Kemerovo, Kemerovo, Russian Federation
Kemerovo, Kemerovo, Russian Federation
Kemerovo, Kemerovo, Russian Federation
Kemerovo, Kemerovo, Russian Federation
Kemerovo, Kemerovo, Russian Federation
Kemerovo, Russian Federation
Kemerovo, Kemerovo, Russian Federation
Introduction. Modern scientific research into the biochemical composition and medicinal value of plants makes it possible to use them as functional ingredients in food technology. The research objective was to test rose root (Rhodiola rosea L.) and scullcap (Scutellaria galericulata L.) for biologically active substances and their potential use in functional dairy products. Study objects and methods. The research featured biologically active substances (BAS) obtained from rose root and scullcap that grow in mountain areas or on rock outcrops along Siberian rivers. The BAS content was determined using high performance liquid chromatography (HPLC). The biologically active substances were screened and identified using HPLC, thin-layer chromatography (TLC), and infra-red identification (IR). The new functional products were based on whey and cottage cheese made from processed whole milk. Results and discussion. The analysis of Rhodiola rosea rhizomes and roots showed the following BAS content (mg/g): rosavin – 16.9, salidroside – 14.3, rosin – 5.04, rosarin – 2.01, and methyl gallate – 6.8. The roots of Scutellaria galericulata had the following BAS content (mg/g): scutellarein – 22.27, baicalin – 34.37, baicalein – 16.30, apigenin – 18.80, chrysin – 6.50, luteolin – 5.40, and vogonin – 3.60. Whey served as a basis for a new functional whey drink fortified with BAS isolated from Rhodiola rosea 100 mL of the drink included 50 mL of whey, 20 mL of apple juice, 0.1 mL of rose root concentrate, 3 g of sugar, 0.5 g of apple pectin, 04 g of citric acid, and 30 mL of ionized water. The content of phytochemical elements ranged from 0.11 ± 0.001 to 0.49 ± 0.08 mg/100 g. Cottage cheese served as a basis for another dairy product fortified with BAS obtained from Scutellaria galericulata. The formulation included 81 g of cottage cheese, 10 mL of cherry jam, 9 g of sugar, and 0.025 mL of scullcap concentrate. The content of biologically active substances in the finished product varied from 0.09 ± 0.02 for luteolin to 0.48 ± 0.11 for baicalin. The whey drink fortified with the BAS extracted from Rhodiola rosea and the cottage cheese product fortified with the BAS isolated from Scutellaria galericulata satisfied 40–45% and 55–60% of the reference daily intake for phenolic compounds, respectively. The obtained data made it possible to recommend the new functional foods for commercial production. Conclusion. A set of experiments was performed to isolate biologically active substances from Rhodiola rosea and Scutellaria galericulata. The research developed and tested formulations of two new functional products based on whey and cottage cheese.
Medicinal plants, Functional food, biologically active substances, whey, cottage cheese
INTRODUCTION
Public nutrition attracts attention of medical science
and affects the development of biotechnology in food
industry [1]. As a result, most solutions lie in the sphere
of functional products designed for particular groups
of population [2]. Miners, geologists, polar explorers,
astronauts, submariners, athletes, and programmers
are prone to various diseases as a result of adverse
working conditions. Unsocial working hours make
them vulnerable to diseases of digestive system, liver,
thyroid gland, cardiovascular system, and bones. Lowincome
families still experience the consequences of
unhealthy diet that lacks natural meat, dairy products,
and fresh vegetables. As a result, a lot of people suffer
from deficiency of proteins, vitamins, and other
biologically active substances. Functional products can
compensate for the missing elements as they are fortified
with biologically active substances of plant origin,
e.g. minerals, macro- and microelements, bioactive
peptides, enzymes, etc. [3, 4, 20].
As a rule, food habits are as old as the nations or
states they belong to. However, they were shaped not
only by the local flora, fauna, climate, soil fertility, water
availability, national traditions, and culture, but also by
the genetic ability of the people to digest certain types of
food [14–16]. Some researchers recommend introducing
ancient Eastern traditions to the achievements of
Western medicine. In fact, European diet includes less
than 2–3% of edible plants while in the East people
enjoy a variety of 1000 different edible species [17]. The
Japanese, whose life expectancy is one of the longest
in the world, consume equal amounts of meat and
vegetables [24].
