IDENTIFICATION OF TOTAL AROMAS OF PLANT PROTEIN SOURCES
Рубрики: RESEARCH ARTICLE
Аннотация и ключевые слова
Аннотация (русский):
Introduction. Due to the deficit and high cost of complete animal protein, the search and analysis of alternative sources is an actual scientific trend. Lentils is a good alternative to animal protein, but the pronounced bean smell and taste limit its full or partial use in food production. The aim of the work was to determine the total aromas of lentils when germinated to eliminate the bean taste and smell. Study objects and methods. The object of the study was brown lentil beans germinated under laboratory conditions. Samples of the equilibrium gas phase formed over samples of wet and sprouted beans were investigated. The analysis of total aromas was carried out on a laboratory odor analyzer MAG-8 (“electronic nose”) by the method of piezoelectric quartz micro-weighing with an array of sensors. Results and discussion. The study results showed qualitative and quantitative differences in the equilibrium gas phase over samples of wet and germinated grain. The quantitative analysis showed that the content of volatile compounds over sprouted grain is 12% less than over wet. The qualitative composition of the samples of wet and sprouted grain differed by 60%, which confirmed the influence of germination on the composition of the equilibrium gas phase and the possibility of eliminating bean odor. Testing showed that the use of pre-processed lentil grains allows to replace up to 50% of raw meat in minced products (minced food, chopped food) without changing the smell of the products. Conclusion. According to the results obtained, preliminary processing of lentils by germination will allow using this bean culture as an alternative source of animal protein to expand the range, and improve the quality of meat and dairy products.

Ключевые слова:
Lentils, germination, amino acid composition, biological value, total flavors, total analytic signal, the equilibrium gas phase
Текст
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INTRODUCTION
The lack of animal protein resources on the planet
arose a long time ago, and a global protein deficit in
human nutrition continues to grow. According to some
experts, for example, over next 20 years, the shortage of
meat will remain one of the global problems of mankind.
At the same time, in foreign and domestic
technologies of obtaining food products, there is
a positive experience in the complete or partial
replacement of expensive and difficultly reproducible
animal protein resources with vegetable ones, including
meat and dairy products [1–3].
The use of alternative sources of protein allows
manufacturers to simultaneously solve economic and
technological problems, such as reducing production
costs, stabilizing and improving the quality of meat
systems, increasing product yield, etc.
Currently, a consumer begins to find benefits
of consuming products with alternative sources of
protein, namely a lower cost and the ability to provide
themselves with healthy food in the required amount.
The experience of using vegetable proteins on an
industrial scale is mainly associated with imported
soy [4]. However, at the moment, during the unfolding
of measures to implement the provisions of the Food
Security Doctrine, either ensuring healthy nutrition, or
the development of domestic technologies, and rejection
of imports become relevant. Russian scientists have proved the benefits of legume proteins traditionally
grown in Russia (Saratov, Samara, Orenburg and Penza
regions and Altai Krai), the potential of which can be
significantly increased by pre-germination [1].
Among alternative sources of protein, legumes
occupy priority positions in terms of protein quantity
and quality. That is why they are preferred when
creating, for example, meat food systems [5, 6].
Among them, lentils, known for its healing properties,
should be distinguished. By the quantity and quality
of amino acids, the lentil protein is closest to the
first grade beef protein. Compared to other legumes,
lentils have a more balanced amino acid composition,
containing an increased amount of valuable vitamins,
macro-, microelements (including iodine), and less
oligosaccharides that cause intestinal flatulence.
Lentils is one of few cultures containing only one
digestive enzyme inhibitor that acts on trypsin (Table 1).
Lentils have good botanical properties, develop
and fruit well in Central Russia (Chernozemye, Volga
region).
The choice of lentils as an object of the study is
justified by the results of chemical analysis obtained by
Howell, according to which this culture has a number
of obvious advantages compared to other legumes [9].
However, the test results achieved in the conditions of
the “Kalacheevsky” meat factory showed the limited
use of lentils even in small quantities (5–8%) due to its
bean taste and smell. In this regard, germination is of
practical interest, not only to improve the balance of
the amino acid composition, and to increase the content
of micronutrients, but also to assess the possibility of
eliminating undesirable sensory properties.
Many researchers confirm that, as a result of
germination, a decrease in the oligosaccharide fraction
is observed (Fig. 1), additional vitamins are synthesized,
increasing the total nutritional and biological value
of the product [10–17]. However, the information
concerning change in organoleptic properties is
extremely insufficient, and ungeneralized [13–16]. Lentil
products (flour, concentrate, isolate) combine well and
replace food systems of animal origin (meat, dairy).
Our previous results proved that germinating
significantly improved the biological value of raw
materials, namely the content of proteins, vitamins, and
minerals. The amount of amino acids increased by a
factor of 1.5–2 (Fig. 1).
The amino acid composition of the protein becomes
more balanced, with the score close to the score of the
reference protein (Fig. 2). A significant increase in lysine
and tryptophan, the most valuable amino acids, can
be mentioned. Lysine is a deficient amino acid, which,
combined with vitamins, strengthens the immune
system, promotes calcium absorption from the intestine,
as well as contributes to cellular protein and bone tissue
formation. Tryptophan is involved in the serotonin (the
hormone of happiness) formation; mood, sleep quality,
level of pain threshold and susceptibility to various
irritants and inflammations depend on its concentration.
An increase in the content of minerals and vitamins,
as well as a decrease in the number of oligosaccharides,

