Ulan-Ude, Ulan-Ude, Russian Federation
Ulan-Ude, Ulan-Ude, Russian Federation
Moscow, Moscow, Russian Federation
Ulan-Ude, Ulan-Ude, Russian Federation
Ulan-Ude, Ulan-Ude, Russian Federation
Introduction. Modern meat scientists are currently interested in combining meat raw materials with plant ingredients to enrich products with biologically active substances of natural origin, including antioxidants. In this regard, we studied the antioxidant capacity of a dry lingonberry marc extract introduced into meat paste and analyzed its effects on the product’s color and stability during storage. Study objects and methods. Our objects of study were a dry lingonberry marc extract originating in the Republic of Buryatia, forcemeat, and a ready-made paste in a casing. The extract was obtained by water-alcohol extraction using microwave irradiation. We investigated the physicochemical characteristics of the dry extract, including its contents of phenolic compounds, benzoic acid, and antioxidants. Results and discussion. During the experiment, we analyzed the extract’s effect on the paste’s total antioxidant capacity, coloring, and shelf life. The results showed that increasing the extract’s amount from 0.1% to 0.4% changed the color of the paste from gray-brown to purple-brown, respectively, due to anthocyanins. In further tests, we used a 0.2% concentration of lingonberry extract – the optimal amount that retained the usual brown color of the paste while increasing the content of antioxidant substances. Then, we analyzed the degree of fat oxidation in the paste samples made with and without sodium lactate during storage. According to the results, the lingonberry marc extract used without the acidity regulator and with it inhibited lipid oxidation by 12.7% and 20%, respectively, by neutralizing free radicals. Finally, we tested the presence of pathogenic microorganisms in the end products. We detected no E. coli bacteria in the samples and found an inhibited growth of mesophilic anaerobic and facultative anaerobic microorganisms due to the extract’s bactericidal effect established in Our earlier studies. Conclusion. Thus, our results indicated that the dry lingonberry marc extract introduced into meat paste increased the product’s total antioxidant capacity and improved its stability during storage.
Meat products, berry extract, lingonberry, paste, phenolic compounds, antioxidants, oxidation, peroxide value
INTRODUCTION
Introducing slaughter by-products into food
formulations and technology is a promising direction
in the meat industry that ensures a rational use of
protein raw materials. Modern meat scientists are
developing new products using heat-treated offal of farm
animals [1–4].
Such products include liver sausage, paste, head
cheese, and jellied meat. Meat pastes are especially
popular. They have a spreading consistency and can
be packaged in a casing or container. According to
the standards, pastes are classified into “meat pastes”
(category A) with at least 20% of muscle tissue and
“meat-containing pastes” (category B) with 0 to 20%
of muscle tissue. Pastes are affordable meat products
due to a lower cost of liver, skirt, lungs, kidneys, and
meat trimmings compared to meat. Our meat market
traditionally offers pastes in a casing that are popular
among students, schoolchildren, tourists, and passengers
on trains and planes. These products are made from
inexpensive protein-containing ingredients with a high
nutritional value and are packaged in small portions. According to literature, modern meat scientists
are interested in combining meat products (including
pastes) with plant ingredients to enrich the product with
biologically active substances of natural origin (microand
macroelements, vitamins, amino acids, antioxidants,
etc.) and increase its functional, technological, and other
properties.
For example, Gurinovich et al. formulated a meat
paste by combining animal protein with that of pine nut
oilcake. This way, the authors improved the functional
properties of meat systems and enriched the end product
with a plant-origin ingredient [5].
Bazhenova et al. mixed forcemeat with wheat flour
containing selenium, an essential trace mineral. The
authors described how they selected their method of
introducing selenium-enriched flour into the forcemeat.
They concluded that a 10–15% protein-fat emulsion
with selenized flour increased the functional and
technological parameters of forcemeat and provided
50–70% of our daily requirement of selenium [6].
Giro and Chirkova proposed enriching paste with
iron [7]. They aimed to develop functional meat-based
products for people predisposed to, or suffering from,
iron-deficiency anemia. Their study showed that offalbased
pastes enriched with chickpea could be used
to prevent disturbed hematopoiesis caused by iron
deficiency. These products contain highly bioavailable
microelements that help the body to quickly mobilize its
compensatory reactions.