European scientists believe that saturated fats and
cholesterol in meat can be reduced by introducing safe
fibers into processed foods [5, 6]. Nitrites and polycyclic
aromatic hydrocarbons (PAH) are often found in
processed meat products and can have a disastrous effect
on human health [7]. Functional ingredients extracted
from medicinal plants can significantly improve meat,
fish, and dairy products [8, 9, 16]. Russian food science
has achieved great success in developing new functional
dairy products based on whey, cottage cheese, and
buttermilk [14–16].
The relevance of the present research lies in the fact
that a few plant species are actually used in functional
products, including the rose root (Rhodiola rosea L.).
According to scientific sources, it is usually used in
herbal tea mixes, water tinctures, or wine products.
As for the scullcup (Scutellaria galericulata L.), this
plant is protected by law, and this is the first time it
has become focus of the attention of food science. Its
properties and prospects for functional food industry
remain understudied. Thus, the research objective was
to identify the biologically active substances that can be
extracted from these plants and study their potential for
the production of new functional foods based on whey
and cottage cheese.
STUDY OBJECTS AND METHODS
The present research featured biologically active
substances (BAS) extracted from two plants: the rose
root (Rhodiola rosea) and the skullcap (Scutellaria
galericulata).
The rose root can be found all over Russia, from its
European part to the Far East. It is especially abundant
on the fragmental soil of the Altai-Sayan mountain
systems. The plant proliferates on the variety of local
minerals and macro- and microelements. They add
unique medicinal properties to the phytochemical
composition of the plant organs [21, 22, 24]. In fact, the
rose root has nearly become extinct due to uncontrolled
herborization. As a result, it is now listed in the regional
endangered-species lists and in the Red Book of Russia.
The scullcap is endemic to Eastern Siberia: it grows
in the Tomsk and Kemerovo regions, in the Republic of
Tuva, in Khakassia, and in the Mongolian areas of the
Altai Mountains [13]. The plant prefers moist forest
woodlands, steep river banks, and sandy terraces.
The scullcap is a popular medicinal plant with unique
adaptogenic, antioxidant, apoptotic, and antiviral
properties. It is also known for its ability to inhibit
the development of free radicals in cells [23–26]. The
research featured aerial parts, rhizomes, and roots.
The content of BAS was determined using Shimadzu
LC-20 Prominense chromatography unit. The device
was equipped with a Shimadzu SPD20MA diode array
detector and a RID refractometric detector with a
Kromasasil C-18 250 × 4.6 mm column.
The TLC chromatography was performed
using Sorbfil PTCX-AF-A plates with subsequent
densitometry on a TLC Sorbfil plate. The experiment
involved a densitometer with a Sony photofixation
system (Handycam HDR-CX-405) purchased from
IMID LLC, Russia. Sulfuric acid and 25% ethanolic
solution of phosphoric-tungsten acid were used for
targeted derivatization. After that, photofixation was
performed at wavelengths of 254 and 365 nm in the
visible range. Elution was conducted in mobile phase
systems: chloroform – methanol – water (62:32:6) and
ethyl acetate – formic acid – glacial acetic acid – water
(100:11:11:26).
During the preparative stage, the chromatographic
zones were excised and subjected to further analysis. The
targeted BAS were screened and identified using HPLC,
TLC, and IR. The obtained statistical data were processed
using the Microsoft® Excel program. The tables show the
arithmetic mean values. All experiments were performed
in triplicates. The quantitative content of the BAS was
determined using calibration curves constructed in the
concentration range of 0.05–200 μg/mL.
The new functional products were based on whole
milk whey and cottage cheese.
RESULTS AND DISCUSSION
In the industrially developed regions of Siberia,
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Zaushintsena A.V. et al. Foods and Raw Materials, 2020, vol. 8, no. 1, pp. 163–170
public health is especially vulnerable. Its maintenance
requires an active use of biological resources in food
biotechnology [22, 23]. The present research featured
the content of BAS in the rhizomes and roots of the
rose root (Rhodiola rosea) harvested in the subalpine
zone of the Kuznetsk Alatau mountains (Figs. 1 and 2).
The BAS were isolated using chromatographic methods
(Figs. 1 and 2). Rosavin (peak 1) and salidroside (peak 3)
appeared to be the most abundant substances. Methyl
gallate (peak 5), rosin (peak 2), and rosarin (peak 4) also
proved significant. Rosavin, rosarian, and rosin belong
to phenylpropanoids.
