AgroLife Scientific Journal - Volume 10, Number 2, 2021
ISSN 2285-5718; ISSN CD-ROM 2285-5726; ISSN ONLINE 2286-0126; ISSN-L 2285-5718
FUNCTIONAL BIOPRODUCT "FucoSTEM" - EVALUATION
OF THE PRODUCT ON ORGANOLEPTIC, PHYSICOCHEMICAL,
BIOCHEMICAL AND MICROBIOLOGICAL INDICATORS
Kamelia LOGINOVSKA, Nikolay SOLAKOV, Maria DONEVA, Alexandar VALCHKOV
Institute of Cryobiology and Food Technologies, 53 Cherni Vrah Blvd, 1407, Sofia, Bulgaria
Corresponding author email: aleksandar.valchkov@abv.bg
Abstract
The authors present the results of organoleptic, physicochemical, biochemical and microbiological studies of a new
lyophilized functional bioproduct "FucoSTEM". The product is a nutritious concentrate developed on the basis of
buffalo colostrum. The inclusion in the composition of fruit (chokeberry) leads to an increase in the content of total
phenolic substances, anthocyanins and high antioxidant activity of the product. The obtained results prove that the new
product "FucoSTEM" has a balanced composition, preserved quality characteristics, high biological and energy value.
It is microbiologically pure and safe to use. These features define it as suitable for use by a wide range of users.
Key words: lyophilized bioproduct, colostrum, chokeberry, antioxidant activity, phenolic substances.
antimicrobial and others (Ćujić et al., 2018;
Sosnowska et al., 2018; Denev et al., 2014;
Hawkins et al., 2020). In the last few years,
research interest in the chemical composition
and healing potential of chokeberry fruits, their
potential health benefits and their clinical
efficacy has increased (Kulling and Rawel,
2008; Valcheva-Kuzmanova and Belcheva,
2006; Chrubasik et al., 2010). Chokeberry
fruits are used in the food industry for the
production of juices, nectars, various food
products and food supplements, and in
pharmacy as traditional medicines based on
healthy actions against influenza and immune
enhancer (Sidor et al., 2019; Kitryte et al.,
2017). Sidor et al. (2019) show that a diet
enriched with black chokeberry juice or extract
improves the lipid balance of people with
metabolic syndrome. Yamane et al. (2017) in
an experiment with feeding hypertensive rats
on a diet with 10% lyophilized black
chokeberry fruit, found that it causes a decrease
in their blood pressure.
Fucose (L-Fucose), as a rare monosaccharide,
finds applications in cosmetics, pharmaceuticals and as a dietary supplement. Fucose
acts as an immunomodulator, increases cellular
communication, suppresses tumor growth,
protects
against
respiratory
infections,
suppresses allergic skin reactions, kills bacteria
and helps the body in the fight against
INTRODUCTION
The increased interest in healthy and
environmentally friendly nutrition stimulates
the production and regular inclusion in the
daily menu of functional products. The
production of foods with a favorable impact on
health human status implies the use of
biologically active ingredients in the basic
formulations for these foods. Extensive
scientific and clinical research has highlighted
the role of colostrum from large horned cattle
as a functional ingredient due to its health
benefits. Interest in colostrum is growing due to
the high content of biologically active peptides,
immunological components and growth factors.
Scientists predict that colostrum-based products
may play a significant role in the functional
products market in the future (Mehra et al.,
2021; Bartkiene et al., 2020; Dzik et al., 2017;
Altomare et al., 2016).
The chokeberry (Aronia melanocarpa) is one
of the richest sources of dietary polyphenols
with
healthy
effects.
Anthocyanins,
proanthocyanidins, flavan-3-ol and flavonol
glycosides and phenolic acids are the main
polyphenols present in chokeberry. Chokeberry
extracts and fruits show various positive health
properties in animal and human studies, such as
antihyperlipidemic, hypoglycemic, hepatoprotective, antiproliferative, anti-inflammatory,
116
organoleptic, physicochemical,
and microbiological indicators.
infections (Roca, 2005; Garcia-Vaquero et al.,
2018; Vanhooren, 1999).
Xylitol represents a natural birch sugar, which
is applied as a substitute for ordinary sugar
(sucrose) in various products such as food
supplements, medicines, toothpaste and more.
