The American Journal of Medical Sciences and Pharmaceutical Research
78
https://www.theamericanjournals.com/index.php/tajmspr
TYPE
Original Research
PAGE NO.
78-81
10.37547/tajmspr/Volume07Issue01-10
OPEN ACCESS
SUBMITED
23 October 2024
ACCEPTED
25 December 2024
PUBLISHED
27 January 2025
VOLUME
Vol.07 Issue01 2025
CITATION
Askarov Ibragim Rakhmonovich, Muminjonov Mirjalol Muqimjon ogli, &
Aliqulova Irodaxon Maxmudovna. (2025). Chemical analysis of polyphenols
in camellia sinensis, their health benefits, and possibility for application in
eye diseases. The American Journal of Medical Sciences and
Pharmaceutical Research, 7(01), 78
–
81.
https://doi.org/10.37547/tajmspr/Volume07Issue01-10
COPYRIGHT
© 2025 Original content from this work may be used under the terms
of the creative commons attributes 4.0 License.
Chemical analysis of
polyphenols in camellia
sinensis, their health
benefits, and possibility
for application in eye
diseases
Askarov Ibragim Rakhmonovich
Doctor of Chemical Sciences, Professor, Andijan State University, Andijan,
Uzbekistan
Muminjonov Mirjalol Muqimjon ogli
Doctor of Chemistry, Docent, Andijan State University, Andijan,
Uzbekistan
Aliqulova Irodaxon Maxmudovna
Master's student, Andijan State University, Andijan, Uzbekistan
Abstract:
This article presents the results of a study on the
determination of polyphenols in Black tea by high-
performance liquid chromatography (HPLC). Polyphenols
are bioactive compounds in plants, whose antioxidant
properties have a therapeutic effect against various
diseases. During the study, the main polyphenols (e.g.
flavonoids and phenolic acids) in Black tea were identified
and their quantitative indicators were determined. The
effective effect of these compounds on eye diseases, in
particular, age-related macular degeneration, glaucoma,
and dry eye syndrome, was studied. The results showed
that polyphenols in Karachay play an important role in
protecting eye tissues from oxidative stress and reducing
inflammatory processes. The results of this study serve to
increase the pharmacological value of Karachay and
expand the possibilities of its application in the healthcare
sector.
Keywords:
Polyphenol, phytochemicals, Camellia
sinensis, Karachoy.
Introduction:
Black tea (Camellia sinensis) contains
many biologically active compounds, in particular
The American Journal of Medical Sciences and Pharmaceutical Research
79
https://www.theamericanjournals.com/index.php/tajmspr
The American Journal of Medical Sciences and Pharmaceutical Research
polyphenols, which are of great importance for human
health.
Polyphenols
have
antioxidant,
anti-
inflammatory, and cardioprotective properties and
play an important role in the prevention and treatment
of various diseases [1]. This study aimed to determine
the amount of polyphenols in black tea and their
health benefits using high-performance liquid
chromatography (HPLC).
Black tea is translated from various languages as red
tea - black tea, which has a more diverse variety and a
stronger taste than other teas. The name of the drink,
which first appeared in China, is hong cha ( "red tea")
due to the color of the leaves that oxidize when
properly processed. Today, it is widely consumed and
harvested, including in China, Japan, Korea, and
Singapore. While green tea usually loses its flavor
within a year, black tea retains its flavor for several
years [2].
A study by Chinese scientists at National Chiayi
University, led by Chih-Yu Lo and Min-Hsiung Pan,
investigated the nutritional value of tea, mainly its
polyphenols and antioxidant properties. A meta-
analysis of observational studies concluded that black
tea consumption does not affect the development of
oral cancer, Asian esophageal or prostate cancer, or
lung cancer in Asian or Caucasian populations. In
addition, black tea contains polyphenols - antioxidants
that help maintain the balance of intestinal microflora
and eliminate pathogenic bacteria in the div [3].
Cleans the mouth According to scientific research, the
polyphenols in black tea protect against oral diseases,
rashes, and stomatitis. Benefits for the heart European
scientists have proven that those who drink 3-4 cups of
tea a day have a 21% lower risk of heart disease. True,
it is not recommended to drink black tea in a bitter
form. But if it is consumed in moderation, there is no
need to worry [4]. Cancer prevention According to
reports, the polyphenols and catechin antioxidants in
black tea act as a preventive measure for certain types
of cancer. A scientific study conducted by European
scientists showed that women who drink black tea
have a lower risk of developing uterine cancer than
others. Strengthens bones The phytochemicals in this
drink protect against bone fragility. Therefore, arthritis
is less common among black tea lovers. Improves
immunity Alkyloamine antibodies stimulate the
immune system, while antioxidants provide resistance
to viruses. So, hot black tea is also good for colds, flu,
or coughs [5].