an anti-nutritional factor of legumes, were also noted
(Table 2).
Identification of total aromas by sensory methods,
taking into account the nutritional value of lentils, is
especially important. All these indicators belong to
food chemistry, the most important section in the study
of food products, providing important information in
the selection of raw materials, and it also means for a
consumer a better applicability of the product in a food
system [17, 18]. The information obtained will allow
us to construct a product with properties (chemical
composition, appearance, taste, and smell) close to those
of meat.
The aim of the study was to assess the total aromas
of lentils during germination, to eliminate extraneous
odor by partial or complete replacement of animal
proteins in food systems of animal origin.
STUDY OBJECTS AND METHODS
The object of study was brown lentil beans
(State Standard 7066-77I) germinated in laboratory
conditions at Voronezh State University of Engineering
Technologies.
Grains of untreated lentils were germinated at 21–
23°C for 3–4 days, preventing their complete drying out.
The general chemical composition of sprouted lentils
is presented in Table 2, and the amino acid composition
of proteins in Fig. 1. It can be seen from the data that
germinated lentil grains have significant advantages in
the content of the most important nutrients.
The study of total aromas was carried out on a
MAG-8 laboratory (experimental) odor analyzer (Fig. 3)
with the electronic nose methodology, by the method
of piezoelectric quartz weighing with an array of
sensors [20–22]. We analyzed three samples: wet grain,
germinated grain and water.
I State Standard 7066-77. Lentil for human consumption (plateshaped).
Requirements for state purchases and deliveries. Moscow:
Izdatelʹstvo standartov; 2003. 8 p.
The sensor array consisted of eight sensors based
on BAW-type piezoelectric quartz resonators with an
oscillation frequency of 10.0 MHz and with diverse film
sorbents on electrodes.
Coatings are selected in accordance with the test
objective (possible emission from samples of various
organic compounds):
Sensor 1 – Multilayer Carbon Nanotubes, MCNT;
Sensor 2 – Polyethylene glycol succinate, PEGS;
Sensor 3 – Polyethylene glycolsebacinate, PEGSb;
Sensor 4 – Polyethylene Glycol Adipate, PEGA;
Sensor 5 – Polyethylene glycol-2000, PEG-2000;
Sensor 6 – Dicyclohexane-18-Crown-6, DCH18C6;
Sensor 7 – Twin-40, Tween;
Sensor 8 – Polyethylene Glycolphthalate, PEGP.
Grain samples were placed in glass tubes (10 g
in each), tightly closed, kept at room temperature
(20 ± 1°C) for at least 20 min to saturate the
equilibrium gas phase over the samples. Then, we
determined moisture content, which amounted
to 51.2%. 3 cm³ of the equilibrium gas phase
was taken through a membrane with individual
syringes and introduced into the detection cell.
The background of the array of sensors was from
15 to 30 Hz·s. The measurement time was 60 s,
the mode for recording sensor responses was uniform
with a step of 1 s, the optimal algorithm for presenting
Figure 2 Effect of germination on lentil amino acid score
Table 2 Effect of germination on chemical composition
of lentil seeds
Indicators Сontent in 100 g of product
Before germination After germination
Proteins, g 26.15 29.56
Fat g 1.2 1.1
Carbohydrates, g 53.7 41.06
including glucose 8.45 13.64
Oligosaccharides:
raffinose 0.9 0.5
stachyose 2.7 2.1
verbascose 1.4 0.8
Starch 33.8 24.12
Cellulose 3.65 3.04
Ash 3.65 3.31
Moisture 12.33 18.1
Minerals, mg
calcium 84.23 84.62
phosphorus 401.16 400.3
magnesium 78.9 76.3
iron 12.06 12.32
sodium 56.12 55.91
potassium 659.18 659.51
Vitamins mg
В1 0.5 0.78
В2 0.21 0.48
РР 1.8 2.21
С – 0.04
β-carotene 0.03 0.08