Another study by Okuskhanova et al. looked into
the composition and properties of maral deer pastes
fortified with beans and protein. The authors developed
three formulations with varying amounts of the protein
fortifier and beans: no protein fortifier or beans; 15%
protein fortifier and 20% beans; and 25% protein
fortifier and 10% beans. The study showed that the
third formulation had a higher content of essential and
non-essential amino acids compared to the first two
variants [8].
Pastes from hypoallergenic horse meat and lamb
were formulated by Lyakh et al. with the addition of
dried dill and Polisorbovit-95, a biologically active
dietary supplement. According to their results, this
combination of ingredients improved the product’s
sensory and physicochemical properties [9].
As we can see, meat scientists have created various
formulations of pastes with plant ingredients rich in
biologically active substances.
Further, modern scientific literature shows increased
interest in studying antioxidant capacities of natural
plant ingredients in order to introduce them into food
products to improve their functional properties and
inhibit fat oxidation processes [10–21]. Antioxidants can
neutralize the destructive effects of free radicals on a
human body. Our antioxidant system is one of the main
mechanisms for stabilizing our adaptive potential. This
is especially important for people who live in adverse
environmental conditions and have an unbalanced diet
containing synthetic ingredients.
For example, Lisitsyn et al. studied the antioxidant
activity of aromatic plant extracts (black pepper,
rosemary, sage, and thyme) at the GORO Research
Center for Ecological Resources (Rostov-on-Don,
Russia). The scientists commercialized a new technology
for processing aromatic raw materials – supercritical
CO2 extraction. This technology produces extracts
with a significantly different composition from those
obtained in traditional ways. Supercritical extracts
contain a variety of terpene compounds, as well as
waxes, pigments, high molecular weight saturated
and unsaturated fatty acids, alkaloids, vitamins, and
phytosterols. These substances have high biological,
antimicrobial, and antioxidant activities. According
to the results, the highest and the lowest contents of
antioxidants were found in sage and black pepper
extracts (3.1% and 0.07%, respectively). It is generally
accepted that natural extracts with an antioxidant
content of at least 0.1% can be considered as a dietary
supplement with antioxidant properties. Therefore,
the authors recommended using sage, rosemary, and
thyme extracts as antioxidant ingredients for meat
products [10].
Another group of researchers, Zabalueva et al.,
looked at antioxidant contents in water-alcohol infusions
of medicinal plants, depending on the method of their
preparation. They found that the concentration of watersoluble
antioxidants in infusions obtained by maceration
did not differ significantly from those prepared by
ultrasound and an ultra-high frequency electromagnetic
field. The study showed the potential of using wateralcohol
infusions from rose hips and barberry fruits
as antioxidant supplements in the production of meat
products [15].
A wide range of plant materials (vegetables, fruits,
berries, and herbs), including wild plants, are introduced
into meat products in the natural form or as extracts,
infusions, and decoctions treated in various ways.
Edible and medicinal plants are collected, processed,
and utilized almost without waste. However, waste from
processing wild plants is not always used rationally,
being an environmentally friendly, renewable raw
material that could be used as a source of biologically
active natural substances. These wild plants include
lingonberries growing in Transbaikalia (east of Lake
Baikal) that are rich in biologically active compounds
with medicinal properties. Lingonberry leaves contain
phenolic glycosides (arbutin and methylarbutin),
vaccinine, lycopene, hydroquinone derivatives, acids
(ursulic, tartaric, gallic, quinic, and ellagic), tannin,
hyperoside, and other flavonoids. Lingonberries are rich
in sugars, ascorbic acid, carotene, and organic acids [22].
The chemical composition of lingonberry leaves and
fruits indicates a high antioxidant capacity of respective
products. In fact, lingonberries are processed in large
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quantities for juice production. However, their byproducts
– such as husks, pulp or marc – could also be
used as a source of biologically active substances. Some
authors propose enriching meat products (e.g. liver
paste) with fresh or dried lingonberry and cranberry
pulp [18, 19].
For example, Bitueva and Ayusheeva introduced
dried cranberry or lingonberry pulp, pre-crushed and
reconstituted, into ground meat products. The powdered
pulp was added at the stage of forcemeat preparation,
replacing 13–15% of bread. This method enriched the
meat products with biologically active substances [18].
