Визначення кратності екстракції листя зеленого чаю антиоксидантним методом
Aim. To determine the optimal extraction frequency of green tea leaves with 60 % ethanol by the antioxidant method.Materials and methods. Chun Myn green tea leaves were the object of the study, the raw material was collected in Anhui province (China) from March to April. Dry green tea leaves were st...
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| Published in: | Журнал органічної та фармацевтичної хімії |
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| Date: | 2022 |
| Volume: | 20 |
| Issue: | 1(77) |
| Pages: | 28-34 |
| ISSN: | 2518-1548 |
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| Language: | English |
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| author | Maslov, Oleksandr Yu. Komisarenko , Mykola A. Kolisnyk, Serhii V. Golik, Mykola Yu. Tsapko, Yevhen O. Akhmedov, Elshan Yu. |
| author_facet | Maslov, Oleksandr Yu. Komisarenko , Mykola A. Kolisnyk, Serhii V. Golik, Mykola Yu. Tsapko, Yevhen O. Akhmedov, Elshan Yu. |
| author_institution_txt_mv | [
{
"author": "Oleksandr Yu. Maslov",
"institution": "National University of Pharmacy of the Ministry of Health of Ukraine",
"orcid": ""
},
{
"author": "Mykola A. Komisarenko ",
"institution": "National University of Pharmacy of the Ministry of Health of Ukraine",
"orcid": ""
},
{
"author": "Serhii V. Kolisnyk",
"institution": "National University of Pharmacy of the Ministry of Health of Ukraine",
"orcid": ""
},
{
"author": "Mykola Yu. Golik",
"institution": "National University of Pharmacy of the Ministry of Health of Ukraine",
"orcid": ""
},
{
"author": "Yevhen O. Tsapko",
"institution": "National University of Pharmacy of the Ministry of Health of Ukraine",
"orcid": ""
},
{
"author": "Elshan Yu. Akhmedov",
"institution": "National University of Pharmacy of the Ministry of Health of Ukraine",
"orcid": ""
}
] |
| author_sort | Maslov, Oleksandr Yu. |
| baseUrl_str | https://ophcj.nuph.edu.ua/oai |
| collection | OJS |
| container_end_page | 34 |
| container_issue | 1(77) |
| container_start_page | 28 |
| container_title | Журнал органічної та фармацевтичної хімії |
| container_volume | 20 |
| datestamp_date | 2026-08-24T13:41:04Z |
| description | Aim. To determine the optimal extraction frequency of green tea leaves with 60 % ethanol by the antioxidant method.Materials and methods. Chun Myn green tea leaves were the object of the study, the raw material was collected in Anhui province (China) from March to April. Dry green tea leaves were standardized according to the European Pharmacopeia 9.0. Spectrophotometry was used to quantify biologically active substances. The antioxidant activity was determined by the potentiometric method. Potentiometric measurements were performed on a HANNA 2550 pH meter (Germany) with a combined platinum EZDO 50 PO electrode (Taiwan). A UV-1000 spectrophotometer (China) was used to measure the optical density.Results and discussion. The total content of phenolic compounds was 9.60 ± 0.17, 1.30 ± 0.03 and 0.12 ± 0.002 %, catechins – 9.20 ± 0.18, 1.20 ± 0.02 and 0.07 ± 0.002 %, flavonoids – 0.27 ± 0.005, 0.04 ± 0.001, 0.005 ± 0.001, hydroxycinnamic acids – 0.49 ± 0.01, 0.07 ± 0.002 and 0.007 ± 0.001 %, dry residue – 10.75 ± 0.11, 1.59 ± 0.02 and 0.15 ± 0.002 %, the antioxidant activity was 474.08 ± 9.48, 67.70 ± 1.35 and 7.01 ± 0.14 mmol-equiv mdry res-1 for the first, second and third extraction, respectively. According to the results obtained, the optimal number of extractions of the raw material with 60 % ethanol was found to be two.Conclusions. The dynamic of extractions of biologically active substances of green tea leaves has been studied by triple extraction of the raw material to find the optimal extraction frequency; for the first time, a method for determining the extraction frequency based on the antioxidant activity of the extracts has been developed and proposed. It has been found that the optimal extraction rate is 2 times. The results obtained will be used in the further production of herbal medicines, dietary supplements, and cosmetic products with a green tea extract. |
| doi_str_mv | 10.24959/ophcj.22.252320 |
| first_indexed | 2025-07-17T13:01:19Z |
| format | Article |
| fulltext |
ISSN 2308-8303 (Print) / 2518-1548 (Online) 28
Original research
http://ophcj.nuph.edu.ua
UDC 663.951:581.45:54.062:543.42
O. Yu. Maslov, M. A. Komisarenko, S. V. Kolisnyk, M. Yu. Golik, Ye. O. Tsapko,
E. Yu. Akhmedov
National University of Pharmacy of the Ministry of Health of Ukraine,
53, Pushkins’ka str., Kharkiv, 61002, Ukraine
Determination of the extraction frequency of green tea leaves
by the antioxidant method
Abstract
Aim. To determine the optimal extraction frequency of green tea leaves with 60 % ethanol by the antioxidant method.
