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The use of orange peel in the technology of

drinks fermentation
Olga Ivanchenko 1*, Anna Gargalyk2, Petr Balanov2, Irina Smotraeva2, and Nadezhda
Zhilinskaya1,3
1Peter the Great St. Petersburg Polytechnic University (SPbPU), 29 Politekhnicheskaya str., St.
Petersburg, 195251, Russia
2Faculty of Biotechnologies (BioTech), ITMO University, 49 Kronverksky Ave., Saint Petersburg,

197101, Russia
3Laboratory of Cancer Chemoprevention and Oncopharmacology, N.N. Petrov National Medical

Research Center of Oncology, St. Petersburg, Russia

Abstract. Nutrition is one of the most important factors determining

human health. Enrichment of food products with minerals, antioxidant
compounds, and vitamins are relevant today. In this work, dry orange peel
was used to enrich a fermented drink based on kvass wort with phenolic
compounds and vitamin C. The following indicators were determined: the
content of vitamin C, the amount of phenolic compounds and
anthocyanins, the antioxidant activity of orange peel dried to a moisture
content of 18%. The drink fermentation lasted 12 hours at 30 °C.
Secondary fermentation was carried out for 48 hours. At this stage, orange
peel was added in the amount of 10 g/l. During the experiment, the
concentration of dry matter content, the acidity of the medium, and the
content of ethyl alcohol were measured. Organoleptic indicators were
determined during the tasting of the finished product. In the finished drink,
the content of ethyl alcohol was 0.98%. It was found that the finished drink
with peel contains 4 times more phenolic compounds and 1.5 times more
vitamin C.

1 Introduction
Achievements of modern medicine indicate that nutrition is one of the most important
factors determining human health, endurance, immune status, and the rate of organism
adaptation. The problem of nutrition is important in the sports industry, since deviation
from the principles of optimal nutrition, deficiency or excess of any nutrients or
biologically active substances in the athlete's diet can lead to a decrease in physical
performance, slow recovery processes, metabolic disorders, and the appearance of
alimentary-dependent diseases.
Human nutritional ingredients are used in the formulations of common food products
such as yoghurts, candies, crispbreads, and other baked goods, not only in special and
healthy products. The Russian manufacturers have a considerable experience in introducing

* Corresponding author: [email protected]

© The Authors, published by EDP Sciences. This is an open access article distributed under the terms of the Creative Commons
Attribution License 4.0 (http://creativecommons.org/licenses/by/4.0/).
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plant-based additives into recipes and food production technologies [1,2]. In addition, many
people began to pay more attention to improving their own health. Therefore, food products
and drinks that have a beneficial effect on the human body came to the fore, and the
Russian food industry was reoriented. For example, now there are many beverage
formulations with various natural plant extracts [3-5].
Orange peel is a secondary raw material obtained during the industrial processing of
orange fruits in juice production technology. Although the peel has many useful substances
in its composition, it is mainly used for livestock feed [6,7]. The chemical composition of
the orange peel is very diverse. It includes a complex of essential substances, such as B
vitamins, various essential oils, phytoncides. The peel is also the richest source of vitamin
C, phenolic compounds, and carotenoids. The human body cannot store vitamin C, so one
should constantly get it from food.
The aim of this work was to develop a fermentation drink based on kvass wort enriched
with orange peel. The objectives of the study included:
1. Determination of the antioxidant activity of orange peel.
2. Determination of the content of vitamin C, polyphenolic compounds, and anthocyanins
in orange peel.
3. Development of a fermentation technology for a drink enriched with orange peel.
4. Assessment of physical and chemical parameters and organoleptic characteristics of the
finished product.

2 Materials and methods

2.1 Materials

The following materials were used in the work:

1. Kvass wort concentrate (Rospak, Russia);
2. Dry yeast "Safmoment" ("SAF-NEVA" Ltd., Russian enterprise of the Lessafre
Group France, St. Petersburg);
3. Oranges (Turkey).