In another study, Ivanova and Izosimova proposed
a formulation of meat paste with 19% polyfunctional
additives – lingonberry or cranberry marc. The marc
contains citric and malic acids that shift the medium’s
pH away from the isoelectric point, enhancing the
dissociation of the main and acid groups of protein, as
well as increasing bound moisture and the yield of the
end product. A high content of low-ester pectins in the
berry marc also contributed to the system’s stabilization.
Biologically active substances and antioxidants
increased the microbiological resistance of the meat and
plant paste. The product also had an improved vitamin
and mineral composition [19].
In our previous work, we prepared a water-alcohol
extract of lingonberry marc which was then dried [23].
The dry extract is a powder rich in biologically active
nutrients that is easy to store and transport. Due to a
high concentration of dry substances, the extract can
be introduced in small amounts into the formulation
of meat products, providing them with functional
properties and eliminating negative effects on the
product’s sensory and physicochemical characteristics.
Thus, it is extremely relevant to create products
based on a combination of meat and plant materials
to enrich them with micro- and macroelements,
vitamins, amino acids, and antioxidants. Many types
of plant materials contain a variety of compounds
with an antioxidant effect. Even low concentrations
of antioxidants in the human body can slow down or
prevent oxidation processes which are known to cause
premature aging and disease. Thus, we can inhibit fat
oxidation by introducing antioxidants into food products.
In view of the above, we aimed to develop a meat
paste formulation with an extract from lingonberry marc
(originating in Transbaikalia), as well as evaluate the
total content of antioxidants and effect on the color and
stability of the paste in the casing during storage.
STUDY OBJECTS AND METHODS
The objects of the study included a dry lingonberry
marc extract (DLME), forcemeat, and a ready-made
paste in the casing.
A dry extract of lingonberry marc was obtained in
accordance with Patent No. 2626565. Lingonberries were
pressed for juice and the remaining marc was placed on
a baking sheet in a 5–8 mm layer and then dried in an
oven with infrared radiation at 35–40°C for 40–50 min
to 10–15% moisture. The dried marc was crushed to a
powder state and subjected to extraction with a wateralcohol
solution using a microwave electromagnetic field
(700 W, 2450 MHz).
The water-alcohol extracts were filtered and
concentrated on a rotary evaporator under vacuum in
a water bath at temperature below 45°C until a syrup
consistency was reached (40–50% dry matter). The
syrup was then vacuum-dried at a temperature below
50°C for 3.5–4.5 h to obtain a powder with a residual
moisture of < 5%. Such process parameters do not cause
any qualitative changes in thermolabile substances.
They preserve maximum biological activity of active
substances and ensure high quality of the extract.
The main ingredients in the formulation of paste
samples were cheek meat, beef liver, meat trimmings,
soy isolate, and semolina. To evaluate the DLME effects
on forcemeat characteristics, samples were made with
0.1, 0.2, 0.3, and 0.4% DLME previously dissolved in
water in a ratio of 1:5. The paste was made in a casing
according to the traditional technology.
A DLME-free paste sample was used as a control.
To evaluate the shelf life of the paste, we conducted
two experiments. For the first experiment, we prepared
control and DLME samples and stored them for 14 days.
For the second experiment, we used a 0.2% acidity
regulator – sodium lactate (E325) and stored the samples
for 18 days.
To evaluate the samples’ antioxidant activity, we
performed amperometric measurement of the total
content of antioxidants in terms of quercetin. The test
samples were subjected to extraction with bidistilled
water to isolate water-soluble compounds with an
antioxidant effect. The total content of antioxidants was
measured on a Tsvet Yauza-01-AA analyzer. Quercetin
solutions were used to construct calibration graphs [24].
The extraction efficiency was determined by the amount
of phenolic compounds isolated spectrophotometrically
using the Folin-Ciocalteu reagent. The content of
benzoic acid was measured by the HPLC method.
The optical density of the colored aqueous extracts
of the test and control samples was determined by
the photocolorimetric method on a KFK-3-01 ZOMZ
photometer. This method is based on measuring the
polychromatic radiation of the visible part of the
spectrum. The dependence between light absorption
and the radiation wavelength is expressed by a curve
(spectrum) of light absorbed by this solution. In the
graph, wavelengths are plotted along the abscissa, while
optical densities are plotted along the ordinate.