Materials and methods. Chun Myn green tea leaves were the object of the study, the raw material was collected in Anhui
province (China) from March to April. Dry green tea leaves were standardized according to the European Pharmacopeia 9.0.
Spectrophotometry was used to quantify biologically active substances. The antioxidant activity was determined by the
potentiometric method. Potentiometric measurements were performed on a HANNA 2550 pH meter (Germany) with a com-
bined platinum EZDO 50 PO electrode (Taiwan). A UV-1000 spectrophotometer (China) was used to measure the optical
density.
Results and discussion. The total content of phenolic compounds was 9.60 ± 0.17, 1.30 ± 0.03 and 0.12 ± 0.002 %, catechins –
9.20 ± 0.18, 1.20 ± 0.02 and 0.07 ± 0.002 %, flavonoids – 0.27 ± 0.005, 0.04 ± 0.001, 0.005 ± 0.001, hydroxycinnamic acids –
0.49 ± 0.01, 0.07 ± 0.002 and 0.007 ± 0.001 %, dry residue – 10.75 ± 0.11, 1.59 ± 0.02 and 0.15 ± 0.002 %, the antioxidant activity
was 474.08 ± 9.48, 67.70 ± 1.35 and 7.01 ± 0.14 mmol-equiv mdry res
-1 for the first, second and third extraction, respectively.
According to the results obtained, the optimal number of extractions of the raw material with 60 % ethanol was found to be two.
Conclusions. The dynamic of extractions of biologically active substances of green tea leaves has been studied by triple extrac-
tion of the raw material to find the optimal extraction frequency; for the first time, a method for determining the extraction
frequency based on the antioxidant activity of the extracts has been developed and proposed. It has been found that the
optimal extraction rate is 2 times. The results obtained will be used in the further production of herbal medicines, dietary
supplements, and cosmetic products with a green tea extract.
Keywords: green tea; leaves; analysis; extract; extraction; antioxidant activity
О. Ю. Маслов, М. А. Комісаренко, С. В. Колісник, М. Ю. Голік, Є. О. Цапко, Е. Ю. Ахмедов
Національний фармацевтичний університет Міністерства охорони здоров’я України,
вул. Пушкінська 53, м. Харків, 61002, Україна
Визначення кратності екстракції листя зеленого чаю антиоксидантним методом
Анотація
Мета. Визначити оптимальну кратність екстракції листя зеленого чаю 60 % етанолом антиоксидантним методом.
Матеріали та методи. Об’єктом дослідження було листя зеленого чаю сорту Чун Мін, зібране в провінції Анхуй (Китай)
з березня до квітня. Сухе листя було стандартизовано відповідно до Європейської фармакопеї 9.0. Для кількісного ви-
значення біологічно активних речовин використовували спектрофотометрію. Для визначення антиоксидантної актив-
ності використовували потенціометричний метод. Потенціометричні вимірювання виконували на pH-метрі HANNA
2550 (ФРН) із комбінованим платиновим електродом EZDO 50 PO (Тайвань). Для вимірювання оптичної густини ви-
користовували спектрофотометр UV-1000 (Китай).
Результати та їх обговорення. Сумарний вміст фенольних сполук становив 8,60 ± 0,17, 1,30 ± 0,03 і 0,11 ± 0,002 %, кате-
хінів – 9,20 ± 0,18, 1,20 ± 0,02 і 0,07 ± 0,002 %, флавоноїдів – 0,27, 0,04 і 0,005 %, гідроксикоричних кислот – 0,49 ± 0,01,
0,07 ± 0,002 і 0,007 ± 0,001 %, сухий залишок – 10,75 ± 0,11, 1,59 ± 0,02 і 0,15 ± 0,002 %, антиоксидантна активність ста-
новила 474,08 ± 9.48, 67,70 ± 1.35 і 7,01 ± 0.14 ммоль-екв. mсух.зал.
-1 для першої, другої і третьої екстракції відповідно.
Одержані результати свідчать, що двократна екстракція досліджуваної сировини 60 % етанолом є оптимальною.