Xylitol is metabolized insignificant by insulin,
so it has little effect on blood sugar. It is
absorbed more slowly than sucrose, has a low
glycemic index and is suitable for diabetics. It
has been found to be effective in improving the
symptoms of chronic rhinosinusitis (Riley et
al., 2015; Weissman et al., 2011).
Mannitol is used as a sweetener and medicine.
Characteristically, it raises blood glucose to a
lesser extent than sucrose and is poorly
absorbed by the intestine. It has anti-edema
effect, stimulates the functions of the urinary
system and intestinal peristalsis. It is suitable
for diabetics (Grenby, 2011; Lawson, 2007).
Beta-glucans represent complex soluble fibers
derived from the cell wall of bacteria, fungi,
yeast, seaweed and the aleurone layer of
cereals. β-glucans are well-known immune
modulators that activate the immune system.
They increase the body's resistance to bacterial
and fungal pathogens. Beta-glucans possess
antioxidant, antidiabetic, hypocholesterolemic
activity and probiotic function (Wani et al.,
2021; Colosimo et al., 2021).
Fructooligosaccharides (FOS) are low-calorie,
indigestible carbohydrates, prebiotics, which
encourage the growth of beneficial bacteria in
the host's intestinal tract while suppressing the
spread of potentially pathogenic bacteria. FOS
stimulate the health of the host by intensifying
peristalsis and digestion, increase the absorption
of essential minerals and ions (Mg2+ and Ca2+)
in the small intestine, regulate blood glucose
metabolism, lower the levels of serum triglycerides, phospholipids and cholesterol, etc.
Fructooligosaccharides are present in various
inulin-rich raw materials such as chicory,
Jerusalem artichoke and others (Singh et al.,
2020; Ojwach et al., 2020; Han et al., 2021).
In response to the increased consumer interest
in functional foods with additional positive
effects for prophylactic and therapeutic
purposes the lyophilized product "FucoSTEM"
has been developed and adapted for use by a
wide range of users. The main goal of the
present study is its analytical determination by
biochemical
MATERIALS AND METHODS
1. Organoleptic evaluation
The evaluation of the organoleptic qualities of
the new lyophilized product "FucoSTEM" was
performed on the indicators: appearance,
consistency, aroma and taste.
2. Physicochemical methods
water content (%) - on a hygrometer
“Sartorius”;
dry matter (%) - calculated as the
difference from the water content;
total protein content (%) - (BDS
6231);
content of total lipids (%) - (BDS EN
16932-3: 2018);
active acidity (pH) - with pH-meter
"Hanna";
total ash (%) - (BDS 9373-80);
reducing sugars (%) - by Schoorl
(Razola-Díaz et al., 2020).
The energy value per 100 g of product (kcal/
kJ) was calculated based on the chemical
composition.
3. Biochemical methods
Determination of antioxidant activity of
functional bioproduct "FucoSTEM" - The
method is spectrophotometric and is based on
analysis of radical scavenging activity with
DPPH to Trolox. Toward 0.3 ml solution on 0.2
mM DPPH is added 0.5 ml of MeOH (HPLC
99.5%) 0.5 ml of the test samples. The solution
thus obtained is homogenized and allowed to
stand in the dark at room temperature for 120
minutes. Absorption (A) is measured at 517 nm
(UV-VIS Spectrophotometer, Biochrom). The
total antioxidant activity was calculated from a
calibration curve (R = 0.9906) using Trolox as
a standard. The results were expressed relative
to Trolox concentration (mg/g).
Determination of total content of phenolic
substances
in
functional
bioproduct
"FucoSTEM" - A modified spectrophotometric method by Singleton and Rossi was
applied to quantitative assessment the total
content of phenols (Alhakmani et al., 2013;
Valyova et al., 2012). The total phenolic
content (TPC) was determined with FolinCiocalteu reagent against a gallic acid standard.
117
TPC is expressed as Gallic Acid (GAE)
equivalents in mg/g or mg/ml. The results of
the studies were expressed in GAE mg/g.
Determination of anthocyanins in a
functional bioproduct "FucoSTEM" - The
content of anthocyanins in the product was
quantified by a differential method (Lee et al.,
2005). The amount of anthocyanins is
presented as cyanidin-3-glucoside equivalents,
expressed in cyd eq mg/l and calculated by the
following formula:
Anthocyanin pigment (cyd eq mg/l) = A x
Mw x DF x 103 / Ɛ x L,
where:
A = (A520 nm - A700 nm) pH 1.0 (A520 nm - A700 nm) pH 4.5;
Mw (molecular weight) = 449.2 g/mol
for cyanidin-3-glucoside;
DF = dilution factor;
L = thickness of the cuvette layer in cm;
Ɛ = 26900 molar extinction coefficient
for cyanidin-3-glucoside;
103 = conversion factor from g to mg.
