successful fat replacer in cake, with no changes to the sensory qualities at
100% fat replacement (Kim et al. 2017). Overall, the novel approach of structuring
liquid oils would be one of the most promising ways to develop healthy lipid meat
products, which makes it possible to create a solid-like material rich in monounsat-
urated fatty acid (MUFA) and polyunsaturated fatty acid (PUFA) and with reduced
saturated fatty acid (SFA) levels and zero trans fatty acids (Jimenez-Colmenero et al.
2015). In addition, as solid-fat replacer, the constructed structured oil systems can be
used in both water-free (shortenings, chocolates, and chocolate pastes) and water-
containing (cooked meat products, margarine, and spreads) products.

3.2 Food Structure Design

While fat replacers have been applied in the development of reduced-fat products, an
attractive strategy for fat reduction is created based on the fundamental structure–
function relationship of food ingredients (Fig. 12.8a) (Campbell et al. 2017). Struc-
tural design principles can be used to mimic some of the desirable physicochemical,
sensory, and physiologic attributes normally associated with fat droplets. A variety
of approaches based on structural design principles that can be used in emulsion-
based products are highlighted in Fig. 12.8b (McClements 2015). For example, the
control of microstructure and physical properties of biopolymer hydrogel particles
can be achieved through modulation of electrostatic interactions, which could be
used to manipulate food formulations to achieve desirable physicochemical or
sensory properties (Chung and McClements 2015).
The potential of controlled aggregation has been used to improve texture prop-
erties for reduced-fat products since the aggregation of emulsion droplets forms a
three-dimensional network that inhibits droplet movement and leads to increased
viscosity and even gel-like structure (Mao and McClements 2011, 2012). The
principle to control emulsion aggregation is through regulating interfacial properties
of the emulsion droplets and induce electrostatic attraction between the colloidal
particles (Mao and McClements 2013). By carefully controlling the pH below,
above, or equal to the protein’s isoelectric point (pI), respectively, the protein-
stabilized emulsion droplets carry positive, negative, or neutral charges. Thus,
droplets self-aggregation can be induced due to the oppositely charged biopolymers
(Wu et al. 2013a). The interactions between negatively charged polysaccharide and
positively charged protein interactions allow efficient approaches to construct food
 12
Replacement of Fat or Starch

Fig. 12.8 (a) The relationships among food structure, oral processing, sensory perception and (b) structural design principles to create reduced-fat emulsion-
based products with physicochemical attributes similar to conventional products. (a) Reproduction with permission from (Campbell et al. 2017), Copyright 2017
Elsevier. (b) Reproduction with permission from (McClements 2015), Copyright 2015 Oxford University Press
429
430 C. Sun and Y. Fang

structures and improve the stability and textural properties of semi-solid food
colloids (Le et al. 2017). Based on controlled aggregation, model reduced-calorie
food emulsions consisting of fat droplets (5 wt%), starch granules (4 wt%), and
xanthan gum (0–0.02 wt%) were developed with desirable textural and optical
properties at pH 3, and the structural organization of the fat droplets could be
regulated by altering xanthan levels (Wu and McClements 2015a). Alternatively,
ions such as calcium can be added to induce aggregation of negatively charged
droplets, and paste-like materials were produced when the fat droplets formed a
three-dimensional network at a high calcium concentration (Wu et al. 2013b).
Casein–maltodextrin conjugates produced smaller fat globule mimetics and
increased the desirable turbidity and viscosity of skim milk (Qu and Zhong 2017).

4 Strategies for Starch Replacement

High consumption of digestible starch is linked to a number of diet-related diseases.

There is an increasing interest in the development of starch mimetics. Based on the
important physicochemical properties and sensory attributes of foods, starch
mimetics should have at least two essential attributes, being capable of effectively
enhancing the viscosity of solutions and giving a desirable mouthfeel to foods such
as thickness and creaminess (Rao 2014). Food hydrocolloids with a relatively low
calorie density are suitable for creating reduced-calorie starch mimetics as they often
contain large quantities of water, therefore increase the viscosity of starch pastes,
influence the retrogradation rate, and prevent the syneresis of starch (Dolz et al.
2006).

4.1 Non-starch Polysaccharides

Non-starch polysaccharides can interact with starch and impart desired functionality
to the resultant blend for oriented application (Yoshimura et al. 1999; Funami 2009;
BeMiller 2011; Mahmood et al. 2017). Mesona chinensis polysaccharide (MCP) can
improve the thermal stability in the early stage of pasting and enhance the rheolog-
ical properties of wheat starch (Liu et al. 2018c). The addition of gums (xanthan
gum, flaxseed gum, konjac glucomannan, or tamarind seed gum) to starch resulted in
softer binary gels, which are effective in retarding retrogradation of starches
(Pongsawatmanit et al. 2013; Liu and Xu 2019). The presence of basil seed gum
(BSG) led to greater water binding capacity and greater water absorption index of the
starch compared to the free-gum systems and also led to a rise in the viscoelasticity
(G0 and G00 ) and hardness of the final gels (Matia-Merino et al. 2019). The addition of
pectin increased the storage modulus (G0 ) and loss modulus (G00 ) of corn starch while
resulted in a decrease in the starch susceptibility to α-amylase and promoted a
remarkable reduction in the fraction of rapidly digested starch (Ma et al. 2019).
12 Replacement of Fat or Starch 431

Modified starch can be replaced by inulin as prebiotic encapsulant matrix of lipo-

philic bioactive compounds (Zabot et al. 2016). Inulin at low concentrations can
effectively restrain the retrogradation of wheat starch (Luo et al. 2017). Rice starch
(RS) can be also partially replaced by inulin because it affects the pasting, thermal,
and rheological properties of RS (Wang et al. 2019). Barley sourced beta glucan
(βG) and microcrystalline cellulose (MCC) could replace starch in meat emulsions.
The maximum inclusion level of MCC and βG that has been previously tested
without detrimentally affecting colour and textural properties of meat emulsions
was 2% (Schuh et al. 2013) and 3% (Mejia et al. 2018), respectively. The combina-
tion of βG (1.5%) and MCC (1.5%) to replace starch resulted in beef emulsions with
less calories, greater insoluble fibre content, and appropriate technological properties
(Mejia et al. 2019).

4.2 Hydrocolloid Microgels

Hydrocolloid microgels have been attracting much attention to produce low-calorie

foods (Norton et al. 2006). Hydrocolloids microgels fabricated by complexation of
cationic proteins and anionic polysaccharides through electrostatic attraction showed
a strong potential to be starch mimetic. Gelatin-pectin-based starch mimetics have
been developed, and the size and morphology of such starch mimetic could be
controlled through manipulation of gelatin/pectin ratio. For example, gelatin at a
fixed concentration of 0.5 wt% can form micro-sized translucent spheroids when
interacting with 0.01 wt% pectin at pH 5, and these hydrogel particles showed
similar dimensions, shape, and rheological properties as swollen starch granules
(Wu et al. 2014). The ionic strength should also be controlled during gelatin–pectin
complex formation, as a too high salt content perturbed the gelatin–pectin interac-
tions through electrostatic screening and ion binding effects (Wu and McClements
2015b). The cross-linking altered the microstructure and rheology of the microgels
under simulated oral processing conditions. The melt-in-the-mouth behaviour of the
hydrocolloid microgels could be made to be similar to that of starch granules by
controlling the degree of cross-linking (Wu and McClements 2015c). In addition, the
gelatin–pectin microgels designed to dissociate around body temperature may be
useful for imitating the melting properties and/or thickening properties of starch
granules in the mouth, which is also particularly helpful for the development of
elderly foods with improved swallowing ease under oral conditions. Electrostatic
complexation of gelatin and modified (OSA) starch shows potential to modify
texture of food products, suggesting their feasibility to replace starch granules in
foods (Wu and McClements 2015d).
432 C. Sun and Y. Fang

5 Commercially Available Fat Replacers

Limited studies have been specifically focused on the development of starch

mimetics though there are lots of reports about the design of biopolymer-based
hydrogel particles, so commercial starch mimetics are limited. As a result, commer-
cial fat replacers are mainly introduced in this part.
Fat-reduction ingredients fall into three categories: carbohydrate-based, protein-
based, and lipid-based, in which carbohydrate-based fat replacers are the most
common. Examples of commercially available fat replacers and their applications
and functional properties are shown in Table 12.1 (Mattes 1998). In some cases, the
Food and Drug Administration (FDA) has approved fat-reduction ingredients as
food additives, including carrageenan, olestra, and polydextrose. In other instances,
fat-reduction ingredients are ‘generally recognized as safe’ (GRAS). Most
carbohydrate-based fat replacers are GRAS substances. For example, oatrim gel
made from whole oat flour behaves like shortening, being solid at room and body
temperatures and liquid at cooking temperatures, and imparts fat-like qualities such
as creaminess, moisture retention, bulking, and texture. Oatrim is heat stable in
cooking and baking applications and can replace fat in foods such as frozen desserts,
salad dressings, soups, cheeses, baked goods, meats, and skim milk. Besides, Oatrim
also contains beta glucan, so it offers double health benefit by replacing the fat and
increasing the soluble fibre content of foods (Hahn 1997).
Protein-based fat replacers are not as many as carbohydrate-based ingredients, but
they have a wide range of applications. Commercially available protein-based fat
replacers are Simplesse and Dairy Lois, which are derived from whey protein
concentrates and are generally regarded as safe (Yazici and Akgun 2004). This
category of fat mimetics is suitable for use in dairy products, salad dressings, frozen
desserts, and table spreads. For instance, Dairy Lois incorporated at 5% by weight
has been used to develop an ice cream containing 1% fat.
Lipid-based fat replacers often provide the closest taste and cooking properties of
fat. Salatrim belongs to a group of structured triacylglycerols and has been used as a
fat mimetic in reduced-calorie food products for many years because it provides
approximately half of the calories and has similar physicochemical and organoleptic
properties as those of conventional fats (Smith et al. 1994). A great concentration of
undigested fat within the lower gastrointestinal tract (GIT) remained, indicating that
Salatrim may be an effective fat replacer due to its ability to suppress hunger and
increase fullness (Sørensen et al. 2008). Olestra is a sucrose polyester, in which the
ester bonds are not hydrolysed by lipase in the human GIT because of steric
hindrance effects. As a result, Olestra is not absorbed by the body and is specially
designed to be a zero-calorie fat substitute to replace triglyceride oils in products
such as fried foods, snacks, breads, and fillings (Bimal and Guonong 2006). How-
ever, Olestra may disturb the absorption of fat-soluble vitamins and may be linked to
undesirable GIT symptoms, so its used amount is limited by FDA (Prince and
Welschenbach 1998).
12 Replacement of Fat or Starch 433

Table 12.1 Commercially available fat replacers and their applications

Fat replacer Trade name Applications Functions
Protein-based Simplesse, Dairy Lois, Dairy products, salad- Mouthfeel,
(microparticulated K-Blazer, Veri-lo, dressing, margarine- and creaminess,
protein, modified Power-pro, Versapro, mayonnaise-type prod- viscosity
whey protein Ultra-Baketm, Ultra- ucts, baked goods, coffee
concentrate) Freezetm, Lita creamer, soups, and
sauces
Lipid-based Caprenin, Salatrim, Confections, baked Mouthfeel,
Dur-Lo, ECT-25, Olestra goods, dairy products stability
Cellulose Avicel cellulose gel, Dairy products, sauces, Water retention,
Methocel, Solka-Floc, frozen desserts, salad texturizer, stabi-
Just Fiber dressings lizer, mouthfeel,
clouding agents
Dextrins Amylum, N-Oil, Stadex Salad dressings, pud- Gelling, thick-
dings, spreads, dairy ening, stabiliz-
products, frozen desserts ing, texturizer
Maltodextrins CrystaLean, Lorelite, Baked goods, dairy Gelling, thick-
Lycadex, Malitrin, products, salad dress- ening, stabiliz-
Oatrim, Stadex, nu-trim ings, sauces, spreads, ing, texturizer
frostings, fillings,
beverages
Gums (guar gum, Kelcogel, Keltrol, Salad dressings, formu- Water retention,
gum arabic, locust Viscarin, Novagel, Jag- lated foods such as des- texturizer,
bean gum, xanthan uar, Fibrex, Slendid, serts and processed thickener,
gum, carrageenan, Splendid, Grindsted meats mouthfeel, gel-
pectin) ling, stabilizer
Inulin Raftiline, Fruitafit, Yoghurt, cheese, frozen
Fibruline desserts, baked goods,
fillings, whipped cream,
fibre supplements
Fibre Opta TM, Oat Fiber, Baked goods, meats, Heat stable
Snowite, Ultracel TM, spreads, extruded
Z-Trim products
Starch and modified Amalean I and II, N-Lite, Dairy products, Bodying agents,
starch Fairnex TMVA15 and processed meats, salad gelling, thicken-
VA 20, Instant dressings, baked goods, ing, texture
StellarTM, Pure-Gel, frozen desserts modifiers
Sta-SlimiTM,
OptaGread
Reproduction with permission from (Mattes 1998), Copyright 1998 Elsevier

6 Future Perspective

Nowadays, healthiness and beauty are common interests among people. Reducing
calorie consumption is a major goal for the consumer. Consequently, both food
manufactures and researchers are sparing no effort to develop reduced-fat/starch
food products. The key point is that the replacement of fat or starch should contribute
434 C. Sun and Y. Fang

low or even zero calorie to food products and should be nondetrimental to their
organoleptic qualities.
Many low-fat products have been based on the use of a wide variety of bio-
polymers, especially hydrocolloids. More basic knowledge of physical, rheological,
chemical, and sensory characteristics, functionality, and fat or starch interactions
with other ingredients is required as to formulating these bases. Further studies are
needed on new food-grade ingredients as potential replacers of fat or starch. Other
approaches are available based upon the use of manufacturing and preparation
procedures that can help to achieve desired product properties such as colour,
texture, and water- and fat-holding abilities. A thorough understanding of the
physicochemical properties and molecular interactions of food-grade ingredients is
necessary for developing innovative fat reduction strategies such as utilizing struc-
tural design approach to control the macroscopic properties.
Currently, the consumer’s unwillingness to give up high-energy foods suggests
that there is a considerable potential market for frequently consumed foods such as
meats which have been reformulated to produce health benefits. It is believed that in
near future, people would adapt to a low-fat or low-starch diet based on the
development of novel advanced technologies for the replacement of fat or starch.

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10942910009524643
Chapter 13
Structuring for Elderly Foods

Makoto Nakauma and Takahiro Funami

Abstract For the elderlies with reduced eating capability, food texture should be
modified from rheological, colloidal, and tribological aspects to make it comfortable
to eat for them. In aged society, formulation/structure design for food texture for the
elderlies is now one of the most important tasks in the food industry. This chapter
aims to provide readers with relevant knowledge for the industrial implementation of
the texture design of elderly foods. Firstly, usefulness of polysaccharides as a texture
modifier will be summarized, and with presenting commercial examples in Japan, it
is indicated that the manipulation of polysaccharides is the most practical way for the
product development. Secondary, mechanical criteria for texture of elderly foods in
Japan will be reviewed with comparison to the global standards. Although Texture
Profile Analysis is the base for the mechanical measurements in Japanese standard-
ization, some cautions should be required in the test operation and data analysis. As
advanced texture assessment, the progress of mechanical and physiological tests will
be also explained. Thirdly, since the intake of too much salt/sugar is the major
concern for consumers including the elderlies, strategy for release control of aroma
and taste through food structure design will be shown. Interesting phenomenon from
our findings is that inhomogeneous spatial distribution of aroma can enhance not
only aroma perception but also the eating behavior, which can be favorable for the
elderlies with reduced sensing and eating capability.

Keywords Elderly foods · Dysphagia · Texture design · Criteria · Release

M. Nakauma (*) · T. Funami

San-Ei Gen F.F.I., Toyonaka, Osaka, Japan
e-mail: m-nakauma@saneigenffi.co.jp

© Springer Nature Singapore Pte Ltd. 2021 445

Y. Fang et al. (eds.), Food Hydrocolloids,
https://doi.org/10.1007/978-981-16-0320-4_13
446 M. Nakauma and T. Funami

1 Introduction

Texture is a sensory property (Szczesniak 2002) as defined by the International

Organization for Standardization (1994) and thus should be evaluated by human. As
defined by ISO, texture is associated with all the mechanical, geometrical, and
surface attributes of a food. Hence, food texture is a multiplex of physicochemical
properties perceived by human in a series of food oral processing.
Texture is the most important element of food palatability (Szczesniak and Kleyn
1963), especially for staple foods such as rice, noodle, bread, and meat, where
texture dominates more than 30% of palatability (Nishinari 1996). Texture can
also modify the flavor perception (Clark 2002; Morris 1993) because how a food
is broken down in the mouth and how the broken food is mixed with saliva both
relate to the flavor release from the food and influence its perception through
retronasal pathway (Baek et al. 1999; Boland et al. 2006; Repoux et al. 2012).
Also, texture can modify the digestion and absorption behaviors of nutriments and
minerals on a similar principle to the flavor release (Nakauma et al. 2012, 2014).
Texture is important for safety of eating and is emphasized in recent aged society
(Nishinari 2009). Changes in human physiology with aging can reduce some eating
capabilities through the depression of muscle and organ activities (Mioche 2004;
Sako 2005), saliva secretion (Sako 2005; Shern et al. 1993; Shin et al. 2015), and
sensitivity to taste and aroma (Arey et al. 1935; Doty 1989; Sako 2005). The design
of the elderlies-friendly food formulation is required to the food industry, and safe
swallowing for dysphagia patients should be a priority as aspiration in such patients
can cause death in the worst case through pneumonia (Nishinari et al. 2011). As for
reduction of the swallowing capability with aging, prolonged swallow response due
to a delay in laryngeal closure (Clavé et al. 2006) may cause the misleading. To
prevent the misleading, food or food bolus should flow at moderate velocity through
the pharyngeal phase, not at very high or very low one, as one cohesive bolus with
low adhesiveness and high lubricity. Dysphagia is managed in most cases using
texture-controlled foods, which are thickened or gelled (Clavé et al. 2006;
Logemann 1998; Quinchia et al. 2011). To the author’s understanding, food texture
design has been done empirically in both food manufactures and hospital/nursing-
care facilities, and more scientific-based approach should be necessary for the future.
This chapter covers essential elements for food texture design for the elderlies,
followed by the illustration of industrially feasible way for the goal and the trend
of production development in Japan. New approach for texture evaluation will be
also elaborated for more advanced strategy in the future.
13 Structuring for Elderly Foods 447

2 Use of Polysaccharides as a Texture Modifier

in Elderly Foods

Polysaccharides are frequently used as a texture modifier in processed food products,

and this is also the key technology for the development of elderly foods. Poly-
saccharides used in the food industry are all natural as their sources are found in
seeds, tree sap, seaweeds, microorganism, etc. in nature (Funami 2011). Polysac-
charides serve a variety of purposes in processed food products due to their capa-
bilities of thickening, gelling, water holding, dispersing, stabilizing, film forming,
and foaming (Funami 2011). Through these functions, mechanical, geometrical, and
surface properties can change, and food texture is modified as a result.
In Japan, various types of processed food products are being marketed for the
elderlies. Representatives are semi-solidified (either thickened or gelled) enteral
nutrition, water-supply jelly, and nutrition-supply jelly as a ready-to-eat product.
Also, instant gelling agent for pasted/pureed foods and dysphagia thickener are both
for supplement purpose working through small addition to existing foods and
distributed commercially in a dry powder form. The terms “jelly,” gelling,” and
“thickener” all relate directly to the polysaccharide functions.
Semi-solidified enteral nutrition can reduce the complication risks encountered in
feeding of thin liquid enteral nutrition via an enteral tube or percutaneous endoscopic
gastrostomy. These risks include the secondary aspiration caused by gastroesopha-
geal reflux, local skin infection by the leakage from gastrostomy, bedsore by
prolonged feeding time, and diarrhea by rapid absorption and dumping (Goda
2008). By selection and combination of polysaccharides, the intensity and quality
of viscosity can be modified to achieve smooth flow through the tube and controlled
feeding speed for the prevention of diarrhea and reflux (Tanishima et al. 2009). This
type of enteral nutrition uses representatively agar, pectin, carrageenan, and xanthan
gum and is gaining the market share in Japan (Nakauma et al. 2012, 2014).
Regarding water-supply jelly, the creation of new texture has been tried by poly-
saccharide technology to mimic not only texture but also appearance of real fruits
(Funami 2013). Appearance can enhance human appetite as one of the dominant
attributes for food palatability. It has been reported recently that the visual food
recognition can influence on swallowing function (Kamiya et al. 2015). This study
demonstrates that a larger amount of saliva secretion was found when subjects saw
the pictures of normal diets compared to those of corresponding blended foods.
Another advantage of using polysaccharides instead of real fruits lies in the feasi-
bility of calorie and nutrition controls. For example, control of potassium intake is
essential for patients with kidney dialysis treatment, and this type of food products is
flexible in changing the formulation. Instant gelling agent is used for shape retention
and the prevention of syneresis for pasted and pureed foods (Funami 2013). Foods,
including cooked rice, fish, meat, and vegetable, are pasted together with the gelling
agent and water or soup if necessary. By remolding to the original shape, people can
recognize what they are eating. Polysaccharides which hydrate or dissolve in water
without heating are preferably used in this type of gelling agent.
448 M. Nakauma and T. Funami