The sensory evaluation of the paste samples was
carried out on a nine-point scale according to State
Standard 9959. The peroxide value was determined by
a method based on the interaction between fat oxidation
products (peroxides and hydroperoxides) and potassium
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iodide in a solution of acetic acid and chloroform,
followed by the quantification of iodine released in a
sodium thiosulfate solution by the titrimetric method
(State Standard R 51487-99I).
The microbiological parameters of the paste samples
were assessed according to State Standards R 50454-92II
and 9958-81III.
The experiments were performed in triplicate.
Statistical processing of the data was carried out in
Microsoft Excel.
RESULTS AND DISCUSSION
First, we analyzed the chemical composition of the
lingonberry marc extract originating in Transbaikalia
(Table 1).
As we can see, the main component of the
lingonberry marc extract is a group of phenolic
compounds (6.63%), including water-soluble pigments,
anthocyanins, and benzoic acid (1.34%). In a preliminary
study [25], we used the disk diffusion method and found
that the extract had antimicrobial activity, partly due to
the presence of benzoic acid with bactericidal properties.
Thus, introducing the lingonberry marc extract into
food products, namely meat, can inhibit the growth of
microorganisms.
Table 1 also shows a high total content of
antioxidants, including polyphenols, anthocyanins,
vitamin C, and other compounds (382.6 mg/g).
Anthocyanins (3.58%), accounting for half of all
phenolic compounds, give lingonberries bright
red or burgundy coloring. They include malvidins
and peonidins (polyphenolic compounds from the
flavonoid group) which contain mono- and diglycosides
decomposing into sugar and aglycon (anthocyanidins)
upon hydrolysis.
I State Standard R 51487-99. Vegetable oils and animal fats. Method
for determination of peroxide value. Moscow: Standartinform;
2008. 6 p.
II State Standards R 50454-92. Meat and meat products. Detection
and enumeration of presumptive coliform bacteria and presumptive
Escherichia coli (Reference method). Moscow: Standartinform;
2010. 7 p.
III State Standards 9958-81. Sausage products and meat products.
Methods of bacteriologikal analysis. Moscow: Izdatelʹstvo standartov;
2001. 14 p.
Anthocyanins are widely used in the food, medical,
pharmacological, and cosmetic industries. A daily intake
of brightly colored berries (160–2000 mg) leads to the
absorption of anthocyanins (0.005–0.1%), which can
have an antioxidant effect. Solutions of anthocyanins
neutralize almost all radical forms of oxygen and
nitrogen four times as efficiently as ascorbate or
α-tocopherol. Even low concentrations of antioxidant
substances can slow down or prevent oxidative
processes. For example, adding only 0.001–0.01% of
antioxidants to oil can slow down its oxidation for a long
time [26].
When dissolved, the lingonberry marc extract
retains its dark red color. When it is added to gray
non-nitrite forcemeat, the latter acquires a purple hue.
Anthocyanins are known to act as pigments and the
color of plants depends on their concentration, as well
as the medium pH. They are red in acidic media, purple
in neutral and blue in alkaline media. In this regard, we
studied how the concentration of the dry lingonberry
extract affected the forcemeat pH (Fig. 1).
The forcemeat pH decreased with the introduction
of the dry extract, while remaining closer to the neutral
region. The extract’s acidity was quite high (3.24, see
Table 1) due to the use of marc, whose biologically active
substances are better extracted into the solution than
those of the fruit juice. Therefore, the marc extract is
rich in acids (about 2.5%) – citric, malic, benzoic, oxalic,
acetic, glyoxylic, pyruvic, hydroxypyruvic, ketoglutaric,
ascorbic, and others, with the highest content of benzoic
acid (1.34%). This high concentration of acids provides
the extract with a low pH, so even very small amounts
of the dry extract can significantly decrease the
forcemeat pH.
Before the extract was introduced into the forcemeat,
it was pre-hydrated for uniform distribution. The DLME
concentrations of 0.1, 0.2, 0.3, and 0.4% reduced the
forcemeat pH by 1.49, 2.98, 3.7, and 4.47%, respectively.
However, the absolute value of the forcemeat pH
remained close to neutral, which did not affect the
functional and technological properties of the forcemeat
system.