Determination of reducing sugars (glucose
and fructose) of product "FucoSTEM" - The
determination of reducing sugars (glucose and
fructose) is performed by the method of Luff
Schoorl (Taufik and Guntarti, 2016). The total
amount of sugars is determined by hydrolyzing
sucrose to glucose and fructose in the sugar
extract with acid. The results were expressed in
% content by weight of the sample.
4. Microbiological assessment
A general analysis of microbiological
insemination of the lyophilized product was
performed according by main microbiological
indicators:
Total number of mesophilic aerobic and
facultative anaerobic microorganisms CFU/g (BDS EN ISO 4833-2013);
Escherichia coli in 0.1 g of the product
(ISO 16649 - 1,2,3);
Pathogenic
microorganisms,
incl.
Salmonella (BDS EN ISO 6579: 2003)
in 25.0 g of the product;
Coagulase-positive staphylococci in 1.0
g of the product (BDS EN ISO 6888-12005);
Sulphite-reducing clostridia in 0.1 g of
the product (ISO 15213-2003);
Spores of microscopic molds and
yeasts, CFU/g (BDS ISO 6611-2006).
5. Statistical processing of results
The statistical processing of the results and
their graphic image was performed using the
software product MS Office Excel 2007.
RESULTS AND DISCUSSIONS
The
composition
of
the
bioproduct
"FucoSTEM" includes sources of constructive
and energy elements, as well as physiologically
active substances of different origin: buffalo
colostrum; chokeberry; fucose; natural birch
sugar (xylitol); mannitol; β-glucans; FOS
(fructooligosaccharides); sodium citrate.
The data on the organoleptic characteristics of
the new lyophilised product are presented in
Table 1.
Table 1. Organoleptic indicators of functional bioproduct
"FucoSTEM"
Product
Functional
bioproduct
"FucoSTEM"
Appearance
Lyophilic
product with
pale purple
color
Consistency
Aroma
Fine, quickly
soluble
Pleasant,
compositionspecific
Taste
Pleasant,
lactic
sour with
chokeberry
aroma
The lyophilized bioproduct "FucoSTEM" has a
fine texture, with a pleasant, compositionspecific aroma and color, quickly soluble,
prepared for direct consumption in the form of
a sachet.
Table 2 presents the main physicochemical
parameters of the lyophilized product.
Table 2. Physicochemical indicators of the lyophilized
bioproduct (n = 3)
Indicators
Water content, %
Dry matter, %
Total protein, %
Total lipids, %
Total ash, %
Reducing sugars, %
Active acidity - pH
Energy value / 100 g
product, kcal/kJ
Values for analysis
2.72 ± 0.02
97.28 ± 0.03
7.04 ± 0.01
4.38 ± 0.01
4.53 ± 0.02
73.0 ± 0.01
6.05 ± 0.01
per 100 g: 29.6 / 125.8
per 2 g:
0.6 / 2.55
The obtained data show that the lyophilized
product represents a food concentrate with a
residual moisture content of 2.72% after
lyophilization within limits of the norm (2-5%),
which is proof of an optimally conducted
technological process. The active acidity is
6.05, which largely predetermines the
preservation of enzymatic and biological
118
Table 4. Total content of phenolic substances in
functional bioproduct "FucoSTEM" (n = 6)
Sample
GAЕ, mg/g
Aronia (lyophilized juice)
7.04 ± 0.06
Bioproduct "FucoSTEM"
1.55 ± 0.07
activity in the product. The content of total
protein is mainly due to the main raw material
in the product - colostrum. Relatively low for
concentrated foods is the quantitative value of
total fat in the product - 4.38%.
The high content of reducing sugars is due to
the
imported
additional
nutrients
of
polysaccharide type. The high water-binding
ability of the fructooligosaccharide allows
interaction in a characteristic way with the
protein of the product. This interaction brings
mainly electrostatic in nature, as a result of
which a strong bond is formed between the
individual particles of the product and after
rehydration, the lyophilized product receives
the required density.