3 Usefulness of Xanthan Gum as Dysphagia Thickener

Low viscosity liquids can be aspirated by dysphagia patients (Nishinari et al. 2011).
To prevent this, dysphagia thickener, called toromi in Japan, is used at hospital and
nursing-care facilities. Dysphagia thickener contains polysaccharides as a main
ingredient, where xanthan gum has been recently replacing modified starch (Funami
et al. 2006, 2009). Dysphagia thickener is usually stirred manually to disperse them
in liquid foods like water, tea, juice, soup, etc. Dysphagia thickener should have
functions of no lumping, rapid viscosity increase, stable viscosity upon storage, and
usability in a wide range of liquid foods even under weak stirring conditions. The
thickener should also provide preferable organoleptic properties with liquid foods,
including cohesive texture and no off-flavor. Xanthan gum meets these requirements
and must be practically the best choice (Funami et al. 2006).
Most polysaccharides, not limited to xanthan, hydrate quickly in water and thus
form lumps easily. Lumping is the most unfavorable for dysphagia thickener. When
this happens, textural homogeneity cannot be ensured, and this adverse effect against
good dispersion should be eliminated by powder processing, mostly granulation.
Here, the relationship between dispersibility and viscosity enhancement can be
described as a seesaw in which when one is improved, the other is deteriorated
(Funami et al. 2006). For good balance of these two attributes, technical know-how
in a granulation process should be necessary, including optimization of certain
operation conditions (e.g., inlet air temperature and volume, liquid spray rate and
amount) or usage of certain co-agents in a liquid spray, either of which aims to
control the speed of polysaccharide hydration.
Xanthan solutions are fluid, but from rheological point of view, they behave like a
solid or gel (Nakauma et al. 2011). As shown in Fig. 13.1, xanthan solutions have

Fig. 13.1 Elastic stress (storage modulus
strain) of xanthan gum and locust bean gum solutions
at 20  C plotted as function of strain. Concentrations of polysaccharide: 0.3, 0.45, 0.6, 0.75 and
0.9% for xanthan gum (a) and 0.5, 0.6, 0.7, 0.75, and 0.8% for locust bean gum (b), represented by
open circles, closed triangles, open triangles, crosses, and closed circles in increasing order for each
polysaccharide. The yield stress and strain were estimated from the maximum point in the curve.
For the experimental details, see Nakauma et al. (2011), Food Hydrocolloids 25, 1165–1173.
Reproduction with permission from (Nakauma et al. 2011), Copyright 2010 Elsevier Ltd
13 Structuring for Elderly Foods 449

elastic character represented by yield stress in contrast to locust bean gum. Yield
stress was detected as the peak in Fig. 13.1, in which the data from strain-
dependence of dynamic viscoelasticity were replotted. The yield stress is a stress
at which the flow begins (Bourne 2002a), relating to the degree of the internal
binding force. Thus, the yield stress can present cohesiveness of food bolus. It has
been believed for a long time that the flow speed of liquid food bolus through the
pharyngeal phase is moderated by thickening and that shear viscosity can only
determine the flow behavior; the higher the viscosity, the slower the flow speed
can be through the pharyngeal phase. Whether it is always true or not and which
shear rate should be used for viscosity measurement to obtain the best correlation
with physiological flow behavior should be of research interest. For the first ques-
tion, the answer must be NO when the results from our recent studies (Nakauma et al.
2011; Funami et al. 2017) are considered. Elasticity should not be negligible or
rather important in explaining physiological flow behavior of food bolus. This is
supported by Chen and Lolivret (2011), who concluded that extensional stretch
ability should be one of the most important mechanical parameters for swallowing
ease. For the second question, the maximum shear rate was expected to range
between 400 s1 (Meng et al. 2005) and 3000 or even larger 180,000 s1 (Nicosia
and Robbins 2001) depending on the type of simulation. However, it is still quite
difficult to estimate such an in vivo parameter in silico due to the irregularity of oral
geometry.
As a non-invasive in vivo measurement, acoustic analysis was performed to
investigate the correlation with sensory evaluation (Funami 2011; Nakauma et al.
2011). From the acoustic analysis, time required for food bolus to transfer through
the pharyngeal phase (t2) decreases with increasing xanthan concentration although
steady shear viscosity is increased (Fig. 13.2). This may be consistent with the
ultrasonic pulse Doppler measurement, elucidating that velocity spectra are narrower
in the distribution range with increasing concentration of food polysaccharide
thickeners (Kumagai et al. 2009). Also, t2 is well correlated with perceived intensi-
ties of cohesiveness and swallowing ease (Fig. 13.3). Bolus from xanthan solutions
flows through the pharyngeal phase as one coherent bolus with small variation of
flow velocity, and this flow behavior leads to a greater sensation of swallowing ease.
For swallowing ease, “structured fluid” or “weak gel” should be a preferable food
structure from rheological point of view (Nakauma et al. 2011). This food structure
can be represented rheologically by yield stress of approx. 7.0–9.0 Pa and steady
shear viscosity of approx. 0.9–1.2 Pas at 10 s1 (Nakauma et al. 2011). Also, it is
reported that non-Newtonian fluids, representatively xanthan solutions with showing
shear-thinning behavior (Nishinari et al. 2011), are safer to swallow than Newtonian
fluids with lower aspiration risk (Meng et al. 2005). The preference of
non-Newtonian fluids for safe swallowing refers to the need of elasticity to ensure
the mechanical cohesiveness of food bolus rather than the issue of shear rate
dependence (Funami et al. 2012).
450 M. Nakauma and T. Funami

Fig. 13.2 Duration t2 in swallowing polysaccharide solutions as a function of steady shear

viscosity h at 10 s1. Both mechanical and in vivo measurements were carried out at 20  C. Closed
circles: xanthan gum; open circles: locust bean gum. Serving volume of polysaccharide solutions
was 15 mL. Each datum was standardized with that for control (water). Data with asterisk are
significantly different between xanthan gum and locust bean gum at p < 0.05. For the experimental
details, see Nakauma et al. (2011), Food Hydrocolloids 25, 1165–1173. Reproduction with
permission from (Nakauma et al. 2011), Copyright 2010 Elsevier Ltd

4 Working Mechanical Criteria for Elderly Foods in Japan

and Comparison to the Global Standards

Representative Japanese standards to define the mechanical criteria of elderly foods

will be presented. As a national standard issued by the Consumer Affairs Agency,
“Foods for swallowing difficulty” has three-level classification and defines the range
of mechanical properties for each classification (Nishinari et al. 2013, 2019a).
“Universal Design Foods” (http://www.udf.jp/about_udf/section_01.html) is an
industry standard issued by the Japan Care Food Conference, in which food manu-
factures and food ingredient/package companies are involved. It has four-level
classification differed by mastication difficulty. “Dysphagia Foods Pyramid” is a
standard proposed by a nutrition manager of a famous hospital and has five-level
classification by swallowing difficulty. It is similar to the national standard in that
both use hardness, adhesiveness, and cohesiveness as a mechanical parameter (Sakai
2007). Mechanical criteria presented in these standards are served as a guideline
mainly for food manufactures in producing processed foods, not for hospital and
nursing-care facilities. This is because the measurement apparatus is neither inex-
pensive nor suitable for on-site use.
Mechanical hardness, adhesiveness, and cohesiveness, adopted in “Foods for
swallowing difficulty” and “Dysphagia Foods Pyramid,” are measured by two-bite
analyses called TPA (Texture Profile Analysis). TPA uses a universal testing
machine such as Instron, and in TPA profiling, a food sample is compressed twice
in a reciprocal manner for mimicking human jaw action during food consumption.
Based on the resultant force-time curve, hardness is determined by the force peak in
13 Structuring for Elderly Foods 451

Fig. 13.3 Correlation between the duration t2 and the score of each sensory attribute in swallowing
polysaccharide solutions. Serving volume of polysaccharide solutions was 15 mL. (a) perceived
adhesiveness; (b) perceived cohesiveness; (c) perceived swallowing ease. Duration t2 was stan-
dardized with the corresponding data for water. For the experimental details, see Nakauma et al.
(2011), Food Hydrocolloids 25, 1165–1173. Reproduction with permission from (Nakauma et al.
2011), Copyright 2010 Elsevier Ltd

the first bite cycle, adhesiveness by the negative force area during the first bite, and
cohesiveness by the ratio of the positive force areas under the first and the second
bites (Bourne 2002b). This was originally developed for measurements of solid or
semi-solid foods (Szczesniak and Hall 1975), but now the TPA measurement is
expanded to less hard and more fluid foods. This generates one major difference
from the original; a support of a container or cup to “pour” the food sample is needed
for TPA measurements, which is not the case of solid or semi-solid foods as they can
stand alone. The size of plunger is smaller than that of test sample, which is another
difference from the original. Due to these differences, some difficulties in operation
and data analysis have been highlighted (Nouchi et al. 2012; Nishinari et al. 2013;
Brenner and Nishinari 2014). In principle, TPA parameters cannot be compared
among food samples of different viscoelastic natures, and the TPA founder
concerned about the misinterpretation of TPA parameters obtained from the mea-
surement of liquid foods in a cup (Szczesniak and Bourne 1998). Although there is
no doubt as to the utility of TPA parameters in presenting food texture, we should be
452 M. Nakauma and T. Funami

Table 13.1 New grouping of thickening led by the Japanese Society of Dysphagia Rehabilitation
Level 1 Level 2 Level 3
Mildly thick Moderately thick Extremely thick
Features • Appropriate to • Viscus to be • Apparently viscus to
(in swallowing) express by “drink” detected be detected
• Spreadable in the • Appropriate to • Easy to form bolus
mouth express by “drink” • Appropriate to
• Not concerned for • Smooth but not express by “eat” using a
some liquids depending on so spreadable in the spoon
the kind, taste, and tem- mouth • Difficult to swallow
perature • Cohesive on through a straw
• Large force unneces- the tongue
sary in swallowing • Perceiving a
• Easy to swallow resistance to swal-
through a straw low through a straw
Feature • Easy to slide on spoon • Slidable on • Shape retention and
(in appearance) surface when tilted spoon surface when resistant to slide on spoon
• Flowing rapidly tilted surface when tilted
through in-between forks • Flowing • Cannot flow through
• Trace of flow on cup slowly through in-between forks
surface when tilted to pour in-between forks • Not flowing out of
off • Thin film for- cup when tilted or
mation on cup sur- flowing slowly as one
face when titled to cohesive bolus
put off
Viscositya 50–150 150–300 300–500
(mPas)
LST valueb 36–43 32–36 30–32
(mm)
a
Measuring at 50s1 at 20  C
b
Pouring 20 mL sample into a cylinder of 30 mm in diameter, drawing it up, and measuring the
length of the spread after 30 s for determination of Line Spread Value. Reproduction with
permission from (Funami 2016), Copyright 2016 Wiley Periodicals, Inc

cautious about their validities or physical meanings in discussing TPA results

(Nishinari and Fang 2018; Nishinari et al. 2019b).
As shown, multiple standards are working in parallel in Japan, showing some
complexity. Under the leadership of the Japanese Society of Dysphagia Rehabilita-
tion (JSDR), these standards were integrated. A novel classification of dysphagia
foods has been proposed by JSDR, in which new five classes are identified with no
mechanical criteria. Interchangeability of these new five classes with previous three
standards has been also indicated in the JSDR standard. JSDR has also proposed a
new grouping of thickening (Table 13.1). Viscosity should be measured at a fixed
shear rate of 50 s1, which is harmonized with American standard National Dys-
phagia Diets; NDD (Cichero et al. 2013), and the choice of this shear rate is based on
Wood’s report (Wood 1968). Line Spread Test, which is similar to USDA
Consistometer (Bourne 2002c) and converts rheological measures for fluids
expressed in viscosity mPas to distance mm (Cichero et al. 2013), is also adopted
13 Structuring for Elderly Foods 453

Table 13.2 International terminology of thickened liquids (cite from Curr Phys Med Rehabil Rep
(2013) 1:280–291
Country < “Water-like” “Pudding-like” >
USA Thin Nectar-Like Honey-like Spoon-thick
(NDD)[45] (1-50 cPa) (51-350 cPa) (351-1750 cPa) (>1750cPa)
United Thin Naturally thick fluid Thickened fluid – stage Thickened fluid 1 Thickened fluid – Stage 3
Kingdom[44] 2
Australia[6] Regular - Level 150 – Level 400 – Level 900 –
Mildly thick moderately thick Extremely thick
Ireland[40] Regular Grade 1 – Grade 2 – Grade 3 – Grade 4 –
Very mildly thick Mildly thick Moderately thick Extremely thick
Japan Less mildly thick Mildly thick Moderately thick Extremely thick Over Extremely thick
(JSDR; (< 50 mPa.sa) (50-150 mPa.sa) (150-300 mPa.sa) (300-500 mPa.sa) (> 500 mPa.sa)
scheme)[41]
Canada Regular/ Thin/ Clear Nectar / Stage 1 / Level Honey / Stage 2 / Pudding / Spoon thick /
1/ >250cP / 51-350 cP Level 2/ > 800 cP / Stage 3 / level 3 / > 2000 cP
351-1750cP / / > 1750 cP
Default Thick
Denmark[46] Normal Chocolate milk Syrup Jelly

Spain Thin Medium Full protection/thick/pudding

Netherlands Thin ‘Thickened’ Pudding-like
Brazil Normal or thin Thicker liquid Nectar or Honey Paste or Creamy
(Homogenous or
Heterogenous)
Sweden[43] Liquids Thickened liquids

a
Shear rate 50 s-1, both cP and mPas are used in the literature as the unit of viscosity, 1 cP ¼ 1 mPas.
Reproduction with permission from (Cichero et al. 2013), Copyright the Authors 2013. This article
is published with open access at Springerlink.com

as more convenient and less expensive method. JSDR can be the most advanced
specification which provides measurable indication. We can see both similarities and
differences when compared to the terminology and viscosity guideline used in other
countries (Table 13.2). Regarding the terminology, real fluid foods, including nectar,
honey, pudding, chocolate milk, syrup, and jelly are used for viscosity classification
in some standards like USA, Canada, Denmark, etc. In Australia, Ireland, and also
JSDR standards, predicative presentation of thickness such as mildly thick, moder-
ately thick, or extremely thick are used. However, these terms mean ambiguously if
the definition is lacking as the image of thickness perceived by the terms depends on
personal cognition. Therefore, subjective measures with physical meaning are nec-
essary to get consensus and to remove the bias caused by the terminological
ambiguity. As a subjective measure, NDD, JSDR, and Canada (maybe) use shear
viscosity at 50 s1, but this is not in the case for other countries. It is necessary to
install the instrument like viscometer for measurement of viscosity. This may cost a
lot for hospitals and care homes and can be one of the reasons for this inhomoge-
neity. To secure the patient safety and to enhance inter-professional collaboration,
international initiative was led by an independent and non-profit entity; the Interna-
tional Dysphagia Diet Standardisation Initiative (IDDSI).
As a noticeable activity, IDDSI developed a new set of standardized terminology
and definition for the description of texture of modified foods and thickened fluids in
2016 (Lam and Cichero 2016; Cichero et al. 2017). The IDDSI framework consists
of a continuum of 8 levels (0–7), identified by numbers, text labels, and color codes.
Levels 0–4 represent fluid foods, levels 3–7 represent solid or semi-solid foods, and
454 M. Nakauma and T. Funami

levels 3 and 4 at the intermediate and apply to both. The IDDSI flow test was
proposed for the measurements of fluid foods, in which a 10-ml syringe is used. In
this test, the syringe is filled with 10 ml fluid food, and the fluid food is allowed to
flow freely by gravity for 10 s. The IDDSI level of the fluid food is determined by
reading the remaining volume after the gravity flow for 10 s. The larger the volume,
the thicker the sample is. This test is for categorizing the texture of fluid foods (i.e.,
thickness) in simple and practical manner and can be utilized universally with low
cost (Su et al. 2018; Yokote et al. 2017; Steele et al. 2018). Although the IDDSI test
requires the measurements under ambient conditions where foods and beverages are
usually consumed, it is desirable from scientific point of view to perform the test
under a temperature-controlled condition because the viscosity of fluid foods
depends strongly on temperature. Texture of solid or semi-solid foods is often
represented by hardness, and hardness is usually determined mechanically on a
texture analyzer, which compresses or extend test sample coaxially. However, this
may cost a lot for hospitals and care homes as in the case of viscometer, leading to
the difficulty in installation. A practical test using a fork or spoon has been previ-
ously recommended as part of the United Kingdom dysphagia diet standard for
assessment of foods with the IDDSI Levels 5–7. In this test, force or pressure is
applied to a food sample with a fork, and deformation behavior of the food sample is
observed. In order to standardize the force applied, the IDDSI fork test recommends
that the fork should be pressed onto the food sample by placing the thumb onto the
bowl of the fork (base of the prongs), and pressing just hard enough to cause
blanching of the thumbnail. Blanching occurs when the pressure overcomes mean
arterial blood pressure and has been quantified at approx. 17 kPa. Details are
described in the following URL;
http://ftp.iddsi.org/Documents/Testing_Methods_IDDSI_Framework_Final_31_
July2019.pdf
This pressure corresponds closely to a typical tongue pressure during swallowing
(Fei et al. 2013; Youmans and Stierwalt 2006). Descriptions and testing methods
have been also developed using chopsticks (indirect as in the case of fork) and
fingers (direct) instead of fork.