2.2 Methods

2.2.1 Assessment of antioxidant potential

The amperometric method is used to determine the mass concentration of antioxidants. It is

based on measuring the amperage. When antioxidant molecules are oxidized, current
appears on the surface of the working electrode, which is converted into a digital signal.
Under the conditions of amperometric measurement, compounds containing hydroxyl
groups are well oxidized [8,9].
The total content of antioxidants was measured using a calibration plot for the
relationship between the intensity of the output signal on the concentration of quercetin.
The total content of antioxidants in terms of quercetin was determined by expression
(1):
CAk ·Vn ·n
CA=
mn
, (1)
where CA is the total content of antioxidants in terms of quercetin (mg/l);
CAk is the quercetin concentration, mg/dm3;
Vn is the sample volume, dm3;
n is the dilution factor;

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mn is the weight of the product sample, g.

2.2.2 Determination of the content of ascorbic acid

This analysis was carried out by the volumetric method, based on the state standard. First,
vitamin C was extracted with a hydrochloric acid solution. Then the solution was titrated
with sodium 2,6-dichlorophenolindophenolate until the color changed to light pink [10].

2.2.3 Determination of the content of polyphenolic compounds

For this analysis, the Folin-Denis method was used. It is based on the formation of colored
products during the oxidation of phenolic compounds with a phosphorus-molybdenum-
tungsten reagent in an alkaline medium. The alkaline medium was created with a saturated
sodium carbonate solution [11].
The total content of phenolic compounds was calculated by expression (2):
a∙V0 ∙P
X= , (2)
M
where X is the amount of phenolic compounds, mg/g;
a is the content of chlorogenic acid, determined by the calibration curve, mg/ml;
V0 is the total extract volume, ml;
P is the degree of dilution;
M is the sample weight, g.

2.2.4 Determination of the content of anthocyanins

This analysis was carried out by the spectrophotometric method. Firstly, 1 g of crushed raw
material was placed into a 100 ml flask with a thin section. Then, 30 ml of 70% ethanol
acidified with hydrochloric acid (1% by volume) was added. Next, the flask with the test
solution was attached to a reflux condenser and heated on a boiling water bath for 40 min.
After cooling to a temperature of 25±2 °C, the flask was weighed, its content was brought
to the initial mass with 70% alcohol, stirred, and filtered through a paper filter. 2 ml of the
studied filtrate was transferred into a 25 ml volumetric flask and adjusted with 70% alcohol
to the mark on the flask. Next, the optical density was measured using the UNICO-2802S
spectrophotometer (USA) [12].
The quantitative content of anthocyanins in terms of cyandin-3-o-glucaside and
absolutely dry raw material was calculated by expression (3):
A∙25∙30∙100
X=
100∙m∙1∙(1-W)
, (3)
where X is the amount of anthocyanins, mg/100 g;
A is the optical density of the test solution;
m is the mass of raw materials, g;
W is the weight loss during drying of raw materials, %;
100 is the specific absorption rate of cyanidin-3-O-glucoside

2.2.5 Acidity of the drink

The method for determining acidity is based on the titration of all acids in the analyzed
drink [3]. The total acidity of the drink (Kvol) was expressed in cm3 of 0.1 normal NaOH
solution per 100 cm3 and was calculated using expression (4):
Kvol = V·k, (4)

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where V is the volume of 0.1 normal NaOH solution that was used for titration;
k is dilution.

2.2.6 Determination of the mass fraction of ethyl alcohol

The analysis was carried out using the analyzer of alcoholic beverages "Kolos-2" (NPP
"Biomer", Russia). This analyzer is designed to measure the mass fraction of ethyl alcohol
in alcoholic beverages, low-alcohol products, and water-alcohol solutions.

2.2.7 Determination of the mass fraction of dry substances

The analysis was carried out by the refractometric method. This method is based on the
determination of the mass fraction of dry matter content on the scale of the IRF-454 B2M
refractometer at a temperature of 25±2 °C (KOMZ JSC, Russia).

2.2.8 Development of technology for a fermented drink with orange peel Citrus
sinensis

The fermentation drink recipe was developed based on kvass wort concentrate. Kvass wort
was prepared with dry matter content concentration of 3 and 4% (Table 1). All components
were added to water and mixed well.