These compounds possess a lot of beneficial
properties. First of all, they have scientifically proven
adaptogenic and antioxidant properties [17]. Phenylpropanoids
(rosavin, rosin, rosarin) are known to have
tonic, antiviral, and immunomodulatory properties.
Salidroside is regarded as one of the most promising
substances for solving gerontology problems. This fact
confirms the hypothesis that BAS extracted from rose
root can be used in functional food industry. The actual
value of rosavin was 16.9 mg/g, which exceeded other
BAS by 15.4–88.2%.
Baicalin has good antioxidant properties. It also
neutralizes oxidation processes and prevents the
formation of free radicals. Scutellarein and vogonin
exhibit mutual synergism and have anticonvulsant
and antitoxic properties. Luteolin and vogonin
have apoptotic, anti-inflammatory, and other useful
properties. The BAS complex obtained from the
Scutellaria genus is actively used for the treatment and
recovery of cancer patients.
The analysis of scullcap roots showed high
concentrations of the following BAS: baicalin (peak 13),
scutellarein (peak 6), baicalein (peak 17), apeginin (peak
18), chrysin (peak 14), luteolin (peak 16), and vogonin
(peak 7) (Figs. 3 and 4).
As for quantification, the content of BAS within this
group varied from 5.4 to 34.4 mg/g. Baikalin had the
biggest share compared with other BAS: 34.37 mg/g. Its
advantage over other components was 35.2–84.3%. The
Figure 1 Chromatogram of ethanol extract from rhizomes
and roots of Rhodiola rosea L.
Figure 2 Content of biologically active substances in Rhodiola
rosea L., mg/g
Figure 3 Chromatogram of ethanol extract from roots
of Scutellaria galericulata L.
Figure 4 Content of biologically active substances
in Scutellaria galericulata L., mg/g
30
25
20
15
10
5
0
mAU
0.0 2.5 5.0 7.5 10.0 12.5 15.0 17.5
min
1
2
3
4
5
Rosavin Salidroside Rosin Rosarian Methyl Gallate
800
600
400
200
0
mAU
0.0 5.0 10.0 15.0 20.0 25.0 30.0 35.0 40.0
min
Scutellarin Baicalin Baicalein Apeginine
Chrysin Luteolin Vogonin
Table 1 Formulation of the whey drink fortified with Rhodiola
rosea L. concentrate
Component
Amount
1 2 3 4 5 6
Whey, mL 70.0 60.0 50.0 70.0 60.0 50.0
Apple juice, mL 30.0 40.0 50.0 20.0 20.0 20.0
Sugar, g 3.0 3.0 3.0 3.0 3.0 3.0
Apple pecin, g 0.5 0.5 0.5 0.5 0.5 0.5
Concentrate of
Rhodiola rosea, mL 0.1 0.1 0.1 0.1 0.1 0.1
Lemon acid, g 0.04 0.04 0.04 0.04 0.04 0.04
Drinking water, mL – – – 10.0 20.0 30.0
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Zaushintsena A.V. et al. Foods and Raw Materials, 2020, vol. 8, no. 1, pp. 163–170
obtained data prove that both plants have good prospects
for functional food industry.
Whey and cottage cheese are high-protein dairy
products and are beneficial for human health. They
served as bases for formulations of two new functional
products: a whey drink fortified with BAS extracted
from rose root concentrate and cottage cheese fortified
with BAS extracted from scullcap roots.
The formulation of the whey drink included cottage
cheese whey, apple juice, sugar, rose root concentrate,
and drinking water. Citric acid served as a regulator
of acidity, while apple pectin was used as a stabilizer
(Table 1). We tested six formulations of the new
product. The first three samples had a different amount
of apple juice. The remaining three samples differed in
the amount of water, while the volume of apple juice
remained the same. Water affects sensory properties and
regulates the acidity of the finished product.
In order to determine the optimal formulation, the
drink underwent a sensory evaluation for appearance,
consistency, flavor, and color on a five-point
scale (Fig. 5).
Variants 3 and 6 received the highest score. When
they were compared with each other, preference was
given to variant 6. It had the highest sensory evaluation
both in terms of flavor and color. Therefore, variant 6
was selected for the production of the functional product.
The technological process for the whey drink
fortified with rose root concentrate included the
following stages: raw material delivery and sensory
evaluation, mixing the components, pasteurization,
cooling, bottling, packaging, and storage (Fig. 1).