5 Mechanical Simulation of Palatal Reduction

Two oral strategies for food size reduction in the mouth are known; mastication
(or chewing) and palatal reduction. Palatal reduction involves squeezing and tongue-
(hard) palate compression, and the elderlies like to use palatal reduction, particularly
those with mastication difficulty. Solid or semi-solid foods are usually reduced to
suitable particle sizes by mastication and/or palatal reduction, followed by incorpo-
ration of the right amount of saliva to form coherent bolus (Chen 2009; Hutchings
and Lillford 1988; Prinz and Lucas 1995). Rheological properties change dramati-
cally from the original food (prior to oral processing) during formation of food bolus
(Chen 2009). Bolus rheology is gaining interest in relation to food texture perception
13 Structuring for Elderly Foods 455

50 mm

10 mm/s Metal probe

(Aluminum)

20 mm
Gellan gel
10 mm

Artificial tongue 10 mm
Metal stage

Fig. 13.4 Setup and standard operation condition of instrumental compression test using artificial
(simulated) tongue as a substrate. Reproduction with permission from (Ishihara et al. 2014),
Copyright 2014 Wiley Periodicals, Inc

(Ishihara et al. 2011b). Easiness to form bolus and suitability of bolus to the
elderlies’ oral physiology should be the key for the formulation or texture design
of elderly foods.
In a series of food oral management, a food is deformed slightly by tongue-palate
compression at the initial stage for recognition of texture (Arai and Yamada 1993)
and for determination of subsequent oral strategy for size reduction. That is, when a
food is soft enough to be fractured by tongue-palate compression, the tongue works
continuously for size reduction, whereas mastication is initiated when a food is too
strong to be fracture by tongue-palate compression. To mimic tongue-palate com-
pression, an in vitro food texture assessment system has been developed, where a
food sample (in a cylindrical shape of 20 mm in diameter and 10 mm in height) is
compressed between hard non-deformable plate (50 mm in diameter) and artificial
tongue from a soft material (i.e., silicone rubber of the same shape and size as the
food sample) on a conventional uniaxial compression apparatus (Ishihara et al. 2013)
(Fig. 13.4). Young’s modulus of artificial tongue corresponds to apparent elastic
modulus determined at approx. 20% strain. It is concluded from their study using
agar gels as food samples that when an artificial tongue with the Young’s modulus of
approx. 55 kPa is used as substrate, the fracture profile of a food sample predicts the
human oral strategy for size reduction. That is, when a food sample is broken down
by compression on the system, tongue-palate compression will be the main oral
strategy for size reduction, whereas mastication will be when the food sample is not
broken down on the system (Table 13.3). The assessment system uses the compres-
sion (crosshead) speed at 10 mm/s and the same size of crosshead as the food sample.
Young’s modulus of artificial tongue lies between that of human (healthy young
456 M. Nakauma and T. Funami

Table 13.3 Fracture profile Artificial tongue

of agar gels during compres-
S40 S50 S60
sion on artificial tongue in
relation to human oral strategy Agar gel (18.3 kPa) (54.9 kPa) (113.0 kPa)
for size reduction A3 F F F
(14.7 kPa) 8/0 8/0 8/0
A4 F F F
(17.5 kPa) 7/1 7/1 7/1
A5 N F F
(53.5 kPa) 3/5 3/5 3/5
A6 N N F
(97.9 kPa) 0/8 0/8 0/8
Symbols F and N represent that agar gels did not fracture upon
compression on each artificial tongue up to 50% strain of the
combination of agar gel and artificial tongue, respectively. Slash
data below the symbol represent the number of subjects who used
the tongue-palate compression for size reduction (upper) and the
number of subjects who used mastication for size reduction
(lower). Figures in parenthesis represent apparent elastic modulus
of either agar gel or artificial tongue. Reproduction with permis-
sion from (Funami 2016), Copyright 2016 Wiley Periodicals, Inc

adults) tongue in relaxation; 12.2 kPa and in a tension state; 122.5 kPa (Ishihara et al.
2013).
Subsequent study confirmed the robustness and the limitation of the assessment
system using gellan gum gels in wider mechanical range (Ishihara et al. 2014). It is
concluded from their study that operation conditions should be modified in some
cases, which may linked to physiological changes in tongue-palate compression
upon food texture. In details, some behavioral modulations may occur during
consumption of highly deformable gels. These include the tongue-palate compres-
sion speed and tongue excitation. The first factor can be recreated by decreasing the
crosshead speed from 10 to 5 mm/s in associated with the stress relaxation, a
phenomenon of energy dissipation increased at lower deformation speed (Luyten
and van Vliet 1995). The second factor can be recreated by increasing the Young’s
modulus of artificial tongue from approx. 55 to approx. 110 kPa. The slope of the
curve from tongue pressure measurement (tongue pressure versus time) during the
first size reduction is almost independent of physical properties (texturally brittle or
deformable) or consistency (soft or hard) of polysaccharide gels (Hori et al. 2015).
Based on this finding, it should be reasonable that decreased speed of tongue-palate
compression gives rise to increased tongue pressure. Decrease in the crosshead
speed or increase in the Young’s modulus of artificial tongue in evaluating highly
deformable samples should be also reasonable. Usage of artificial tongue of larger
size can be an option as surface contact area between the food sample and the tongue
will be enlarged when the food sample is highly deformable. In relation to “highly
deformable,” it is suggested that the mode to decide oral strategy for size reduction
should depend on the fracture strain of food, and its boundary can be at approx.
60%–70% (Ishihara et al. 2014). The oral strategy for size reduction can be decided
13 Structuring for Elderly Foods 457

Fig. 13.5 Mechanical mouth-simulator for model bolus formation. Reproduction with permission
from (Ishihara et al. 2011a), Copyright 2010 Elsevier Ltd

by sensing the difference in strain of food relative to the tongue perceived dynam-
ically during oral processing (Kohyama 2015). Fracture strain is a dominant
mechanical parameter which can govern the decision of oral strategy for size
reduction (Arai and Yamada 1993), and this is the same as the determination of
the biting speed (Mioche and Peyron 1995). These are all in line with the results from
Ishihara et al. (2013, 2014).
Mechanical mouth-simulator is useful to collect simulated food bolus as shown in
Fig. 13.5 (Funami 2011; Ishihara et al. 2011a, 2011b). This simulator, originally
developed for flavor analysis, works in a closed system, allowing the process of a
food sample in the presence of artificial (simulated) saliva. Human saliva is two
orders of magnitude lower than that of water in the boundary friction coefficient
(Bongaerts et al. 2007), and the lubrication of food bolus is determined by the
miscibility with saliva. This is why the saliva incorporation in bolus rheology and
tribology is important (Torres et al. 2019). With regard to deformation, a food
sample is compressed with shear on the simulator, which recreate the tongue
movement during food consumption; 10 cycles of vertical compression at
17.8 mm/s with 2.0 mm-clearance and simultaneous rotational shearing at rate of
12 /s. In studies by Ishihara et al. (2011a, 2011b), four gel samples prepared using
two different gelling agents at two different consistencies were processed mechan-
ically to obtain simulated food bolus. Gels can be base for dysphagia foods due to
their versatile viscoelastic character (Funami 2011), and thus gel samples were used
as a model food. For example, gels from the mixture of low-acylated gellan gum and
psyllium seed gum have more deformable (plastic) texture than that of gellan single
gels due to the effects of psyllium seed gum, and water-holding capacity of the
mixture gels is also increased (Ishihara et al. 2011c). As shown in Fig. 13.6, the
frequency-dependence of dynamic mechanical loss tangent of stimulated bolus from
458 M. Nakauma and T. Funami

Fig. 13.6 Frequency-dependence of dynamic mechanical loss tangent for model bolus. Measure-
ments were carried out at 20  C at a fixed strain of 1%. Open circles: 1.0% SAN SUPPORT®
(a mixture of de—acylated gellan gum and psyllium seed gum); closed circle: 1.5% SAN SUP-
PORT®; open triangle: 0.075% gellan gum; closed triangle: 0.15% gellan gum. Data are presented
as means  SD of triplicate. For the experimental details, see Ishihara et al. (2011a), Food
Hydrocolloids 25, 1016–1024. Reproduction with permission from (Funami 2016), Copyright
2016 Wiley Periodicals, Inc

the mixture gels is almost independent of the addition level of simulated saliva,
whereas the bolus form gellan single gel is dependent on. Here, mechanical loss
tangent presents how a sample is elastic or viscous, and the smaller the parameter,
the more elastic the sample is. The mixture gels are less influenced by the saliva
secretion in bolus formation and exhibit rheologically weak gel property, assumedly
leading to a comfortableness of eating for the elderlies.

6 Progress of In Vivo Physiological Tests as Advanced

Texture Assessment

Although rheological measurements reveal the mechanical and related microstruc-

tural properties of foods, the reality and the complexity of oral experience can be
oversimplified (Peleg 2006). The deviation is due to the lack of salivation (Stokes
2012) and the use of stiff materials (Peleg 2006), for instance. Limitations of
rheological measurements in identification of food texture, causing inconsistency
between both results, are explained by at least two reasons; temporal difference in
judgement and the complexity/simplicity of sensing between instrument and human
(Kohyama 2015). Another explanation should be from Peleg (2006) indicating that
the sensory response to a mechanical (or acoustic) stimulus should be expressed not
as a characteristic ‘value’, i.e. as a mean relative or an absolute score, but as a
distribution of the terms used by those who sense them. Therefore, in vivo human
physiological measurements should be introduced to reconcile instrumental
13 Structuring for Elderly Foods 459

measures (Foster et al. 2011; Koç et al. 2013; Wilkinson et al. 2000), where
non-invasive sensors attached to human subjects are used to monitor the organ and
muscle signals related to food oral processing for tracing the dynamic changes of
food texture. A various kinds of in vivo measurements are often used for texture
assessment, involving palatal pressure measurements (Nakazawa and Togashi 2000;
Takahashi and Nakazawa 1991, 1992), electromyography (Kohyama et al. 1998,
2005b, 2005c, 2007), biting force measurements (Kohyama and Nishi 1997;
Kohyama et al. 1997, 2001, 2003, 2005a), and ultrasonic pulse Doppler methods
(Kumagai et al. 2009; Nakazawa et al. 2000). In terms of dynamics, in vivo
measurements lie between instrumental measurements and sensory evaluations,
and understanding on the dynamic changes of foods during oral processing should
be the key for texture studies (Chen 2009). From this perspective, in vivo measure-
ments should be utilized more in this research area. As a model food/beverage for
texture studies, polysaccharide gels/sols are useful because of their controllable
rheological properties without significantly affecting other organoleptic properties,
particularly flavor and appearance (Peleg 2006).

6.1 Tongue Pressure Measurement

As the tongue plays a crucial role in a series of food oral processing, it is physio-
logically reasonable to link its movement to food texture. Two literatures published
recently, relating to tongue pressure measurement during food oral processing and
its usefulness for food texture study, are highlighted. In these two literatures, effects
of food texture on tongue movement were investigated using a novel sensing system
and polysaccharide gels as a model food (Hori et al. 2015; Yokoyama et al. 2014).
They indicated potential usage of tongue pressure measurement for elucidation of
the dynamics of food oral processing and for visualization of food-tongue interac-
tion. T-shaped sensor sheet with five measuring channels is the main part of the
sensing system (Fig. 13.7), which makes it possible to monitor tongue pressure in
real time on an electrical transducer (from resistance to electrical current). This
device has been developed by Osaka University research team, which specializes
the design of the palatal plate particularly for tongue cancer patients (Hori et al.
2009). Their idea to apply this device to food science is novel utilizing its features
beneficial for texture study. The sensor sheet has only 0.1 mm thickness with various
sizes, and its mechanical flexibility realizes easy adaptation to the oral shape and
does not change the oral physiology nor interfere with the occlusion. In addition, the
sensor sheet does not cover the whole area of the palate and thus does not interfere
with taste, aroma, and texture perceptions. The sensor sheet does not interfere with
natural eating behavior of human, and this can be the greatest advantage over a
conventional system using a small balloon-shaped probe called a manometer
(Shaker et al. 1988). Using this sensing system, temporal and spatial distribution
patterns of tongue pressure during food oral processing were elucidated (Hori et al.
2015; Yokoyama et al. 2014). Polysaccharide gels were used as a test sample with
460 M. Nakauma and T. Funami

Fig. 13.7 Tongue pressure sensing system (a) and location of sensor points (b). The lengths
between Ch. 1 and Ch. 3 were 23, 25.5, 28 mm for Small, Medium, Large size of sensor sheet
respectively. Those between Ch. 4 and Ch. 5 were 35, 38, 41 mm for S, M, L size. Reproduction
with permission from (Hori et al. 2015), Copyright 2014 Elsevier Ltd

various textures by using different gelling agents; low-acylated gellan gum, and the
mixture of low-acylated gellan gum and psyllium seed gum. By changing polysac-
charide concentration, gels of three different hardness values (defined here as the
stress at 67% strain); approx. 1600, 5500, and 12,000 Pa were prepared. Texture of
the gellan single gel is brittle (Morris 2006) with detectable syneresis, particularly at
low concentrations. For the mixture gel, syneresis is not detective due to the function
of psyllium seed gum, which also increases textural deformability (Ishihara et al.
2011c). When subjects were asked to eat the gel samples by compressing them
between the tongue and the hard palate (i.e., squeezing), temporal pattern of tongue
pressure during squeezing did not change substantially upon gel texture, presenting
the first onset at the mid-median part (Ch. 2), followed by the anterior-median
(Ch. 1), the posterior-median (Ch. 3) parts, and the circumferential parts (Chs.
4 and 5) in this order for each type of gel sample (Fig. 13.8). However, differences
were noted in offset between the two gel samples. As a result, duration of squeezing
for texturally deformable gels (i.e., the mixture gels) tended to become shorter than
that for texturally brittle gels (i.e., gellan single gels) when the gel hardness was
lower, whereas vice versa when the gel hardness was higher. Spatial distribution
pattern of the tongue pressure during squeezing changed upon gel texture; the
maximum amplitudes at Chs. 1 and 2 were larger than those at Chs. 4 and 5 for
gellan single gels, whereas no marked difference was found between channels for the
mixture gels (Fig. 13.9). Consequently, human eating behavior can be visualized by
tongue movement, which changes upon food texture. Foods in the form of weak
gels, emulsions, and fluids are eaten by squeeze between the tongue and the palate
during oral processing, and in these cases, texture relates to thin film rheological
behaviors of foods as well as the bulk properties (Malone et al. 2003), including
mayonnaise (Giasson et al. 1997) and chocolate (Luengo et al. 1997). The combi-
nation of both shear and squeeze flow is more realistic representation for tongue
movement during food oral processing (Stokes 2012). The influence of polysaccha-
rides’ conformation and structure on lubrication has been presented (Garrec and
Norton 2012, 2013; Malone et al. 2003), and effects of polysaccharides on food
13 Structuring for Elderly Foods 461

Fig. 13.8 Time sequences of tongue pressure during the initial squeeze. The time “0” was set as the
onset of Ch. 1. *: p < 0.05 (Kruskale–Wallis test and post hoc test with Bonferroni correction).
S1–3 presents texturally deformable gel samples from the mixture of low-acylated gellan gum and
psyllium gum with increased consistency in order, whereas G1–3 presents texturally brittle gel
samples from low-acylated gellan gum with increased consistency in order. When the number is the
same, consistency of the gel sample is equivalent between both types of gel sample. Reproduction
with permission from (Hori et al. 2015), Copyright 2014 Elsevier Ltd
462 M. Nakauma and T. Funami

Fig. 13.9 The maximum amplitude of tongue pressure at each channel during the initial squeeze. *:
p < 0.05 (one-way ANOVA test and Tukey’s post hoc test). Reproduction with permission from
(Hori et al. 2015), Copyright 2014 Elsevier Ltd

texture, including creaminess, smoothness, sliminess, and thickness, should be

investigated by tribological approaches (See Chap. 7 in this book). From these
perspectives, it is expected that tongue pressure measurement will be facilitated
and emphasized in food texture study.

6.2 Electromyography

Electromyography (EMG) has a long history of usage in food science area (Dea et al.
1988; Boyar and Kilcast 1986) and has gained the status as one of the most popular
in vivo measurements for food texture (Espinosa and Chen 2012; Funami et al.
2014). Relationship of some EMG variables with mechanical properties from instru-
mental test or texture perception from sensory evaluation has been investigated a lot,
and some representative investigations will be reviewed briefly in this section.
Eight kinds of solid foods in a wide range of physicochemical properties were
selected as a test food (i.e., dry hard bread, elastic konjac gel, dry sausage, soft
candy, raw radish, pickled radish, boiled carrot, and raw carrot), and nine indepen-
dent parameters were chosen from 28 physicochemical parameters (i.e., stress at
10%, 50%, 70%, or 90% compression strain, breaking stress, cohesiveness, adhe-
siveness, density, and moisture content) (Kohyama et al. 2008). It is concluded that
the mechanical properties under larger deformation should correlate well to EMG
variables. Results from other studies using buckwheat noodles (Kohyama et al.
2010) and gummy candies (Hayakawa et al. 2009) support this conclusion. When
the correlation with EMG variables is studied for solid foods which needs mastica-
tion, it would be recommended in general to see the mechanical properties of food
sample under large deformation conditions, sometimes beyond the fracture point.
13 Structuring for Elderly Foods 463

High correlation can be found between EMG variables and mechanical properties
under extremely deformation condition regardless of rheological nature of food
sample; elastic or plastic since the upper and lower teeth almost reach contact during
mastication for each type of sample. EMG can also detect food oral process at the
early stage, it would be necessary to further investigate why there is hardly seen a
good correlation between EMG parameters and sensory evaluation in small defor-
mation ranges. It is not clear whether the sensitivity of EMG is not high enough or
human did not care about small deformation.
All these studies used relatively hard foods consumed by mastication or chewing
for size reduction, but similar principle can be applied to relatively soft foods which
do not necessarily require mastication. From this perspective, EMG measurements
were performed for foods which are soft enough to be processed with the tongue-
palate compression (Ishihara et al. 2011c). In this study, polysaccharide gels from
either low-acylated gellan gum or the mixture of low-acylated gellan gum and
psyllium seed gum were fed to young healthy adult subjects, and the correlation
between EMG variables and mechanical properties was investigated. For each
gelling agent, gels of different hardness; approx. 1000 and 4000 Pa determined as
the compression stress at 67% strain were prepared by changing polysaccharide
concentration, and 15 g for each gel was served to the subjects. As mentioned, there
was a texture difference between both types of gel (i.e., brittle or deformable). It is
concluded that whole sequence of food oral processing is prolonged and that EMG
activity of the suprahyoid musculature increases by increasing hardness for each
type of gel. EMG activity of the suprahyoid musculature is well correlated to the
compression load at extremely large strain conditions (e.g., 90% strain) and to
sensory perceived hardness. This is similar to another study using gelatin gels,
which are different in rheological natures; elastic or plastic and are processed by
mastication, where the duration of the whole sequence and EMG activity of the
masseter and the temporalis increase with Instron hardness represented as the
compression stress at 50% strain (Foster et al. 2006). In summary, mechanical
properties at large strains relate to physiological response during food oral
processing regardless the oral strategy for size reduction; mastication or tongue-
palate compression.

