Eur Food Res Technol (2013) 237:299–307

DOI 10.1007/s00217-013-1991-0

ORIGINAL PAPER

Gelatinization properties of wheat flour as determined

by empirical and fundamental rheometric method
Milica Pojić • Miroslav Hadnad̄ev •

Tamara Dapčević Hadnad̄ev

Received: 6 February 2013 / Revised: 1 April 2013 / Accepted: 2 April 2013 / Published online: 16 April 2013
Ó Springer-Verlag Berlin Heidelberg 2013

Abstract Certain empirical rheological methods are in Keywords Gelatinization properties
Wheat flour

extensive use in wheat and/or flour research to assess starch Empirical rheological method
Fundamental rheological
gelatinization and pasting behavior primarily due to easy method
Vane geometry
performance and good correlation with final product
quality. However, their applications are often associated Abbreviations
with specific drawbacks that could be limiting factors for PCA Principal component analysis
certain applications, such as poor definition of the mea- PC Principal component
sured parameters, time-consuming nature, difficulties in SDr Repeatability standard deviation
interpretation of results, large sample sizes, etc. The listed RSD Relative standard deviation
shortcomings can be overcome by application of funda- r Repeatability limit
mental rheological methods that are based on well-defined Amy Brabender Amylograph
rheological parameters such as stress, strain, viscosity and Slow Slow rheometric procedure
modulus. The objective of this study was to optimize the Rapid Rapid rheometric procedure
fundamental rheological method for determination of the PV Peak viscosity
gelatinization properties of wheat flour that correspondents PT Peak temperature
to the standard widely accepted empirical rheological PST Pasting temperature
method—Amylograph method and to compare them in
order to determine whether they can be interchangeable
depending on different analytical needs. The obtained Introduction
results have shown that the application of fundamental
rheometric procedure for determination of pasting proper- Gelatinization properties of starch determine many food
ties of wheat flour provides reliable determination of the and non-food starch applications [1, 2]. A variety of
gelatinization properties of wheat flour. Moreover, sub- methods for the monitoring of gelatinization of starch are
stantial advantages of fundamental rheometric method over available, such as different microscopic, thermoanalytical,
the empirical one were identified including smaller sample enzymatic and rheological methods [3]. The recent trends
size, ability to set the desirable heating and shear rate, in starch research have indicated the utilization of com-
shorter test duration and better precision. bined measurement techniques as reported by Li et al. [4]
who coupled hot-stage light microscopy and differential
scanning calorimetry (DSC) to study the dynamic changes
of starch during gelatinization process. Rheological meth-
ods are based on the measurement of viscosity changes
during heating and shearing of starch slurries [5]. For this
M. Pojić (&)
M. Hadnad̄ev
T. D. Hadnad̄ev
purpose, different instruments are available including
Institute of Food Technology, University of Novi Sad, Bulevar
cara Lazara 1, 21000 Novi Sad, Serbia Brabender Visco Amylograph (BVA) and Micro Visco
e-mail: milica.pojic@fins.uns.ac.rs Amylograph (MVA), Rapid Visco Analyzer (RVA),

