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com

IN VITRO ANALYSIS OF ANTIOXIDANT ACTIVITY IN

POLYHERBAL PLANT EXTRACT

1
M SASIDHARAN, 2R VIGNESH, 3G VIGNESHWARAN,
4
[Link]*
1Student, Department of Biotechnology, VSB Engineering College,
Karur-639111. Tamil Nadu, India
2Student, Department of Biotechnology, VSB Engineering College,
Karur-639111. Tamil Nadu, India
3Student, Department of Biotechnology, VSB Engineering College,
Karur-639111. Tamil Nadu, India
4AssistantProfessor, Department of Biotechnology, VSB EngineeringCollege,
Karur-639111. Tamil Nadu, India
1Email- sasidharan02012001@[Link] , 2Email- vigneshravi91940@[Link]
3
Email- vigneshwarang2001@[Link] , 4*Email- santhaseelan07@[Link]
*Corresponding Author Email id: santhaseelan07@[Link]

Abstract
Most plants are considered to prevent free radicals associated damages by numerous ways
including direct scavenging of free radicals and inhibition of enzymes involved in free radical
production. The aim of the present study was to examine the antioxidant activities of the hexane from
polyherbal plants of Carica papaya, Punica granatum, Catharanthus roseus and Cympbopogan
citratus which is long been known to be a very important source of pharmaceutical. Different
concentrations of polyherbal extract was tested for anti-oxidant properties using hydrogen peroxide
assay. Similarly, the free radical scavenging activity was determined by DPPH assay. The results
showed that the polyherbal extract showed a strong inhibition value at IC50 of concentration of 170.7
μg/ml, 229.57 μg/ml. This study provides evidence that the polyherbal formulation possesses a strong
antioxidant activity. Therefore, it might be beneficial as medicinal plant as an antioxidant and
anticancer nutraceutical and pharmaceutical sources.

Keywords: Antioxidant activity, DPPH assay, Hexane, Hydrogen peroxide assay

[Link]

Skin acts as a protective barrier for the body which protects the body from the external
environment. Epidermal layer of skin can be damaged due to the wound [1]. Which is defined as the
disruption of the integrity of the skin by various factors such as pressure, trauma, animal or insect bites,
and mechanical abrasions. Invasion of various pathogenic microorganisms at the wounded tissue
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results in severe chronic wound infection. Anatomical and functional integrity of tissue gets disrupted
due to this chronic wound infection. Worldwide, it is estimated that at least 6 million people suffer
from chronic wound infections every year [2]. Wound healing is natural phenomenon by which body
restores the cellular and functional continuity of tissue [3].Appropriate wound healing is necessary to
regain the functional and anatomical status of the damaged tissue that got disturbed due to wound.
Various complex biochemical events are involved in appropriate healing of wound. These events are
divided into three phases: Inflammatory phase, proliferative phase, and remodeling phase [4].
Wound infection with the pathogenic microorganisms such as Staphylococcus aureus,
Escherichia coli, and P. aeruginosa results in delay of physiological events involved in the healing of
wound [5]. Antioxidants also act as radical scavengers, hydrogen donors, electron donors, peroxide
decomposers, singlet oxygen quenchers, enzyme inhibitors, and metal chelating agents. Due to the
effect on immune system, there is a need for natural antioxidants (safe and nontoxic) as compared to
synthetic antioxidants (toxic for human) [6].
Plants contain many constituents with local physical impact on body tissues, and the topical
use of herbal remedies is among the most noticeable in the simplest traditions of health care [7]. To
support the usage of selected plant extracts in Ayurveda, the antioxidant potential of the fruits of
Momordica charantia Linn., bark of Eugenia jambolana Linn., fruits of Ziziphus mauritiana Lam.,
and bark of Acacia catechu Willd. of Indian origin was examined.
Carica papaya, belongs to the family of Caricaceae, and several species of Caricaceae have
been used as remedy against a variety of diseases [8]. Originally derived from the southern part of
Mexico, Carica papayais a perennial plant, and it is presently distributed over the whole tropical area.
In particular, Carica papayafruit circulates widely, and it is accepted as food or as a quasi drug. Lot
of scientific investigations have been conducted to evaluate the biological activities of different parts
of Carica papaya, including fruits, shoots, leaves, rinds, seeds, roots or latex. The leaves of papaya
have been shown to contain many active components that can increase the total antioxidant power in
blood and reduce lipid peroxidation level, such as papain, chymopapain, cystatin, ascorbic acid,
flavonoids, cyanogenic glucosides and glucosinolates [9].
The objective of this work was to assess the antioxidant activity of the combination of extract
(polyherbal formulation [PHF]) by in vitro studies and relate them with ascorbic acid, a known
antioxidant.

