ICASF-2022 IOP Publishing

IOP Conf. Series: Earth and Environmental Science 1110 (2023) 012041 doi:10.1088/1755-1315/1110/1/012041

Environmentally Friendly Beeswax: Properties,

Composition, Adulteration, and its Therapeutic Benefits
Garima Gupta1, a) and Kumari Anjali 1
1
University Institute of Agricultural Sciences, Chandigarh University,
Gharuan, Mohali- 140413, Punjab, India
a)
Corresponding author: garima.e8884@cumail.in

Abstract. Bees produce beeswax, which is used to make honeycombs is biodegradable, sustainable, compostable, eco-friendly. In
addition to honey and pollen, honey bees use comb to store nectar and rear their young. More than 300 things are made with
beeswax, such as candles, shoe polish and vehicle and floor polishes. Beeswax can also be used in the food processing and
packaging, wax crayons, metal casting and modelling, food processing, and cosmetics industries. In addition to these, embalming,
papyrus preservation, and artwork conservation are all possible uses for honey beeswax. Beeswax has numerous industrial,
pharmacological, and medical uses that demand an understanding of its chemical makeup. Similar to honey, beeswax is considered
effective in the treatment of bruising, inflammation and burns. Beeswax's antibacterial properties have recently been the subject of
inquiry, despite the fact that there are currently few studies on the subject. For example, beeswax has been proven to significantly
reduce affectivity of disease casing bacteria like: Staphylococcus aureus, Candida albicans and Salmonella enterica. This study
discusses the characteristics, composition, and adulteration of beeswax, as well as its medicinal effects as an antibiotic and in skin
therapy.
Key words: Beeswax, Antimicrobial, Dermatological, Healing, Apis, Honey bees

INTRODUCTION
Several insects in nature create wax, but bees and other Apoidea are responsible for the production of wax
that is greatly appreciated and utilized by humans. Beeswax is produced by the two most widely cultivated bee species,
Apis mellifera and A. cerana, and is the most widely used wax on the market. Beeswax is produced by younger worker
bees (between the ages of 12 and 18 days, when they have completed their nursing duties) and is a complex material
that is available in liquid form. Beeswax is made by mixing beeswax, pollen, and enzymes with resin from tree buds.
Beeswax can then be used as a substitute for wax. There is a plethora of practical applications for honey beeswax,
which include embalming, the preservation of papyrus, the conservation of artworks, the manufacture of candles and
cult artefacts, as well as its usage as a medicinal component and adhesive ingredient [1]. Also, it acts as a UV radiation
barrier to keep the skin from being corroded or scratched, and it keeps moisture from escaping. Additionally, beeswax
serves as a source of nest-mate identification clues for honey bee colonies because of its unique chemical composition.
Beeswax is making a comeback in the beekeeping industry, thanks to the fact that it is currently predominantly used
to construct comb foundations [2]. Beeswax is commonly used in the food business as a food additive and to extend
the shelf life of goods, and it is also widely employed in the pharmaceutical and nutraceutical sectors. Although there
has been some improvement, concerns regarding contamination and adulteration of high-quality beeswax have
persisted. An adulterant has been introduced into beeswax. For commercial gain, natural or synthetic chemicals are
added to beeswax. This is now one of the most important challenges relating to beeswax quality, accounting for around
20% of all problems [3]. Furthermore, the information gathered in this research on the features, content, and
adulteration of beeswax, as well as its therapeutical effects as an antibiotic and in skin treatments, is useful and
demonstrates beeswax's future usage in related fields.
Structure of Bee Comb:
Many bees work together to build a single cell for different purposes. Cells are hexagonal in structure and are of
different sizes [4]. Despite the fact that bees join combs to the surfaces along their upper lateral sides of frame, they
make little tunnels around the comb's borders. The fundamental nest structure is as follows: honey storage on top,
brood nest on the bottom, and pollen storage in the area between them (Figure 1). There are peculiarities in the comb
structure that are associated with this arrangement. Brood nest combs are frequently darker in colour than honey
storage combs, and the breadth and cell structure of the combs are more uniform. The drone comb is situated on the
outskirts of the brood nest [5].