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Ottawa starch viscometer [6, 7] and viscometer [8]. Given spite of the difference in heating protocol, geometry and
the variety of instruments, it is not surprising that a large examined system has shown good correlation with Brab-
number of comparative studies have been performed ender Amylograph [22].
between certain instruments as being applied for determi- The objective of this study was to optimize the funda-
nation of pasting properties of starch and starch-containing mental rheological method for determination of the gela-
products [9–14]. Considering the role of starch as a func- tinization properties of wheat flour that correspondents to
tional ingredient in a diversity of food formulations, those the standard, widely accepted, Brabender Amylograph test
comparative studies have been mainly performed on native and to compare them in order to determine whether they
or modified starches of different botanical origin [15]. can be interchangeable depending on different analytical
Wheat flour, with 75–80 % of starch content on a dry needs. In order to avoid different constraints associated
weight basis, represents significant source of starch for the with concentric cylinder measuring geometry, such as
majority of population. The functional properties of starch starch granules sedimentation [23] and/or wall slippage
such as gelatinization, gelation and viscosity are of great effects [24], a two-bladed shaped rotor was used which
importance because they affect processing flour quality and represents a type of vane geometry.
final quality of baked products [1, 16]. The Brabender
Amylograph (BA) has been in extensive use in wheat and/
or flour research to assess starch gelatinization and pasting Materials and methods
behavior primarily due to easy performance, standardized
procedure and good correlation with final product quality. Samples
Taking into account the nature of flour as a testing material,
it must be noted that maximum viscosity attained during A total of 20 wheat samples were collected in 2008 from
the Amylograph test indicates both—the flour gelatiniza- the wheat-growing localities in Serbia. Wheat samples
tion behavior and the a-amylase activity present in the flour were milled to straight-grade flour using Bühler MLU-202
[17]. However, the use of Brabender Amylograph is laboratory mill (Bühler, Switzerland) according to AACC
characterized by certain methodological, geometrical and method 26–31 [25]. The samples for the study were chosen
technical shortcomings such as complex geometry, large on the basis of peak viscosities obtained by Brabender
sample size, inability to program the temperature profiles, Amylograph in order to cover as much variability as pos-
poor definition of the measured parameters, non-uniform sible (Table 1).
shear rate within the sample, the difference in heat transfer
inside the measurement cup and time-consuming nature of
testing. Therefore, the development and the use of more
flexible devices and tests based on well-defined rheological Table 1 The characteristics of selected sample set in terms of peak
viscosity as measured by Brabender Amylograp (Amy)
parameters (stress, strain, viscosity and modulus) have
been initiated [5, 18]. Among them, a Rapid Visco Ana- Sample Amy (BU) Sample Amy (BU)
lyzer (RVA) has been of a particular interest among cereal S1 90.0a S11 877.5k
researchers, due to smaller sample sizes required and S2 182.5b
S12 922.5l
similar pasting patterns it provides when compared to other S3 247.5c S13 1,005.0m
similar instruments [5]. However, the RVA results are
S4 320.0d S14 1,022.5mn
expressed as viscosity in centipoise (cP) or Rapid Visco e
S5 422.5 S15 1,042.5n
Units (RVU), being a disadvantage compared with the f
S6 555.0 S16 1,280.0o
fundamental rheological methods. Rosell et al. [19] g
S7 650.0 S17 1,325.0p
reported the use of a constant stress rheometer equipped h
S8 672.5 S18 1,590.0q
with cone-and-plate measuring geometry for determination
S9 707.5i S19 1,640.0r
of pasting properties of starch–hydrocolloid slurries j
S10 835.0 S20 1,725.0s
(8.0 %, w/w). Concerning starch gelatinization in the
Mean 855.6
water-limited dough system, there have been a number of
SD 485.2
examples of using rheometer to perform temperature sweep
Min 90.0
test which is especially useful in breadmaking where
Max 1,725.0
changes that occur during the fermentation and baking
processes could be explained [20, 21]. In attempt to sim- CV 0.6
ulate breadmaking process (mixing and baking), where Figures followed by the different letters are significantly different
water amount for starch gelatinization is limited, Chopin (p \ 0.05)
Technologies developed a device called Mixolab, which in SD Standard deviation, CV coefficient of variation

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Amylograph tests Results and discussion

The Amylograph tests were performed according to ICC Determination of pasting and peak temperature
standard No. 126/1 [17], which involved heating the sam-
ples from 30 to 95 °C at a constant rate of 1.5 °C min-1. Pasting temperature indicates temperature at which the
The peak viscosity and the peak temperature were deter- viscosity begins to increase during the heating process [27].
mined from the Amylogram. Three parameters were Pasting temperatures of selected wheat flours differed with
recorded in duplicate: pasting temperature, peak viscosity flour sample, instrument type and heating rate as previously
and peak temperature. indicated by Suh and Jane [14]. The pasting temperature
variation between the samples and observed method is
Fundamental rheometric tests shown in Fig. 1a. The pasting temperatures as determined