2. Materials and Methods

2.1. Collection of plant material
Carica papaya, Cymbopogon citratus, Punica granatum L., Cantharanthus roseus the
plant species were collected from the wild source which was available in Trichy District, Tamil Nadu,
India.

2.2. Hydrogen peroxide scavenging assay

2.2.1. Principle
Hydrogen peroxide is a weak oxidizing agent and it can inactivate some enzymes directly
by oxidation of essential thiol groups (-SH). H2O2 can cross cell membranes rapidly, once it goes inside
the cell, mostly react with Fe2+, and possibly Cu2+ ions to form hydroxyl radical. This may be the origin
of many of its toxic effects. It is therefore biologically beneficial for cells to control the amount of
H2O2 that is allowed to accumulate.

2.2.2. Materials Required

Hydrogen peroxide solution and Sodium phosphate buffer.
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2.2.3. Procedure
Ability of plant extracts to scavenge hydrogen peroxide was estimated according to the method
reported by Ruch et al. with minor modification. A solution of H2O2 (43 mM) is prepared in phosphate
buffer (1 M pH 7.4). Different concentration of polyherbal extract sample (500 µg/ml, 250 µg/ml, 100
µg/ml, 50 µg/ml and 5 µg/ml) was added to H2O2 solution (0.6 ml, 43 mM). Absorbance of hydrogen
peroxide at 230 nm was determined after 10 minutes against a blank solution containing phosphate
buffer without hydrogen peroxide. Ascorbic acid was used as standard. The free radical scavenging
activity was determined by evaluating % inhibition as above.
% inhibition = [(Control- Test)/control] ×100.

2.3. DPPH Radical scavenging activity

2.3.1. Principle
The DPPH assay is popular in natural product antioxidant studies because it is very simple and
sensitive. DPPH assay is based on the concept that a hydrogen donor is an antioxidant. This assay
measures compounds which are radical scavengers. Figure 1, below, shows the mechanism by which
DPPH accepts hydrogen from an antioxidant. DPPH is one of the few stable as well as commercially
available organic nitrogen radicals. The antioxidant effect is proportional to the disappearance of
DPPH in test samples. Monitoring DPPH with a UV spectrometer has become the most commonly
used method because it is very simple and accurate. DPPH shows a strong absorption maximum at 517
nm (purple). The color changes from purple to yellow followed by the formation of DPPH upon
absorption of hydrogen from an antioxidant. This is a stoichiometric reaction with respect to the
number of hydrogen atoms absorbed. Therefore, the antioxidant effect can be easily evaluated by
following the decrease of UV absorption at 517 nm.

2.3.2. Materials Required

0.1mM DPPH solution, Ascorbic acid, Methanol
0.1 mM DPPH Solution
Dissolve 39 mg of DPPH in 100 ml of methanol and store at -20o C until needed.
Ascorbic acid (Standard)
1mg/ ml of Ascorbic acid
2.3.3. Procedure
1. Briefly, prepare 0.1 mM of DPPH solution in methanol and add 100 μl of this solution to 300 μl
of the solution of Poly herbal formulation sample at different concentration (500, 250, 100, 50 and
5 μg/mL).
2. The mixtures have to be shaken vigorously and allowed to stand at room temperature for 30
minutes.
3. Then the absorbance has to be measured at 517 nm using a UV-VIS spectrophotometer. (Ascorbic
acid can be used as the reference).
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4. Lower absorbance values of reaction mixture indicate higher free radical scavenging activity.
5. The capability of scavenging the DPPH radical can be calculated by using the following formula.
6. DPPH scavenging effect (% inhibition) = [(absorbance of control - absorbance of reaction
mixture)/absorbance of control] X 100

2.4. Phytochemical analysis

2.4.1. Detection of resins
To 0.5 of plant extract, 3ml of CuSo4 solution is added. Shake for about 1-2 min formation of
green colour ppt indicates the presence of resins.