Висновки. Для визначення оптимальної кратності екстракцій було проведено дослідження динаміки екстракцій біо-
логічних активних речовин листя зеленого чаю шляхом трикратної екстракції сировини, а також уперше розроблено
ISSN 2308-8303 (Print) / 2518-1548 (Online) 29
Journal of Organic and Pharmaceutical Chemistry 2022, 20 (1)
й запропоновано метод визначення кратності екстракції, заснований на визначенні антиоксидантної активності екс-
трактів. Виявлено, що двократна екстракція є оптимальна. Отримані результати будуть використані для створення
фітопрепаратів, дієтичних добавок та косметологічної продукції з екстрактом зеленого чаю.
Ключові слова: зелений чай; листя; аналіз; екстракт; екстракція; антиоксидантна активність
Citation: Maslov, O. Yu.; Komisarenko, M. A.; Kolisnyk, S. V.; Golik, M. Yu.; Tsapko, Ye. O.; Akhmedov, E. Yu. The study of green tea leaves
biological active substances dynamic extraction. Journal of Organic and Pharmaceutical Chemistry 2022, 20 (1), 28 – 34.
https://doi.org/10.24959/ophcj.22.252320
Received: 8 January 2022; Revised: 1 February 2022; Accepted: 10 February 2022
Copyright© 2022, O. Yu. Maslov, M. A. Komisarenko, S. V. Kolisnyk, M. Yu. Golik, Ye. O. Tsapko, E. Yu. Akhmedov. This is an open access
article under the CC BY license (http://creativecommons.org/licenses/by/4.0).
Funding: The work is a part of the research of the National University of Pharmacy on the topic “Organic synthesis and analysis of bio-
logically active compounds, drugs development based on synthetic substances” (the state registration No. 01144000943; the research
period 2019 – 2024).
Conflict of interests: The authors have no conflict of interests to declare.
■ Introduction
Oxidative stress is a part of many diseases,
such as diabetes mellitus, cancer, atherosclero-
sis, Alzheimer’s disease, and chronic obstructive
pulmonary disease [1]. Literature data evidence
represents that oxidative stress is caused by the
imbalance between the production and degrada-
tion of reactive oxygen species (ROS). Examples
of ROS include free radicals, such as the super-
oxide radical (O2
•-), hydroxyl radical (OH•), and
nitric oxide radical (NO•) [2].
The defense system of the human body in-
volves endogenous enzymes inactivating ROS.
The members of this group are superoxide dis-
mutase, catalase, and glutathione peroxidase [3].
However, there are cases when the endogenous
defense system is exhausted by chronic diseas-
es, stress, and not a good diet. As a result, it re-
quires exogenous antioxidants, such as phenolic
compounds, carotenoids, vitamin E, and amino
acids. Among them, the leading position in the
top of antioxidants is taken by catechins – a sub-
group of phenolic compounds [4].
Green tea leaves are the major source of cate-
chins. The content of catechins there varies from
25 to 35 % [5]. Moreover, green tea leaves contain
other subgroups of phenolic compounds. Thus, fla-
vanols (1 – 2.5 %), flavanones (1.5 – 3 %), phenolic
acids (2 – 5 %); caffeine (1.5 – 2.5 %), amino acids
(1 – 5.5 %), organic acids (1 – 1.8 %) have been iso-
lated from green tea leaves [6].
Owing to the chemical composition of green
tea leaves, the extract from this raw material can
be used in developing and creating herbal medi-
cines, dietary supplements, and cosmetic products,
which further can be used by patients with car-
diovascular diseases, diabetes mellitus, cancer,
polycystic ovary syndrome, infectious diseases,
etc. Today, the optimal extraction frequency is
determined by measuring the content of biologi-
cally active substances, as well as the dry resi-
due in the extracts obtained. However, no one
has previously described the determination of the
extraction frequency by measuring the antioxi-
dant activity of the extracts. Our recent study
[3] has revealed that 60 % ethanol is the optimal
solvent for obtaining an extract from green tea
leaves. Therefore, the current work aimed to study
the dynamics of extraction of biologically active
substances from green tea leaves with 60 % etha-
nol to determine the optimal extraction frequen-
cy by the method of measuring the antioxidant
activity.
■ Materials and methods
Chun Myn green tea leaves were the object
of the study, the raw material was collected in
Anhui province (China) from March to April.
Dry green tea leaves were standardized accord-
ing to the European Pharmacopeia 9.0 [7]. All sol-
vents and other chemicals used in the study were
of analytical grade.
Potentiometric measurements were performed
on a HANNA 2550 pH meter (Germany) with a
combined platinum EZDO 50 PO electrode (Tai-
wan). The quantitative analysis of biologically
active compounds was performed on a UV-1000
spectrophotometer (China) with matched 1 cm
quartz cells.
The weighing was done using an АN100 di-
gital analytical balance (AXIS, Poland) with
d = 0.0001 g.