Adding the DLME in concentrations from 0.1 to
0.4% affected the forcemeat color. To select the optimal
Table 1 Qualitative indicators of dry lingonberry marc extract
Indicator Meaning
Appearance loose mass
Taste and smell sweet and sour, tangy, with
lingonberry flavor
Color burgundy
Acidity, pH units 3.24 ± 0.08
Moisture, % 4.52 ± 0.08
Phenolic compounds, %
including anthocyanins
6.63 ± 0.04
3.58 ± 0.04
Benzoic acid, % 1.34 ± 0.02
Total antioxidants, mg/g 382.60 ± 8.70
Figure 1 Amounts of dry lingonberry marc extract
vs. forcemeat acidity
6
6.2
6.4
6.6
6.8
0 0.1 0.2 0.3 0.4
Forcemeat pH
Dry extract, %
0
30
60
90
0 Total antioxidants, mg/100 g
0.4
0.8
1.2
1.6
Optical density D
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concentration, we analyzed the color characteristics
of ready-made pastes in the casing after heat treatment
(Table 2).
As we can see, 0.1% and 0.2% DLME concentrations
did not change the habitual sectional color of paste, graybrown
or slightly darker. However, larger amounts of
the extract (even 0.3%) gave the paste a purple hue. This
change is associated with the presence of anthocyanins,
water-soluble plant pigments, in the extract.
Lingonberries may contain such anthocyanins as
cyanidin-3-galactoside, peonidin-3-galactoside, cyanidin-
3-arabinoside, peonidin-3-arabinoside, cyanidin-
3-glucoside, and others. However, their qualitative
composition depends on the growth conditions. Of great
importance for the color of plant pigments is the pH of
vacuoles where pigments accumulate. The same pigment
in different media can exhibit varying colors: yellowgreen
in an alkaline medium, purple in a neutral, and red
in an acidic medium [27].
In the DLME, just like in lingonberries, the medium
is strongly acidic (pH 3.24), so the color is bright red.
When the extract is introduced into the forcemeat,
whose medium is close to neutral, it acquires a purple
tint from water-soluble anthocyanins. The color,
however, depends on the concentration of pigments in
the forcemeat.
For the quantitative and qualitative analysis of watersoluble
compounds, we determined the optical density of
the paste samples with different DLME concentrations,
using the spectrophotometric method (Fig. 2).
Optical density is known to be directly proportional
to the concentration of compounds in a solution. The
scale of the abscissa did not show a significant difference
in the samples. However, a thorough analysis indicated
that the highest peaks for the control sample (curve 1),
0.1% DLME sample (curve 2), 0.2% DLME sample
(curve 3), and 0.3% DLME sample (curve 4) were at
wavelengths of 554 nm, 557 nm, 557 nm, and 559 nm,
respectively (Fig. 2). As we know, the spectral range
from 500 to 560 nm corresponds to purple, while that
from 560 to 575 nm to purple. Our study showed the
same results: the sample with 0.3% DLME had a violet
hue (Table 2).
Optical density along the ordinate axis characterizes
the color intensity. It means that the height of the peaks
corresponds to the concentration of dissolved substances
(polyphenols) in the samples. As we can see, the optical
density values for the control sample (curve 1), 0.1%
DLME sample (curve 2), 0.2% DLME sample (curve
3), and 0.3% DLME sample (curve 4) were 0.86, 0.92,
0.93, and 0.93, respectively. The results show that larger
amounts of the extract led to higher concentrations of
water-soluble compounds, having reached a maximum
on curve 4 (0.3% DLME sample).
Thus, we found that increasing the DLME
concentration to 0.3% provided the paste with a high
content of antioxidants, but added a purple hue to its
color due to the presence of anthocyanins, which might
spoil the product’s appearance.
Antioxidants, including phenolic compounds,
neutralize lipid peroxidation, all radical forms of oxygen
and nitrogen. Therefore, we analyzed the samples for the
total content of antioxidants (Fig. 3).