responses was based on the maximum responses of
individual sensors. The measurement error was 10%.
The total analytical signal is generated by using the
integrated signal processing algorithm of eight sensors
in the form of a “visual imprint”. To determine the total
composition of sample smell, we used the full “visual
imprints” of the peaks (the largest responses of eight
sensors), constructed from the maximum responses of
the sensors in the equilibrium gas phase of the samples
during the measurement time (no more than 1 min).
The similarity and difference in the composition of the
volatile odor fraction over the analyzed samples was
established [20]. Slight differences in the composition
of the gas mixture were established by comparing
the kinetic “visual imprints” constructed from the
responses of all the sensors recorded at different times
over the entire measurement interval. The nature of the
components mixture is more apparent in such analytical
signals. Both types of signals, as well as the area of the
figures are calculated automatically in the instrument
software.
The following criteria for assessing differences in the
smell of the analyzed samples are selected:
A qualitative characteristic - the form of a “visual
imprint” with characteristic distributions along the
response axes, was determined by the set of compounds
in the equilibrium gas phase.
Quantitative characteristics:
1) The total area of the full “visual imprint”
(SΣ, Hz·s) was used to estimate the total intensity of
the aroma proportional to the concentration of volatile
substances, including water. This parameter was
constructed from all signals of all sensors for the full
measurement time;
2) The maximum signal of sensors with the most
active or specific sorbent films (ΔFmax, Hz) was
applied to assess the content of individual classes of
organic compounds in the EGP by the normalization
method [21, 22];
3) The identification parameter (Aij) was used to
identify individual classes of compounds in a mixture.
This parameter was calculated from the signals of
the sensors in the analyzed samples and for standard
compounds.
Sensor responses were recorded, processed and
compared in the software of the MAG Soft analyzer.

RESULTS AND DISCUSSION
In the course of experimental studies (Table 3), the
total content of volatile compounds in the EGP was
found to correlate with the total analytical signal of the
“electronic nose” – the area of the “visual imprint” of
the response peaks.
Insignificant differences were found in the total odor
intensity over samples of wet and germinated grains,
however, the contribution to the total sorption response
of different classes of compounds is not equal. To
establish differences in the composition (qualitative and
quantitative) of the volatile odor fraction, we analyzed
the total content of readily volatile components in the
equilibrium gas phase over the samples (Fig. 4).
The shape of the “visual imprint” of the sensor
responses in the array showed insignificant differences
in the chemical composition of the equilibrium gas
phase over samples of wet and germinated grain. The
content of volatile compounds in the equilibrium gas
phase over germinated grain was less by 12% than over
wet grain.
Additionally, we noted the change in the quantitative
composition of the odor above the samples according
to the relative content of the main classes of volatile
compounds, evaluated by the normalization method
(Table 4).
Figure 3 A general view of the workplace with the MAG-8
analyzer
Table 3 “Visual imprint” area of the sensor signals (S1-S8) in the equilibrium gas medium above the samples
Samples S1-MCNT S2-PEGS S3-PEGSb S4-PEGA S5-PEG-2000 S6-DCH18 C6 S7-Twееn S8-PEGP Ssum, Hz·s
Wet grain 9 13 9 14 14 9 11 11 353
Germinated
grain
8 13 8 12 13 9 11 11 312
Water 18 21 15 24 23 17 21 22 1135
MCNTs – multilayer carbon nanotubes, PEGS – polyethylene glycol succinate, PEGSb – polyethylene glycol sebacinate, PEGA – polyethylene
glycol adipate, PEG-2000 – polyethylene glycol, DCH18C6 – dicyclohexane-18-Crown-6, Tween – Twin 40, PEGP – polyethylene glycolphthalate

Figure 4 “Visual prints” of the maximum sensor signals in
the equilibrium gas phase above the samples. The rotary axis
indicates the numbers of the sensors in the array (experimental
part), and the vertical – the responses of the sensors at a
particular point in time (ΔFmax, Hz)