Based on the chemical composition, the energy
value of the obtained lyophilisate was
calculated (for 100 g: 29.6 kcal/125.8 kJ and
for 2 g: 0.6 kcal/2.55 kJ), as the main energy
carrier is the carbohydrate-protein complex.
The antioxidant activity of the product was
determined spectrophotometrically on the basis
of an analysis of the radical scavenging activity
with DPPH relative to Trolox. Trolox
Equivalent Antioxidant Capacity (TEAC) is a
measure of the antioxidant power based on
Trolox, measured in units called Trolox
equivalents (TE). The antioxidant activity of
chokeberry juice was also determined experimentally, as it is assumed that it is he who
imports in the final product the antioxidant
potential. According to the prescription composition of the product, chokeberry juice is 15%
of the total amount, which fully corresponds as
a ratio between the two tested samples. The
results are pronounced relative to Trolox concentration (mg/g) and are presented in Table 3.
As with antioxidant activity, so in the results
for the content of total phenols in the product,
the ratio between the indications of pure
lyophilized chokeberry juice and its percentage
content in the finished product is preserved.
The assumption that the main carrier of
phenolic substances is the added amount of
fruit (chokeberry) is confirmed.
Quantitative analysis of the content of
anthocyanins in the product and in chokeberry
juice was performed spectrophotometrically.
The results of the studies performed are
presented in Table 5 in Anthocyanin pigment,
cyd eq mg/l.
Table 5. Quantitative content of anthocyanins in
functional bioproduct “FucoSTEM” (n = 6)
Sample
Aronia (lyophilized juice)
Bioproduct “FucoSTEM”
The high content of anthocyanins in chokeberry
juice also determines the content available in
the product, which provides additional health
benefits.
The results of the tests performed to determine
the reducing sugars (glucose and fructose) of
the obtained functional product, expressed as a
percentage content of the sample weight, are
presented in Table 6.
Table 6. Percentage content of reducing sugars in
functional bioproduct "FucoSTEM" (n = 3)
Table 3. Antioxidant activity of functional bioproduct
"FucoSTEM" relative to Trolox (n = 6)
Sample
Aronia (lyophilized juice)
Bioproduct "FucoSTEM"
Anthocyanin pigment,
cyd eq mg/l
6232.9 ± 0.03
1022.8 ± 0.04
Sample
Bioproduct
"FucoSTEM"
TEAA mg/g
1.629350 ± 0.05
0.295321 ± 0.09
Glucose, %
Fructose, %
7.3 ± 0.02
7.8 ± 0.01
Table 7 presents the data from the performed
microbiological tests of the lyophilized
product.
The results of the conducted microbiological
analyzes prove that the indicators of the studied
lyophilized bioproduct “FucoSTEM” are in the
admissible norms for microbial contamination
and meet the requirements and criteria of BDS
ISO.
Quantitative assessment of the total content of
phenols in the product and in chokeberry juice
was performed. The total phenolic substance
(TPC) content was determined by a FolinCiocalteu reagent spectrophotometric method
relative to the gallic acid standard. The results
of the studies performed are presented in Table
4 in GAE mg/g.
119
Table 7. Values of microbiological contamination of
functional bioproduct "FucoSTEM"
Indicators
Total number of
mesophilic aerobic and
facultative anaerobic
microorganisms - CFU/g
Escherichia coli in 0.1 g
of product
Pathogenic
microorganisms,
including Salmonella in
2.0 g of product
Coagulase-positive
staphylococci in 1.0 g of
product
Sulphite-reducing
clostridia in 0.1 g of
product
Spores of microscopic
molds, CFU/g
control of purified colostrum as food supplement.
Journal of Chromatography B, 1028, 130-144.
Bartkiene, E., Lele, V., Sakiene, V., Zavistanaviciute, P.,
Ruzauskas, M., Stankevicius, A., Grigas, J.,
Pautienius, A., Bernatoniene, J., Jakstas, V., Zadeike,
D., Viskelis, P., Juodeikiene, G. (2020). Fermented,
ultrasonicated, and dehydrated bovine colostrum:
Changes
in
antimicrobial
properties
and
immunoglobulin content. Journal of Dairy Science,
103(2), 1315-1323.
Chrubasik, C., Li, G., Chrubasic, S. (2010). The clinical
effectiveness of chokeberry: a systematic review.