The flour suspension for fundamental rheological tests was

prepared with 12.4 g (calculated on 14 % moisture basis)
of flour and 70 ml of distilled water in order to obtain the
same flour–water ratio as in the standard Amylograph
method. Fundamental rheometric measurements were
taken using the HAAKE MARS—Modular Advanced
Rheometer Systems (Thermo Scientific, Germany). The
measuring geometry consisted of measuring cup Z40
(43.4 mm diameter, 8 mm gap) and FL2B paddle-shaped
rotor with 2 blades. In order to prevent moisture evapora-
tion from the sample, the solvent trap Z40 DIN was used.
Two methods for determination of pasting properties of
wheat flour were developed: so-called slow and rapid
procedure. Slow procedure comprised the complete simu-
lation of Amylograph test with a heating rate of 1.5 °C
min-1 from 30 to 95 °C and maintaining the temperature of
95 °C for a period of 600 s. A shear rate of 10 s-1 was
applied, which corresponded to 95 rpm. The rapid proce-
dure comprised the tempering of flour suspension at 50 °C
for 180 s, heating from 50 to 95 °C at a heating rate of
3 °C min-1 and subsequent maintenance of temperature of
95 °C for 300 s to ensure starch gelatinization. Three
parameters were recorded in duplicate: pasting tempera-
ture, peak viscosity and peak temperature.

Data analysis

All samples were analyzed in duplicate except for method

precision determination, where eight replicates were per-
formed. The obtained results were analyzed by descriptive
statistics (mean, standard deviation, minimum and maxi-
mum, coefficient of variation), Pearson’s correlation coef-
ficients, one-way analysis of variance (ANOVA) and
Duncan’s new multiple range tests to determine the sta-
tistical significance of differences, using software Statistica
10. The principal component analysis (PCA) was carried
out using the Software XLSTAT, version (2012.2.02).
Determination of the method repeatability comprised
determination of repeatability standard deviation (SDr),
Fig. 1 Pasting temperature (a), peak temperature (b) and peak
relative standard deviation (RSD) and repeatability limit viscosity (c) as measured by Brabender Amylograph, slow and rapid
(r = 2.8*SDr) [26]. rheometric procedure