Figure 3 shows a correlation between increased
amounts of lingonberry marc extract and higher total
antioxidant capacity of the paste. According to the
results, the total antioxidants in the test samples with 0.1,
0.2, 0.3%, and 0.4% DLME was higher than that of the
control by 1.3, 1.5, 1.8, and 2.05 mg/g, respectively. The
extract antioxidant complexes were rich in polyphenols
Table 2 Paste color with different concentrations of dry
lingonberry marc extract
Paste samples Color in section
Control (without DLME) Gray-brown
Test (with DLME):
0.1% Gray-brown
0.2% Dark gray-brown
0.3% Brown with a light purple hue
0.4% Brown with a light violet hue
1 – control, 2 – 0.1% DLME sample, 3 – 0.2% DLME sample, 4 – 0.3% DLME sample
Figure 2 Optical density of paste samples with different concentrations of dry lingonberry marc extract
6
6.2
6.4
6.6
6.8
0 0.1 0.2 0.3 0.4
Forcemeat pH
Dry extract, %
0
30
60
90
0 0.1 0.2 0.3 0.4
Total antioxidants, mg/100 g
Dry extract concentration, %
0
0.4
0.8
1.2
1.6
700 680 660 640 620 600 580 560 540 520 500
Optical density D
Wavelength, nm
1 2 3 4
0
1
2
3
4
Peroxide value, mmol O/kg
0
1
2
3
0 7 10 15 18
Peroxide value, mmol O/kg
1
2
3
4
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(6.63%; Table 1). They also contained organic acids,
vitamins, lycopene, and other antioxidant compounds.
Further, we performed a sensory evaluation of the
paste on a nine-point scale to establish how the extract
affected the product’s consumer appeal (Table 3).
According to the results, small amounts of the dry
lingonberry marc extract did not significantly affect the
texture, smell, or taste of the end product. However, its
concentrations above 0.2% had a negative effect on the
paste color in section. As we could see in Table 2, the
sample with 0.3% DLME acquired a light purple hue and
that with 0.4% DLME, a light purple tint. The sensory
evaluation showed a concentration of 0.2% as optimal
since it did not spoil the characteristics of the end
product while enriching it with antioxidant compounds
(Fig. 3). Therefore, we used this concentration in further
studies. For a physicochemical analysis, we prepared a
control and a test sample with 0.2% DLME (Table 4).
The results showed that the dry lingonberry marc
extract did not affect the quality of the paste, but it
doubled the total content of antioxidants. The content of
phenolic compounds in the extract is the most important
indicator of its biological value, which determines its
antioxidant activity.
To assess the extract’s antioxidant capacity,
we studied the process of fat oxidation. For this,
we prepared a control and a test paste samples and
determined the peroxide value which characterizes the
accumulation of primary lipid decomposition products
during storage.
Storage periods were selected in accordance with
State Standard R 55334-2012IV (10 days for pastes in
polyamide casings and 15 days for pastes with acidity
regulators). Our experiment consisted of two tests. In
the first test, the control and the test samples (without
DLME and with 0.2% DLME, respectively) were
made without acidity regulators and stored for 14 days
(Fig. 4). In the second test, the control and the test
samples (without DLME and with 0.2% DLME,
respectively) contained 0.2% sodium lactate as an
acidity regulator and were stored for 18 days (Fig. 5).
The reason for this experiment was that acidity
regulators are necessarily used in production, especially
in summer, to increase the shelf life of perishable meat
products. Thus, the experiment could show the DLME
role in the inhibition of peroxidation of animal lipids.
The samples were stored under identical conditions, in
the dark at 2 ± 2ºС.
Figure 4 shows the effect of DLME on the process of
fat oxidation in the paste.
In Fig. 4, we can see an irreversible process of
fat oxidation with the accumulation of primary fat
decomposition products. Animal fats contained in the
paste undergo auto-oxidation or peroxidation. The
polyamide casing cannot completely prevent oxidation,
since it is caused by a complex of factors: oxygen, light,
positive temperature, unsaturated fatty acids, etc.
According to regulatory documents, the peroxide
value for a high-quality fat product, low-oxidized raw
materials, and fat raw materials should not exceed
0.5, 3.5, and 10 mmol of active oxygen per 1 kg of fat.