Black tea is widely used as a natural remedy for eye
diseases. The tannins and antioxidants in black tea
have antimicrobial effects. This is especially useful for
mild eye infections (conjunctivitis, etc.). Applying cold
black tea bags to the eyes reduces swelling and redness.
This is especially useful for tired or sleep-deprived eyes.
A cooled infusion of black tea reduces redness in the
eyes and soothes the eyes [6]. It can also be used in case
of allergies or irritation. Black tea bags or compresses
stimulate blood circulation, which has a positive effect
on the skin around the eyes. The light hydrating
properties of black tea help moisturize the eyes.
Washing or compressing with tea leaves reduces eye
dryness.
Experience part
Reagents and equipment used. Gallic acid was obtained
from Macklin (China), Salicylic acid from Rhydburg
Pharmaceuticals (Germany), quercetin, apigenin, and
kaempferols from Regal (China), and rutin were isolated
from natural sources by extraction and column
chromatography. Water, acetonitrile, acetic acid of the
chemically pure brand, and sodium hydroxide were
used as reagents of HPLC purity.
The content of polyphenols in the plant was determined
using an LC-40 Nexera Lite high-performance liquid
chromatograph manufactured by Shimadzu, Japan.
Preparation of standard solutions
. Gallic acid (5.2 mg),
salicylic acid (5.2 mg), rutin (5 mg), quercetin (5 mg),
apigenin (5 mg), kaempferol (5 mg) were dissolved in
96% ethanol for 20 minutes in an ultrasonic bath and
transferred to a 50 ml flask and made up to the mark
with ethanol. 200 μl of each solution was taken, mixed,
and diluted to prepare a total of 4 different solutions.
Each solution was poured into a vial and used for
analysis.
Preparation of plant extract
. For the extraction of
phenolic compounds, 1 g of the test sample was
weighed with an accuracy of 0.01 g on an NV222 balance
manufactured by OHAUS (USA), placed in a 50 ml conical
flask, and 25 ml of 96% ethanol was added. The mixture
was extracted in an ultrasonic bath of the GT SONIC-D3
brand (China) at a temperature of 60 C for 20 minutes.
Then the mixture was cooled, filtered, and made up to
25 ml with ethanol in a volumetric flask. 1.5 ml of the
extract was centrifuged at 7000 rpm in a Mini-7 brand
(BIOBASE, China) centrifuge and filtered through a 0.45
μm syringe filter, and used for analysis.
Chromatographic conditions
Determination of phenolic compounds
. Standard
solution, sample extract Shim pack GIST C18 reversed-
phase column (150 × 4.6 mm;
5 μm, Shimadzu, Japan)
and a gradient mobile phase consisting of acetonitrile
(A) and 0.5% acetic acid in water (B) (Table 1) were used.
The injection volume was 10 μl, the flow rate was 0.5
ml/min, and the column thermostat was set to 40 °C.
The analytical signal (peak area) of phenolic compounds
The American Journal of Medical Sciences and Pharmaceutical Research
80
https://www.theamericanjournals.com/index.php/tajmspr
The American Journal of Medical Sciences and Pharmaceutical Research
was recorded at 300 nm (Figure 1).
Table 1. Mobile phase gradient program.
Time
Acetonitrile (A), %
0,5% acetic acid (B), %
0
5
95
5
5
95
17
40
60
22
40
60
22,1
5
95
40
Termination
Figure 1. Chromatogram of standards at 300 nm.
RESULTS
Determination of the amount of phenolic compounds
in the sample extract of Black tea. A chromatogram of
a sample extract weighing 1 g was obtained (Figure 2),
and based on the results, the amount of phenolic
compounds in 100 g of the sample was calculated using
the following formula and presented in Table 3.