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by Amylograph varied between 54.6 and 61.0 °C, with the temperature values were not affected by the position of
highest value being observed for sample S20, while the temperature sensor within the measuring cup. Conversely,
pasting temperature of selected wheat flour samples as the MARS’s temperature measuring system is based on the
determined by slow rheological method varied between feedback from the block temperature, but not from the
57.7 and 65.6 °C with the highest value being observed for temperature of the sample. The registered higher pasting
sample S17. The pasting temperatures as determined by temperatures within the slow procedure could be attributed
Amylograph were significantly lower than those obtained to the existing lag behind that of the heating block [30].
within the slow rheometric method (p \ 0.05). Although Also, the temperature throughout the sample of the larger
these two methods were characterized by the same heating size in the Brabender Amylograph (cca. 530 g) probably
rate, it must be noted that other test conditions such as changed at a slower rate in comparison with the 82.4-g
measuring geometry were different [9]. sample within the slow and rapid rheometric procedure.
The pasting temperature of selected wheat flour samples
as determined by rapid rheometric method varied between Determination of peak viscosity
62.1 and 67.1 °C with the highest value being observed for
sample S19, being significantly higher than those obtained The peak viscosity is the highest viscosity reached during
within the slow procedure (p \ 0.05). The differences in starch gelatinization when the balance between granules
the pasting temperatures obtained within the slow and swelling and granules breakage supported by stirring and
rapid procedure could be attributed to the differences in heating is reached shortly before their physical breakdown
starch granules swelling rates affected by variable heating and leaching of amylose [3]. The selected sample set
rates. At slower heating rate, starch granules were allowed covered wide range (from 90 to 1,725 BU) and uniform
to longer swelling which resulted in lower pasting tem- distribution of samples with significantly different peak
perature [14, 28]. There is usually a difference in the viscosities (p \ 0.05) (Table 1). The peak viscosity mea-
gelatinization temperature range between the starch and sured by the slow and rapid fundamental rheometric pro-
flour, in that is shifted toward higher temperatures in the cedures ranged from 3.64 to 80.35 Pa s and from 7.54 to
wheat flour suspension compared with the starch–water 94.52 Pa s, respectively (Fig. 1c). The principal compo-
suspension [29]. nent analysis was used to visualize the variability of
The peak temperature is the temperature at which selected flour samples with regard to their gelatinization
swollen starch granules in water dispersion under shear properties, where each sample is presented as a point in a
force tend to disintegrate from the crystalline state to a gel two-dimensional plane—the PCA score plot (Fig. 2a).
[5]. The peak temperatures differed with flour sample, Samples with distinctive features appeared completely
instrument type and heating rate as indicated by Suh and separated from each other, while samples with similar
Jane [14]. The peak temperature variation between the gelatinization properties appeared close to each other.
samples and observed method is shown in Fig. 1b. The Moreover, samples that appeared close to the origin were
peak temperature as determined by Amylograph varied characterized by such gelatinization properties that were
between 67.4 and 90.2 °C, with the highest value being similar to those of their mean values [31]. In order to
observed for sample S9. The peak temperature of selected visualize the correlation between the selected gelatinization
wheat flour samples as determined by slow rheometric properties of wheat flour as determined by Brabender
method varied between 71.8 and 94.3 °C with the highest Amylograph, slow and rapid rheometric procedure, as well
value being observed for sample S20. The same heating as the underlying structure in experimental data and rela-
rate applied within those two methods, resulted in signifi- tionships between data and samples, the PCA loading plot
cantly lower peak temperatures as determined by Amylo- is presented (Fig. 2b). The first principal component (PC1),
graph (p \ 0.05). By increasing the heating rate within the accounted for 76.12 % of the total variability in the data
rapid procedure, the peak temperature increased and varied set, correlated highly with the peak viscosity as determined
between 77.6 and 93.9 °C with the highest value being by rapid, Brabender Amylograph and slow rheometric
observed for sample S18. method, and peak temperature as determined by slow
The observed pasting and peak temperatures variation rheometric method (PC1 [ 0.90). The second and third
between standard Brabender Amylograph and slow rheo- PCs (PC2 and PC3) accounted for 9.09 and 6.29 % of the
metric method might be attributed to the differences in the total variance in the data, respectively. PC2 correlated
temperature measuring system between these two devices. highly with pasting temperature as determined by Brab-
The temperature sensor with Brabender Amylograph is ender Amylograph method.
located in one point inside the measuring bowl, thus Direct comparison of the peak viscosities obtained by
allowing measuring actual sample temperature. Hazelton the Amylograph and fundamental rheometric procedures
and Walker [30] previously proved that measured was difficult due to the difference in the measurement units

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swelling and amylose solubility which resulted in increased