2.4.2. Detection of Carboxylic acid

To 1ml plant extract, 2ml of sodium bicarbonate solution is added. Colour changes occur
indicates the presence of carboxylic acid.

2.4.3. Detection of Tannins

To 2ml of plant extract, 2-3ml of 10% HCL is added and boiled for 5-6 min. Formation of red
colour indicates the presence of tannins.

2.4.4. Detection of Steroids

To 0.5 ml extract, 5ml of chloroform is added and equal amount of conc. H2SO4 is added. In
the upper layer formation of red colour is and in the lower layer, yellow with green colour is formation
indicates the presence of steroids.

2.4.5. Detection of Flavanoids

To 0.5 ml extract, add 4ml of 1% ammonia and to this add 1ml of conc. H2SO4. Formation of
yellow colour indicates the presence of flavonoids.

2.4.6. Detection of Carbohydrates

To 0.5ml of extract, 0.5ml of Benedict reagent is added ad boiled for 2 min. Colour changes
and precipitate is formed. It indicates the presence of carbohydrate.

2.4.7. Detection of Glycosides

To prepare Hydrosalyte :To 50mg of extract, 2ml of conc. HCL is added and kept in water
bath for 1 hour and then filteration methods. The filtrate is hydrated.

2.4.8. Born- Trageru’s Test

Take 2ml of hydrolysate, add 3ml of chloroform, shake vigorously, then the chloroform
layer gets separated. To formation 10% ammonia solution of pink colour indicates the presence of
glycosides.

2.4.9. Saponification test

To 1 or 2ml of normal sodium hydroxide, 2mlof extract is added and boiled for 2 minutes.
Formation of soap or fat indicates the positive test for saponification.
2.4.10. Detection of Proteins
Estimation of Brad Ford Method: To 500 and 1 of extract, add 5mlof brad ford reagent, Take

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OD at 575nm.

2.4.11. Detection of Phenol

Ferric Chloride Test : To 50 mg of extract, 5ml distilled water, few drops of 5% ferric
chloride solution, dark green colour. Indicates the presence of phenol.

2.4.12. Biuret Test :

To 2ml of extract, 1 drop of 2% CuSo4 solution. Add 1 ml of 95% ethanol, then add 2 to 3
sodium hydroxide [Link] of pink colour indicates the test is positive.

2.4.13. Sapon Test :

To 50 mg of extract,20 ml of distilled water. Shake vigorously for 15 min, at 2 cm layer of
foam formation indicates the presence of saponins.

2.4.14. Gum Test:

To 100 mg extract. Dissolved in 2 ml of distilled water. 2ml of absolute alcohol with constant
stirring. White colour cloudy ppt indicates gums & mucilage’s.

2.4.15. Detection of Flavanoglycoside:

50 mg extract is dissolved in 5ml ethanol. Add few drops of magnesium sulphate & few drops
of [Link] Formation of pink colour. Presence of Flavanoglycoside.

3. Results and Discussion

3.1. H2O2 assay

In recent years much attention has been devoted to natural antioxidants and their health
benefits. Antioxidant-based drug formulations are used for the prevention and treatment of many
complex diseases. Plants are a major source of natural antioxidants; they produce a wide range of
secondary metabolites with antioxidative activities that have therapeutic potential. Polyphenols are the
most abundant antioxidant compounds of plant raw material. Their antioxidant activity is based on to
their redox properties, which facilitate their activity as reducing agents, hydrogen donors, singlet
oxygen quenchers, metal chelators and reductants of ferryl hemoglobin. The reducing ability is
generally associated with the presence of reductants which exert antioxidant action through breaking
the free radical chain by donating a hydrogen atom or preventing peroxide formation. In the present
study, the polyherbal extract showed excellent antioxidant activity by H2O2 assay. The IC50
concentration of polyherbal extract was found to be μg/ml. The polyherbal formulation inhibited 46.63
% of free radicles at the concentration of 500 μg/ml.