Content from this work may be used under the terms of the Creative Commons Attribution 3.0 licence. Any further distribution
of this work must maintain attribution to the author(s) and the title of the work, journal citation and DOI.
Published under licence by IOP Publishing Ltd 1
ICASF-2022 IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1110 (2023) 012041 doi:10.1088/1755-1315/1110/1/012041

Fig.1. Structure of Bee Comb

Pollen cells

Brood cells

Honey cells

Drone cells

Queen cells

BEESWAX
Honeybee Apis mellifera is the only honeybee that produces beeswax commercially available. Because of its long
history of usage in the manufacture of other natural products (such as ozocerite), beeswax is considered a highly
important resource [6]. According to Faostat, the worldwide production is 64,000 tonnes in 2011, with Asia (mostly
India) being the leading producer with 31,000 tonnes [7]. As a by-product of honey harvesting and refinement, wax
can be recovered [8]. Accurate knowledge of the beewax's chemical composition is essential for numerous industrial,
pharmaceutical, and medical applications. In order to conduct reliable analyses, the composition of the waxes has been
fairly characterized. Beeswax is chemically composed of around 300 distinct compounds. Beeswax contains a variety
of free fatty acids, monoesters, diesters, and hydroxyl monoesters, as well as alkanes and alkenes. The majority of
them are fatty acid esters (67%), hydrocarbons (14%), and free fatty acids (13 %) [6]. Although their concentrations
fluctuate based on the honeybee species and geographical origin, very minor changes in the concentration of particular
components and substance groups are identified [9]. In beeswaxes from various bee species, a total content of 27 to
40 percent monoesters, 9 to 23 percent hydroxymonoesters, 7 to 16 percent diesters, 3.9 percent hydroxydiesters, 11
to 28 percent hydrocarbons, 1 to 18 percent saturated, unbranched free fatty acids, 4 to 8 percent other substances, and
0.3 percent free fatty alcohols has been reported [9].

PROPERTIES OF BEESWAX
The newly created beeswax was white at first and subsequently became yellow. Propolis and pollen dyes are used to
create this golden tint. Beeswax has an unusual odour and is made from bees, honey, propolis, and pollen. Beeswax
smell is sweet, warm, and balsamic, gourmet with a honey aroma, and a highly sought product. Although its perfume
is one of its most cherished characteristics, it also has a hay-like undertone, and no research into the volatile ingredients
has previously been published [10].

2
ICASF-2022 IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1110 (2023) 012041 doi:10.1088/1755-1315/1110/1/012041

It darkens as it is used in the hive because pollen, silk, and larvae waste are mixed in with the honey. During
cold weather conditions, natural beeswax gets harden. Because the composition of beeswax changes a lot depending
on where it came from, the melting point is not the same. Its temperature ranges from 61 to 66 °C. It has a relative
density of 0.958–0.970 g/cm3 at 15°C and an electrical resistance range of 51012 to 201012 m. It has a thermal
conductivity value of 2.5103 J cm/s °C cm2. Because beeswax can be used to make soap, its saponification value is
between 85 and 100. Beeswax is inert and very strong. While it is insoluble in water and resistant to a wide variety of
acids, it is soluble in the majority of organic solvents, as well as alcohol and fatty oils, when heated [2].
The fundamental basic elements for the synthesis of wax are sugars, fructose, glucose, and sucrose. In
general, the sugar-to-wax ratio ranges from 3 to 30:1. In Central Europe, the usual ratio is around 20:1. There is a
direct correlation between a colony's strength and its ability to produce wax. Only 100 grammes of wax in a Langstroth
frame may hold up to 4 kilos of honey [1].
Process of Beeswax production
Wax is required by bees to construct their honeycombs. They create wax glands in 12–18-day workers, which are
completely matured. The wax glands of aged bees become less active, yet they may reactivate wax production in an
emergency. The development stage of the bee colony will create a lot of wax in the moderate environment of April to
June. They get their four pairs of beeswax glands on the ventral side of their abdomen. Wax is made by bees in these
glands. These glands convey liquid wax, which is then cooled to make fine white wax flakes. Bees collect beeswax
from tree buds. The scales on the hind legs are removed and processed using mouthparts. During this process, scales,
which the bees use to make honeycombs with their mandibles and fill them with pollen and propolis, start to form.
The color of beeswax changes after it is exposed to honey and pollen. It turns brown after about four years. This is
because beeswax is made from the exuvae of pupae, which is what makes the wax look different shades of yellow and
brown. Because a gramme of wax weighs around 1 milligram, one kilogram of wax requires approximately one million
scales [1].
Wax collection and processing
During the process of extracting honey, wax is often removed from the cap. Wax that is good quality and has a light
color comes from this method. By separating white honeycombs from dark ones, different wax grades can be made.
There will be more wax in the honey (10–15 percent) than there will be in frame hive beekeeping, where only 1–2
percent of the honey is made from wax. Most of the beeswax in the world comes from the honeybee A. mellifera. This
bee makes a lot of beeswax. Other honeybee species, such as A. dorsata and A. florea, have the ability to produce
beeswax. During the buildup of honey combs, bees use wax from glands in their abdomen to make them. Bees and
their larvae live in the combs, and honey and pollen are stored there, as well as honey and pollen [2].
Wax Extraction Method
There is a big difference in the quality of pure beeswax that can be made by how it is made. Melting and chemical
extraction are two of the most common ways to get wax out of things. Boiling water, steam, electricity, or even the
sun can be used to melt wax. When producing wax on a small scale, chemical solvent extraction is only practical in
the lab. When beeswax is melted and cleaned, it usually turns out to be a bright yellow colour. Once it becomes darker
for any reason, such as when it becomes too hot or has a lot of metal in it, it can be illuminated by exposing it to sun
or by using chemicals. Complexing agents, which make metals bind together, have also been considered, however
these compounds are toxic and detrimental to the environment. After the wax has cured, store it in a cold place. It's
possible to keep them in wrapping paper, on a shelf, or in a stainless steel, glass, or plastic container to keep their
color and scent the same [1].