Extraction procedure
Five (5.0) g of the grinded leaves was mixed
with 100 mL of 60 % ethanol. The extraction was
carried out within 1 hour on a water bath with a con-
denser, then it was repeated two times with a new
portion of the solvent. After that, the extracts
ISSN 2308-8303 (Print) / 2518-1548 (Online) 30
Журнал органічної та фармацевтичної хімії 2022, 20 (1)
obtained were filtrated and concentrated to 10 mL
using a rotary evaporator.
The dry residue of the extracts was determi-
ned by the gravimetric method according to the
State Pharmacopeia of Ukraine (SPhU) [8].
Preparation of the standard solutions
To plot the calibration curve of the depend-
ence of absorbance on the amount of gallic acid,
the stock solution (250 mg mL-1) was prepared
by dissolving 50.0 mg (accurate weight) of gallic
acid in 96 % ethanol, and the solution was dilut-
ed to 200.0 mL with the same solvent. The stock
solution was diluted with the solvent to prepare
the model solutions 1 – 5 with concentrations of
1.0; 2.0; 3.0; 4.0; 5.0 μg mL-1, respectively.
To plot the calibration curve of the depend-
ence of absorbance on the amount of (−)-epigallo-
catechin gallate, the stock solution (10 mg mL-1)
was prepared by dissolving 250.0 mg of (−)-epi-
gallocatechin gallate in 96 % ethanol, and the so-
lution was diluted to 25.0 mL with the same sol-
vent. The stock solution was diluted with the sol-
vent to prepare the model solutions 1 – 5 with the
concentrations of 100; 150; 200; 300; 400 μg mL-1,
respectively.
The standard solution of rutin was prepared
by weighing 10 mg (accurate weight) of rutin,
transferred in a 25 mL volumetric flask, dissolved
in 70 % ethanol while heating on a water bath,
and diluted to the volume with the same solvent.
1.0 mL of the solution prepared was transferred
into a 25 mL volumetric flask and diluted to the
volume with the same solvent.
The total content of phenolic compounds
was measured by the Folin-Ciocalteu assay, the
optical density was measured at 760 nm [9].
The calibration curve was plotted with interval
concentrations of 1.0 – 5.0 μg mL-1, the calibra-
tion equation Y = 0.1055X + 0.1745 (R2 = 0.9951).
The total content of phenolic compounds with
reference to gallic acid was calculated according
to the following equation:
X(%) � Cx × Kdil × 100
V
where Сx – is the concentration of gallic acid ac-
cording to the calibration curve, С × 10-6, g mL-1;
V – is the volume of the extract, mL; Kdil – is the
dilution coefficient.
The vanillin reagent assay was applied to de-
termine the total amount of catechins [5], and the
absorbance was measured at 505 nm. The cali-
bration curve was plotted within interval concen-
trations of (100 – 400) × 10-6 g mL-1, the calibra-
tion equation Y = 0.0025X – 0.0851 (R2 = 0.9951).
The total catechins content in the extracts with
reference to (−)-epigallocatechin gallate was cal-
culated according to the following equation:
X(%) � Cx × Kdil × 100
V
where Сx – is the concentration of (−)-epigallo-
catechin gallate according to the calibration curve,
С × 10–6 g mL-1; V – is the volume of the extract, mL;
Kdil – is the dilution coefficient.
The total flavonoids were determined using
the assay of the complex formation with 2 % AlCl3,
the absorbance was measured at 417 nm [10].
y = 0 1055x. + 0.1745
R² = 0 9951.
0 0.
0 1.
0 2.
0 3.
0 4.
0 5.
0 6.
0 7.
0 8.
А
C × 10
–6, g mL–1
0 1 2 3 4 5 6
Figure 1. The calibration curve of the absorbance against the concentration of gallic acid
ISSN 2308-8303 (Print) / 2518-1548 (Online) 31
Journal of Organic and Pharmaceutical Chemistry 2022, 20 (1)
The total content of flavonoids in the extracts with
reference to rutin was calculated according to the
following equation:
X(%) =
A × Kdil × 100
Ast × V
where A – is the absorbance of the solution ana-
lyzed; Ast – is the absorbance of the standard so-
lution of rutin; V – is the volume of the extract,
mL; Kdil – is the dilution coefficient.
The total content of hydroxycinnamic acids was
measured by the assay of complex formation with
NaNO2-Na2MoO4, and the absorbance was meas-
ured at 505 nm [11]. The total content of hydroxy-
cinnamic acids in extracts with reference to chlo-
rogenic acid was calculated according to the fol-
lowing equation:
X(%) =
188 × V
A × Kdil
where A – is the absorbance of the solution ana-
lyzed; 188 – is the specific absorption coefficient
of chlorogenic acid; V – is the volume of the ex-
tract, mL; Kdil – is the dilution coefficient.