3. 1. 1. OD Value at 230 nm
Control Mean OD value: 1.681

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TABLE 1: OD values of tested sample concentration at 230nm in triplicates forH2O2 assay

S. No Tested sample concentration (μg/ml) OD Value at 230 nm (in triplicates)

1. Control 1.705 1.648 1.691

2. 500 μg/ml 0.779 0.940 0.972
3. 250 μg/ml 0.552 0.598 0.611
4. 100 μg/ml 0.114 0.140 0.119
5. 50 μg/ml 0.122 0.117 0.101
6. 5 μg/ml 0.094 0.109 0.105

2.0
Control
OD value at 230 nm

1.5 Polyherbal extract g/ml

1.0

0.5

0.0
0 200 400 600
Concentration of polyherbal extract (g/ml)

FIGURE 1: OD value graph of tested sample at various concentrations at 230nm in triplicates

for H2O2 assay

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3.1.2 Percentage of inhibition

TABLE 2: Percentage of inhibition in the tested sample at given concentrations for H2O2 assay

Percentage of inhibition Mean value

S. No Tested sample concentration (μg/ml)
(in triplicates)

1. Control 100 100 100 100

2. 500 μg/ml 53.65 44.08 42.17 46.63

3. 250 μg/ml 67.16 64.42 63.65 65.07

4. 100 μg/ml 93.21 91.67 92.92 92.6

5. 50 μg/ml 92.74 93.03 93.99 93.25

6. 10 μg/ml 94.40 93.51 93.75 93.88

Percentage of inhibition (%)

150

100

50

0
l l l l l
r ol /m /m /m /m /m
o nt g g g g g
C 0 0 0 50 5
50 25 10
Concentration of polyherbal extract (g/ml)

FIGURE 2: Percentage graph of inhibition in the tested sample at given concentrations

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3.1.3 IC50 Value of tested sample: 229.5 μg/ml

TABLE 3: IC50 Value tabulation for the H2O2 assay
log(inhibitor) vs. normalized response -- Polyherbal
Variable slope extract μg/ml
Best-fit values
LogIC50 2.361
HillSlope 5.295
IC50 229.5
Std. Error
LogIC50 0.01837
HillSlope 2.073
95% CI (asymptotic)
LogIC50 2.321 to 2.400
HillSlope 0.8166 to 9.772
IC50 209.4 to 251.4
Goodness of Fit
Degrees of Freedom 13
R squared 0.9841
Sum of Squares 401.1
Sy.x 5.554

Number of points
# of X values 15

FIGURE 3: H2O2 assay for the polyherbal extract

3.2. DPPH Radical scavenging activity

Free radicals which are delivered as a consequence of typical biochemical responses in

the body are involved in cancer, ischemic heart disease, inflammation, diabetes, aging,
atherosclerosis, immunosuppression, and neurodegenerative disorders. The human body has
characteristic barrier system to counter free radical as proteins, for example, catalase,
superoxide dismutase, and glutathione peroxidase. Selenium, vitamin C, carotene, vitamin E,
lycopene, lutein, and different carotenoids have been utilized as supplementary antioxidants.
Hence, the secondary metabolites of the plant as flavonoids and terpenoids act an important
role in the defense against free radicals. Here in this study, the polyherbal extract exhibited an
potential anti-oxidant activity by DPPH assay at the IC50 concentration.
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3.2.1. OD Value at 517 nm

Control Mean OD value: 3.005

TABLE 4: OD values at 517nm for tested sample at various concentrations for

DPPH assay

S. No Tested sample concentration (μg/ml) OD Value at 517 nm (in triplicates)

1. Control 2.970 3.168 2.879

2. 500 μg/ml 0.970 0.870 0.888
3. 250 μg/ml 0.807 0.674 0.797
4. 100 μg/ml 0.659 0.746 0.707
5. 50 μg/ml 0.666 0.625 0.653
6. 5 μg/ml 0.583 0.606 0.612
7. Ascorbic acid 0.08 0.11 0.12

4
Control

3 Polyherbal Extract g/ml

OD at 517 nm

0
0 200 400 600
Polyherbal Extract g/ml

FIGURE 4: OD values graph at 517nm for tested sample at various concentrations for
DPPH assay

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3.2.2. Percentage of inhibition

TABLE 5: Percentage of inhibition in the tested sample at given concentration for
DPPH assay

Mean
Tested sample Percentage of inhibition
S. No value (%)
concentration (μg/ml) (in triplicates)