QUALITY CONTROL
Quality management of beeswax necessitates a high degree of specialized knowledge and experience. Wax comes in
white and yellow, with white beeswax being described as bleached yellow wax. There are two types of wax: white
and yellow. The bleached wax lacks beeswax's inherent colourants and scent. Beeswax in solid blocks should always
be clean and have the proper colour and odour. While adulteration is straightforward (typically accomplished with
cheap paraffin waxes), detection necessitates chemical testing [2]. Paraffins, calf tallow, stearic acid, and carnauba
wax were among the adulterants found in commercial beeswax [11].

3
ICASF-2022 IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1110 (2023) 012041 doi:10.1088/1755-1315/1110/1/012041

Different methods can be employed to detect wax adulteration. Pure beeswax was tested for its specific
density, saponification, acid-ester ratio number, refractive index, hydrocarbons percentage, melting point, and ash
content (Table 1) [12]. Sensory and physicochemical criteria calculated in conformity with the Pharmacopeia do not
guarantee that the wax is not adulterated, but they may offer indications of potential adulteration in some cases.
Adulteration is frequently detected today by analysing the wax's components using gas chromatography (GC) or liquid
chromatography. The components of beeswax are also identified using gas chromatography coupled with
spectrophotometry detection (GC–MS) [13, 9]. Some hydrocarbons and monoesters' molecular weights are affected
by contaminated beeswax. For a long time, measuring physical-chemical characteristics, such as melting point and
saponification number, has been recognized as a method of determining the purity of beeswax based on the
standard value [14].

Table.1. Quality parameters for routine beeswax testing

Parameters Permissible Range of Value

Melting point 61℃ to 65℃

Refractive index, 75℃ 1.4398 to 1.4451
Ester/Acid ratio 3.3 to 4.3
Saponification number 87 to 102
Hydrocarbons Max. 14.5%*
Water content <1%

THERAPEUTIC EFFECTS OF BEESWAX

When used with therapeutic pharmaceuticals or dangerous baits, beeswax aids in the preservation of the active
ingredients and their gradual release without causing any harm to people or bigger animals [2]. However, its indirect
effects can be rather significant, since it aids in the preservation of active compounds and their gradual release. It may
be used to cover a wide range of surfaces, including human skin, with thin, non-corrosive, non-allergenic coating
materials. As a result of its anti-inflammatory and antioxidant properties, it protects against rusting and scratches,
along with moisture loss. This may be related to the presence of certain propolis inclusions in beeswax. When beeswax
is saponified with borax, a very stable and smooth emulsion for cosmetics can be created [15].