7 Release Control of Flavor and the Effects on Human

Eating Behavior

Using food gels as a feeding sample, it was found that perceived flavor (including
strawberry aroma and sucrose) intensity increased as their fracture strain decreased
(Morris 1993). Another study showed that overall flavor (including aroma and
sweetener but not specified) intensity decreased linearly with increasing hardness
(i.e., rupture force) of food gels (Clark 2002). However, a highly cohesive gel was
exceptional and positioned below the linear regression line when compared at
464 M. Nakauma and T. Funami

equivalent hardness. Whereas gelatin gel was positioned upward the linear regres-
sion line when compared at equivalent hardness, representing increased flavor
intensity due to lower melting temperature of the gels. Thermal property and
water-holding capacity (i.e., syneresis) are also essential attributes in determining
flavor release (Nishinari 2006), while the effect of free diffusion may be relatively
low compared to the effect of external deformation through compression and
fracture (Wang et al. 2014). As a rheological parameter, fracture strain presents
perceived flavor intensity better than fracture stress, which should be reasonable
when food breakdown during oral processing is considered. Foods with smaller
fracture strain and more brittle texture collapse at an early stage of oral processing
and disintegrate into smaller particles. This rupture behavior of foods can lead to
increased surface exposed area of food bolus in the mouth, stimulating perceived
intensity of flavor via contact with saliva. It has been reported (Weel et al. 2002) that
significant changes in aroma intensity are observed between different concentrations
of whey protein gels though nosespace aroma concentration is independent of their
hardness; the lower the gel hardness, the higher the aroma intensity is. Based on this
finding, it can be said that aroma perception is determined by texture rather than the
aroma release although biological mechanism is not still clear. It has been also
confirmed that perceived aroma intensity is more influenced by the release rate than
by the release amount of the aroma compound (Baek et al. 1999). Using gelatin gels
sweetened with sucrose and flavored with furfuryl acetate, no significant differences
are found in the maximum in-nose volatile concentration between different concen-
trations of gelatin gels. The amount of volatile present does not correlate with the
sensory analysis, while the rate of volatile release shows good correlation. As these,
release rate of aroma is associated with the degree of food breakdown in the mouth,
demonstrating the importance of dynamic (not static) analysis of aroma release for
high correlation with sensory data.
Regarding flavor perception, It has been indicated that inhomogeneous spatial
distribution of tastants like sweetness (Holm et al. 2009; Mosca et al. 2010, 2013)
and saltiness (Mosca et al. 2013; Noort et al. 2010, 2012) for solid foods increase
perceived taste intensity, and similar effect can be expected also for liquid foods.
Adaptation occurs when exposed continuously to taste stimulus for extended period
of time, leading to elevated threshold particularly at high doses, and in this context,
the effect of inhomogeneous spatial distribution can be attributed to prevention of
adaptation (Meiselman 1972). That is, discontinuous taste stimulus helps recovery
from adaptation and enhances taste perception during oral processing. In contrast,
there are few reports found on the similar effects of aroma inhomogeneous distri-
bution. Using polysaccharide gels as a feeding sample, effects of inhomogeneous
spatial distribution of aroma on its perceived intensity and also eating behavior have
been investigated (Nakao et al. 2013). The sample architecture used in their study is
illustrated in Fig. 13.10, where the degree of aroma inhomogeneity is variable by
changing aroma concentrations in both the matrix and the dispersed gels with
keeping the overall aroma concentration constant within one whole gel sample.
This situation presents inhomogeneous spatial distribution of aroma. In their study
(Nakao et al. 2013), there was no difference in the mechanical properties between the
13 Structuring for Elderly Foods 465

Fig. 13.10 Schematic drawings of structured gels used. SNO: homogeneous spatial of aroma, where
both the dispersed and the matrix gels contained 0.3% aroma compounds; SLOW: the lower degree
of inhomogenous spatial distribution of aroma, where all dispersed gels contained 0.6% aroma
compounds (0.3% in total); SHIGH: the higher degree of inhomogenous spatial distribution of aroma,
where 40% of the dispersed gels contained 1.5% aroma compounds (SHIGH Fr1) and remaining 60%
of the dispersed gels did not contain aroma compounds (SHIGH Fr2) (0.3% in total). Reproduction
with permission from (Nakao et al. 2013), Copyright 2013 Wiley Periodicals, Inc

gel samples tested, and also all gel samples were soft enough to be processed by
tongue-palate compression for size reduction without chewing. Under these exper-
imental conditions, perceived aroma intensity increased with increasing degree of
aroma inhomogeneity. Also, increase in the degree of aroma inhomogeneity
increased both duration of oral processing and activity of the suprahyoid muscula-
ture, a group of muscles active for jaw opening and also tongue movement
(Kohyama et al. 2010; Palmer et al. 1992; Shiozawa et al. 1999a, 1999b; Taniguchi
et al. 2008), decreased particle size of expectorated bolus before swallowing, and
increased saliva content in the bolus (Fig. 13.11). These results indicate the impor-
tance of food structure design for enhancement of perceived aroma intensity and
human eating behavior. In relation to the interaction between texture and aroma
during food oral processing, a hypothesis is that texture perception on consistency or
hardness and eating behavior can be changed by the degree of aroma inhomogeneity
(Funami et al. 2016). That is, the higher the aroma inhomogeneity, the less the
perceived hardness of foods can be even if a similar effort should be necessary for
consumption, and eating behavior can be less consistency-dependent as aroma
inhomogeneity is greater. In their study (Funami et al. 2016), the same sample
architecture as Nakao et al. (2013) was used but with three different consistency
levels of the gel samples. Their finding from surface EMG is that EMG variables;
duration and activity of the suprahyoid musculature generally increase with consis-
tency at each degree of aroma inhomogeneity, accompanied with decreased intensity
of aroma and taste perception. Another important finding is that as aroma inhomo-
geneity is greater, increases in the EMG variables are less consistency-dependent,
and the intensity of perceived hardness is lower for eating effort (Fig. 13.12). No
interaction is found in the EMG variables between consistency and aroma inhomo-
geneity. From these results, change of aroma inhomogeneity can be a strategy for
food product development which promotes oral processing without loading too
much for eating, particularly for elderly and dysphagia patients.
466 M. Nakauma and T. Funami

a b c
Perceived aroma intensity (VAS score)

120 2.5 2.5

* *

Suprahyoid musculature activity

* * *

Duration of oral processing

100 2 2

(standardized with N0)

(standardized with N0)
80
1.5 1.5
60
1 1
40

20 0.5 0.5

0 0 0
SNO SLOW SHIGH SNO SLOW SHIGH SNO SLOW SHIGH
d 3
e
5
* *
Mean particle size of bolus (mm2)

2.5 *
Saliva content in bolus (g)

2
3
1.5
2
1

0.5 1

0 0
SNO SLOW SHIGH SNO SLOW SHIGH

Fig. 13.11 Effect of the degree of aroma inhomogeneity on variables associated with food oral
processing. (a): Perceived aroma intensity; (b): electromyography duration of oral processing; (c):
electromyography suprahyoid musculature activity; (d): mean particle size of bolus; (e): saliva
content in bolus. Six subjects of 32.0 years old on average participated in tests. Data were presented
means  SD of the six subjects. Data with asterisk are significantly different ( p < 0.05). Repro-
duction with permission from (Nakao et al. 2013), Copyright 2013 Wiley Periodicals, Inc

8 Conclusion

In this chapter, essential elements for formulation design of elderly foods were
described along with introduction of recent product developments in Japan. Use of
food polysaccharides can be the most practical and reasonable way for texture
creation and optimization of elderly foods, but this should be evidenced scientifi-
cally. For texture design of elderly foods, both in vivo measurements and bolus
rheology should be novel approach by taking human physiological response during
eating and drinking into account. For the goal, food scientists/technologists are
urgently required to work more closely with medical doctors, dentists, dietitians,
and therapists to contribute to improved quality of life for the elderlies from dietary
aspect.
13 Structuring for Elderly Foods 467

Fig. 13.12 Relationship between sensory and EMG variables. Liner regression analysis was done
for the same degree of aroma inhomogeneity with various consistencies. (a) Electromyography
duration of oral processing as a function of VAS score of perceived hardness; (b) electromyography
activity of the suprahyoid musculature as a function of VAS score of perceived hardness. Closed
circle: homogeneous spatial distribution of aroma; open triangle: the lower degree of inhomoge-
neous spatial distribution of aroma; closed squares: the higher degree of inhomogeneous spatial
distribution of aroma. Reproduction with permission from (Funami et al. 2016), Copyright 2015
Elsevier Ltd

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Chapter 14
Bioactivities

Kang Liu, Xue-Ying Li, Jian-Ping Luo, and Xue-Qiang Zha

Abstract Polysaccharides and proteins are representative natural

biomacromolecules existing in animals, plants, and microorganisms. They are
attracting a great attention of scholars worldwide due to their various healthy
functions, such as immunomodulation, anti-tumor, anti-oxidative, hypoglycemic,
and hypolipidemic activities. Besides the strong bioactivity, these natural polysac-
charides and proteins are non-toxic and show no side effects. In recent decades, a
large number of bioactive polysaccharides and proteins with different structure and
bioactivity from natural resources have been extracted, purified, and characterized.
The aim of this chapter is to summarize the bioactivities, active mechanisms,
structure features, structure–activity relationships of natural polysaccharides, pro-
teins, and their derivatives. Moreover, this chapter also presented the applications of
some active natural biopolymers in foods and medicines.

Keywords Polysaccharide · Protein · Bioactivity · Structure–activity relationship

1 Introduction

The food hydrocolloid is an edible soft matter system, which determines the texture
and flavor characteristics of food products (Van der Sman and Van der Goot 2009).
In food processing, various food materials such as polysaccharides, proteins, lipids,
emulsifiers, sugars, minerals, and water are often mixed and fabricated. Among these
ingredients, polysaccharides and proteins are the most used materials, which not
only acting as “building blocks” for designing food hydrocolloids, but also provid-
ing interface-stabilizing properties via the interaction with other molecules (Wijaya
et al. 2017).

K. Liu · X.-Y. Li · J.-P. Luo · X.-Q. Zha (*)

School of Food and Biological Engineering, Hefei University of Technology, Hefei, China
e-mail: zhaxueqiang@hfut.edu.cn

© Springer Nature Singapore Pte Ltd. 2021 473

Y. Fang et al. (eds.), Food Hydrocolloids,
https://doi.org/10.1007/978-981-16-0320-4_14
474 K. Liu et al.

Polysaccharide is defined as carbohydrate polymers consisting of different mono-

saccharide linked by glycosidic bonds (Xie et al. 2016). Protein is a macromolecular
compound, which is formed by the binding of peptide chains composed of amino
acids. In recent decades, many natural polysaccharides and proteins have been
extracted and purified from plants, animals, and microorganisms. In addition to the
properties of food hydrocolloids, these natural macromolecules also possess many
bioactivities, such as anti-tumor, immunomodulation, anti-oxidative, hypoglycemic,
and hypolipidemic (Cho et al. 2015). Therefore, this chapter mainly introduces the
bioactivities of natural hydrocolloids, including anti-tumor, immunomodulation,
anti-oxidation, antimicrobial, hypoglycemic, and hypolipidemic effects. Moreover,
the applications of these natural biomacromolecules in functional foods and medi-
cines are presented in this chapter.

2 Bioactivities

In recent years, natural polysaccharides and proteins extracted from different mate-
rials have attracted increasing attention because of their wide bioactivities, such as
anti-oxidation, immunomodulation, anti-tumor, antimicrobial, hypoglycemic, and
hypolipidemic effects. Moreover, more and more evidence indicated that most of
these bioactivities of polysaccharides and proteins are related to the immune system.

2.1 Anti-Tumor

Cancer is a group of diseases involving abnormal cell growth with the potential to
invade or spread to other parts of the body. According to the report released by the
World Health Organization (WHO) 2018, cancer is one of the main causes of human
death worldwide. Although there are many different types of antineoplastic drugs in
clinic, these drugs not only have limited efficacy, but also have strong side effects.
Since Lentinan was first recognized to have anti-tumor efficacy (Chihara et al. 1969),
more and more studies on natural polysaccharides used in cancer treatment have
been carried out in vitro and in vivo.
Up to date, a series of human carcinoma cell lines have been employed to
investigate the anticancer activity of polysaccharides, such as the lung cancer cell
line (A549 cell), the cervical carcinoma cell line (Hela cell), the gastric carcinoma
cell line (BGC-823 cell), the breast carcinoma cell line (MCF-7 cell), the colon
cancer cell line (HCT116 cell and HT29 cell), and the liver cancer cell line (HepG2
cell). In addition, some mouse-derived cancer cell lines were also used to evaluate
the activity of polysaccharides. It has been suggested that the anti-tumor mechanisms
of polysaccharides were possibly attributed to their inhibition of tumor cell prolif-
eration, initiation of tumor cell apoptosis, and activation of immune system to kill
tumor cells (Zong et al. 2012).
14 Bioactivities 475

It has been reported that polysaccharides from Dendrobium (Yu et al. 2018),
Astragalus (Zhai et al. 2018), Lentinus edodes (Ya 2017), Ganoderma lucidum
(Mohan et al. 2017), and Portulaca Oleracea L. (Zhao et al. 2016) exhibited good
inhibitory effects on HeLa cells proliferation. These anti-proliferation effects might
be related to the increase in autophagic activity of HeLa cells via regulating the
expression of some key proteins in mitochondria-mediated signaling pathway, such
as beclin1, LC3, and p62 (Zhai et al. 2018). Polysaccharides extracted from
Houttuynia cordata (Han et al. 2018), Tremella (Shi et al. 2018), Sargassum
integerrimum (Liu et al. 2016), Pleurotus nebrodensis (Cui et al. 2016), Auricularia
polytricha (Yu et al. 2014) exhibited strong activity to resist the proliferation of
human A549 cells. Lin et al. (2018) reported that Hedyotis diffusa polysaccharides
could induce the apoptosis of A549 cells via regulating caspase-3-dependent mito-
chondrial pathway. Wu et al. (2017) found that polysaccharide from Glehnia
littoralis could inhibit A549 cell proliferation and migration via decreasing the
expression of PCNA, leading to cell cycle arrested in S and G2/M phase. Luo
et al. (2016) also found that coix polysaccharides had the function to inhibit the
migration and invasion of A549 cells via down-regulating the expression of
S100A4. S100A4, a member of the S100 family, is a sort of calcium binding protein
with EF double helix domain. The S100A4 expresses in kinds of tumor and stem
cells of human rather than normal somatocytes.
HepG2 is an immortalized cell line consisting of human liver carcinoma cells. It
has been reported that polysaccharides extracted from Phormidium versicolor
(Belhaj et al. 2018), Ganoderma lucidum (Yang et al. 2017), Lentinus edodes
(Zhao et al. 2017), Antrodia camphorata (Li et al. 2009a, b), Grifola frondosa
(Wang et al. 2013) showed strong ability to prevent the proliferation of HepG2 cells.
Li et al. (2013a) reported that polysaccharide from Phellinus linteus could induce
S-phase arrest in HepG2 cells via decreasing calreticulin expression and activating
the P27kip1-cyclin A/D1/E-CDK2 pathway. Shen et al. (2014) found that polysac-
charide from Ganoderma lucidum mycelia could induce HepG2 cells apoptosis via
regulating the expression of miRNAs. Some algae polysaccharides have been
proved to possess broad-spectrum antineoplastic effects. For instance, polysaccha-
rides from Sargassum plagiophyllum and Sargassum pallidum showed strong inhib-
itory effects on the proliferation of HepG2 cells, A549 cells, and MGC-803 cells
in vitro (Ye et al. 2008; Suresh et al. 2013).
β-glucans, a type of the most abundant polysaccharides in the cell wall of bacteria
and fungus, are glucose polymers linked by 1!3 linear β-glycosidic bond (Chan
et al. 2009). Over the last half-century, fungi-derived β-glucans have received great
attention because of the potential medical and edible value all over the world.
Lentinan is a representative β-glucan. It has been widely proved to have therapeutic
effect on many kinds of tumors. In clinical trials, compared to chemotherapy alone,
the addition of lentinan to standard chemotherapy could relieve the pain and prolong
survival in patients with gastric cancer (Oba et al. 2009), pancreatic cancer (Shimizu
et al. 1999), colorectal cancer (Hazama et al. 2009), liver cancer (Ina et al. 2016),
breast Cancer (Taguchi 1983). A large number of cell and animal experiments have
also proved these anti-tumor effects. For instance, lentinan has ability to inhibit
476 K. Liu et al.

proliferation and differentiation of cancer cells, such as human autologous tumor cell
line (K562 cell) (Tani et al. 1993), human gastric cancer cell line (BGC823 cell)
(Zhao et al. 2013), human pancreatic cancer cell line (BXPC-3 cell) (Qian et al.
2018), human cervical cancer cell line (Hela cell) (Qian et al. 2018), human breast
cancer cell line (MCF-7 cell) (Yi et al. 2018), non-small cell lung cancer (Wang et al.
2020), human bladder cancer cell line (T24) (Bao et al. 2015), liver cancer cell
line (H22 cell) (Yamamoto et al. 1989). Animal experiments demonstrated that
lentinan could inhibit colitis-associated cancer (CAC) development via regulating
TLR4/NF-kappaB signaling-mediated inflammatory responses in model mice (Liu
et al. 2018).
β-glucans have also been reported to kill cancer cells directly. The anticancer
mechanisms of these polysaccharides are mainly dependent on the ability of
enhancement of host immune system, increase in the antioxidant capacity of host,
up-regulation of phase I and phase II enzymes in metabolic transformation, and the
detoxification of mutagenic compounds (Vannucci et al. 2013). Masuda et al. (2013)
reported that both oral administration and intraperitoneal injection of β-glucans from
Grifola frondosa could inhibit tumor growth via regulating the systemic immune
response. Moreover, the possible mechanism was revealed that the Grifola frondosa
β-glucans can induce systemic tumor-antigen specific T cell response via dectin-1-
dependent activation of DCs, enhance the infiltration of the activated T cells into the
tumor, and decrease number of tumor-caused immunosuppressive cells such as
myeloid-derived suppressor cells and regulatory T cells, thus leading to the anti-
tumor activity. Yeast β-glucans, extracted from by-product yeast of beer production,
have been known to exhibit anti-tumor activities by potentiating host immunity
(Suphantharika et al. 2003).
In recent years, a large number of anticancer peptides have been identified from
plant-derived proteins. For instance, corn peptides can induce the apoptosis of
HepG2 cells by increasing caspase-3 expression (Díaz-Gómez et al. 2018). The
lunasin peptide from soybean has the ability to resist skin cancer (Hernandez-
Ledesma et al. 2009). The potato protein was found to suppress the proliferation
of mouse melanoma B16 cells (Sun et al. 2013). Kannan et al. have extracted a
pentapeptide (Glu-Gly-Arg-Pro-Arg) from rice bran and proved that it had the ability
against the proliferation of colon cancer cells (Kannan et al. 2010). It has been
reported that peptides derived from fish proteins have the inhibitory effect on MCF-7
cells in a dose-dependent manner (Hsu et al. 2011). Nongonierma and FitzGerald
(2016) also found that milk protein-derived peptides exhibited anti-proliferative
activity to tumor cells.

2.2 Immunoregulation

The immune system is a complex network of cells, tissues, and organs that work
together to protect the body from harmful substances and organisms and defend
against disease. However, when the immune system is disorder, autoimmune
14 Bioactivities 477

Table 14.1 Immunomodulatory effects of some polysaccharides on macrophages

Effects on immune
Source Mice type Cell type cells References
Ganoderma – RAW264.7 Phagocytosis ", NO ", Yu et al.
atrum TNF-α", IL-1β " (2013)
Dendrobium – RAW264.7 Phagocytosis ", NO " Xia et al.
officinale (2012)
Astragalus H22 tumor-bearing mice H22 IL-2", IL-12" Yang et al.
membranaceus TNF-α ", IL-10# (2013)
Mushroom Male BALB/c mice and – IL-13", IL-17", Wong
sclerotia athymic BALB/c nude IFN-γ" et al.
mice (2011)
Grifola – HepG-2 NO ", TNF-α", IL-1β Mao et al.
frondosa " (2015)
Inonotus – SGC-7901 TNF-α", Fan et al.
obliquus (2012a, b)
Porphyra A BALB/c murine RAW264.7 Phagocytosis ", Liu et al.
haitanensis DC Tregs TNF-α ", NO", (2017a, b)
IL-10", IL-6"
Dictyophora – RAW TNF-α", NO",(IL)-6" Liao et al.
indusiata 264.7 (2015)
Ganoderma – CD – Lai et al.
lucidum (2010)
Prunella – RAW NO", TNF-α", (IL)- Li et al.
vulgaris 264.7 6" (2015)
Longan pulp – Splenic Proliferation" Yi et al.
cells (2012)
Cordyceps – RAW TNF-α", (IL)-6", Wu et al.
sinensis 264.7 IL-10", IL-1α" (2014)
Panax – DC Proliferation", IL-12 Kim et al.
Ginseng ", TNF-α" (2010)
Laminaria – RAW264.7 NO", TNF-α", Fang et al.
japonica IL-1β", IL-6", IL-10 " (2015)

diseases, inflammatory diseases, and cancer will happen in the body. Many natural
polysaccharides have exhibited the ability to affect the immune system via modu-
lating the immune functions including ROS production, cytokine/chemokine pro-
duction, cell proliferation, and so on (Table 14.1). Therefore, polysaccharide is
considered as a potential immunomodulator with great development prospects
(Schepetkin and Quinn 2006). Figure 14.1 shows the possible signaling pathways
involved in macrophage activation by polysaccharides (Schepetkin and Quinn
2006). It is known that the anti-tumor activity of polysaccharide is partly related to
the enhancement of immune system.
The host defense mechanism consists of innate immunity and adaptive immunity,
where the innate immunity is the first line of defense mediated the initial protection
against infections. It is known that the innate immune system mainly contains
macrophages, monocytes, granulocytes, and humoral elements. Among these
478 K. Liu et al.

Fig. 14.1 Schematic model of potential signaling pathways involved in macrophage activation by
polysaccharides. Reproduction with permission from (Schepetkin and Quinn 2006), Copyright
2006 Elsevier

components, macrophages are reported to exhibit various biological functions, such

as chemotaxis, surveillance, phagocytosis, and destruction of targeted organisms,
indicating the macrophages activation might be a hopeful strategy to resist diseases
(José et al. 2007). It has been reported that Juniperus scopolorum polysaccharides
could increase macrophage cytotoxic activity against tumor cells and microorgan-
isms, activate phagocytic activity, and enhance the secretion of cytokines and
chemokines, such as tumor necrosis factor (TNF-α), interleukin (IL)-1, IL-6, IL-8,
IL-12, interferon gamma (IFN-γ), and IFN-2 (Chen et al. 2010a, b; Schepetkin et al.
2005). Polysaccharide from Lycium barbarum could activate macrophages via
regulating the transcription factors AP-1 and NF-κB to induce TNF-α production
and up-regulating the expression of MHC class II costimulatory molecules, resulting
in the enhancement of innate immunity (Chen et al. 2008). Those phenomenons
indicated that macrophage activation is required for the activation of innate immune
system (Plüddemann et al. 2011). For these events, the pattern recognition receptors
(PRRs) are required for these cells to recognize stimulators, triggering the activation
of signaling pathways and the synthesis of pro-inflammatory cytokines (Kumar et al.
2011). Toll-like receptors (TLRs), the important PRRs, are existed on plasma
membrane (Kawai and Akira 2010). It has been reported that macrophage activation
14 Bioactivities 479

induced by polysaccharides involves TLRs mediated recognition (Li et al. 2011).