𝑋 =
𝐶
𝑝ℎ𝑒𝑛
∙ 𝑉
𝑒𝑘𝑠𝑡𝑟𝑎𝑘𝑡
𝑚
𝑒𝑥𝑎𝑚𝑝𝑙𝑒
∙ 100 𝑔
Here, X
–
the amount of phenolic compounds in 100
grams of plant material, mg;
C
phen
–
the concentration of phenolic compounds in
the extract determined by the HPLCH method, mg/l;
V
extract
–
the volume of the sample extract, l;
example
–
the mass of the sample taken for extract
preparation.
Figure 2. Chromatogram of polyphenols in the sample extract.
The American Journal of Medical Sciences and Pharmaceutical Research
81
https://www.theamericanjournals.com/index.php/tajmspr
The American Journal of Medical Sciences and Pharmaceutical Research
Table 2. Amount of polyphenols in the extract and retention times.
Phenol compound name
Catch time,
sec
Concentration,
mg/l
Amount in 100 g
of sample, mg
Gallic Acid
7,765
61,433
153,583
Rutin
18,906
4,935
12,338
Salicylic Acid
22,404
12,472
31,180
Quercetin
23,724
0,175
0,438
Apigenin
25,898
58,004
145,010
Kaempferol
Not specified
0
0,000
From Table 1 above, it can be seen that the content of
gallic acid (153.583 mg/100g) in the “Black tea” plant
is significantly higher than that of others. It is also
known that the content of apigenin, rutin, and salicylic
acid in “Black tea” is also signi
ficantly higher.
CONCLUSION
Determining the content of polyphenols in Karachoy
using the HPLC method is a reliable tool for studying
the biological significance of these substances. The
results obtained show that polyphenols in Karachoy
can play an important role in supporting human health,
in particular, in the prevention and treatment of eye
diseases. The beneficial effects of gallic acid and
apigenin in eye diseases are manifested through their
biologically active properties.
Gallic acid reduces oxidative stress by neutralizing free
radicals in the eye. Oxidative stress is a major factor in
eye diseases associated with clouding of the vision,
such as macular degeneration, cataracts, and diabetic
retinopathy. Gallic acid has the ability to reduce
inflammation, which is beneficial in uveitis, dry eye
syndrome, and other inflammatory diseases. Its ability
to repair blood vessels and improve microcirculation
helps improve blood circulation in diabetic retinopathy
and glaucoma. Apegnin has a healing effect on uveitis,
conjunctivitis, and dry eye syndrome by reducing
inflammation. This substance is effective in blocking
the activity of inflammatory agents. Apegnin helps to
prevent cataracts and macular degeneration by
neutralizing free radicals, and apegenin helps to
preserve vision in glaucoma by reducing intraocular
pressure and improving blood circulation.
Apigenin and gallic acid, when used in combination,
effectively reduce oxidative stress and inflammation in
eye tissues. They can be used to: slow the
development of cataracts, prevent macular
degeneration and glaucoma, improve visual function,
and eliminate dry eye syndrome and inflammatory
diseases. Timely and targeted use of these substances is
important in the prevention and therapy of eye
diseases. The combination of these two substances can
be effective in treating and preventing many health
problems for humans. Since the consumption of Black
tea is significant in supporting a healthy lifestyle, it is
recommended to include it in the regular diet. In the
future, it is advisable to study polyphenols in more
depth using other biochemical methods.
REFERENCES
Дидманидзе О. Н., Дидманидзе Р. Н. Повышение
эффективности
процессов
производства
и
реализации chая. –
2003.
Cheung, L., PC Cheung and VE Ooi. 2003. Antioxidant
activity and total phenolics of extracts. Food Chem., 81:
249-255.
Engelhardt U.H. Chemistry of tea. Comprehensive
natural products II. Chemistry and Biology: eds. L.
Mander & H.W. Liu. UK, Elsevier Ltd., 2010, pp. 1000-
1027. doi: 10.1016/b978-0-12-409547-2.02784-0.
Fedoseeva A.A., Lebedkova O.S., Kanibolotskaia L.V.,
Shendrik A.N. Chemistry of plant raw materials, 2008,
no. 3, pp. 123-127 (in Russian).
Гурева К. Б., Тарасова Е. А. Влияние вида упаковки на
каchество chерного chая при хранении //Товаровед
продоволственных товаров. –
2015.
–
№. 11. –
С. 49
-
52.
Татарченко И. И., Славянский А. А., Макарова С. А.
Показатели качества черного чая, зависящие от
переработки
чайного
листа
//Технология
и
товароведение инновационных пищевых продуктов.
–
2013.
–
№. 5. –
С. 76
-80.