The antioxidant activity of the extracts was
measured by the potentiometric method [12] cal-
culated according to the following equation, and
expressed as mmol-equiv mdry res.
-1:
AOA =
Cox – α × Cred
1 + α × Kdil × 10
3
×
m1
m2
where α = Cox/Cred × 10(∆E – Eethanol)nF/2.3RT; Сox – is the
concentration of K3[Fe(CN)6], mol L-1; Cred – is the
concentration of K4[Fe(CN)6], mol L-1; Еethanol –
0.0546 × С % – 0.0091; С% – is the concentration
of ethanol; ∆E – is the change of the potential;
F = 96485.33 C mol-1 – is the Faraday constant;
n = 1 – is the number of electrons in the elec-
trode reaction; R = 8.314 J mol-1 К-1 – is the uni-
versal gas constant; T – 298 K; Kdil– is the dilu-
tion coefficient; m1 – is the mass of a dry residue;
m2 – is the mass of a dry residue in 1.0 mL of the
extract.
For all experiments, five samples were ana-
lyzed, and all assays were performed 5 times.
The results were expressed as mean values with
confidence intervals. MS EXCEL 7.0 and STATI-
STIKA 6.0 were used to perform the statistical
analysis.
■ Results and discussion
The total content of phenolic compounds
was determined by the Folin-Ciocalteu method.
According to the results obtained given in Table,
the total content of phenolic compounds was
9.60 ± 0.17 % in the first extraction, whereas in the
second extract the amount of phenolic compounds
decreased by 86 %, and in the case of the third
extraction by 99 %. The summary content of phe-
nolic compounds was equal to 11.02 % in the to-
tal extract.
The amount of catechins was measured using
the vanillin reagent assay. Table represents that
in the first extraction the content of catechins
was 9.20 ± 0.18 %, in the second extraction –
1.20 ± 0.02 %, and in the third one – 0.07 ± 0.002 %.
The total catechins were equal to 10.47 % in the
total extract. Hence, the contribution of the first
y = 0 0025x 0.0851. –
R² = 0 9951.
0
0 1.
0 2.
0 3.
0 4.
0 5.
0 6.
0 7.
0 8.
0 9.
1
0 50 100 150 200 250 300 350 400 450
A
C × 10
–6, g mL–1
Figure 2. The calibration curve of the absorbance against the concentration of (−)-epigallocatechin gallate
ISSN 2308-8303 (Print) / 2518-1548 (Online) 32
Журнал органічної та фармацевтичної хімії 2022, 20 (1)
extraction to the overall extract was 87.87 %, the
second one – 11.46 %, and the third one – 0.67 %.
The total amount of flavonoids in extracts was
relatively low compared to other biologically acti-
ve substances. In the first extract their content was
0.27 ± 0.005 %, in the second one – 0.04 ± 0.001 %,
and the third one – only 0.005 ± 0.001 %.
Table also shows that the total content of hy-
droxycinnamic acids was 0.56 % in the total ex-
tract. The first extraction provided 0.49 ± 0.01 %,
the second one – 0.07 ± 0.002 %, and the last one
– 0.007 ± 0.001 % to the total extract. Among all
phenolic compounds, hydroxycinnamic acids took
second place after catechins.
The dry residue of the extracts analyzed was
determined by the gravimetric method of analy-
sis. Table shows that the dry residue in the first
extraction was 10.75 ± 0.11 %, in the second one –
1.59 ± 0.02 %, and the third one – 0.15 ± 0.005 %.
The total dry residue of three extractions was
12.49 %.
Today the main method of finding the extrac-
tion frequency from various types of the raw ma-
terial is based on determining the dry residue and
the amount of biologically active substances in
extracts. According to the data obtained (Table,
Figure 3), the optimal extraction frequency equals
two. The third extraction with a new portion of
the solvent does not significantly increase the
amount of biologically active substances in the
total extract.
However, in our view, this way of determin-
ing the extraction frequency is not rather suita-
ble due to several disadvantages of the method.
Among others, one should note that the gravi-
metric method and the measuring amount of all
Table. The total content of phenolic compounds, catechins, flavonoids, and hydroxycinnamic acids in green tea extracts
1st extraction 2nd extraction 3rd extraction Sum
Dry residue, % 10.75 ± 0.11 1.59 ± 0.02 0.15 ± 0.005 12.49
Total phenolic
compounds, % 9.60 ± 0.17 1.30 ± 0.03 0.12 ± 0.002 11.02
Total catechins, % 9.20 ± 0.18 1.20 ± 0.02 0.07 ± 0.002 10.47
Total flavonoids, % 0.27 ± 0.005 0.04 ± 0.001 0.005 ± 0.001 0.32
Total hydroxycinnamic
acids, % 0.49 ± 0.01 0.07 ± 0.002 0.007 ± 0.001 0.56
Antioxidant activity,
mmol-equiv mdry res.