Table 1. Drink components

Ingredient Quantity, g/ dm3
Kvass wort concentrate, 4%
of dry matter content:
Total 45.0
Added before fermentation 31.5
Sugar 40.0
Water 26.7
Sugar syrup, 60% of dry matter
content:
Total 66.7
Before fermentation 16.7
For blending 50
Based on the dry matter
Water content of the initial
wort
Orange peel 10

The yeast was removed by filtration after 12 hours of fermentation. The blending of the
drink was carried out at the stage of secondary fermentation. The rest of the sugar was
added to the kvass wort in the form of sugar syrup after the main fermentation stage. At the
same time, the orange peel in an amount of 10 g/l and the remaining kvass wort concentrate
were also added. The drink was left at a temperature of 4±2 °C for 48 hours. During this
time, the extraction of substances from the peel took place, the taste and aroma of the drink
were finally formed.

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3 Results and discussion

The peel is the outer pigmented layer of the citrus fruits, which contains many essential
compounds. The useful area of orange peel is relatively small, although it is known that
orange peel contains a lot of substances beneficial to the human body [13-16].
Firtsly, fresh oranges were washed with a brush under running warm water. Then, strips
of peel about 1–2 cm wide were cut with a knife. As it is known, humidity is one of the
indicators which determines the long-term storage of the product. Therefore, orange peel
was crushed and dried at 25 °C and 70 °C until the moisture content reached 18-20%.
Antioxidant activity was determined in the obtained dried samples (Table 2).

Table 2. Antioxidant activity of orange peel, mg/dm3

Antioxidant activity,
Drying temperature, °C
mg/dm3
25 18.3± 0.7
70 9.7±0.4

The study showed that a higher antioxidant activity was recorded in the sample dried at
25 °C, so this peel was used in further studies.
Natural antioxidants are vitamins C, E, P, A, carotenoids, flavonoids, aromatic hydroxy
acids, anthocyanins and other compounds [17,18].
A series of experiments was carried out to determine substances from the group of
natural antioxidants in the considered samples. This was a necessary step to study the
components that determine the antioxidant activity of the peel.
Vitamin C is a powerful antioxidant. It is hydrophilic, so it functions well in an aquatic
environment. It plays an important role in redox processes, has a protective effect in case of
colds, normalizes vascular permeability, and increases the body's resistance to various
infections. Theoretical and experimental studies prove the importance of vitamin C intake
in preventing the development of cancer [19].
Anthocyanins are a group of water-soluble pigments found in most plant species. They
are part of plant cells and cause pink, red, purple color of flowers and fruits of plants.
Anthocyanins have pronounced antioxidant, capillary-strengthening, and bactericidal
properties [20]. Therefore, the next step in this work was to determine the amount of
anthocyanins in the studied peel.
The results of peel examination are presented in table 3.

Table 3. The content of vitamin C, anthocyanins, phenolic compounds in the orange peel
Indicator Quantity
Vitamin C, mg/100 g 94.3 ±0.3
Anthocyanins, %. 0.23±0.03
Phenolic compounds, mg/g. 4.73±0.04
Antioxidant activity mg/l 18.3± 0.7

Considering the therapeutic and prophylactic properties of individual antioxidants, it

should be noted that their principal functions are performed in close synergy and complex
interaction. For example, fat-soluble antioxidants (vitamins A and E) perform the biological
function in membrane lipids and lipoproteins; water-soluble antioxidants (vitamin C and
bioflavonoids) perform the function in biological fluids of the body; trace elements are
indispensable cofactors of enzymes. Thus, it is impossible to discuss the isolated properties
of individual antioxidants. It is necessary to consider them as part of the entire antioxidant
system and in interaction with other compounds.

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3.1 Development of technology for a fermented drink with orange peel Citrus
sinensis

Two types of kvass wort were prepared during the development of the new drink. The first
sample contained 3% of dry matter, and the second one contained 4%. The fermentation of
the drink took place within 12 hours. Then a secondary fermentation was carried out. At
this time, orange peel was added in an amount of 10 g/l.
Further the drink acidity was determined. Acidity is one of the most important
indicators of the finished drink, as it affects its taste characteristics. The drink acidity
increases due to the organic acids (citric, succinic, fumaric, malic, etc) synthesized by
yeast. The dynamics of changes of the drink acidity with orange peel is presented in table 4.