Figueiredo et al. (2012) have evidenced that TLR2 and TLR4 were the receptors
involved in the recognition of fungal polysaccharides. Ferwerda et al. (2008)
reported that the saccharomyces cerevisiae cell wall polysaccharide (Zymosan)
has function to induce macrophages to release cytokines by the recognition of
TLR2, TLR4, and Dectin-1. Nuclear factor κB (NF-κB) and mitogen-activated
protein kinases (MAPK) are the key proteins in the downstream signaling pathway
of TLR, which play important role in the host defenses via regulating the expression
of multiple inflammatory and immune genes (DiDonato et al. 2012). With respect to
MAPKs, mammalian cells expressed three representative MAPK pathways,
containing C-Jun-N-terminal kinase (JNK), extracellular signal regulated kinase
(ERK1/2), and p38. In recent years, the immunostimulatory activity mechanisms
of polysaccharides have been widely studied. Results suggested that the regulation
of intracellular signaling pathways is essential for the activation of macrophages
(Diao et al. 2014; Zhang et al. 2014; Maeda et al. 2012). Extracellular polysaccharide
LBP32 from Bacillus sp. strain was reported to inhibit LPS-induced production of
pro-inflammatory cytokines via attenuating the phosphorylation of P38 and JNK, but
not ERK1/2 (Diao et al. 2014). Lycium barbarum polysaccharide (LBPF4-OL) was
found to have the ability to promote the secretion of TNF-α and IL-1β via inhibiting
JNK and ERK1/2 MAPK phosphorylation and increasing the phosphorylation of
p38-MAPK (Zhang et al. 2014). The sulfated polysaccharide SP1 from Caulerpa
lentillifera had the function to activate macrophages and enhance NO production via
regulating NF-κB and P38 MAPK signaling pathways (Maeda et al. 2012). These
results demonstrated that various polysaccharides can exert their biological activities
through regulating different signaling pathways. The inflammatory response has
been reported to be highly dependent on MAPK signaling pathways via activating
its downstream cytosolic proteins and nuclear transcriptional factors (Arthur and Ley
2013). NF-κB is a ubiquitous transcription factor, which plays a critical role in the
host defenses via regulating the expression of multiple inflammatory and immune
genes (DiDonato et al. 2012). In unstimulated cells, NF-κB locates in cytoplasm and
combines with inhibitory proteins to form an inactive trimer (p50-p65-IκB). When
cells are stimulated, IκBs will be phosphorylated by IκB kinase, leading to IκB
degradation and translocation of NF-κB to the nucleus for binding to its cognate
DNA in the regulation region of a variety of genes (He et al. 2013). It has been
reported that the ability of some polysaccharides to activate macrophages is depen-
dent on their level to the activation of NF-κB pathway (Zhang et al. 2011; Yu et al.
2013).
Lymphocyte is considered as a mediator of innate and adaptive immunity. Shriner
et al. (2010) reported that pneumococcal polysaccharide could stimulate the prolif-
eration of IL-7-driven B lymphocytes, regulate their cytokine production, and restore
impaired T cell by immune response. Among the specialized cell subsets of the
innate immune system, DCs are the critical sensors via expressing various pattern
recognition receptors (Steinman and Banchereau 2007). In particular, TLRs and
cytosolic sensors for DNA and RNA recognition expressed by DCs use endogenous
host elements carrying microbial components (such as the alarmin HMGB1),
480 K. Liu et al.

pathogen associated molecular patterns, and/or nucleic acids to stimulate intrinsic

apoptotic pathways to generate protective immune responses (Peng et al. 2005;
Poeck et al. 2008; Besch et al. 2009). During this process, polysaccharide was
found to regulate the immunity via inducing DC maturation. For example, Astrag-
alus polysaccharides could induce the differentiation of DCs to
CD11chighCD45RBlow DCs by shifting of Th2 to Th1, resulting in the enhancement
of T lymphocyte immune function in vitro (Liu et al. 2011a). Achyranthes bidentata
polysaccharide was reported to enhance DC maturation and function, supplying
extra IL-12 and MHC class II molecules to up-regulating antigen presentation,
activating CD4+ T cell, and thus leading to an enhancement of DC-CD4+ T cell
(Zou et al. 2011). Meng et al. (2011) reported that polysaccharides from Ganoderma
lucidum could promote effective activation of murine DCs in the immune response
via up-regulating the expression of CD86, CD40, and MHC II and down-regulation
of acid phosphatases.
In the past decades, the structure–activity relationships of immunomodulatory
polysaccharides have been studied, indicating the interaction of immunostimulatory
polysaccharides with cell receptors may trigger signaling pathways and thereby
result in the induction of gene transcription (Ferreira et al. 2015a, b). A Houttuynia
cordata pectic polysaccharide (HCP-2) with a linear chain of 1,4-linked α-D-
galacturonic acid residues has been reported to increase the secretion of MIP-1α,
MIP-1β, TNF-α, IL-1β, and RANTES in human peripheral blood mononuclear cells
via regulating TLR-4 mediated signaling (Cheng et al. 2014). Bose et al. (2014)
reported that 1,3-linked β-D-glucans could activate innate immune functions via
regulating Dectin-1 and CR3 mediated signaling pathways. SR has been shown to
be the pattern recognition receptor of fucoidan. Guo et al. (2009) found that a
1,3-linked glucan from spores could be recognized by dectin-1 on macrophages
and thereby possess the biological activities. These results suggested that what is
polysaccharide’s pattern recognition receptor might be determined by the structure
of polysaccharide. Lo et al. (2007) suggested that galactose, mannose, xylose, and
arabinose played an important role in the stimulation of macrophages, but not
glucose. The residues of 1,4-lined β-D-Rhap and 1,5-lined α-L-Araf were reported
to be important for lymphocytes activation (Yang et al. 2012). The 1,4-linked
mannose and glucose was reported to be the important elements for macrophages
activation by a purified Laminaria japonica polysaccharide LJP-31 (Fang et al.
2015).
In recent years, some immunomodulatory peptides have been prepared from food
proteins (Agyei and Danquah 2012). Otani et al. (2003) reported that
phosphopeptides from casein could stimulate gastrointestinal tracts of mice to
release immunoglobulin A. Pan et al. (2013a) revealed that peptide from milk
protein exhibits immunomodulatory property in ICR mice. After modification with
dicarbonyl methylglyoxyl, ovalbumin has the ability to stimulate immune cells to
release tumor necrosis factor (TNF) alpha (Fan et al. 2003). The immunogenic
ovalbumin peptides have been employed to enhance the immune response of
different cancer patients (Vidovic et al. 2002; Goldberg et al. 2003). Some fish
protein-derived immune peptides have also been identified in recent years (Yang
14 Bioactivities 481

et al. 2009; Hou et al. 2016). Sheu et al. (2004) have separated an immunomodula-
tory protein from the Jew’s Ear mushroom Auricularia polytricha. Some peptides
extracted from macroalgae were also reported to exhibit immunomodulatory activity
via regulating the nuclear factor kappa B (NF-κB) pathway (Ahn et al. 2011).

2.3 Anti-Oxidation

Oxidative damage of biomolecules triggers not only physiological process of aging,

but also causes various physiological functional disorders, leading to serious health
problem ultimately (Harman 1993). In theory, antioxidants might have a positive
effect on our health because they have ability to clear free radicals from human body.
As is well known, free radicals can attack macromolecules such as proteins, mem-
brane lipids, and DNA, leading to many health problems (i.e., cancer, neurodegen-
erative diseases, and diabetes mellitus) via damaging cells and tissues (Lim et al.
2014). Reactive nitrogen species (RNS) and reactive oxygen species (ROS) are free
radicals that are formed during the normal metabolism of cells, which can be
removed by cellular anti-oxidative defense systems, such as glutathione peroxidase
(GSH-Px) and superoxide dismutase (SOD). Under normal physiological condi-
tions, the generation and elimination of RNS and ROS are balanced. Once this
balance is broken, either by the overproduction of ROS and RNS, or by the damage
in anti-oxidative system, oxidative stress will occur (Klaus et al. 2011; Sun et al.
2010). In food industries, some synthetic commercial antioxidants such as
tertbutylhydroquinone (TBHQ), butylated hydroxytoluene (BHT), butylated
hydroxyanisole (BHA), and propyl gallate (PG) have been extensively used to
reduce the oxidation and peroxidation damage. However, these antioxidants have
potential hazards to human health (Nagaoka et al. 2010). Therefore, screening
antioxidants from natural resources is always a hot topic (Peña-Ramos and Xiong
2002). Natural polysaccharides have attracted extensive attention and are proposed
to be the potential resource of novel antioxidants due to their low toxicity and
excellent anti-oxidation. Algal polysaccharides have been demonstrated as a scav-
enger of free radicals for the prevention of oxidative damage in vivo (Cristina Diaz
et al. 2017).
In general, the polysaccharide eliminates free radicals through four aspect,
including: (1) Hydrogen atoms on the structure of polysaccharides react with free
radicals to form water, and the single electrons generated by the reaction can be
further reduced. (2) Polysaccharides capture free radicals produced in lipid reactions
or chelate with metal ions, which are important factors for the formation of free
radicals. (3) Polysaccharides enhance the activity of some antioxidant enzymes.
(4) Polysaccharides indirectly achieve antioxidant effect by regulating immunity.
As shown in Table 14.2, β-glucan extracted from mushrooms and yeast have been
reported to be the potential antioxidants. Three polysaccharides isolated from
Ganoderma lucidum (GLP-H, GLP-V, and GLP-F) were found to possess the
stronger radical scavenging activities (Fan et al. 2012a, b). Astragalus
482 K. Liu et al.

Table 14.2 Antioxidants activity of β-glucan

Source Antioxidant activity References
Jinqian ABT radical scavenging activity was 63.96% at 5 mg/mL Liu et al.
mushroom DPPH scavenging ratio was 89.84% at 5 mg/mL (2014a, b)
Iron chelating effect was 14.06% at 5 mg/mL
Hydroxyl radical scavenging activity was 24.30% at 5 mg/
mL
Polyporus Hydroxyl radicals inhibition was 96% at 267 μg/mL Dore et al.
dermoporus Lipid peroxidation inhibition was 42.9% at 67 μg/mL (2014)
Superoxide inhibition was 83.3% at 67 μg/mL
Saccharomyces Decreasing the formation of RBARS in LPS stimulated Saluk et al.
cerevisiae human blood platelets (2013)
Decreasing the formation of O2 in LPS stimulated human
blood platelets
Geastrum Inhibition of the formation of hydroxyl radicals in a dose- Guerra Dore
saccatum dependent manner et al. (2007)
mushroom
Pleurotus The antioxidant enzymes activity, ferric reducing activity, Pietrzycka
ostreatus and ascorbate concentration in human red blood cells et al. (2006)
hemolysates were markedly increased
Lentinus edodes Inhibition of lipid peroxidation, as well as a strong Feng et al.
hydroxyl radical scavenging activity and superoxide radi- (2010)
cal scavenging activity
Yeast The level of glutathione was replenished and Sener et al.
myeloperoxidase activity was suppressed in a rat model of (2005)
sepsis
Reproduction with permission from (Nie et al. 2018), Copyright 2018 Elsevier

polysaccharides were reported to inhibit the generation of ROS via suppressing the
NF-κB signal pathway (Xue et al. 2015). When the mice were orally administrated
with extracellular polysaccharides of Morchella esculenta, the activity of SOD and
GSH-Px were elevated in the blood, heart, liver, spleen, and kidney of mice (Meng
2010).
It has been reported that some compounds such as proteins, peptides, pigments,
polyphenols, and flavones could bind to polysaccharides. Compared to the poly-
saccharides, these complexes have stronger antioxidant activity. For instance, the
antioxidant effects of tea polysaccharides (TPS)-protein conjugates are dose-
dependent on the protein content (Nie et al. 2008). In addition, the in vivo and
in vitro antioxidant activities of crude tea polysaccharides were found to be better
than that of tea polysaccharide fraction, which can be interpreted by the relatively
higher proportion of tea pigments, vitamins, tea polyphenols, and other antioxidant
components in the cruder fractions (Zhou et al. 2007). Moreover, Zhang et al.
(2016a, b, c) suggested that the antioxidant activity of polysaccharides from
Ganoderma atrum (PSG) was depended on the content of phenolic compounds/
proteins.
It is a fact that the antioxidant activity of polysaccharides is closely correlated
with their structural parameters, such as solubility, degree of substitution, degree of
14 Bioactivities 483

branching, molecular weight, monosaccharide composition, solution conformation,

and functional groups. Jing et al. (2009) reported that phosphorylated modification
could enhance the ability of fucoidan to scavenge hydroxyl and superoxide radicals.
The possible mechanism might be that the phosphate is a polyelectrolyte group and
could activate the hydrogen atom of the anomeric carbon.
The acetylated modification was reported to enhance the ability of mushroom
Inonotus obliquus polysaccharides to inhibit lipid peroxidation via affecting the
conformation of polysaccharides (Ma et al. 2012). The carboxymethylated modifi-
cation can increase the water solubility and antioxidant activity of β-(1,3)-glucan
from the sclerotium of Poria coco via changing the flexibility of polysaccharides
chain (Wang and Zhang 2006). Molecular weight was an important factor that can
influence the antioxidant ability of polysaccharides. Yan et al. (2009) used an acidic
solution to hydrolyze the exopolysaccharide from Cordyceps sinensis, giving a
degraded exopolysaccharide. Results showed that the degraded products had much
higher antioxidant activity than that of original polysaccharide.

2.4 Antimicrobial

β-glucans are ubiquitously found in both bacterial and fungal cell walls and have
been implicated in the initiation of antimicrobial immune response. It has been
proved that the (1!3)-β-D glucan with (1!6)-β-D branches could act as antimi-
crobial agent in vivo (Ferreira et al. 2015a, b). Hetland et al. (2000) obtained a
soluble branched β-(1,3)-glucan (SSG) from the culture broth of the fungus
Sclerotinia sclerotiorum IFO9395. Results exhibited that the oral administration of
SSG could help the mice to resist the infection of Streptococcus pneumonia sero type
4 and 6B. Faccin et al. (2007) reported that β-glucan from the fruiting body of
Agaricus brasiliensis mushrooms exhibited an antiviral activity against poliovirus
typ1 in HEp-2 cells. Peter et al. found that oral administration of β-glucan (glucan
phosphate, scleroglucan, and laminarin) at a dose of 1 mg/kg/day could enhance the
survival of mice infected with S. aureus or Candida albicans. In Sharma’s study
(Sharma et al. 2015), it was found that both extracellular polysaccharide (EPS) and
intracellular polysaccharides (IPS) extracted from Cordyceps species exhibited
obvious antimicrobial activities against all pathogenic microorganism tested. More-
over, IPS showed stronger antimicrobial activity than that of EPS. The polysaccha-
rides extracted from Ganoderma species have also been proved to exhibit spectral
antimicrobial activity (Table 14.3). It has been found that the acidic polysaccharide
CS-F2 from green tea had a selective anti-adhesive activity against some pathogenic
bacteria and strongly inhibited the growth of gastric and skin pathogenic bacteria.
More and more evidences exhibited that some proteins and their derived peptides
from plants, mammals, insects, and bacteria also have antimicrobial activity against
eukaryotes, fungi, bacteria, and viruses (Zhu et al. 2019). The first antimicrobial
peptide was found from the moth Hyalophora cecropia in 1981 (Steiner et al. 1981).
A lectin-like peptide from red lentil seeds exhibited antimicrobial activity against
484 K. Liu et al.

Table 14.3 The antimicrobial activity of polysaccharides

Type
of
Specie Microorganisms used assay References
Ganoderma Bacillus subtilis, Bacillus cereus, Erwinia In Bai et al.
lucidum carotovora, Escherichia coli, Penicillium digitatum, vitro (2008)
Botrytis cinerea
Ganoderma Acrobacter aerogenes, Acitenobacter aerogenes, In Bhattacharyya
applanatum Arthrobacter citreus, Bacillus brevis, B. subtilis, vitro et al. (2006)
Corynebacterium insidiosum, Clostridium
pasteurianum, Escherichia coli, Micrococcus
roseus, Mycobacterium phlei, Proteus vulgaris,
Sarcina lutea St
Ganoderma Listeria monocytogenes In vivo Wang et al.
formosanum (2011)
Glucans Staphylococcus aureus or Candida albicans In Rice et al.
vitro (2005)
Schizophyllum Salmonella enterica serovar In vivo Chen et al.
(2008)
Oat Herpes simplex virus 1 In vivo Murphy et al.
(2009)

Mycosphaerella arachidicola (Wang and Ng 2007). The defensin PDC1 peptide, a

fermentation product of corn using Pichia pastoris and Escherichia coli as the mixed
strain, has an ability to inhibit the growth of Fusarium graminearum (Kant et al.
2009). Using Bacillus subtilis sck-2 to ferment soybean paste, a peptide was
obtained and exhibited antibacterial activity against Bacillus cereus (Yeo et al.
2012). These antimicrobial peptides can be considered as the good candidates for
food antisepsis.