-1 474.08 ± 9.48 67.70 ± 1.35 7.01 ± 0.14 548.79
0
2
4
6
8
10
12
1 2 3
C
o
n
te
n
t,
%
The extraction frequency
Dry residue Total content phenolic compounds
Total content of catechins Total content of flavonoids
Total content of hydroxycinnamic acids
Figure 3. Curves of the dry residue, total phenolic compounds, catechins, flavonoids, hydroxycinnamic acids versus the extraction
frequency
ISSN 2308-8303 (Print) / 2518-1548 (Online) 33
Journal of Organic and Pharmaceutical Chemistry 2022, 20 (1)
biologically active substances in extracts are rath-
er time-consuming processes.
Therefore, herein we suggest determining the
optimal extraction frequency using the antioxi-
dant activity of the extracts obtained and the
total antioxidant activity of the raw material.
There are some reasons for that. Firstly, the po-
tentiometric method of the antioxidant activity
determination is simple and not time-consuming.
Secondly, the accuracy of the method is not infe-
rior to that one of gravimetry. Moreover, it does
not need expensive reagents and equipment for
analysis. In addition, in many scientific studies
the antioxidant activity correlates with the anti-
inflammatory effect, this gives an idea of other
possible effects of the extracts under research [14].
In our previous study [15] it was found that
the total antioxidant activity of green tea leaves
equaled 660.75 mmol-equiv mdry res.
-1. In our opin-
ion, the most optimal value for the acceptance
criterion of the antioxidant activity is 80 % of the
total antioxidant activity of the raw material, which
is due to the fact that, depending on the type of
green tea, the antioxidant activity of its extracts
may vary. Thus, in our case, the acceptance cri-
terion equals 528 mmol-equiv mdry res.
-1. Table re-
presents that the antioxidant activity of the 1st ex-
tract was 474.08 mmol-equiv mdry res.
-1, the 2nd ex-
tract – 67.70 mmol-equiv mdry res.
-1
and the 3rd ex-
tract – 7.01 mmol-equiv mdry res.
-1. The antioxi-
dant activity of the total extract was equal to
548.79 mmol-equiv mdry res.
-1 as shown in Table.
The 1st extraction contributes 86 % to the total
antioxidant activity, the 2nd extraction – 12.73 %,
and the 3rd one only 1.27 %. According to the re-
sults, the total antioxidant activity is equal to
542 mmol-equiv mdry res.
-1 after two extractions of
green tea leaves, and it meets the requirement
of the acceptance criteria as it is not less than
528 mmol-equiv mdry res.
-1. Therefore, the 3rd ex-
traction was redundant and only led to overspend-
ing of the solvent, energy, and labor resources,
and decreasing the labor productivity.
To compare the methods of determining the
extraction frequency, the correlation between an-
tioxidant activity and the content of biologically
active substances in green tea extracts was as-
sessed. For this purpose, the linear regression
analysis and Pearson’s coefficient (R) were used.
As a result, it was found that there was a very high
correlation between the antioxidant activity and
the amount of phenolic compounds (R = 0.9991),
catechins (R = 0.9995), flavonoids (R = 0.9991),
and hydroxycinnamic acids (R = 0.9991). Thus,
there is no difference in the results and the me-
thod of the antioxidant activity can be applied to
determine the optimal extraction frequency from
green tea leaves. The results obtained are shown
in Figure 4.
■ Conclusions
The dynamic of extractions of biologically ac-
tive substances of green tea leaves has been studied
y = 0.0184x + 0.1471
R² = 0.9977
R = 0.9991
y = 0.0199x + 0.0506
R² = 0.9991
R = 0.9995
y = 0.0006x + 0.0049
R² = 0.9983
R = 0.9991
y = 0.0011x + 0.0064
R² = 0.9984
R = 0.9991
0
1
2
3
4
5
6
7
8
9
10
0 50 100 150 200 250 300 350 400 450 500
C
o
n
te
n
t,
,%
Antioxidant activity, mmol-equiv mdry res.
-1
total content of phenolic compounds total content of catechins
total content of flavonoids total content of hydroxycinnamic acids
Figure 4. The correlation between the antioxidant activity and the content of total phenolic compounds, catechins, flavonoids,
hydroxycinnamic acids in extracts of green tea leaves
ISSN 2308-8303 (Print) / 2518-1548 (Online) 34
Журнал органічної та фармацевтичної хімії 2022, 20 (1)
by triple extraction of the raw material to find
the optimal extraction frequency; for the first
time, a method for determining the extraction
frequency based on the antioxidant activity of
the extracts has been developed and proposed.