4
ICASF-2022 IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1110 (2023) 012041 doi:10.1088/1755-1315/1110/1/012041

Figure 2. Benefits of Beeswax

Cosmetic and Dermatological Property

Beeswax is used to treat a range of dermatological diseases, including boils, wounds, atopic dermatitis, psoriasis, and
paediatric dermatitis. Ointments based on beeswax are mentioned in the Ebers Papyrus, by the Greek-Roman physician
Galen, and in historical manuscripts of traditional Chinese medicine, such as the "Shen Nong Book of Herbs"
[16]. Numerous studies have demonstrated that a mixture of equal parts honey, olive oil, and beeswax is effective for
the treatment of dermatitis, psoriasis, and bacteria and fungi infections of the skin [17]. Because of its minimal
irritating and comedogenic properties, beeswax is frequently utilised as a thickening, emollient, and emulsifier in
modern cosmetics and makeup. Beeswax protects the skin from UV rays, increases its flexibility, and does not cause
allergic reactions [18]. Beeswax has long been used as an Ayurvedic treatment for inflammation, bruising, burns, and
cracked heels [19]. Beeswax is used in cosmetics as a stiffener, a material that imparts flexibility and plasticity to the
skin, as well as an emulsifier. Beeswax is a foundation ingredient of lipsticks, lip glosses, and lotions. Beeswax
lubricates and softens the skin, as well as decreasing trans-epidermal water loss. Squalene, 10-hydroxy-trans-2-
decenoic acid, and flavonoids (chrysin) included in beeswax give this product antibacterial characteristics and protect
the skin from harmful microbes. Beeswax contains -carotene, a good source of vitamin A, which slows collagen
breakdown, accelerates mitotic division in the epidermis, and protects the skin from a variety of external influences
by producing a film on the surface. This results in faster skin regeneration following injury [20].

Antimicrobial activity
Because of its antimicrobial properties, beeswax is being used in Asian and European traditional therapies since
ancient times. Its usage in embalming and mummification methods by ancient Egyptians and Persians, perhaps to
imitate ancient Romans' death masks, might be attributed to its preservative properties [21]. Antimicrobial action of
natural goods, particularly hive products, is receiving attention, and unlike other bee products, beeswax has been
investigated just lately [22]. Beeswax in its natural state shown antibacterial action against a variety of bacterial strains
including the yeast C. albicans. Beeswax has been demonstrated to be specifically effective against Brevibacterium

5
ICASF-2022 IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1110 (2023) 012041 doi:10.1088/1755-1315/1110/1/012041

alvei, Proteus vulgaris, S. gallinarum, and B. subtilis. Its efficiency was halved when S. pullorum, S. dublin, E. coli,
and Bacillus larvae were used [23]. A combination of honey, beeswax, and olive oil when mixed in equal
concentrations is effective at inhibiting the growth of S. aureus and C. albicans isolated from infected people
[24]. Honey, beeswax, and olive oil were also evaluated in vivo for antifungal effectiveness against Pityriasis
versicolor, Tinea cruris, T. corporis, and T. faciei [24]. A crude extract of beeswax inhibited S. aureus, S. enterica,
C. albicans, and Aspergillus niger [23]. Whereas methanol and ethanol extracts inhibited pathogenic bacteria and
microscopic fungus [25]. This type of impact might be due, at least in part, to plant-derived beeswax components
[26]. Propolis and beeswax in combinations (1:1, v/v) have also been examined for their antibacterial efficacy against
S. aureus, S. epidermidis, B. subtilis, and Candida albicans [27].

CONCLUSION
Beeswax is the substance derived from the sugars found in honey. It has a crystalline structure that is good for building
hives with. Chemically, it's used in a broad range of industrial items and operations, including food preservation and
packaging, candle production, and cosmetics manufacturing. It is employed in pharmaceutical preparations as a
thickener, binder, drug carrier, and release retardant. For ages, beeswax has been used in traditional European and
Asian therapies because to its antimicrobial characteristics. In conclusion, additional research should be done on the
manufacturing, characteristics, composition, quality control, and antibacterial activity of beeswax, as well.
Investigations into the antibacterial activity of the many components that distinguish beeswax produced under various
climatic and geographical settings would also be interesting to consider.