2.5 Hypoglycemic

Diabetes mellitus (DM) is a group of metabolic disorders characterized by high

blood sugar levels over a prolonged period. Symptoms of high blood sugar mainly
include frequent urination, increased thirst, and increased hunger. If not treated in
time, diabetes will cause many complications. Acute complications include
hyperosmolar hyperglycemic state, diabetic ketoacidosis, or death. Serious long-
term complications include cardiovascular disease, chronic kidney disease, foot
ulcers, stroke, and damage to the eyes. According to the International Diabetes
Federation (IDF) in 2017, an estimated 425 million individuals are living with
DM. At present, the main treatment of diabetes is oral hypoglycemic drugs and
insulin injection. However, long-term use of these drugs could lead to insulin
resistance and other side effects. Thus, it is quite necessary to develop helpful,
innoxious, and inexpensive drugs for DM patients. As the non-toxic biological
14 Bioactivities 485

Table 14.4 The anti-diabetic effects of polysaccharides

Source Model Effects and mechanisms References
Opuntia STZ mice Protecting the liver from peroxidation damage, Zhao et al.
dillenii maintaining tissue function, improving the sen- (2011)
sitivity and response of target cells in diabetic
mice to insulin
Tremella KK-Ay mice Reducing levels of insulin, total cholesterol and Kiho et al.
aurantia triglyceride in the mice blood, decreasing the (2001)
level of plasma lipoperoxide
Phellinus NOD mice Inhibiting the development of autoimmune dia- Kim et al.
linteus betes by regulating cytokine expression (2010)
Morus alba T2DM mice Repairing of damaged pancreatic tissues of Jiao et al.
fruit diabetic rats. (2017)
Lycium T2DM mice Increasing insulinogenic index Cai et al.
barbarum (2011)
Pleurotus STZ mice Reducing the risk of oxidative damage by Zhang et al.
ostreatus increasing catalase (CAT), glutathione peroxi- (2016a, b, c)
dase (GSH-Px) and superoxide dismutase
(SOD) activities and decreasing malonaldehyde
(MDA) level
Portulaca Alloxan- Controlling blood glucose, and modulating the Li et al.
oleracea L. induced dia- metabolism of glucose and blood lipid in diabe- (2009a, b)
betic mice tes mellitus mice
Corn silk T2DM Regulating the levels of serum lipid profile, Pan et al.
decreasing the levels of glycated serum protein, (2017)
non-esterified fatty acid
Hedysarum Alloxan- Increasing insulin secretion, inhibiting lipid Hu et al.
polybotrys induced dia- peroxidation, promoting the sensitivity to insu- (2010)
betic mice lin, suppressing gluconeogenesis, and reducing
the biosynthesis fatty acid, cholesterol, and cell
cytokines related to insulin resistance
Taxus STZ mice Increasing the body weight of diabetic mice, and Zhang et al.
cuspidata reversing the decrease of SOD and the increase (2012)
of thiobarbituric acid reactive substances
(TBARS) in kidney and liver of diabetic mice
Ganoderma T2DM Down-regulation of the hepatic glucose regu- Xiao et al.
lucidum lated enzyme mRNA levels via AMPK activa- (2016)
tion, improvement of insulin resistance and
decrease of epididymal fat/BW ratio

macromolecules, natural polysaccharides were observed to have positive effects to

treat DM (Huie and Di 2004; Zhou et al. 2007). Table 14.4 presents the anti-diabetic
effects of different polysaccharides.
A polysaccharide (CSP-1) from Cordyceps sinensis consists of glucose, mannose,
and galactose in a molar ratio of 1:0.6:0.75, and exhibited a significant drop in blood
glucose level in both normal and streptozotocin (STZ)-diabetic animals. These
results could be related to the increase in blood insulin level via the release of insulin
from the residual pancreatic cells and/or CSP-1-induced the reduction of insulin
486 K. Liu et al.

metabolism in body (Li et al. 2006a, b). Another two purified polysaccharides
(CS-F30 and CS-F10) from Cordyceps sinensis mycelia was also proved to have
hypoglycemic effects on mice (Kiho et al. 1996, 1999). Compared to CSP-1, these
two polysaccharides have different monosaccharide composition. In recent years,
the hypoglycemic effect of tea polysaccharides (TPS) has also attracted much
attention. Chen et al. (2010a, b) reported that the oral administration of TPS at
150 mg/kg/day can significantly reduce the blood glucose level in non-obese
diabetic (NOD) mice. From TPS, two water-soluble polysaccharide fractions of
TFP-1 and TFP-2 were obtained. The average molecular weight was determined to
be 15.9  104 and 1.12  104 Da, respectively. Results showed that continuous
administration of TFP-2 could dose-dependently decrease the blood glucose level in
alloxan-induced diabetic mice by inhibiting α-amylase and α-glucosidase (Han et al.
2011a, b). Zhou et al. (2007) isolated a crude tea polysaccharides (CTP) and a tea
polysaccharide fraction (TPF) from green tea. It was found that CTP and TPF have
hypoglycemic effects on alloxan-induced diabetic mice. The hypoglycemic action of
polysaccharides from oolong tea and black tea has also been investigated. Results
exhibited that these tea polysaccharides could alleviate the diabetic mice and
improve diabetes symptoms, indicating tea polysaccharides have good effects on
the prevention of hyperglycemia (Nie et al. 2011).
Food-derived peptides have been reported to have antagonistic effects against
glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-1
(GLP-1) in type 2 diabetes via inhibiting the activity of Dipeptidyl peptidase IV
(DPP-IV) (Matteucci and Giampietro 2009). So far, this active peptide has been
isolated from corn, milk, soybean, rice, and other foods (Zhu et al. 2019; Mochida
et al. 2010; Hira et al. 2009; Sanjukta and Rai 2016). The dipeptides and tripeptides
isolated from whey proteins have been used as competitive inhibitors to interact with
DPP-IV substrates (Gunnarsson et al. 2006). Recently, more attention have been
paid on the application of marine bioactive peptides to treat type 2 diabetes. The
peptides from Porphyra dioica were observed to have the inhibitory effect on
DPP-IV (Stack et al. 2017). Three polypeptides (ILAP, LLAP, and MAGVDHI)
with similar bioactivity were also isolated from macroalga Palmaria palmata
(Harnedy et al. 2015).

2.6 Hypolipidemic

Hyperlipidemia is a lipid metabolic disorder characterized by the enhancement of

blood total cholesterol (TC), low-density lipoprotein (LDL), very low-density lipo-
protein-cholesterol (VLDL-C) and triglyceride (TG), with a concomitant decrease in
the level of high-density lipoprotein-cholesterol (HDL-C) in the plasma. With the
improvement of people’s living standard and the change of dietary structure, the
incidence of hyperlipidemia is increasing year by year (Chan et al. 2009). Hyper-
lipidemia is one of the main risk factors for inducting cardiovascular diseases, such
as hypertension, atherosclerosis, and coronary heart disease (Rosenson et al. 2002).
14 Bioactivities 487

In a meanwhile, lipid accumulation in liver (steatosis) can result in oxidative stress

and inflammation, leading to the damage of liver (Esposito et al. 2002). Thus, how to
reduce lipid level is one of most important clinical problems. At the present time,
statins, nicotinic acid, and its derivatives are the most common lipid-lowering
prescribed drugs. Although these drugs help in lowing lipids, their side effects are
also enormous, such as headache, muscle pain, and nausea. Moreover, the long-term
use of these drugs can increase the risk of type 2 diabetes (Hsu et al. 2015). In recent
decades, more and more evidences exhibited that natural polysaccharides had the
function to low lipids. For instance, Enteromorpha prolifera polysaccharides
exhibited a high hypolipidemic action in high fat rats via decreasing the plasma
LDL-C, TC, and TG levels and increasing HDL-C level (Teng et al. 2013).
Ganoderma lucidum β-glucan was also reported to decrease the TC, TG, and
LDL-C levels in the serum of diabetic mice, whereas the HDL-C level was increased
(Li et al. 2011). Recently, significant in vivo studies testing, the efficacy of cereal
β-glucans using animal and human subjects has led to health claims for this material
in many industrialized countries. The soluble dietary fibers (i.e., β-glucan from
cereal grains) are well accepted to be the polysaccharides with the ability to lower
lipids (Pomeroy et al. 2001). Therefore, adding β-glucan-rich fiber products in the
daily diet is now considered as an effective approach to lower lipids. Extensive
efforts have been made to establish the relationship between the structure of cereal
β-glucans and reduction of LDL-C levels (Lazaridou and Biliaderis 2004; Li et al.
2006a, b). Cereal β-glucans are linear homopolysaccharides formed by the linkage of
β-D-glucopyranosyl units via (1!4)-β-linkage and separated by single
(1!3)-β-linkages. They are the main component of water-soluble dietary fibers
from cereals. The physiological activity of soluble β-glucan from cereal is closely
related to its unique structure of (1!3)(1!4)-β-D-glucan. It was concluded that the
LDL-C lowering effect and the ability to control blood glucose of cereal β-glucan
may depend on its viscosity in solution, which is controlled by the MW, structure,
and concentration in the intestine (Wood 2007).
It has been reported that β-glucan can bind to the bile acid in intestinal lumen.
This combination would reduce the circulation of bile acid liver and further stimulate
the production of more bile acids from cholesterol. Moreover, β-glucan could be
fermented in the large bowel by colonic bacteria, and producing the short-chain fatty
acids. The short-chain fatty acids could be absorbed by portal vein, and inhibit
hepatic cholesterol synthesis via regulating the activity of HMG-CoA reductase
(a rate-limiting enzyme required for cholesterol biosynthesis), or increasing catabo-
lism of LDL-cholesterol. Another phenomenon was observed that β-glucan reduced
the concentration of postprandial serum insulin by delaying gastric emptying,
leading to the inhibition of hepatic cholesterol production. It was also reported that
β-glucan could interfere with the absorption of dietary fat via increasing intestinal
viscosity (Bell et al. 1999).
488 K. Liu et al.

2.7 Other Activities

Besides the healthy benefits mentioned above, some other bioactivities of poly-
saccharides and protein-related compounds have been studied in recent years,
including antidiarrheal, anti-fatigue, anti-tussive, anti-analgesia, anti-allergic, and
anti-dyszoospermia activities. Baek et al. (2010) evaluated the anti-diarrhea effect of
ginseng polysaccharides in vitro using the model of rotavirus infection. Results
showed that two pectic ginseng polysaccharides dose-dependently rescued the cell
viability from rotavirus infection. Moreover, the prevention of polysaccharide on
rotavirus was attributed to the inhibitory effects of rotaviral attachment to cells. In
recent years, the anti-fatigue effect of polysaccharides extracted from different
materials is gradually accepted by people (Jing et al. 2009). Pimentel et al. (2019)
demonstrated that macroalgae-derived peptides and enzymes have protective effects
on skin via eliminating free radicals and promoting moisture.

3 Structure Features

Chemical structure is the basis of polysaccharides and to exert their biological

activity, including monosaccharide composition, monosaccharide arranging order,
anomeric carbon configuration, glycoside bond types, branches, substituted groups,
and the spatial conformation. It has been reported that most bioactive polysaccha-
rides are mainly composed of glucose, fucose, galactose, arabinose, mannose,
xylose, ribose, glucuronic acid, and galacturonic acid. According to the published
literatures, the fungal derived polysaccharides have been found to be α-mannan,
β-glucans and hetero-β-glucans, α-mannan-β-glucan complexes, heteroglycans, gly-
copeptides or glycoprotein and proteoglycan. The polysaccharides extracted from
plants mainly include glucans, glucomannans, heteroglycans, arabinans,
arabinogalactan, pectins, rhamnogalacturonan, and their sulfated or acetylated
forms. Animal polysaccharides are mainly composed of glycosaminoglycan and
sulfated glycosaminoglycan.
Although it is quite difficult to determine the structural variability of polysaccha-
rides from different resources, a series of analytical methods have been established to
achieve this. The high performance liquid chromatography (HPLC) is often
performed to determine the molecular weight of polysaccharides. The infrared
spectroscopy (IR), ultraviolet spectroscopy (UV), gas chromatography-mass spec-
trometry, NMR spectroscopy, periodate oxidation, partial acid hydrolysis, methyl-
ation, and periodate oxidation-Smith degradation are the common methods to
analyze structural features of polysaccharides (Yang et al. 2009).
The chemical structure of natural polysaccharides is very complex. Table 14.5
showed the structure features of some polysaccharides extracted from different
species. In most cases, one material could contain a variety of polysaccharides
with different structures. Moreover, the same species growing in different places
14 Bioactivities 489

Table 14.5 Structure features of polysaccharides from various sources. Reproduction with per-
mission from (Nie et al. 2018), Copyright 2018 Elsevier
Monosaccharide
Source Mw (Da) composition Backbone References
Pleurotus florida 180,000 Glucose (1!6)-linked-β-D-Glcp Maji et al.
(1!3,6)-linked-β-D-Glcp (2012)
Auricularia 120,000 Glucose 1,3-β-glucan Song and
polytricha 1,3-α-glucan Du (2012)
1,4-α-glucan
Cistanche 10,000 Glucose 1,4-linked-α-D-glucan Dong et al.
Deserticola Y. (2007)
Ma
Cordyceps sinensis – Glucose, Man- (1!4)-linked-α-D-Glcp Nie et al.
nose, Galactose (2011)
Ganoderma lucidum 83,000 Rhamnose, 1,4-linked-α-D-Glcp Bao et al.
Galactose, 1,6-linked-β-D-Glcp (2002a, b)
Glucose
Ganoderma lucidum 200,000 Glucose, 1,3-,1,4-, 1,6-linked-β-D- Bao et al.
Mannose Glcp (2002a, b)
1,6-linked-β-D-Manp
Ophiopogon 35,200 Arabinose, Glu- 1,4-linked-Glcp Chen et al.
japonicus cose, Galactose 1,6-linked-Glcp (2011)
1,4,6-linked-Glcp
Lentinus 196,000 Galactose, Glu- (1!4)-linked-α-D-Glcp Bhunia
squarrosulus (Mont.) cose, Fucose (1!6)-linked-β-D-Glcp et al.
Singer (1!4,6)-linked-D-Glcp (2010)
(1!3,6)-linked-D-Glcp
Dendrobium 73,000 Glucose, Galac- 1,4-linked-β-D-Glcp Pan et al.
huoshanense tose, Xylose 1,6-linked-β-D-Glcp (2013b)
1,4,6-linked-β-D-Glcp
Radix Astragali 1334,000 Rhamnose, Glu- 1,4-linked-α-Glcp Yin et al.
cose 1,4- (2012)
Arabinose, linked-α-GalAp6Me1,2,4-
Galactose, linked-Rhap
Galactose acid 1,3,6-linked-β-Galp

could contain different polysaccharides. It is also reported that the polysaccharides in

different organs of the same species is also different in the structures. It has been
observed that polysaccharides extracted from the spores of Ganoderma lucidum
possessed a backbone of (1–3)-β-linked glucans (Bao et al. 2001). However, the
backbone of fruit bodies polysaccharides was composed of 1,3-linked glucose,
1,6-linked glucose, 1,6-linked mannose, 1,4-linked glucose, and 1,6-linked galac-
tose (Bao et al. 2002a, b).
For the preparation of natural polysaccharides, the extraction methods and
extraction parameters are also critical factors affecting polysaccharide’s structures
and properties. For instance, Palacios et al. compared the effects of different extrac-
tion methods on the structure of polysaccharides from Pleurotus ostreatus mush-
room. Results showed that the polysaccharides extracted by cold-water mainly
490 K. Liu et al.

consisted of 1–3-α-linked and 1–6-α-linked galactose. The polysaccharide extracted

by hot-water was mainly composed of 1–4-α-linked glucose. However, the fraction
extracted by hot-aqueous NaOH was mainly composed of 1–3-β-linked and 1–-
6-β-linked glucose (Palacios et al. 2012).
In addition to the natural factors, chemical modification can also change the
polysaccharide’s structure, leading to the variation of physicochemical properties
and bioactivities (Fiorito et al. 2018). According to the published literatures, the
acetylation, carboxymethylation, sulfation, phosphorylation, alkylation, and
selenization have been employed to modify the structure of polysaccharides
(Prashanth and Tharanathan 2007). Moreover, some modified polysaccharides
have been developed into drug delivery systems (Shah et al. 2011).
In addition to the natural factors, polysaccharide derivatives also contribute to the
structural diversity, which can also be classified as a semisynthetic polysaccharide. It
has been reported that the effective chemical modification of this natural structure
could improve the bioactivities and some key parameters, including solubility,
bioavailability, and pharmacokinetics (Fiorito et al. 2018). Chemical modification
can control the final structure of polysaccharides, and thus determining the specific
biological functions. In addition, the chemical modification of polysaccharide struc-
ture mainly utilizes the polysaccharide’s reactive groups, such as hydroxyl, car-
boxyl, and amino groups, to chemically introduce new functional groups. Chemical
modification of polysaccharide includes sulfation, carboxymethylation, acetylation,
alkylation, phosphorylation, and selenization. Some semisynthetic polysaccharides
have been developed into various drug delivery systems.
In recent years, a large number of studies have been carried out on the structure–
activity relationship of polysaccharides. Results exhibited that the activity of poly-
saccharides was mainly related to the molecular weight, chemical structure, and
physical properties (Jin et al. 2012). The (1,3)-β-D-glucan from Poria cocos sclero-
tium is a water-insoluble polysaccharide and exhibited low bioactivity. After
carboxymethylation, both the water solubility and bioactivity were enhanced
(Wang et al. 2009). Di et al. (2017) reported that sulfated polysaccharides from
Gracilaria rubra exerted immunologic activity by promoting the proliferation of
RAW264.7 cells. Moreover, the activity was improved with the decrease of poly-
saccharide molecular weight. Chen et al. (2015) demonstrated that sulfated polysac-
charide from Ganoderma atrum has the strongest immunological activity when the
molecular weight was intermediate (4.0  106 Da). Zhang et al. (2005) found that
the physicochemical property and steric conformation of polysaccharides from
Poria cocos mycelia were changed by the introduction of sulfate groups, leading
to the changes in the bioactivity of polysaccharides. Similarly, sulfated modification
changed the structure and conformation of polysaccharides from Hypsizygus
marmoreus, enhancing the ability of anticancer and immunity (Bao et al. 2010).
The substitution degree of substituent groups also has a great influence on the
biological activity of polysaccharides. For instance, chitosan with different substi-
tution degree of sulfuric acid group exhibited different strength of immunoregulation
ability (Yang et al. 2018). Liu et al. (2017a, b) investigated the structure–activity
relationship of selenium-containing polysaccharide. Results showed that the
14 Bioactivities 491

anti-diabetic activity was improved with the increase of selenium content of

Catathelasma ventricosum polysaccharide with triple helical structure. However,
when the tri-helical structure was damaged, the anti-diabetic effect was decreased.
With respect to the structure–activity relationship of proteins, it has been reported
that the bioactivity of proteins was highly related to its structure characteristics, such
as amino acid composition, amino acid sequence, molecular weight, and hydropho-
bicity (Silva et al. 2017).

4 Application

Because natural polysaccharides have wide biological activities and low toxicity,
some of them have been successfully applied in fields of drugs and foods
(Table 14.6). In the field of drugs, some polysaccharides such as Lentinan, Astrag-
alus polysaccharide, Ginseng polysaccharide, Poria polysaccharide, and Chondroi-
tin have been developed into injections, tablets, and capsules. In the field of foods,
some polysaccharides have been used as the additives to endow new nutritional and
healthy functions of foods. For instance, Lycium barbarum polysaccharides (LBPs)
have been processed into different forms of functional foods, such as Goji beverage,
Goji wine, Goji tea, Goji oral liquid, Goji tablet, Goji capsule, and Goji granules.
Among these healthy foods, the Goji capsule and Goji oral liquid were popular in the
public. Long-term use of Goji capsule and Goji oral liquid can enhance the immu-
nity, improve sleep, protect liver, and reduce fatigue (Wu et al. 2018).
Proteins are the important components in foods. Since soybean protein isolate
was produced in large-scale in 1958, the application of food protein becomes more
and more pluralistic. In the field of traditional foods, the soybean protein, milk
protein, egg protein, meat protein, and nut protein have been applied in beverage
food, baby food, baked food, pastry, and meat products. In the field of
non-traditional foods, the protein has been used as a main component to prepare
reproduced foods and simulated foods, such as soybean protein beef, plant protein
chicken, vegetarian ham, vegetarian sausage, and imitation meat hamburger. More-
over, food protein widely applied in cosmetics and biomedicine.

5 Conclusions and Future Prospects

In recent decades, the research of bioactive polysaccharides has made some impor-
tant progress and has been widely used in pharmaceutical, biochemical cosmetic,
and functional food industries. However, due to the limitations of existing experi-
mental methods and the complexity of their structure, research about polysaccha-
rides is still far behind than that of proteins and nucleic acids. Although the
functional properties of polysaccharides have been widely studied, the mechanism
of their actions is still unknown. One of the main reasons is that the structural
492 K. Liu et al.

Table 14.6 Practical Applications of Polysaccharides

Applications Practical
areas Polysaccharides applications Bioactivities References
Clinical Astragalus Astragalus poly- Immunomodulation; Xu (2012)
drugs and polysaccharide saccharide Antioxidant; Zhang et al.
medicines injection Anti-hypertensive (2016a, b, c)
Ginseng Ginseng polysac- Anti-tumor Xu (2015)
polysaccharide charide injection immunity
Lentinan Lentinan injection; Immunomodulation; Wang
polysaccharide Lentinan capsules Anti-tumor (2012)
Wang et al.
(2013)
Poria Poria polysaccha- Anti-gastric cancer Yang et al.
polysaccharide ride oral solution (2017)
Hou and
Luo (2017)
Chondroitin sulfate Chondroitin sul- Anti-arthritis; Gacci et al.
fate tablets; chon- Anti- (2015)
droitin sulfate angiocardiopathy Liu et al.
capsules (2014a, b)
Fucoidan Active pharma- Hypolipidemic; Wu and
ceutical ingredi- Antivirus Yang (2010)
ent; Mandal
Antivirus drugs et al. (2007)
Ganoderma atrum Hypoglycemic Hypoglycemic Zhu et al.
polysaccharide drugs (2013)
Heparin Anticoagulant Anticoagulant Bai and
drugs Ahsan
(2009)
Dong and
Fang (2001)
Food Exopolysaccharides Fermented dairy Anti-ulcer; Yang et al.
industry products Immunomodulation; (2010)
Anti-tumor
Soybean soluble Yogurt; Milk Anti-hypertensive; Ron et al.
polysaccharide beverage Reduce weight; (2010)
Hypoglycemic Nakamura
et al. (2006)
Red ginseng Noodles, bread, or Immunomodulation; Yu et al.
polysaccharide cake making Anti-tumor (2012)
Carrageenan Desserts (ice Antivirus Prajapati
cream and et al. (2014)
puddings)
Cosmetics Hyaluronic acid Sodium Moisture absorption Zhang et al.
industry hyaluronate and moisture (2008)
injection retention
Aloe Aloe Vera gel Anti-aging; Takahashi
polysaccharides Versatile skin care et al. (2009)
Marine algae Marine algae deep Anti-radiation; Moore
polysaccharides moisturizing Whitening and (2002)
cream moisturizing
14 Bioactivities 493

diversity and heterogeneity of natural polysaccharides hamper the research and

product development. Moreover, the polysaccharides extracted from the same raw
material by different preparation methods often have different compositions. There-
fore, the preparation process of polysaccharides needs to be standardized. To reveal
the structure–activity relationship is always the focus of polysaccharides research,
which will disclose the structural basis for polysaccharides to perform their healthy
functions. This is not only necessary to screen and design high-active polysaccha-
rides, but also have important theoretical guiding significance to study the medicinal
mechanism of polysaccharides. In addition, because polysaccharides have a wide
range of biological activities, they will have broad application prospects in func-
tional foods in the future. Therefore, it is also necessary to strengthen the research on
the activity development and mechanism of polysaccharides.