Table 4. The dynamics of changes in the acidity of the drink

Initial
concentration of
Experimental option Stage Acidity, a.u.
dry matter content
in the wort
Starting 0.4±0.02
Control 12h. 2.2±0.02
After fermentation 2.7±0.02
3%
Starting 0.5±0.02
Sample with peel 12h. 2.0±0.02
After fermentation 2.5±0.02
Starting 0.6±0.02
Control 12h. 1.7±0.02
After fermentation 2.5±0.02
4%
Starting 0.7±0.02
Sample with peel 12h. 1.5±0.02
After fermentation 2.3±0.02

The concentration of ethyl alcohol was determined at the end of fermentation and gave
the following results.
1. Sample with an initial wort concentration of 3%.
The alcohol content was 0.95% in the control sample and 0.94% in the sample with
peel.
2. Sample with an initial wort concentration of 4%.
The alcohol content was 1.05% in the control sample, and 0.98% in the sample with
peel.
The intensity of fermentation is determined by the change in dry matter content. A
decrease in dry matter content from 3% to 1.7-2.2% was observed in the experimental and
control option at the fermentation stage. An increase in the dry matter content up to 3.3%
was registered after the addition of peel at the secondary fermentation stage. This can be
explained by the extraction of peel substances into kvass wort. A further decrease in the dry
matter content to 1.95% was observed in the control sample after 36 hours of the entire
technological process.
The content of vitamin C and the concentration of phenolic compounds were assessed in
the test samples. The results are presented in Table 5.

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Table 5. The amount of phenolic compounds and vitamin C in finished drinks

Content of Vitamin C
Initial wort
Experiment phenolic content,
concentratio
al option compounds, mg/100
n, %
mg/ml ml
3 1.10±0.07 5.0±0.1
Control
4 1.15±0.08 5.1±0.1
Sample with 3 4.55±0.10 6.1±0.09
peel 4 4.76±0.09 7.3±0.1

At the end of the study it was found that the concentration of kvass wort has a slight
effect on the change in the content of phenolic compounds, while orange peel showed a
significant influence on their amount.
A tasting assessment allows one to identify all the disadvantages and advantages of a
new product. Sometimes this is the only way to judge the quality of a sample. An important
contribution to the aroma and taste is made by its components. The stages and conditions of
the technological process also matter. 10 respondents participated in the tasting assessment
of kvass drinks. The scoring of the quality of finished drinks is given in Table 6.

Table 6. Organoleptic assessment of the drink in points

Initial wort concentration
3% 4%
Indicator Sample
Sample with
Control with Control
peel
peel
Sweetnes
4.3 4.5 4.3 4.7
s
Acidity 4.1 4.1 4.1 4.1
CO2
saturation 2.9 3.5 3.1 4.5
degree
Fullness
2.8 3.6 3.4 4.8
of taste
Bread
3.4 3.1 4.1 3.3
tone
Lack of
3.2 5.0 3.9 5.0
yeast tone
Orange
0 3.1 0 3.2
flavor

Table 6 shows that the introduction of orange peel affects not only the physicochemical
parameters of the product but also its organoleptic characteristics. The strongest influence
was registered in such indicators as fullness of taste, orange flavor, and degree of CO2
saturation, that strongly contrasts the finished drink with the control sample.
So, the sample with the initial dry matter content of 4% and orange peel concentration
of 10 g/dm3 was chosen as the best. This sample is enriched with the beneficial substances
of orange peel and has the most attractive characteristics for the consumers.

4 Conclusion
Currently, the most relevant development direction in the food industry is the creation of
products for systematic use that preserve and strengthen human health due to the presence

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of functional food ingredients in their composition. Such products are suitable for most
diets of all age groups of the healthy population, as they reduce the risk of developing many
diseases.
Based on literature data, the prospects of using Citrus sinensis orange peel in food
production technology were shown. It was determined that the content of vitamin C in the
studied orange peel was 94.3±0.3 mg/100g; the amount of phenolic compounds was
4.73±0.4 mg/g; the content of anthocyanins was 0.23±0.03% in terms of cyanidin-2-O-
glucoside and absolutely dry matter. It was studied how different modes of orange peel
drying affect the preservation of its antioxidant activity. The antioxidant activity of peel
dried at 25 °C and 70 °C was 18.3 ± 0.7 mg/dm3 and 9.7 ± 0.4 mg/dm3, respectively. The
tasting assessment of drink enriched with orange peel in an amount of 10 g/l showed that
the sample with an initial wort concentration of 4% was the most harmonious drink.

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