Phytother Res, 24, 1107-1114.
Colosimo, R., Mulet-Cabero, A. -I., Cross K.L., Haider,
K., Edwards, C.H., Warren, F.J., Finnigan, T.J.A.,
Wilde, P.J. (2021). β-glucan release from fungal and
plant cell walls after simulated gastrointestinal
digestion. Journal of Functional Foods, 83, 104543.
Ćujić, N., Savikin, K., Miloradovic, Z., Ivanov, M.,
Vajic, U.-J., Karanovic, D., Grujic-Milanovic, J.,
Jovovic, D., Mihailovic-Stanojevic, N. (2018).
Characterization of dried chokeberry fruit extract and
its chronic effects on blood pressure and oxidative
stress in spontaneously hypertensive rats. Journal of
Functional Foods, 44, 330-339.
Denev, P., Kratchanova, M., Ciz, M., Lojek, A., Vasicek,
O., Nedelcheva, P., Vojtek, L. (2014). Biological
activities of selected polyphenol-rich fruits related to
immunity and gastrointestinal health. Food
Chemistry, 157, 37-44.
Dzik, S., Miciński, B., Aitzhanova, I., Miciński, J.,
Pogorzelska, J., Beisenov, A., Kowalski, I.M. (2017).
Properties of bovine colostrum and the possibilities
of use. Polish Annals of Medicine, 24 (2), 295-299.
Garcia-Vaquero, M., Rajauria, G., Tiwari, B., Sweeney,
T., O’Doherty, J. (2018). Extraction and Yield
Optimisation of Fucose, Glucans and Associated
Antioxidant Activities from Laminaria digitata by
Applying Response Surface Methodology to High
Intensity Ultrasound-Assisted Extraction. Mar.
Drugs, 16 (8), 257.
Grenby, T.H. (2011). Advances in Sweeteners. Springer,
66.
Han, S., Pan, L., Zeng, W., Yang, L., Yang, D., Chen
G., Liang, Z. (2021). Improved production of
fructooligosaccharides (FOS) using a mutant strain of
Aspergillus oryzae S719 overexpressing βfructofuranosidase (FTase) genes. LWT, 146, 111346.
Hawkins, J., Hires C., Baker, C., Keenan, L., Bush, M.
(2020). Daily supplementation with aronia
melanocarpa (chokeberry) reduces blood pressure
and cholesterol: a meta analysis of controlled clinical
trials. Journal of Dietary Supplements, Aug 14, 1-14,
doi: 10.1080/19390211.2020.1800887.
Kitryte, V., Kraujaliene, V., Sulniute, V., Pukalskas, A.,
Rimantas Venskutonis, P. (2017). Chokeberry,
pomace valorization into foodingredients by enzymeassisted extraction: Processoptimization and product
characterization. Food Bioprod. Process, 105, 36-50.
Kulling, S.E., Rawel, H.M. (2008). Chokeberry (Aronia
melanocarpa) - a review on the characteristic
components and potential health effects. Planta Med,
74, 1625-1634.
Functional bioproduct
"FucoSTEM"
CFU/g
Log
3.1 х102
2.49
Not
established
-
Not
established
-
Not
established
-
Not
established
-
1.2 х101
1.07
CONCLUSIONS
The
composition
of
the
bioproduct
"FucoSTEM" includes sources of constructive
and energy elements, as well as physiologically
active substances of different origin colostrum, mono, polysaccharides, betaglucans, natural antioxidants and others.
High levels of content of total phenolic
substances, anthocyanins and high antioxidant
activity of the product have been established.
The assumption that the main carrier of these
properties is the added amount of fruit
(chokeberry) is confirmed.
Summarizing the results of the analytical
studies, it is established that the new product
"FucoSTEM" is a lyophilized concentrate with
preserved
quality
characteristics,
high
biological and energy value, microbiologically
pure and safe for use. These characteristics
define it as suitable for use for prophylactic and
curative purposes by a wide range of users.
REFERENCES
Alhakmani, F., Kumar, S., Khan, S.A. (2013).
Estimation total phenolic content, in-vitro antioxidant
and anti-inflammatory activity of flowers of Moringa
oleifera. Asian Pac J Trop Biomed, 3(8), 623-627.