Thus, it has been found that the optimal extrac-
tion rate is 2 times. The results obtained will be
used in the further production of herbal medi-
cines, dietary supplements, and cosmetic products
with a green tea extract.
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Authors information:
Oleksandr Yu. Maslov (corresponding author), Teaching Assistant of the Department of Analytical Chemistry and Analytical
Toxicology, National University of Pharmacy of the Ministry of Health of Ukraine; https://orcid.org/0000-0001-9256-0934;
e-mail for correspondence: alexmaslov392@gmail.com; tel. +380 66 4257224.
Serhii V. Kolisnyk, D.Sc. in Pharmacy, Professor, Head of the Department of Analytical Chemistry and Analytical Toxicology,
National University of Pharmacy of the Ministry of Health of Ukraine; https://orcid.org/0000-0002-4920-6064.
Mykola A. Komisarenko, Ph.D. in Pharmacy, Teaching Assistant of the Department of Pharmacognosy, National University
of Pharmacy of the Ministry of Health of Ukraine; https://orcid.org/0000-0002-1161-8151.
Mykola Yu. Golik, D.Sc. in Pharmacy, Associate Professor, Head of the Department of Inorganic and Physical Chemistry,
National University of Pharmacy of the Ministry of Health of Ukraine; https://orcid.org/0000-0003-3134-9849.
Yevhen O. Tsapko, Ph.D. in Pharmacy, Associate Professor of Inorganic and Physical Chemistry Department, National University
of Pharmacy of the Ministry of Health of Ukraine; https://orcid.org/0000-0003-0735-1797.
Elshan Yu. Akhmedov, Ph.D. in Pharmacy, Associate Professor of the Department of Analytical Chemistry and Analytical Toxicology,
National University of Pharmacy of the Ministry of Health of Ukraine; https://orcid.org/0000-0001-6727-8259.
|
| id | oai:ojs.journals.uran.ua:article-252320 |
| institution | Journal of Organic and Pharmaceutical Chemistry |
| issn | 2518-1548 |
| keywords_txt_mv | |
| language | English |
| last_indexed | 2026-08-25T01:02:40Z |
| publishDate | 2022 |
| publisher | National University of Pharmacy |
| record_format | ojs |
| resource_txt_mv | ophcjnupheduua/3f/5141219edca00059c4b100b78e078a3f.pdf |
| spelling | oai:ojs.journals.uran.ua:article-2523202026-08-24T13:41:04Z Determination of the extraction frequency of green tea leaves by the antioxidant method Визначення кратності екстракції листя зеленого чаю антиоксидантним методом Maslov, Oleksandr Yu. Komisarenko , Mykola A. Kolisnyk, Serhii V. Golik, Mykola Yu. Tsapko, Yevhen O. Akhmedov, Elshan Yu. green tea leaves analysis extract extraction antioxidant activity зелений чай листя аналіз екстракт екстракція антиоксидантна активність Aim. To determine the optimal extraction frequency of green tea leaves with 60 % ethanol by the antioxidant method.Materials and methods. Chun Myn green tea leaves were the object of the study, the raw material was collected in Anhui province (China) from March to April. Dry green tea leaves were standardized according to the European Pharmacopeia 9.0. Spectrophotometry was used to quantify biologically active substances. The antioxidant activity was determined by the potentiometric method. Potentiometric measurements were performed on a HANNA 2550 pH meter (Germany) with a combined platinum EZDO 50 PO electrode (Taiwan). A UV-1000 spectrophotometer (China) was used to measure the optical density.Results and discussion. The total content of phenolic compounds was 9.60 ± 0.17, 1.30 ± 0.03 and 0.12 ± 0.002 %, catechins – 9.20 ± 0.18, 1.20 ± 0.02 and 0.07 ± 0.002 %, flavonoids – 0.27 ± 0.005, 0.04 ± 0.001, 0.005 ± 0.001, hydroxycinnamic acids – 0.49 ± 0.01, 0.07 ± 0.002 and 0.007 ± 0.001 %, dry residue – 10.75 ± 0.11, 1.59 ± 0.02 and 0.15 ± 0.002 %, the antioxidant activity was 474.08 ± 9.48, 67.70 ± 1.35 and 7.01 ± 0.14 mmol-equiv mdry res-1 for the first, second and third extraction, respectively. According to the results obtained, the optimal number of extractions of the raw material with 60 % ethanol was found to be two.Conclusions. The dynamic of extractions of biologically active substances of green tea leaves has been studied by triple extraction of the raw material to find the optimal extraction frequency; for the first time, a method for determining the extraction frequency based on the antioxidant activity of the extracts has been developed and proposed. It has been found that the optimal extraction rate is 2 times. The results obtained will be used in the further production of herbal medicines, dietary supplements, and cosmetic products with a green tea extract. Мета. Визначити оптимальну кратність екстракції листя зеленого чаю 60 % етанолом антиоксидантним методом.