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Chapter 15
Dietary Fibers: Structural Aspects
and Nutritional Implications

Bin Zhang, Shaokang Wang, Santad Wichienchot, Qiang Huang, and

Sushil Dhital

Abstract Along with the brief outline of the definition evolution of dietary fibers,
this chapter focuses on how the physical and chemical structures of fibers are related
to the health effects. The role of fiber in the human digestive tract including oral,
gastric, small intestinal, and large intestinal phases is also discussed. Our further
emphasis is on the colonic microbial fermentation performance and health implica-
tions of dietary fibers. Fibers with higher water holding capacity or viscosity are
desirable for slowing down the metabolic response (e.g., post prandial blood glucose

B. Zhang (*)
School of Food Science and Engineering, Guangdong Province Key Laboratory for Green
Processing of Natural Products and Product Safety, South China University of Technology,
Guangzhou, China
Department of Applied Biology and Chemical Technology, The Hong Kong Polytechnic
University, Hung Hom, Kowloon, Hong Kong, China
Overseas Expertise Introduction Center for Discipline Innovation of Food Nutrition and Human
Health (111 Center), Guangzhou, China
e-mail: zhangb@scut.edu.cn
S. Wang
School of Food Science and Engineering, Guangdong Province Key Laboratory for Green
Processing of Natural Products and Product Safety, South China University of Technology,
Guangzhou, China
S. Wichienchot
Center of Excellence in Functional Foods and Gastronomy, Faculty of Agro-Industry, Prince of
Songkla University, Hat Yai, Songkhla, Thailand
Q. Huang
School of Food Science and Engineering, Guangdong Province Key Laboratory for Green
Processing of Natural Products and Product Safety, South China University of Technology,
Guangzhou, China
Overseas Expertise Introduction Center for Discipline Innovation of Food Nutrition and Human
Health (111 Center), Guangzhou, China
S. Dhital
Department of Chemical Engineering, Monash University, Clayton Campus, Clayton, VIC,
Australia

© Springer Nature Singapore Pte Ltd. 2021 505

Y. Fang et al. (eds.), Food Hydrocolloids,
https://doi.org/10.1007/978-981-16-0320-4_15
506 B. Zhang et al.

level), whereas the fibers with slow fermentation throughout the colon and higher
level of butyrate/propionate production are considered ideal for colonic health. Thus
foods that have the balance of viscous and slowly fermentable fibers are considered
beneficial in terms of overall nutrition and health. Although role of fibers in small
intestine is well studied and known, how the fiber specificity can modulate the gut
microbial ecology for desired health benefits still needs further research
investigation.

Keywords Dietary fiber · Definition · Chemical structure · Fermentation

performance · Health implications

1 Definition, Structure, and Analysis of Dietary Fiber

1.1 What Is Dietary Fiber?

Definition of dietary fiber has a very long history. Several progressions are made on
defining the dietary fiber, yet no single consensus definition of dietary fiber is agreed.
Hipsley (1953) first introduced the term “dietary fiber (DF).” In 2001, the American
Association of Cereal Chemists International (now Cereals & Grains Association)
proposed a definition and is most widely accepted. DF is defined as “the edible parts
of plants or analogous carbohydrates that are resistant to digestion and absorption in
the human small intestine with complete or partial fermentation in the large intes-
tine.” Thus, the term included non-starch polysaccharides (e.g., cellulose, pectin,
gums), resistant oligosaccharides (e.g., fructooligosaccharides,
galactooligosaccharides), and other carbohydrates (e.g., resistant starch (RS), resis-
tant dextrin). Moreover, the non-carbohydrate-based polymers (e.g., lignin) and
some animal origin carbohydrates (e.g., chitin, hyaluronan, chondroitin sulfate) are
also considered in this definition. In 2006, the Japanese Association for Dietary Fiber
Research defined as “dietary components which are not digested and/or absorbed in
the human small intestine and which exert physiological effect that are useful in
maintaining good health via the gastrointestinal tract”(Kiriyama et al. 2006). In
2009, Codex Alimentarius Commission, an international body that sets guidelines
for national regulatory authorities, defined DF as carbohydrate polymers with at least
ten monomer units, which cannot be hydrolyzed by the enzymes in the small
intestine. Three categories were set as follows: (1) Edible carbohydrate polymers
existing in food as ingredients, (2) Carbohydrate polymers prepared from raw food
ingredients by different processing methods, including physical, enzymatic, and
chemical methods; beneficial metabolic effects were tested and approved. (3) Car-
bohydrate polymers which are synthesized but proven by scientific evidence with
healthy effects to the host (Alimentarius 2010). Most recently, US Food and Drug
Administration (FDA) published the official definition for the first time that defines
the DF as “non-digestible soluble and insoluble carbohydrates (with 3 or more
monomeric units) and lignin that are intrinsic and intact in plants; isolated or
15 Dietary Fibers: Structural Aspects and Nutritional Implications 507

synthetic non-digestible carbohydrates (with 3 or more monomeric units) deter-

mined by the FDA to have physiological effects that are beneficial to human health.”
In the recent rule, USFDA (2016) clearly mentioned that fibers that are naturally
present in the food and associated with food matrices (intrinsic) with no relevant
components removed or destroyed during processing (intact) do not need to be
approved to be quantified as dietary fiber. However, the externally added dietary
fiber needs to be approved by FDA before listing them as dietary fiber or making any
nutrient content or health claim. This definition is dramatically different from the
previous definition, in a sense, the definition specifies that the added dietary fiber
(except that are approved by FDA), though technically is fiber but do not necessarily
provide the physiological benefits so are excluded from the approved list of fibers.
The FDA has identified psyllium husk, beta-glucan soluble fiber, guar gum, locust
bean gum, cellulose, pectin, and hydroxypropyl methylcellulose as the
non-digestible carbohydrates that can be added into the food or food formulations
as dietary fiber. However, widely accepted fiber such as “inulin” is not in the list of
approved fiber, so foods that have additional inulin will not be able to make fiber
claims (Dhital et al. 2018).

1.2 Structure and Physicochemical Properties of Dietary

Fibers

Dietary fibers can be categorized in different ways such as structure and solubility.
According to solubility, dietary fibers are normally identified as either soluble or
insoluble DFs. Insoluble dietary fiber (IDF) mainly consists of cell wall components,
such as cellulose, lignin, hemicellulose. While soluble dietary fiber (SDF) mainly
contains non-cellulosic polysaccharides, such as beta-glucans, gums, pectin, pento-
sans, and mucilage (Chawla and Patil 2010). The molecular/chemical structure is the
key to understanding whether the dietary fiber is soluble in water. Chemical struc-
tures of common dietary fibers are given in Table 15.1, which shows that dietary
fibers are composed of linked monosaccharide units. Physical properties (e.g.,
solubility) of dietary fiber is determined more by the inter- and intra-chain linkages
than the nature of the monosaccharide units, which can be defined by comparing two
forms of the water-soluble β-glucan and insoluble poly-D-cellulose. Cellulose is a
homopolymer of β-1-4-linked glucose, whereas β-glucans have mixedβ-(1-3) and
β-(1-4) linkages. The regularity of cellulose can be realized by adopting crystalline
structures with hydrogen bonds and thus formed the ordered insoluble structure. On
the other hand, the irregular structure of the β-glucans prevents the formation of
ordered crystalline structures, so the water soluble fibers could be achieved. Besides,
branched structures are unable to adopt ordered crystalline structures, and they are
also soluble (arabinoxylans in cereals). However, the ester bond between ferulic acid
moieties “cross-links” the multiple chains making them insoluble. Thus based on the
degree of crosslinking, the arabinoxylan (AX) can be soluble (e.g., in endosperm cell
508 B. Zhang et al.

Table 15.1 Chemical structure of selected dietary fibers

Chemical
Dietary fiber structure Dietary fiber Chemical structure
Pectin Hemicelluloses
Homogalacturonan Linear α-(1,4) Arabinoxylan β-(1,4) Xyl units as the
GalA units par- backbone with side chains
tially methylated of Ara units via α-(1,2),
and acetylated α-(1,3), and α-(1,5). Gal,
GluA, and FerA units may
also be present in the
branched points
Linear α-(1,4) Xylan β-(1,4)-D-xylose
Rhamnogalacturonan- GalA and α-(1,2)
I Rha units
Linear α-(1,4) Xyloglucan β-(1,4) Glu units as the
Rhamnogalacturonan- GalA units backbone with single unit
II branched with side chains of α-(1,6) Xyl.
Api, Ara, AceA, Gal, and Fuc may also
Dha, Fuc, Gal, present in the branched
GluA, Kdo, Rha, point
Xyl
Pectic galactan β-(1,4) Gal, Glucomannan Linear or slightly branched
β-(1,6) backbone chain of β-(1,4)
Gal, α-(1,4) Ara Man and β-(1,4) Glu
unit
Arabinogalactan- β-(1,4)-D- Galactomannan Linear or slightly branched
type I galactose backbone chain of β-(1,4)
Man moreover, β-(1,4) Glu
with side chains of α-(1,6)
Gal
Arabinogalactan- β-(1,3)- and β-Glucan Repeating linear polymer
type II β-(1,6)-D- of two β-(1,4) Glu alter-
galactose nated with β-(1,3) Glu units
Arabinan α-(1,5), (1,3) β-(1,4)-D-mannose,
Moreover, α-(1,2) Galactoglucomannan β-(1,4)-D-glucose
Ara units
β-1,4-D-Galactan β-1,4-D- Mannan β-(1,4)-D-mannose
galactose
β-1,3-D-Galactan β-1,3-D- Arabinogalactan β-(1,6)-D-galactose.
galactose (proteoglycan) Linked to peptides
Cellulose Linear β-(1,4) Gums
Glu units
Polyfructans Carrageenan Sulfated galactans, units of
β-(1,3) Gal and α-(1,4)
linked 3,6 anhydro Gal
Inulin β-(2,1) Fru Alginate Linear β-(1,4) ManA and
α-(1,4) GluA
Levan β-(2,6) Fru Locust bean gum β-(1,4)-D-mannose
Chitin β-(1,4)-N-acetyl- Furcellaran D-galactose, D-galactose-
D-glucosamine sulfate, 3,6-anhydro-D-
(continued)
15 Dietary Fibers: Structural Aspects and Nutritional Implications 509

Table 15.1 (continued)

Chemical
Dietary fiber structure Dietary fiber Chemical structure
galactose, 3,6-anhydro-D-
galactose-2-sulfate
Chitosan β-(1,4)-D-glucos- Agar Heterogenous product:
amine and β- mainlyβ-D-galactose and
(1,4)-N-acetyl-D- 3,6-anhydro-α-Lgalactose,
glucosamine which alternate through 1,4
and 1,3 linkages
Lignin Polyphenols, Gum Arabic β-(1,3)-D-galactose
Syringyl alcohol,
Guaiacyl alcohol,
and p-coumaryl
alcohol
Resistant starch α-(1,4)-D-glu- Gum Guaran β-(1,4)-D-mannose
cose units with (guar)
few α-(1,6)-D-
glucose branches
Xanthan gum β-(1,4)-D-glucose
Abbreviations used: AceA acetic acid, Ara arabinose, Api apiose, Dha 3-deoxy-D-lyxo-2-
heptulosaric acid, FerA ferulic acid, Fru fructose, Fuc fucose, Gal galactose, GalA galacturonic
acid, Glu glucose, GluA glucuronic acid, Kdo 3-deoxy-D-manno-2-octulosonic acid, Man mannose,
Man A mannuronic acid, Rha rhamnose, Xyl xylose

walls of oat and barley) or insoluble (e.g., in wheat bran and wheat endosperm cell
walls) (Gartaula et al. 2018). Dietary fibers with charged groups (e.g., COO
or
SO3
) such as the pectin and carrageenan are also water soluble, due to the
electrostatic repulsion preventing the molecules from being closely packed together
in an ordered structure (Oakenfull et al. 2001).
Solubility is also closely related to other physicochemical properties of dietary
fibers, such as water holding capacity and viscosity. Generally, strongly hydrophilic
properties existed in the polysaccharide constituents (monosaccharides) of dietary
fibers. Water is conserved on the void spaces in the molecular structure or the
hydrophilic sites (Mudgil and Barak 2013). Gel-forming or viscosity ability is linked
with the capacity to absorb water and form a gelatinous mass. It is attributed to the
physical interactions between polysaccharide molecules in solution, and the mole-
cules become entangled. Generally, the viscosity of solution is mainly caused by
water-soluble fibers. Gels were obtained from soluble fibers, and the viscosity and
bulk of the contents of the gastrointestinal tract could increase. It was well reported
that gels have good viscoelastic properties and could respond both elastic behavior
and viscous behavior in human gastrointestinal tract. This viscoelastic behavior is
responsible for the delayed gastric emptying of foods rich in fibers. Soluble fibers
increase bulk (gelling) property, whereas insoluble fibers increase the mass (water
holding or absorption) of the luminal content. However, the classification of dietary
fibers as “soluble” or “insoluble” may not be sufficient to explain the functionality of
dietary fibers (Gidley and Yakubov 2019). For instance, a large amount of dietary
510 B. Zhang et al.

fiber in foods is either soluble (e.g., cereal endosperm flours), or highly hydrated but
insoluble (e.g., fruits and cereal brans).

2 Roles of Dietary Fiber in the Digestive Tract

2.1 The Oral and Gastric Processing of Dietary Fiber

Mouth and stomach play crucial roles in the mechanical and enzymatic breakdown
of food structure as well as in appetite regulation. The mechanical process in mouth
facilitates the structural breakdown of food, which further disintegrates the fiber into
more softer and homogenous boluses, which can be easily swallowed and pass
through the esophagus before reaching to stomach. Only a small amount of macro-
nutrients can be digested, including hydrolysis of starch (Brownlee 2014). However,
the bio-accessibility of nutrients could be improved through the plant cell walls’
mechanical breakdown during mastication (Ellis et al. 2004), and is possibly to
change the taste perception of these foods.
The end-product of oral processing, i.e., food bolus, enters the stomach and mixes
with gastric secretions. Dietary fibers have a significant impact on gastric processing
events, as the physical structure and material profiles of plant-based food bolus could
be determined, hence the dilution, transport along with mixing profile of the stomach
could be affected. For instance, the gastric emptying can be slowed down when
dietary fibers form gel in the acid conditions in the large intestine, and the timescale
of nutrient delivery along with perceived satiety can be prolonged. For example, oat
porridge has a higher satiety index as well as glycemic response compared to white
bread. This is solely due to the bulking property of oats in the stomach increasing the
gastric retention time. Set micellar casein as another example. Through acidification,
micellar casein in milk can be converted from a thin liquid to a solid, thus it can be
delivered from mouth to stomach. This process leads to the less rapidly digested
properties of the casein fraction of milk when compared to whey protein fraction
(without gelation under gastric condition) (Gidley 2013).

2.2 Small Intestine Digestion and Dietary Fiber

The small intestine is the primary site for digestion and absorption of nutrients such
as dietary carbohydrates, lipids, proteins as well as micronutrients. Although a minor
physical degradation on molecular size may occur in the small intestine, dietary
fibers remain intact in the small intestine but play an essential role in modulating
small intestinal behavior of plant-based food, such as passage rate and enzyme
digestion process. It was well suggested that the absorption and digestion as well
as bolus transport in human small intestine can be influenced as a function of
physicochemical profiles belong to soluble DF, including water holding capacity,
15 Dietary Fibers: Structural Aspects and Nutritional Implications 511

organic compound entrapment, and viscosity (Bach Knudsen 2001). Generally,

soluble DFs with high viscosity are linked with slow transit time through stomach
and small intestine. The increase in viscosity and bulky forming is usually associated
with the high molecular size and branching structures, as well as the ability to tied
endogenous compounds such as bile acids and enzymes (Mackie et al. 2016).
Absorption and digestion of dietary substrates could also be slowed down as a
function of the capacities of organic compound entrapment or viscosity of DF. For
example, it may affect the digestion by reducing the nutrient diffusion and bioavail-
ability through the small intestine as a result of the viscosity and diluting of bolus
compounds by adding non-digestible material (Innami et al. 2000).
Insoluble fiber does not dissolve in water but retains its structure. The influence of
insoluble DF on nutrients digestibility in the small intestine is summed up in two
hypotheses: (1) A higher absorption and digestion could occur when the nutrient
properties of the bolus at the start stage of the small intestine are changed due to the
prolonged retention time in the stomach. For example, insoluble DF in the bolus
starts the degradation partially in the stomach and increases the solubilization, which
speeds up the motion of food through the gut and increases the digestibility in the
small intestine (Wilfart et al. 2007). (2) The digestion process could be changed due
to the water and nutrients absorption ability of insoluble DF (Taghipoor et al. 2014).
For example, insoluble DFs were proven to improve the access of the enzymes to the
substrates by enhancing the propulsive contraction effects of the small intestine,
which has a positive impact on digestion and absorption (Choct et al. 2000).
Insoluble DFs also play a significant role by increasing the mass of luminal content
forming feces in the colon.

3 Gut Microbial Fermentation of Dietary Fibers

in the Colon

Because of the restriction of human-specific enzymes, only a few α-glucosidic

linkages (e.g., α-1,2, α-1,3, α-1,4, α-1,6 bonds) present in a subset of carbohydrates
can be degraded in the upper gastrointestinal tract. A complex microbial community,
defined as gut microbiota, inhabits in the human gut. Human gut microbiota consist
of at least 1014 microorganisms belonging to over 1000 species, of which, members
of Firmicutes (~60–65%), Bacteroidetes (~20–25%), Actinobacteria (~3%), and
Proteobacteria (~5–10%) are the most common bacteria at the phylum level. Gut
bacteria have more catabolic enzymes and advanced metabolic systems to ferment
DFs for an energy source. The fermentation rate and end products of the fermenta-
tion are essential in providing the metabolic, immunologic, and protective functions.
Gut microbiota, thus, can be perceived as a “digestive partner” or an “additional
digestive organ” that has offered connection with human host and that provide
healthy benefits.
512 B. Zhang et al.

3.1 Fermentation Rate

During fermentation, the number of metabolites (normally gases and SCFAs) usu-
ally decreased from the proximal colon to the distal colon. Thus, a slow fermentation
and metabolism of fiber are more preferable, and the requirements of energy of the
distal colon could be realized (Rose et al. 2010; Williams et al. 2011a). It was
generally accepted that the critical factors determining fermentation rate contain
physical form and chemical structure of dietary fibers (Gidley 2013). In vitro
fermentation models present a good way to investigate how gut microbiota use
specific fiber and measure the quantity of metabolites and microbiota changes after
fermentation, which has important implications for food nutrition. Here, we com-
pared the in vitro fermentation profiles using human fecal inocula for a wide range of
dietary fibers in Table.15.2. Chemical structure of dietary fibers includes monosac-
charide and linkage compositions, monosaccharide arrangements, molecular size,
complexity, etc., and shows a considerable effect on the fermentation rate, particu-
larly for soluble DFs. In general, soluble DFs are fermented faster and to a greater
extent than insoluble fibers, and over-consumption might positively linked with
some bad effects accompanied by fast fermentation rate, including eructation,
bloating and flatulence (Hamaker and Tuncil 2014). Fermentation rate is also closely
related to the physical form of fibers such as particle size (Day et al. 2012), and
solubility (Williams et al. 2011b).