7. Control 100 100 100 100

8. 500 μg/ml 67.72 71.04 70.44 69.73
9. 250 μg/ml 73.14 77.57 73.47 74.72
10. 50 μg/ml 78.06 75.17 76.47 76.56
11. 10 μg/ml 77.83 79.20 78.26 78.43
12. 5 μg/ml 80.59 79.83 79.63 80.01
13. a Ascorbic acid 97.33 96.33 96 96.55
Percentage of inhibition (%)

150

100

50

0
l

l
id

l
/m

/m

/m

/m
/m
ac

g

g

g

g
g
c
bi

5
50
50

25

10
or
sc
A

Concentration of polyherbal extract (g/ml)

FIGURE 5: Percentage graph of inhibition in the tested sample at given

concentrations for DPPH assay

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3.2.3 IC50 Value of tested sample: 170.7 μg/ml

TABLE 6: IC50 Value tabulation for the DPPH Assay

log(inhibitor) vs. normalized Polyherbal Extract
response -- Variable slope g/ml
Best-fit values
LogIC50 2.232
HillSlope 1.618
IC50 170.7
Std. Error
LogIC50 0.07344
HillSlope 0.4058
95% CI (asymptotic)
LogIC50 2.074 to 2.391
HillSlope 0.7409 to 2.494
IC50 118.5 to 246.0
Goodness of Fit
Degrees of Freedom 13
R squared 0.8228
Sum of Squares 3581
Sy.x 16.60

Number of points
# of X values 15

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FIGURE 6: DPPH Assay for the polyherbal extract

3.3. Phytochemical analysis

Phytochemical characterization of polyherbal formulation was carried out using qualitative

assay method. The results showed the presence of resins, carboxylic acid, steroids, carbohydrates,
glycosides and phenol compounds.

TABLE 7: Results tabulation of the phytochemical analysis

S. No Test Result

1 Resins Present
2 Carboxylic Acid Present
3 Tannins Absent
4 Steroids Present
5 Flavonoids Present
6 Carbohydrates Present
7 Glycosides Present
8 Saponification Absent
9 Protein(OD Value) 0.64
10 Phenol Present
11 Biuret Absent
12 Saponin Absent
13 Gum test Absent
14 Flavanoglycoside Absent
15 Alkaloids Absent

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FIGURE 7

FIGURE 8

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Punica granatum L. Carica papaya L.

Cantharanthus roseus L. Cymbopogon citrates L.

Polyherbal Plant extraction

FIGURE 9: Plant extracts and collected polyherbal mixture

4. Conclusion
The results from this study indicate that they possess antioxidant properties and could serve as
free radical inhibitors, acting possibly as primary antioxidants. Since reactive oxygen species are
thought to be associated with the pathogenesis of AIDS, and HIV-infected individuals often have
impaired antioxidant defenses, the inhibitory effect of the extracts on free radicals may partially
justify the traditional use of these plants in the management of OFIs in HIV patients in South
Africa.

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5. Acknowledgments
The authors wish to express their gratitude to the Management and Department of
Biotechnology, VSB Engineering College, Karur, Tamil Nadu, India, for providing the
required facilities for this study.