As we can see in Fig. 4, the peroxide value in the
control and test samples immediately after preparation
was 0.71–0.75 mmol O/kg. After six days of storage,
it increased 3.17 times in the control and 2.98 times
in the test sample, reaching 2.38 and 2.12 mmol O/kg,
respectively. We found that on day 6, the peroxidation
process in the test sample slowed down by 10.9%
compared to the control. After 10 days (the shelf life
for this type of product), the process of lipid oxidation
continued to intensify and the peroxide value increased
to 3.1 mmol O/kg in the control sample (6.2 as high as
IV State Standard R 55334-2012. Meat and meat containing pate.
Specifications. Moscow: Standartinform; 2014. 34 p.
Figure 3 Total content of antioxidants in paste samples with
different concentrations of dry lingonberry marc extract
0.4
0
30
60
90
0 0.1 0.2 0.3 0.4
Total antioxidants, mg/100 g
Dry extract concentration, %
620 600 580 560 540 520 500
Wavelength, nm
2 3 4
0
1
2
3
0 7 10 15 18
Peroxide value, mmol O/kg
Storage period, days
контроль опыт
Table 3 Sensory characteristics of paste samples with dry
lingonberry marc extract
Characteristic Control Test samples with DLME
0.1% 0.2% 0.3% 0.4%
Appearance 8.5 ± 0.2 8.5 ± 0.1 8.5 ± 0.2 8.3 ± 0.1 8.2 ± 0.1
Texture 8.4 ± 0.2 8.5 ± 0.2 8.4 ± 0.1 8.4 ± 0.2 8.3 ± 0.2
Color and
appearance
in section
8.6 ± 0.2 8.6 ± 0.1 8.6 ± 0.1 8.0 ± 0.1 7.4 ± 0.1
Smell and taste 8.7 ± 0.2 8.7 ± 0.1 8.7 ± 0.2 8.3 ± 0.2 8.1 ± 0.2
Table 4 Physicochemical characteristics of paste in casing
(0.2% DLME)
Characteristics Paste in casing
Test Control
Content, %
moisture 71.5 ± 0.1 71.8 ± 0.03
protein 13.6 ± 0.1 13.3 ± 0.1
fat 11.9 ± 0.05 12.1 ± 0.2
minerals 1.4 ± 0.02 1.2 ± 0.1
salt 1.6 ± 0.02 1.6 ± 0.02
Total antioxidants, mg/100 g 67.84 31.23
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the initial value) and to 2.7 mmol O/kg in the test sample
(3.8 times as high as the initial value). On day 10, the
difference between the control and the test samples
reached 12.9%.
Further analysis of the oxidation process showed that
after 12 days, the peroxide values in the control and the
test samples were 3.55 and 3.1 mmol O/kg, respectively.
After 14 days, the process accelerated and the values
reached 4.1 and 3.7 mmol O/kg, respectively. The
difference between the control and the test samples was
12.7% on day 12 and 9.7% on day 14. Thus, the steadily
lower peroxide value in the test sample, compared to the
control, indicated a decreased rate of lipid peroxidation
reactions throughout the whole storage period. This
result can be explained by the presence of DLME rich in
antioxidant compounds that can neutralize the effects of
free radicals playing a significant role in chain reactions
of lipid oxidation.
The first test showed that introducing DLME into
the paste made without acidity regulators helped to
slow down fat oxidation and increase the shelf life by
two days (total of 12 days without signs of oxidative
damage).
In the second test, the control (without DLME) and
test (0.2% DLME) samples contained 0.2% sodium
lactate as an acidity regulator (Fig. 5).
The growth of peroxide values indicated the
accumulation of fat oxidation products in the paste
samples with sodium lactate throughout storage.
However, we found a certain inhibition of the oxidation
process compared to the first test, in which the samples
were made without an acidity regulator. For example,
on day 10, the peroxide values of the DLME samples
without and with sodium lactate were 2.7 and 2.5 mmol
O/kg, respectively. Thus, we can see a synergistic effect
of sodium lactate and DLME antioxidant compounds
during fat oxidation in the paste.
Further, we compared the peroxide values in the
control and test samples with sodium lactate. We found
that after 15 and 18 days of storage, the peroxide value of
the test samples was 20% lower compared to the control,
which was significantly higher than in the samples
without sodium lactate (12.9%).
At this stage, we concluded that a combination of
lingonberry extract with sodium lactate produced a more
pronounced antioxidant effect. At the end of the storage
period (18 days), the peroxide values of the control and
test samples were 3.5 and 2.7 mmol of active oxygen per
1 kg of fat. This means that the paste’s shelf life could be
extended by three days.