After germination, a decrease in the intensity of
aroma was noted in the universal indicator, as well as
in the “O-containing”, “alcohols, ketones, amines” and
“alcohols, ketones, water” indicators.
Alcohols are the most commonly found compounds
in natural essential oils. As part of essential oils, they
do not only add a peculiar aroma, but also contribute to
the manifestation of antiseptic activity against bacterial
and viral infections, have analgesic, anesthetic and
tonic effects, as well as regulate hormonal activity.
The absence of toxicity is very important, therefore
essential oils with a predominant alcohol content
are relatively safe.
Natural essential oils with a high content of ketones
can cause side effects: neurotoxicity (negatively affect
the functions of the nervous system), embryotropic
effect (dangerous during pregnancy), and hepatotropic
effect (disrupt liver function).
Some amines are very toxic substances. Inhalation
of their fumes and contact with skin are dangerous.
Aliphatic amines affect the nervous system, as well as
cause violations of the permeability of the walls of blood
vessels and cell membranes, liver functions and the
development of dystrophy.
We found that grain samples differed in quality
composition. For a more visual presentation of the
results, a spectrum of identification parameters was
constructed (Fig. 5). The compositions of the equilibrium
gas phase can be considered identical if the spectra
coincide within the error (equal to or greater than 0.1).
Germinated grain contained fewer components in
quality composition than wet grain. To establish such
differences, the distribution by identification parameters
of their absolute differences from water was traced
(Fig. 6).
Differences by more than ± 0.1 units are significant
and indicate a different composition of compounds
in the equilibrium gas phase above the samples. In
Fig. 5 the allowable difference interval is marked with
black lines.
A significant change in the qualitative composition
of the sprouted grain compared to wet grain was
Table 4 Signal ratio of several sensors in the matrix for test samples
Samples S1-MCNT S2-PEGS S3-PEGSb S4-PEGA S5-PEG-2000 S6-DCH18 C6 S7-Twееn S8 -PEGP
Universal
sensor
N-containing Alcohols,
ketones, amines
Alcohols,
ketones, water
O-containing Alcohols, acids Aliphatic
acids
Ether
Wet grain 10 14.4 10 15.6 15.6 10 12.2 12.2
Germinated
grain
9.4 15.3 9.4 14.1 15.3 10.6 12.9 12.9
water 11.2 13 9.3 14.9 14.3 10.6 13 13.7
MCNT – multilayer carbon nanotubes, PEGS – polyethylene glycol succinate, PEGSb – polyethylene glycol sebacinate, PEGA – polyethylene
glycol adipate, PEG-2000 – polyethylene glycol, DCH18C6 – dicyclohexane-18-Crown-6, Tween – Twin, PEGP – polyethyleneglycolphthalate

Figure 6 Absolute differences in identification parameters for
grain samples compared to water. The X axis is the number of
identification parameters

established. Water was also very different from both
grain samples, which suggests that not only water
vapor, but also other organic volatile compounds were
present in the equilibrium gas phase. The qualitative
composition of the EGP above wet and germinated
samples differed significantly (for selected points –
by 60%).
In parallel, an analysis of the sensory characteristics
of the grain was carried out. A significant decrease in
the sharp smell of legumes after germination was found.
This makes it possible to reduce the legume smell, which
is a drawback when added to food products, or when
traditional raw materials are completely replaced.
CONCLUSION
Testing in laboratory and pilot production conditions
showed that the use of pre-processed lentil grains
would allow replacing up to 50% of raw meat in minced
products (ready-to-cook food, cupats) without changing
the smell of the products. Smell is easily masked by
spices and food additives. The products possess juiciness
and attractive appearance.
The conducted studies opened up new prospects for
the creation of meat and vegetable products enriched
with biologically active substances, that have the
possibility of wider use of domestic raw materials and
the development of import-substituting technologies for
healthy nutrition products.
Germinated lentils are supposed to be used both as
part of meat systems and as an independent ingredient
for salads, as well as when creating products that
simulate meat for fasting, or when creating enriched
extruded products for people who lose weight (bread,
bran, etc.).
CONTRIBUTION
L.V. Antipova – 40%, T.A. Kuchmenko – 30%,
A.A. Osmachkina – 20%, N.A. Osipova – 10 %.
CONFLICT OF INTERESTS
The authors declare no conflict of interest.

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