Матеріали та методи. Об’єктом дослідження було листя зеленого чаю сорту Чун Мін, зібране в провінції Анхуй (Китай) з березня до квітня. Сухе листя було стандартизовано відповідно до Європейської фармакопеї 9.0. Для кількісного визначення біологічно активних речовин використовували спектрофотометрію. Для визначення антиоксидантної активності використовували потенціометричний метод. Потенціометричні вимірювання виконували на pH-метрі HANNA 2550 (ФРН) із комбінованим платиновим електродом EZDO 50 PO (Тайвань). Для вимірювання оптичної густини використовували спектрофотометр UV-1000 (Китай).Результати та їх обговорення. Сумарний вміст фенольних сполук становив 8,60 ± 0,17, 1,30 ± 0,03 і 0,11 ± 0,002 %, катехінів – 9,20 ± 0,18, 1,20 ± 0,02 і 0,07 ± 0,002 %, флавоноїдів – 0,27, 0,04 і 0,005 %, гідроксикоричних кислот – 0,49 ± 0,01, 0,07 ± 0,002 і 0,007 ± 0,001 %, сухий залишок – 10,75 ± 0,11, 1,59 ± 0,02 і 0,15 ± 0,002 %, антиоксидантна активність становила 474,08 ± 9.48, 67,70 ± 1.35 і 7,01 ± 0.14 ммоль-екв. mсух.зал.-1 для першої, другої і третьої екстракції відповідно. Одержані результати свідчать, що двократна екстракція досліджуваної сировини 60 % етанолом є оптимальною.Висновки. Для визначення оптимальної кратності екстракцій було проведено дослідження динаміки екстракцій біологічних активних речовин листя зеленого чаю шляхом трикратної екстракції сировини, а також уперше розроблено й запропоновано метод визначення кратності екстракції, заснований на визначенні антиоксидантної активності екстрактів. Виявлено, що двократна екстракція є оптимальна. Отримані результати будуть використані для створення фітопрепаратів, дієтичних добавок та косметологічної продукції з екстрактом зеленого чаю. National University of Pharmacy 2022-05-30 Article Article application/pdf https://ophcj.nuph.edu.ua/article/view/252320 10.24959/ophcj.22.252320 Journal of Organic and Pharmaceutical Chemistry; Vol. 20 No. 1(77) (2022); 28-34 Журнал органической и фармацевтической химии; Том 20 № 1(77) (2022); 28-34 Журнал органічної та фармацевтичної хімії; Том 20 № 1(77) (2022); 28-34 2518-1548 2308-8303 en https://ophcj.nuph.edu.ua/article/view/252320/254343 Copyright (c) 2022 Oleksandr Yu. Maslov, Mykola A. Komisarenko , Serhii V. Kolisnyk, Mykola Yu. Golik, Yevhen O. Tsapko, Elshan Yu. Akhmedov http://creativecommons.org/licenses/by/4.0 |
| spellingShingle | зелений чай листя аналіз екстракт екстракція антиоксидантна активність Maslov, Oleksandr Yu. Komisarenko , Mykola A. Kolisnyk, Serhii V. Golik, Mykola Yu. Tsapko, Yevhen O. Akhmedov, Elshan Yu. Визначення кратності екстракції листя зеленого чаю антиоксидантним методом |
| title | Визначення кратності екстракції листя зеленого чаю антиоксидантним методом |
| title_alt | Determination of the extraction frequency of green tea leaves by the antioxidant method |
| title_full | Визначення кратності екстракції листя зеленого чаю антиоксидантним методом |
| title_fullStr | Визначення кратності екстракції листя зеленого чаю антиоксидантним методом |
| title_full_unstemmed | Визначення кратності екстракції листя зеленого чаю антиоксидантним методом |
| title_short | Визначення кратності екстракції листя зеленого чаю антиоксидантним методом |
| title_sort | визначення кратності екстракції листя зеленого чаю антиоксидантним методом |
| topic | зелений чай листя аналіз екстракт екстракція антиоксидантна активність |
| topic_facet | green tea leaves analysis extract extraction antioxidant activity зелений чай листя аналіз екстракт екстракція антиоксидантна активність |
| url | https://ophcj.nuph.edu.ua/article/view/252320 |
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