At the first stage, the raw material was evaluated
according to the main quality indicators. Raw materials
that met the requirements of regulatory and technical
documentation passed on to the next stage. The initial
mix was made up of the main ingredients, i.e. whey
and drinking water, which entered the tank through a
pipeline. Apple juice and rose root concentrate were
introduced manually. The rose root concentrate was
a dense, homogeneous dark brown mass. The dry
ingredients, i.e. sugar, pectin, and citric acid, were
gradually added to the resulting solution. A continuously
working stirrer prevented lump formation. To suppress
the development of vegetative microorganisms, the mix
was pasteurized at 80–85°C for 15–20 s. The resulting
drink was cooled to 10°C, bottled, and capped in
uniform vessels.
Tables 2 and 3 show the content of BAS in the
finished product and the results of sensory, physicochemical,
and microbiological evaluation. All the
BAS introduced into the formulation of the functional
drink were represented in quantities that were found
sufficient for practical use. State-issued Recommended
(a) (b) (c)
Figure 5 Sensory evaluation of the whey drink: (a) formulations 1–3; (b) formulations 4–6; (c) formulations 3 and 6, which proved
optimal
Figure 6 Flow chart for the whey drink fortified with biologically
active substances extracted from Rhodiola rosea L.
Table 2 Biologically active substances in the functional whey
drink fortified with Rhodiola rosea concentrate L.
Component Content in the
concentrate, mg/g
Content in the finished
drink, mg/100 g
Rosavin 16.89 ± 2.11 0.31 ± 0.077
Salidroside 14.35 ± 2.52 0.49 ± 0.08
Rosin 5.04 ± 0.93 0.11 ± 0.001
Rosarin 2.01 ± 0.37 0.17 ± 0.012
Methyl gallate 6.8 ± 1.05 0.12 ± 0.032
Appearance and consisntency
Flavor Color
Appearance and consisntency
Flavor Color
Appearance and consisntency
Flavor Color
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Practice MP 2.3.1.1915-04 highlights the level of BAS
consumption. According to the data provided in the
document, the new functional drink satisfied 40–45% of
the reference daily intake for phenolic compounds and
phenylpropanoids. The performed evaluation of sensory,
physico-chemical, and microbiological properties of
the drink showed that it corresponded to another stateissued
standard – Technical Requirements 10.51.55-001-
02068309-2019.
The unique properties of skullcap, or Scutellaria
galericulata, have never become an object of food
technology. However, it is rich in flavonoids, and a
functional product fortified with its BAS will have a
beneficial effect on various systems of human body.
Using the above techniques, we obtained another
functional dairy product – cottage cheese fortified with
skullcap concentrate. The experiment involved five
variants: a control sample, two samples with cranberry
jam, and two samples with cherry jam.
Table 4 demonstrates the formulation, while Fig. 7
shows the flow chart for the producing of cottage cheese
enriched with skullcap concentrate.
The sensory analysis of the cherry jam samples
revealed good monogenicity, consistency, and
appearance in both variants. Variant 3 was given the best
scores for flavor (Fig. 7a). This sample contained 81 mL
of cherry jam, 9 g of sugar, and 0.025 mL of scull-cap
concentrate per 81 g of cottage cheese. The samples with
cranberry jam showed no significant differences. After
a comparative analysis of all the options, variant 3 was
announced best according to taste properties.
The technology for the new cottage cheese product
included the following stages: preparation of the raw
material, mixing, heating, homogenization, cooling,
packaging, and storage (Fig. 8). Raw materials were
evaluated according to the main quality indicators and
regulatory documentation. To prepare the mix, cottage
cheese was put into the kneading machine. Jam, sugar,
and scull-cap concentrate were added manually. The
obtained mix underwent a thermal treatment at 62°C
for 15–20 s to suppress the development of vegetative
microorganisms. To obtain a homogeneous texture, the
cottage cheese was homogenized at 62°C. After that,
the finished cottage cheese was cooled to 20°C and
packaged. The product was stored at 4 ± 2°C.
A biochemical analysis of the finished product
revealed sufficient quantities of BAS (Table. 5). Table 6
shows sensory, physico-chemical, and microbiological
indicators of the fortified cottage cheese.
Figure 7 shows the results of the sensory evaluation
of appearance, consistency, flavor, and color on a fivepoint
scale.