peak viscosity [14, 28, 34, 35].
Apart from the difference in measurement units and
homogeneity of temperature, there are other distinctions
between those two methods, with regard to measurement
geometry, shear rate, definition of measured parameters,
the time required for testing and sample size. Brabender
Amylograph is a rotational viscometer that consists of
rotational bowl rotating at 75 rpm with built-in pins and a
stationary upper spindle with sensing pins [1, 17].
Although the role of the pins is to prevent the sedimenta-
tion of particles present in the dispersion, they make
Brabender Amylograph measurement geometry very
complex. Hence, the shear modulus is too complicated to
calculate, and the flow field within the Amylograph mea-
suring bowl is too difficult to describe [1]. Concerning the
fundamental rheological devices applied to characterize the
pasting properties of starch and starch-containing products,
and the rotational rheometer equipped with different
geometries (concentric cylinder, cone-and-plate, parallel
plate) has been the most common choice [1, 23]. Despite
the numerous advantages their applications offer, it must be
noted that a relatively narrow gap with cone-and-plate
geometry could contribute to capillary suction of the par-
affin oil from the edge of the geometry into the tested
dispersion, applied to minimize moisture loss. On the other
hand, with the application of a concentric cylinder, there is
a possibility of starch granules sedimentation in the early
stages of heating [23]. Applicability of the various forms of
vane geometry for measuring rheological properties of
food and non-food systems, especially for coarse and dis-
persed systems, has been reported [23, 36]. However, the
vane-in-cup geometry has been mostly used to obtain yield
stress of food systems and to measure flow properties as it
acts as an effective cylinder. By given review of Barnes
and Nguyen [36] regarding the reported uses of vane
Fig. 2 PCA score plot of PC1 versus PC2 of the sample set selected
for the study (a) and PCA loading plot of PC1 versus PC2 of analyzed geometry for food and non-food systems, there is no
gelatinization parameters ( PST pasting temperature, PT peak reported application of vane geometry for flour systems.
temperature and PV peak viscosity ) as determined by Brabender One example of the application of vane geometry to the
Amylograph (Amy), slow (Slow) and rapid rheometric procedure starch system was reported by Tucker et al. [37] who used
(Rapid)
it for the measurement of viscoelasticity and apparent
used to express viscosity [14]. Although based on the same viscosity of modified starch systems. Genovese and Rao
measurement principle (recording the change in viscosity [38] reported the use of vane geometry for the measure-
of flour–water suspension during a uniform temperature ment of yield stress of cross-linked starch dispersions of
increase under constant stirring), the peak viscosity different botanical origin at different rotational speeds. The
obtained by Amylograph is expressed as torque in the chosen vane geometry consisted of paddle-shaped rotor
arbitrary Brabender Unit (BU), whereas the viscosity with two blades with slits (Fig. 3).
obtained by fundamental rheometric methods is expressed Suh and Jane [14] reported the differences in the peak
in the SI units (Pa for elastic modulus, Pa s for viscosity) viscosity obtained by RVA and MVA that could had been
[32, 33]. When comparing the slow and rapid procedure, attributed to the different spindle geometry between those
the increase in the heating rate influenced the increase in two devices. Paddle-shaped spindle, due to the larger sur-
the peak viscosity, due to enhancement of starch granule face area, affected increased breakdown and consequently

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Fig. 3 Brabender Amylograph

geometry (a) and two-bladed
shaped rotor (b)

the increased shear force with regard to hollow-structured Table 2 Correlation coefficients (r) between the peak viscosity (PV),
spindle [14, 39]. peak (PT) and pasting (PST) temperature of wheat flour as determined
Another important distinction between the Amylograph by Brabender Amylograph (Amy), slow and rapid rheometric
and slow and rapid rheometric method is a sample size. procedure
The sample size which is used with Brabender Amylograph Variable PST-Amy PST-Slow PST-Rapid
has been a limiting factor for certain applications, such as
PST-Amy 1
breeding programs, where small amounts of samples are
PST-Slow 0.53 1
normally available [28, 32]. To overcome this drawback,
PST-Rapid 0.54 0.89 1
the attempts to record the viscosity profile using a reduced
amount of sample have been reported and, new instruments PT-Amy PT-Slow PT-Rapid
based on similar operating principles, such as Rapid Visco PT-Amy 1
Analyzer (RVA), introduced [14, 28, 35]. It was shown that PT-Slow 0.95 1
obtained viscograms were comparable with those obtained
PT-Rapid 0.82 0.91 1
by Brabender Amylograph. Moreover, the reduction in
amount of sample had not disturbed the possibility to PV-Amy PV-Slow PV-Rapid
deduce the a-amylase activity, being in agreement with
PV-Amy 1
that of Brabender Amylograph [28].
PV-Slow 0.99 1
The correlation coefficients between the gelatinization
PV-Rapid 0.99 0.98 1
properties of wheat flour as measured by Brabender Am-
ylograph, slow and rapid rheometric procedure is presented Values in bold are significant at p \ 0.01
in Table 2. The low and not significant values of correla-
tion coefficient (r) were observed between Amylograph
pasting temperature and pasting temperature as determined Amylograph and slow and rapid rheometric procedure
by slow and rapid rheometric procedure (r = 0.53 and regarding the sample size, applied heating rates and mea-
r = 0.54, respectively). However, pasting temperature as surement geometry, peak viscosity as measured by Brab-
determined by slow and rapid rheometric procedure ender Amylograph showed significant positive correlations
exhibited strong mutual relationship (r = 0.89). Moreover, with the peak viscosity measured by slow and rapid
the significant coefficient of correlation (r) was observed rheometric method (r = 0.99, p \ 0.01). The relationship
between peak temperature as determined by empirical and between the peak viscosities obtained by slow and rapid
fundamental rheometric procedures, being the highest rheometric method is described by significant positive
between Amylograph and slow procedure (r = 0.95). coefficient of correlation (r = 0.98, p \ 0.01) (Table 2).
Thiewes and Steeneken [10] previously reported the These observations confirmed the universal nature of
poor correlation coefficients between peak viscosity and rheological measurements which should be independent of
pasting temperature for modified starches, while Limpisut size, shape and how they are measured. Moreover, the
and Jindal [13] observed the same phenomenon for rice obtained results confirmed the objective of rheology to
flour. Despite the listed differences between Brabender determine properties reproducibly in a manner that allows