6. References
[1]
Shevchenko RV, James SL, James SE. A review of tissue engineered skin bioconstructs available
for the skin reconstruction.(2010) J R Soc Interfac, 20:229-40.
[2]
Kumar K, Singh KK, Asthana AK . Ethno therapeutics of bryophyte Plagiochasma appendiculatum
among the Gaddi Tribes of Kangra Valley, Himachal Pradesh, India.(2010) Pharmaceutical Bio,
38:353-6.
[3]
Lien Ai Pham-Huy, Hua He, Chuong Pham-Huy. Free Radicals, Antioxidants in Disease and Health,
International journal of Biomedical science, vol. 4 no. 2(2008).
[4]
Lazarus GS, Cooper DM, Knighton DR . Definitions and guidelines for assessment of wounds and
evaluation of healing.(1994) Arch Dermatol, 130:489-93.
[5]
Hess T. Checklist for factors affecting wound healing (2011). Adv Skin Wound Care, 24:192-4.
[6]
Mertz P, Ovington L . Wound healing microbiology(2011). Dermatol Clin, 24:739-45.
[7]
Gutteridge JM, Halliwell B . Free radicals and antioxidants. A historical look to the future.
Ann N Y Acad Sci, 899:136- 47(2000).
[8]
Zerargui F, Baghiani A, Khennouf S, Arrar L. Antioxidant activity assessment of Tamus communis
L. Roots. Int J Pharm Pharm Sci (IJPPS), 8:64-71(2016).
[9]
Mello, V.J., Gomes, M.T., Lemos, F.O., Delfino, J.L., Andrade, S.P., Lopes, M.T., Salas, C.E.,. The
gastric ulcer protective and healing role of cysteine proteinases from Carica candamarcensis.
Phytomedicine 15, 237–244(2008).
[10]
Seigler, D.S., Pauli, G.F., Nahrstedt, A., Leen, R.,. Cyanogenic allosides and glucosides from
Passiflora edulis and Carica papaya. Phytochemistry 60, 873–882(2002).
[11]
Almokhtar A Adwas,Ata Sedik Ibrahim Elsayed,Azab Elsayed Azab,Fawzia Amhimmid Quwaydir .
Oxidative stress and antioxidant mechanisms in human body, Journal of Applied Biotechnology and
Bioengineering,Volume 6 Issue 1(2019).
[12]
Amit K. Jain, Neelesh K. Mehraand Nitin K. Swarnakar . Role of Antioxidants for the Treatment of
Cardiovascular Diseases: Challenges and Opportunities, Current Pharmaceutical Design, 21(2015).
[13]
Amzad Hossain, M., Hitam, S., and Hadidja Ibrahim Ahmed, S. Pharmacological and toxicological
activities of the extracts of papaya leaves used traditionally for the treatment of diarrhea. Journal of
King Saud University – Science(2019).
[14]
Aneta Acsova, Silvia Martiniakova, Jarmila Hojerova. Selected in vitromethods to determine
antioxidant activity of hydrophilic lipophilic substances, Acta Chimica Slovaca, Vol. 12, No. 2, pp.
200-211(2019).
[15]
Anoop A Shetty, Santoshkumar Magadumand Kalmesh Managanvi. Vegetables as Sources of
Antioxidants, Journal of Food & Nutritional Disorders, J Food Nutr Disor.2:1(2013).
[16]
Anuj Yadav, Rewa Kumari, Ashwani Yadav, J.P. Mishra, Seweta Srivatva and Shashi Prabha
Antioxidants and its functions in human body - A Review, n Research in Environment and Life Sciences,
9(11) 1328-1331(2016).
[17]
Behnaz Hassanbaglou. Antioxidant activity of different extracts from leaves of Pereskia bleo
(Cactaceae). Journal of Medicinal Plants Research, 6(15). doi:10.5897/jmpr11.760(2012).
VOLUME 21 : ISSUE 5 (May) - 2022 Page No:1513
YMER || ISSN : 0044-0477 [Link]

[18]
Jarisarapurin, W., Sanrattana, W., Chularojmontri, L., Kunchana, K., and Wattanapitayakul, S.
K. Antioxidant Properties of Unripe Carica papaya Fruit Extract and Its Protective Effects against
Endothelial Oxidative Stress. Evidence-Based Complementary and Alternative Medicine,
(2019), 1–15.
[19]
Kirti Rani. Role of antioxidants in prevention of diseases, Journal of Applied Biotechnology &
Bioengineering, Volume 4 Issue 1,495-496(2017).
[20]
Malik, Farrukh Afaq, Sami Sarfaraz, Vaqar M. Adhami, Deeba N. Syed, and Hasan Mukhtar.
Pomegranate fruit juice for chemoprevention and chemotherapy of prostate cancer(2005).
[21]
Masria phetheresia sianipar, Edy suwarso, Rosidah . Antioxidant and anticancer activities of
hexane fraction from Carica papaya. Male flower, Asian J Pharm Clin Res, vol 11, issue 3, 81-
83(2018).
[22]
Mohamed A. El-Awady, Nabil S. Awad and Adel E. El-Tarras. Evaluation of the anticancer
activities of pomegranate (Punica granatum) and harmal (Rhazya stricta) plants grown in Saudi
Arabia, International Journal of Current Microbiology and Applied Sciences Volume 4 Number 5, pp.
1158-1167(2015).
[23]
Monika Sain, Vandana Sharma. Catharanthus roseus (An anti-cancerous drug yielding plant) A
Review of Potential Therapeutic Properties, International Journal of Pure and Applied Bioscience, 1
(6): 139-142(2013).

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