3.2 Short-Chain Fatty Acids Production and Related Gut

Bacteria Growth

DFs are major substrates of microbial fermentation leading to SCFA production.

Generally, acetate, propionate, and butyrate are the main SCFAs produced, mainly in
the proximal colon at very high concentrations (70–140 mM) and but distal colon at
lower concentrations (20–70 mM) (Wong et al. 2006). The molar ratios of acetate,
propionate, and butyrate from fiber fermentation normally are 60:25:15, respectively
(Tan et al. 2014). SCFAs show excellent physiological activity, especially propionic
and butyric acids. Except providing energy for the colon epithelial cells, the stim-
ulation of the secretion of PYY and GLP-1 as well as the suppression of the primary
inflammation of endothelial cells is an important function of SCFA (Topping and
Lockett 2016). Acetate, a potential appetite-regulating agent, links to the blood–
brain barrier and could reduce appetite through the function of a central homeostatic
mechanism. Propionate is largely metabolized in the liver where it is possibly used
as a glucogenic substrate, thus the related cholesterol synthesis could also be
suppressed (Hassan Younes et al. 1995). Butyrate is the primary energy source for
colonocytes and could hinder the proliferation of cancer cells, and accelerate differ-
entiation of cancer cell apoptosis without affecting normal epithelial cell differenti-
ation and proliferation (Comalada et al. 2006).
15 Dietary Fibers: Structural Aspects and Nutritional Implications 513

Table 15.2 In vitro fermentation performance of various dietary fibers using human fecal inocula
Microbiota
Fermentation SCFA composition
Dietary fiber rate molar ratio changes References
Resistant starch type 2 Slow Acetate" Bifidobacterium ", (Plongbunjong
(native starch granules) Butyrate" Blautia #, Dorea#, et al. 2017;
Bacteroides# Yang et al.
2013)
Resistant starch type 3 Slow Acetate" Bifidobacterium " (Plongbunjong
(retrograded starches) Butyrate" et al. 2017;
Arcila and
Rose 2015;
Jonathan et al.
2012)
Resistant starch type 4 Slow Acetate" Bifidobacteria" (Bae et al.
(chemically modified Butyrate" 2013;
starches) Thompson
et al. 2011)
Arabinoxylan Fast Acetate" Bacteroides", (Yang et al.
Butyrate" Coprococcus", 2013; Chen
Faecalibacterium" et al. 2017;
Rumpagaporn
et al. 2015)
Xyloglucan Fast Propionate" Lachnospiraceae", (Tuncil et al.
Butyrate" Bacteroides" 2017)
Inulin Fast Acetate" Bacteroides#, (Yang et al.
Butyrate" Bifidobacterium", 2013)
Catenibacterium",
Collinsella",
Dorea#
Pectin Fast Acetate" Bacteroides#, (Yang et al.
Propionate# Bifidobacterium", 2013; Bang
Butyrate" Dorea#, et al. 2018;
Parabacteroides# Ferreira-
Lachnospira", Lazarte et al.
Clostridium", 2018; Jonathan
Sutterella" et al. 2012)
Guar gum Fast Acetate" Roseburia" (Yang et al.
Propionate# 2013; Jonathan
Butyrate" et al. 2012)
Glucomannan Slow Acetate" N/Aa (Jonathan et al.
Propionate" 2012)
Butyrate#
β-Glucan Fast Acetate" Coprobacillus", ,
(Jonathan et al.
Butyrate" Dorea#, 2012; Hughes
Lactobacillus", et al. 2008)
Enterococcus"
Cellulose Very limited Acetate" Bacteroides sp.", (Jonathan et al.
Butyrate" Ruminococcus 2012; Chassard
et al. 2010)
(continued)
514 B. Zhang et al.

Table 15.2 (continued)

Microbiota
Fermentation SCFA composition
Dietary fiber rate molar ratio changes References
sp.", Enterococcus
sp."
Fructooligosaccharides Fast Acetate" Lactobacilli", (Jonathan et al.
Butyrate" Prevotella" 2012; Chen
et al. 2017; Li
et al. 2015)
Galactooligosaccharides Fast Propionate" Bifidobacteria" (Li et al. 2015)
Butyrate"
a
N/A not available

Type and structure of cereal dietary fibers can regulate the yield and molar ratio of
metabolites SCFAs as well as certain bacteria promotion during the gut microbial
fermentation. For instance, it was well documented that β-glucans and grain-derived
xylans could promote the SCFAs concentration during fermentation, particularly
butyric acid (Bach Knudsen 2015), whereas the fermentation of arabinose is asso-
ciated with increased production of acetic acid (Hu et al. 2013). Compared with
non-starch polysaccharides, in vitro and in vivo fermentation of RS (especially RS
type III) usually results in a significant increase in butyrate production
(Plongbunjong et al. 2017; Tan et al. 2014).
In regard to bacterial types, propionic acid was found to be positively correlated
with Ruminococcus obeum, Bacteroides spp., Megasphaera elsdenii, Roseburia
inulinivorans, Phascolarctobacterium succinatutens, Dialister spp., Veillonella
spp., Coprococcus catus, Salmonella spp., etc. (Bindels et al. 2012; Louis et al.
2014). Butyrate can be produced by a series of bacteria, including Eubacterium
hallii, Coprococcus eutactus, Coprococcus catus, Faecalibacterium prausnitzii,
Eubacterium rectale, Anaerostipes spp., Roseburia spp., etc. (Duncan et al. 2002).
The intake of dietary fibers could improve the metabolic interactions among various
bacterial species through the cross-feeding process, which means the SCFAs pro-
duced from fermentation by one bacterial group could provide substrates for the
growth of other bacteria.
The metabolic consequences of fermenting dietary substrates by gut microbiota
may be influenced by single culture composition. Some bacteria may grow through
cross-feeding, whereas pH value and nutrient shifts may inhibit the growth of other
species in the microbial community.
The fermentation of dietary fibers results in the acidic environment in the
gastrointestinal tract, which could change the microbial communities composition.
For example, the acidic conditions are more preferable to the growth of butyrate-
producers such as Roseburia spp., while inhibit the proliferation of the acid sensitive
bacteria such as Bacteroides (Walker et al. 2005; Duncan et al. 2009). The ratio of
Firmicutes/Bacteroides could be used as an indicator which represents health status
for diabetes and obesity.
15 Dietary Fibers: Structural Aspects and Nutritional Implications 515

4 The Role of Dietary Fiber in Health and Disease

Adequate DF intake is often considered an essential component of healthy diets. The

positive influence of healthy diets arises not only from the low-calorie components
but also from the partially or fully fermentability of DFs and their microbiota
response in the colon. Nutritional and potential health benefits can be provided by
diets which contain a wide range of DFs. These include active competition against
pathogenic organisms, regular bowel movement, short-chain fatty acid production
which provides energy for colonocyte cells, avoidance of cancer-promoting metab-
olites as well as toxic from protein fermentation. Some particular DFs stimulate on
specific groups of bacteria such as Akkermansia muciniphila, bifidobacteria, and
lactobacilli with beneficial host effects such as strengthen gut barrier, anti-
inflammation, and immune function (Gidley 2013). In the absence of dietary fibers
or other luminal energy sources, gut microbiota will turn to large intestinal mucus as
an energy source before attacking the underlying mucosa caused by gut-leak
(Brownlee 2011).
The interplay between DF, microbiota, and health conditions is complex. Both
genetic background and individual profiles (e.g., lifestyle, diets) affect microbial
composition. Apart from this, products of microbial fermentation also play a major
role in the microbiota composition shift and the immunological barrier function and
final health effect of the host. The DF metabolism by microbiota is an example of the
symbiotic connection between the host and the microbiota preventing dysbiosis.
Microbiota rely on DFs as a carbon source for bacteria growth, and DFs involved in
regulating the microbiota composition during the fermentation process. The rela-
tionship offers an option for dietary regulation of the microbiota because metabolism
and microbial growth depend on substrate availability, such as the content, chemical/
physical structures of dietary fibers consumed by the host.
Due to the expanded development of molecular techniques, including
metagenomics and 16S rRNA sequencing, the numerous studies focus on the
relationship between diets and gut microbiota. It was reported that diet patterns
contribute a lot to host gut microbiota composition (almost 60%), and dietary fiber as
well as animal diets make up larger proportions. Several reports suggested the
bacterial diversity in our gut is decreasing due to the consumption of westernized
food, which is related to the increase of metabolic diseases as well as allergic and
autoimmune diseases (Bach 2002). The westernized diets are rich in animal protein
compared to complex carbohydrates. Even though the trend of adding fiber to
processed foods has increased, most food manufacturers prefer to add soluble fiber
compared to complex insoluble fiber (Redgwell and Fischer 2005). As explained in
previous sections, the soluble fibers are fermented quickly in the proximal colon.
This leads to a decrease in metabolic and physiological activity in the distal colon
opening the door for colon disorders (Leach 2007). In a recent report, Martínez et al.
(2015) characterized the fecal microbial structure, diversity profiles, and assembly
processes in United States residents and rural Papua New Guineans. The results
showed that Papua New Guineans harbor communities with vastly different
516 B. Zhang et al.

abundance properties, greater bacterial diversity, lower inter-individual difference,

and bacterial lineages undetectable in US residents. It is suggested that a lower intake
of plant-derived carbohydrates and dietary fiber along with the unhealthy lifestyle
are affecting “microbial diversity” in westernized societies. In the short term, the
increase in consumption of diets rich in dietary fiber as well as whole grains
substantially increases the diversity of fecal microbiota in rats (Sonnenburg et al.
2016) and humans (Tap et al. 2015). Thus, switching to a high and complex fiber diet
can help to replenish the lost microbial diversity in our gut, leading to improved
metabolic and immunological health. The following sub-sections describe the rela-
tionship between dietary fiber and some common chronic and metabolic diseases.

4.1 Constipation

Constipation is the passing of hard, dry stools that may be infrequent or difficult to
pass. Up to 30% of people experience constipation which can be chronic, sometimes
severe, and have significant and often debilitating influences on quality of life (Tack
et al. 2011). Constipation can be caused by many different factors. However, a poor
diet and an unhealthy lifestyle are two risk factors for constipation. Lack of fiber in
the diet makes the stool hard that is unable to pass (move) through the colon. The
accumulation of hard stool may lead to more serious illness and often require
immediate medical attention. However, constipation can be alleviated if the diet is
rich in soluble fiber (that softens stool), and insoluble fiber (that adds stool bulk).
Adequate fiber intake (at least 25 g/day) was recommended by the Academy of
Nutrition and Dietetics, as fibers from a wide range of plant foods could contribute to
laxation through a series of ways, such us promotion of stool frequency, increase of
fecal biomass, and reduction in intestinal transit time (Dahl and Stewart 2015).

4.2 Irritable Bowel Syndrome

A most common gastrointestinal disease, named irritable bowel syndrome (IBS),

generally occurred in people no more than 45 years old. A great number of people
(10–15%) are being affected by IBS, and between the ages of 20 and 30 is the age of
high incidence. Also, it was well recorded that IBS is twice as common in males as it
is in females (Lovell and Ford 2012). A person who has had unexplained discomfort
more than 3 times during the past 3 months might be diagnosed as IBS. Four
categories can be listed as follow: IBS with diarrhea, IBS with constipation, mixed
IBS alternating constipation and diarrhea, and unsubtyped with a milder degree of
abnormal stool consistency.
Food ingredients may cause IBS symptoms, and reasonable diet management can
alleviate related symptoms. It is widely believed that increased fiber consumption
may help to promote long-term alleviation of IBS symptoms because of fiber’s
15 Dietary Fibers: Structural Aspects and Nutritional Implications 517

multiple roles in promoting digestive health. However, the report about fibers’
function on IBS showed by the American College of Gastroenterology is still
inconsistent and is specific for each fiber type (Ford et al. 2014). The study showed
that bran (water-insoluble fiber) lowered the risk by only 10%, whereas psyllium
(water-soluble fiber) resulted in a significant 17% reduction in IBS. A systematic
summary and meta-analysis (1299 participants; 4.1–40 g fiber/day; 3–16 weeks)
suggested that soluble fiber plays a pivotal role in alleviating the symptoms of IBS
and reducing the risks of harm (Nagarajan et al. 2015). Highly fermentable and
short-chain soluble dietary fiber (oligosaccharides) could result in rapid gas produc-
tion that might cause abdominal bloating/distension, flatulence, and abdominal pain/
discomfort in patients with IBS. Conversely, moderately fermentable dietary fiber
(long-chain, intermediate viscous, and soluble) such as psyllium can lead to a low
gas production and the inhibition of the associated symptoms.

4.3 Colorectal Cancer

Colorectal cancer accounts for the fourth highest number of deaths of any cancer
worldwide. Except for the genetic factors, diet and lifestyle are still believed to be the
significant factors that can delay or prevent the occurrence of this condition. Ade-
quate fiber in the diet is associated with a lower risk of colorectal cancer.
A dose-response meta-analysis (16 prospective studies; 1,985,552 participants;
4.5–26 years of follow-up; 14,514 colorectal cancer cases) found a significantly
lower risk of colorectal cancer (10%) for each 10 g/day intake of total fiber and cereal
fiber and a 17% reduction for each three servings (90 g/day) of whole grain daily
with further reductions at higher intake (Aune et al. 2011). Several plausible
mechanisms have been proposed to explain the hypothesis, including the increase
in stool bulk, decreasing transit time, and diluting fecal carcinogens, thus reducing
the contact between carcinogens and the lining of the colorectum (Lipkin et al.
1999). Also, bacterial fermentation of fiber generates short-chain fatty acids produc-
tion, which might be linked with the protective effects against colorectal cancer.

4.4 Inflammatory Bowel Disease

Inflammatory bowel disease (IBD) is an umbrella term to describe disorders that

involve chronic inflammation of digestive tract, with Crohn’s disease and ulcerative
colitis being the most dominant prominent types. This disease is accompanied by a
damaged large intestinal epithelium along with a compromised barrier function.
Active inflammatory bowel disease are generally related to dysbiosis and dietary
intake of indigestible fibers, whereas a reduced risk of inflammatory bowel disease is
positively corrected with the high intake of plant-based dietary. However, it is still
unclear whether dietary fiber intake positively or negatively impacts disease
518 B. Zhang et al.

progression and symptomology in IBD patients, and there is evidence that the active
inflammation experienced in IBD is, at least in part, due to the low-fiber intake and
inappropriate immune response to the luminal gut microbiota.
Crohn’s disease patients always related to the high level of the phylum
Proteobacteria and a decreased level of the phyla Firmicutes and Bacteroidetes.
The Crohn’s and Colitis Foundation of America Partners Internet cohort dietary
survey (1619 participants in remission; dietary intake and disease activity index
survey; 6 months) reported that a higher fiber intake during remission is linked to
disease remission (Brotherton et al. 2016).
A meta-analysis of observational studies (two cohort studies, one nested case–
control study and five case–control studies) indicated that higher fiber intake signif-
icantly decreased Crohn’s disease risk (56%) and ulcerative colitis risk by 20% (Liu
et al. 2015). Several mechanisms support increased fiber intake to reduce the risk of
inflammatory bowel disease by (1) improving the health of colonic microflora,
which can regulate colonic immune response and maintain immune homeostasis;
(2) promoting direct anti-inflammatory effects through the production of butyrate by
fermenting fibers, which is known to improve colon endothelial condition; (3) reg-
ulating the protective response of aryl hydrocarbon receptors to the pathogenesis of
inflammatory bowel disease; and (4) reducing C-reactive protein levels associated
with increased risk of Crohn’s disease (Liu et al. 2015; Jiao et al. 2015).

4.5 Obesity and Diabetes

Nowadays, both developing and industrialized countries are troubled by the epi-
demic of obesity. A wide range of factors may influence obesity, but epidemiological
and clinical studies have proved the link of low-fiber consumption and increased risk
of metabolic conditions such as obesity. It has been shown that adequate fiber in the
daily diet could reduce excessive food intake and depot fat storage by increasing the
effort involved in eating, interfering slightly with the efficiency of energy absorp-
tion, promoting intestinal satiety, stowing rate of food ingestion, and decreasing the
caloric density of the diet (Van Itallie 1978). Further, a cycle is established; people
with lower fiber intake have reduced microbial diversity, which is associated with
reduced satiety and increase appetite. It is agreed that individuals with lower
microbial diversity have higher body lipid content, insulin resistance, and lower-
grade systemic inflammation than individuals with higher microbial diversity
(Makki et al. 2018). Overweight and obese people tend to eat more low-fiber refined
foods, which are more metabolic; the number of bacteria producing butyrate
decreases, in particular, F. prausnitzii; increases inflammation-promoting effects,
such as mucus degradation and endotoxin production such as lipopolysaccharides
(LPS) (Brahe et al. 2016; Miquel et al. 2013). On the contrary, a high fiber-rich foods
intake could lead to a more diverse microbiota bacterial genes functions, including
metabolism of cofactors and vitamins, cell motility, and increased abundance of
Bifidobacterium species and butyrate-producing bacteria such as F. prausnitzii, a
15 Dietary Fibers: Structural Aspects and Nutritional Implications 519

symbol of a healthy microbiota due to its anti-inflammatory activity (Brahe et al.

2016; Furet et al. 2010). SCFAs are the most probable fiber-related microbiota
metabolites linked to leanness and obesity. High fiber content could contribute to
protecting against obesity, as the SCFA’s ability to act as ligands of free fatty acids
receptors, which increases secretion and expression of satiety hormones glucagon-
like peptide 1 or peptide YY and leptin from adipocytes (Blaut 2015; Chambers et al.
2015).
Diabetes is the result of an increased level of circulating blood glucose associated
with insufficient insulin to utilize the glucose in cells. Therefore, a diet with high
glycemic index (high digestible starch and low fiber) is associated with an increased
risk of diabetes (Sluijs et al. 2012). Overweight and obesity are the initial risk factors
for diabetes, because low-grade inflammation increases to tissues involved in met-
abolic regulation, such as liver, adipose tissue, and muscle, interfering with cellular
insulin signaling, leading to insulin resistance. This low-grade inflammation and
insulin resistance are associated with an imbalance of colonic microflora.
Proteobacteria are significantly abundant in diabetic in comparison with healthy
persons and positively associated with plasma glucose (Larsen et al. 2010). Two
other studies also reported that diabetic subjects were characterized by a decrease of
the level of Clostridiales bacteria (Faecalibacterium prausnitzii and Roseburia
species), which generate butyrate (Moreno-Indias et al. 2014; Weitkunat et al.
2017). Hence, it is tempting to speculate that whereas the incidence rate of obesity
is resulted by reduced fiber intake, lack of butyrate-producing and fiber-degrading
bacteria may predispose to type 2 diabetes.

5 Conclusions and Future Direction

There is a consensus that dietary fibers have great potential to improve or maintain
balanced health and wellbeing. Dietary fiber’s beneficial effect on health almost
uniquely depends on the modulation of digestive processes, which largely depends
upon the intrinsic property of the fiber. In summary, fiber-enriched foods are less
calorific and often need to more oral processing such as chewing. This slows down
the food intake. On the other hand, fiber increases the bulk in the stomach and lowers
the gastric emptying rate. In the small intestine, fiber interferes with the absorption of
nutrients such as glucose as well as increases the peristalsis (foods moves faster). In
the colon, fiber increases the fecal mass and also provides the carbon source of
proliferation of bacteria. The metabolic products of bacterial fermentation of fiber
are known to affect the immune system and the neuroendocrine system positively.
By now, there is no question on the importance of fiber and food authorities around
the globe are encouraging the public to consume at least 25 g of fiber from a variety
of sources. However, there is still a lack of a universal definition of fiber. Further, the
effect of processing on the nutritional functionality of fiber needs to be clearer. It is
also necessary to establish proper models to speculate microbial composition and
520 B. Zhang et al.

metabolites in both health and disease people with intake of different dietary fibers,
and verify the effectiveness as well as feasibility of the model through in vivo study.

Acknowledgements We thank the National Natural Science Foundation of China (31701546) and
the 111 Project (B17018) for financial support. Bin Zhang thanks the Hong Kong Scholar Program
(XJ2019049), Pearl River Talent Recruitment Program of Guangdong Province (2017GC010229),
and the Pearl River Nova Program of Guangzhou (201906010079).

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