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Fig. 4 Variability in viscosity

curves obtained by the
Brabender Amylograph (a) and
slow and rapid fundamental
rheological procedure (b) of
selected samples

comparison between different samples, test sizes and Precision of methods used for determination
shapes and test methods [18]. of gelatinization properties of wheat flour
Figure 4 shows the variability in viscosity profiles of
the selected samples (S4 and S19) obtained by Brabender The final step of the evaluation of the method character-
Amylograph and slow and rapid rheometric procedure istics comprised the determination of method repeatability.
using vane geometry. Their shape was similar to that Repeatability checks were made for each observed pasting
obtained by the Brabender Amylograph. During the first parameter in eight replicates. As indicated in Table 3, the
stage of heating of flour water suspensions and during the fundamental rheometric method (slow and rapid) was
increase in viscosity associated with starch gelatinization characterized by better repeatability in terms of pasting and
and a-amylase liquefaction, a negligible difference peak temperature and peak viscosity in comparison with
between the slow and rapid fundamental rheometric pro- Brabender Amylograph method. The rapid procedure had
cedure was observed. The more apparent differences in better repeatability of pasting and peak temperature
viscosity profiles obtained by the slow and rapid funda-
mental rheometric procedure were observed in the peak
viscosity being higher for rapid procedure. Weipert [40] Table 3 Repeatability of different methods for determination of
indicated the significance of test duration when describing gelatinization properties of wheat flour
the pasting behavior of wheat starch. The starch granule SDr r RSDr
requires certain time to absorb and bind the water, to swell
and finally to gelatinize, so the rapid tests could result in Amylograph
the higher viscosity. Moreover, the differences in viscosity Pasting temperature (°C) 1.19 3.33 1.82
profiles of selected samples between slow and rapid pro- Peak temperature (°C) 2.25 6.30 2.64
cedure are more apparent for sample S4, which was Peak viscosity (BU) 9.04 25.31 2.10
characterized with lower peak viscosity (and higher a- Slow procedure
amylase activity). This system was subjected to the more Pasting temperature (°C) 0.29 0.81 0.48
efficient breakdown under the longer joint influence of Peak temperature (°C) 0.58 1.62 0.68
shearing and heating. The enzyme acts on (1,4)-bonds of Peak viscosity (mPa s) 0.17 0.48 1.23
starch fractions, resulting in a peak viscosity decrease. Rapid procedure
The lower the alpha amylase activity, the lower the dif- Pasting temperature (°C) 0.23 0.64 0.35
ference in the appearance of curves was, regardless of the Peak temperature (°C) 0.22 0.62 0.24
duration of the test due to the fact that the lower a- Peak viscosity (mPa s) 0.60 1.68 0.93
amylase activity generally contributes to the stability and
SDr Repeatability standard deviation (expressed in corresponding
rigidity to the swollen granule structure and less break- units), r repeatability limit (expressed in corresponding units), RSDr
down with shear [41]. repeatability relative standard deviation (%)

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determination in comparison with the slow procedure. 5. Gunaratne A, Corke H (2004) In: Wrigley C, Corke H, Walker
Moreover, determination of peak viscosity by slow rheo- CE (eds) Encyclopedia of grain science. Elsevier, Oxford,
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