Thus, our experiment showed that although DLME
contributed to the inhibition of lipid oxidation, its
synergism with sodium lactate could significantly slow
down these reactions.
The shelf life of paste containing a large amount
of water (71–72%) is affected by not only oxidative
Figure 4 Effect of dry lingonberry marc extract
on the peroxide value of paste without acidity regulators
during storage
0
700 680 660 640 620 600 580 560 540 520 500
Wavelength, nm
1 2 3 4
0
1
2
3
4
0 6 8 10 12 14
Peroxide value, mmol O/kg
Storage period, days
контроль опыт
0
1
2
3
0 7 10 15 18
Peroxide value, mmol O/kg
Storage period, days
control test контроль опыт
Figure 5 Effect of dry lingonberry marc extract on the
peroxide value of paste with sodium lactate during storage
0.3 0.4
extract, %
0
30
0 0.1 0.2 0.3 0.4
Total antioxidants, Dry extract concentration, %
640 620 600 580 560 540 520 500
Wavelength, nm
2 3 4
12 14
days
опыт
0
1
2
3
0 7 10 15 18
Peroxide value, mmol O/kg
Storage period, days
кcоoнnтtrрoоl л ь о tпesыtт
Table 5 Microbiological indicators of control and test samples during storage
Indicator Storage of paste without sodium lactate, days
DLME test samples Control
0 6 10 12 0 6 10 12
QMAFAnM, CFU/g 1.4×102 4.1×102 6.2×102 7.3×102 1.4×102 5.8×102 7.5×102 8.5×102
Coliforms in 1 g not detected
Indicator Storage of paste with sodium lactate, days
DLME test samples Control
0 7 15 18 0 7 15 18
QMAFAnM, CFU/g 1.4×102 4.5×102 8.4×102 1.6×103 1.4×102 6.1×102 1.0×103 2.1×103
Coliforms in 1 g not detected
257
Bazhenovа B.A. et al. Foods and Raw Materials, 2020, vol. 8, no. 2, pp. 250–258
processes, but also by the growth of microorganisms.
According to Table 1, the dry lingonberry extract is rich
in benzoic acid (1.34%) that has strong antimicrobial,
antiseptic, and bactericidal effects inhibiting decay
and fermentation processes. As a result, lingonberries
last quite a long time without canning. Also, previous
studies have proven the antimicrobial activity of DLME
added to bakery products [25].
In our study, we investigated a possibility of
inhibiting microorganisms in the DLME paste samples
with and without sodium lactate (Table 5).
As we can see, all the samples showed a growth of
microorganisms. However, it was less intensive in the
DLME test samples with and without sodium lactate,
compared to the controls. Thus, the presence of benzoic
acid with strong bactericidal action slowed down the
growth of microorganisms and had a positive effect on
the test samples’ shelf life.
As for pathogens, no E. coli bacteria were detected
in any of the test samples, which might be due to the
preliminary heat treatment of the raw materials and the
use of a polyamide casing that excludes the product’s
contact with air, containers or equipment.
CONCLUSION
Thus, our study showed that 0.2% of dry lingonberry
marc extract was the optimal amount to be introduced
into paste forcemeat. This amount increased the
nutritional and biological value of the paste and
maintained high consumer appeal. We found that the
extract provided the product with a high content of
polyphenols with antioxidant properties, including
anthocyanins. Rich in antioxidant compounds, the
extract inhibited fat oxidation in the paste and, in
combination with sodium lactate, produced a synergistic
effect on lipid peroxidation processes. In addition, the
dry lingonberry marc extract slowed down the growth
of microorganisms due to a high content of benzoic
acid with antimicrobial and bactericidal properties. The
integrated effect of the extract’s components extended
the shelf life of the paste in a casing by two or three
days.
CONTRIBUTION
Authors are equally related to the writing of the
manuscript and are equally responsible for plagiarism.
CONFLICT OF INTEREST
The authors declare that they have no conflict of
interest.
FUNDING
This study was part of State Assignment
No. 19.5486.2017/BCh commissioned by the Ministry
of Science and Highev Education of the Russian
Federation.
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