Table 3 Sensory, physico-chemical, and microbiological
indicators of the functional whey drink fortified with
Rhodiola rosea L. concentrate
Index Property
Appearance and texture Opaque liquid with slight phase
layering
Color Intrinsic, uniform
Taste and smell Characteristic, no extraneous flavors
and odors; tastes a little sour
Mass fraction of solids, % 9.7 ± 0.3
Mass fraction of fat, % 0.02 ± 0.03
Acidity, °Т 47.5 ± 0.8
Release temperature, °C 4 ± 2
Coliform bacteria, per
0,01 cm3
Not detected
Yeast and mold, CFU/cm3 ≤ 1,0×10–1
Pathogens, including
salmonella
Not detected
Table 4 Formulation of the cottage cheese fortified with
Scutellaria galericulata L. concentrate
Component Amount
1 2 3 4 5
Cottage cheese, g 91.0 86.0 81.0 86.0 81.0
Cherry jam, mL – 5.0 10.0 – –
Cranberry jam, mL – – – 5.0 10.0
Sugar, g 9.0 9.0 9.0 9.0 9.0
Concentrate of Scutellaria
galericulata, mL
0.025 0.025 0.025 0.025 0.025
(a) (b) (c)
Figure 7 Sensory evaluation of the cottage cheese: (a) formulations 2 and 3; (b) formulations 4 and 5; (c) control formulation 1 and
formulations 3 and 5, which proved optimal
Appearance and consisntency
Flavor Color
Appearance and consisntency
Flavor Color
Appearance and consisntency
Flavor Color
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Figure 8 Flow chart for the cottage cheese fortified with
biologically active substances extracted from Scutellaria
galericulata L.
Table 5 Main flavonoids in the cottage cheese fortified with
biologically active substances extracted from Scutellaria
galericulata L.
Component Content in the
concentrate, mg/g
Content in the finished
cottage cheese, mg/100 g
Scutellarein 22.27 ± 2.23 0.26 ± 0.019
Baicalin 34.37 ± 3.47 0.48 ± 0.11
Baicalein 16.3 ± 2.19 0.26 ± 0.019
Apigenin 18.80 ± 1.98 0.23 ± 0.019
Chrysin 6.50 ± 1.13 0.14 ± 0.012
Luteolin 5.40 ± 1.00 0.09 ± 0.02
Vogonin 3.60 ± 0.90 0.12 ± 0.014
Table 6 Sensory, physico-chemical, and microbiological
indicators of the cottage cheese fortified with biologically
active substances extracted from Scutellaria galericulata L.
Index Property
Appearance
and consistency
Homogeneous, pasty, soft
Color White, with the hue characteristic
of the introduced components
Flavor Pure, sour-milk, sweet, with
a touch of added ingredients
Moisture content, % 59.3 ± 3.9
Mass fraction
of protein, %
12.4 ± 0.7
Mass fraction of fat, % 3.8 ± 0.7
Acidity, °Т 149.3 ± 10.9
Release temperature, °C 4 ± 2
Lactic acid
microorganisms, CFU/cm3
1×107
Coliform bacteria,
per 0,01 cm3
Not detected
Yeast and mold, CFU/cm3 Not detected
Pathogens, including
salmonella
Not detected
Raw material delivery and quality
evaluation
Mixing
Thermal treatment
t = 62 ± 3°C, τ = 15–20 s
Homogenization
t = 62 ± 3°C
Cooling
t = 20 ± 3°C
Packaging
Storage
t = 4 ± 2°С
CONCLUSION
The present research established the content of
biologically active substances obtained from two
medicinal plants of the Kemerovo Region. It featured
the rhizomes and roots of Rhodiola rosea harvested in
the subalpine belt of the Kuznetsk Alatau mountains and
the roots of the Scutellaria galericulata harvested on the
rocky outcrops along the Tom’ River.
The biomass was tested for biologically active
substances and revealed good pharmacological
prospects, i.e. high antioxidant, anti-inflammatory,
antibacterial, antiviral, and apoptotic properties.
A set of experiments resulted in two formulations of
new functional dairy products: a whey drink fortified
with biologically active substances extracted from
Rhodiola rosea concentrate and cottage cheese fortified
with biologically active substances extracted from
Scutellaria galericulata.
CONTRIBUTION
The authors were equally involved in writing the
manuscript and are equally responsible for plagiarism.
CONFLICT OF INTEREST
The authors declare that there is no conflict of
interest related to the publication of this article.
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