REFERENCES

[1] S. Bogdanov, Beeswax: quality issues today. Bee world, 85(3), 46-50 (2004)
[2] Tinto, W. F., Elufioye, T. O., & Roach, J. Waxes. In Pharmacognosy (pp. 443-455). Academic Press (2017)
[3] L. Svečnjak, L. A. Chesson, A. Gallina, M. Maia, M. Martinello, F. Mutinelli, et al., Journal of Apicultural
Research, 58(2), 1-108 (2019)
[4] D. B. Casteel, The Manipulation of the Wax Scales of the Honey Bee (United States Department of Agriculture,
1912), vol. 161, pp. 1–13.
[5] T. D Seeley and R. A. Morse, The nest of the honey bee (Apis mellifera L.). Ins. Soc 23, 495–512 (1976)
https://doi.org/10.1007/BF02223477
[6] A. P. Tulloch, Beeswax—composition and analysis. Bee World, 61(2), 47-62 (1980)
[7] S. Bogdanov, Beeswax: uses and trade. The Beeswax Book, 1-16 (2009)
[8] J. Menezes, and K. A. Athmaselvi, Report on edible films and coatings. In Food Packaging and Preservation (pp.
177-212). Academic Press. (2018)
[9] R. Aichholz, and E. Lorbeer, Journal of Chromatography A, 855(2), 601-615 (1999)
[10] C. E. Ferber, and H. E. Nursten, Journal of the Science of Food and Agriculture, 28(6), 511-518 (1977)
[11] J. S. Bonvehi and F. O. Bermejo, Food chemistry, 132(1), 642-648 (2012)
[12] J. L. Bernal, J. J. Jiménez, M. J. del Nozal, L. Toribio and M. T. Martín, European journal of lipid science and
technology, 107(3), 158-166 (2005)
[13] J. J. Jiménez, J. L. Bernal, S. Aumente, M. J. del Nozal, M. T. Martı́n, and J. Bernal Jr (2004). Journal of
Chromatography A, 1024 (1-2), 147-154.
[14] J. J. Jiménez, J. L. Bernal, M. J. del Nozal, L. Toribio and J. Bernal (2007). European Journal of Lipid Science
and Technology, 109 (7), 682-690.
[15] R. Krell, Value-added products from beekeeping (No. 124). Food and Agriculture Org. (1996)
[16] T. Rit and R. Behrer Beeswax Through the Ages. Bladel: Koster Keunen Holland BV (1999)

6
ICASF-2022 IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1110 (2023) 012041 doi:10.1088/1755-1315/1110/1/012041

[17] N. S. Al-Waili, Clinical microbiology and infection, 11(2), 160-163 (2005)

[18] K. Münstedt, and S. Bogdanov, Journal of ApiProduct and ApiMedical Science, 1(3), 57-63 (2009)
10.3896/IBRA.4.01.3.01
[19] T. Gokani. Ayurveda–the science of healing. Headache 54, 1103–1106. (2014) 10.1111/head.12363
[20] A. Kurek-Górecka, M. Górecki, A. Rzepecka-Stojko, R. Balwierz, and J. Stojko, Molecules, 25(3), 556 (2020)
[21] L. Cornara, M. Biagi, J. Xiao, and B. Burlando, Frontiers in pharmacology, 8, 412. (2017)
https://doi.org/10.3389/fphar.2017.00412
[22] F. Fratini, G. Cilia, B. Turchi, and A. Felicioli, Asian Pacific Journal of Tropical Medicine, 9(9), 839-843. (2016)
[23] N. Ghanem, Res. J. Biotechnol. 6, 38–43 (2011)
[24] N. S. Al-Waili, Complementary Therapies in Medicine, 12 (1), 45-47. (2004)
https://doi.org/10.1016/j.ctim.2004.01.002.
[25] M. Kacaniova, N. Vukovic, R. Chelibo, P. Haskic, K. Rovna, J. Cubon, M. Dzugan, A. Pasternakeiwicz, Archives
of Biological Sciences 64 (3), 927-934 (2012) 10.2298/ABS1203927K
[26] S. L. Puleo and K. Keunen. Cosmet. Toiletries 106 (2): 83-89 (1991)
[27] S. Castaldo, and F. Capasso, Fitoterapia, 73 (Suppl 1) (2002), pp. S1-S6

7
Reproduced with permission of copyright owner. Further reproduction
prohibited without permission.