Altomare, A., Regazzoni, L., Parra, X.M.P., Selmin, F.,
Rumio, C., Carini, M., Aldini, G. (2016). Set-up and
application of an analytical approach for the quality
120
Lawson, P. (2007). Mannitol. Blackwell Publishing Ltd.,
219–225.
Lee, K.W., Lee, H.J., Kang, K.-S., Lee, C.Y. (2002).
Preventive effects of vitamin C on carcinogenesis.
Lancet, 359, 172.
Mehra, R., Singh, R., Nayan, V., Buttar, H.S., Kumar,
N., Kumar, S., Bhardwaj, A., Kaushik, R., Kumar, H.
(2021). Nutritional attributes of bovine colostrum
components in human health and disease: A
comprehensive review. Food Bioscience,
40,
100907.
Ojwach, J., Kumar, A., Mukaratirwa, S., Mutanda, T.
(2020). Fructooligosaccharides synthesized by
fructosyltransferase from an indigenous coprophilous
Aspergillus niger strain XOBP48 exhibits antioxidant
activity. Bioactive Carbohydrates and Dietary Fibre,
24, 100238.
Razola-Díaz, M.C., Verardo, V., Martín-García, B.,
Díaz-de-Cerio, E., García-Villanova, B., GuerraHernández, E.J. (2020). Establishment of Acid
Hydrolysis by Box–Behnken Methodology as
Pretreatment to Obtain Reducing Sugars from Tiger
Nut
Byproducts.
Agronomy,
10(4),
477,
https://doi.org/10.3390/agronomy10040477.
Riley, P., Moore, D., Ahmed, F., Sharif, M.O.,
Worthington, H.V. (2015). Xylitol-containing
products for preventing dental caries in children and
adults. The Cochrane Database of Systematic
Reviews, 3, CD010743.
Roca, C. (2005). Exopolysaccharides enriched in rare
sugars:
bacterial
sources,
production,
and
applications. Front Microbiol., 6, 288.
Sidor, A., Drożdżyńska, A., Gramza-Michałowska, A.
(2019). Black chokeberry (Aronia melanocarpa) and
its products as potential health-promoting factors An overview. Trends in Food Science & Technology,
89, 45-60.
Singh, R.S., Singh, T., Kennedy, J.F. (2020). Enzymatic
synthesis of fructooligosaccharides from inulin in a
batch system. Carbohydrate Polymer Technologies
and Applications, 1, 100009.
Sosnowska, D., Podsędek, A., Redzynia, M., Kucharska,
A.Z. (2018). Inhibitory effect of black chokeberry
fruit polyphenols on pancreatic lipase – Searching for
most active inhibitors, Journal of Functional Foods,
49, 196-204.
Taufik, I.I., Guntarti, A. (2016). Comparison of
reduction sugar analysis method in cilembu sweet
potato (Ipomoea batatas l.) using luff schoorl and
anthrone method. Indonesian Journal of Medicine
and Health, 7(5), 219-226.
Valcheva-Kuzmanova, S., Belcheva, A. (2006). Current
knowledge of Aronia melanocarpa as a medicinal
plant. Folia Med, 48, 11-17.
Valiova, M., Stoyanov, S., Markovska, Y., Ganeva, Y.
(2012). Evaluation of in vitro antioxidant activity and
free radical scavenging potential of variety of Tagetes
erecta L. Flowers growing in Bulgaria. International
Journal of Applied Research in Natural Products,
5(2), 19-25.
Vanhooren, P.T. (1999). L-fucose: occurrence,
physiological role, chemical, enzymatic and
microbial synthesis, J. Chem. Technol. Biotechnol.,
74(6), 479-497.
Wani, S.M., Gani, A., Mir, S.A., Masoodi, F.A.,
Khanday, F.A. (2021). β-Glucan: A dual regulator of
apoptosis and cell proliferation. International Journal
of Biological Macromolecules, 182, 1229-1237.
Weissman, J.D., Fernandez, F., Hwang, P.H. (2011).
Xylitol nasal irrigation in the management of chronic
rhinosinusitis: a pilot study. The Laryngoscope. 121
(11), 2468-2472.
Yamane, T., Kozuka, M., Imai, M., Yamamoto, Y.,
Ohkubo, I., Sakamoto, T. (2017). Reduction of blood
pressure by aronia berries through inhibition of
angiotensin-converting enzyme activity in the
spontaneously hypertensive rat kidney. Functional
Foods in Health and Disease, 7, 280-290.
121