International Journal of Engineering Science Invention

ISSN (Online): 2319 6734, ISSN (Print): 2319 6726

The Comparative Study of Antioxidant Activity Of

Shivangi Dave , 2,Asma Khan , 3,Yogita Kamble , 4,Dr.Vijay Dabholkar,

Department Of Chemistry and Department of Life Sciences, K.C. College, D.N. Road, Churchgate, Mumbai
400020, India

ABSTRACT: The increasing demands of convenient food have led to rapid growth in the ready-to-eat product
category. Many of the food ingredients contain unsaturated fatty acids that are quite susceptible to quality
deterioration, especially under oxidative stress. To combat this, the best strategy is addition of antioxidants.
Also, natural antioxidants in the daily diet need to be encouraged in order to improve human health and prevent
degenerative diseases like Alzheimers, coronary heart disease, neurogenetic disorders, cancer, atherosclerosis
and inflammations by oxidation of proteins, lipids, nucleic acids, DNA, etc. The DPPH method of free radical
scavenging was carried out and the effect of the commercial, fresh, dried and cooked sample of Clerodendrum
serratum were compared and calculated to check for its antioxidant activity. All samples showed antioxidant
activity, highest being in cooked, followed by commercial, fresh and dried samples. Thus, it is advisable to
consume the vegetable in cooked form at home as it shows highest radical scavenging power. The dried samples
were over-heated, thereby showing minimum antioxidant activity.
KEYWORDS: Antioxidants, Compared, Degenerative diseases, Oxidative stress, DPPH, Clerodendrum
serratum.

I.

INTRODUCTION

LITERATURE REVIEW: The literature review reveals some studies undertaken on the plants of various
geographic zones such as Western Ghats, where plants best known to local tribes, are investigated. (1) References
are also available for some of the monsoon vegetables and the species like Chelodendrum inerme (2) have been
used as an antioxidant drug in various indogenous medicines. (3)Organic and aqueous extractions of
C.colebrookianum(4) showed significant inhibition of lipid peroxidation in vitro and in vivo by FeSO4 ascorbate
in rats. This lends scientific support to the therapeutic uses of the plant leaves in tribal medicines. (5)
Antioxidant activity of some common plants:including Ocimum sanctum(6), Piper cubeba Linn.(7), Camellia
sinensis Linn.(8), Zingiber officinale Roscoe(9) and several Indian and Chinese plants are well studied. The
majority of the antioxidant activity is due to the flavones, isoflavone, flavonoids, anthocyanin, coumarin,
lignans, catechins, and isocatechins. The antioxidant rich foods are natural cleansers of the body, acting also as
anti-aging agents and protection against cancer. They absorb bad cholesterol substantially reducing the risks of
heart diseases (10). Some of the phytoconstituents being used in formulating dietary supplements or health foods
have their basis in the antioxidant activity of that constituent. Clerodendrum serratum Linn (11) local name,
Bharangi, is very widely distributed in tropical and subtropical regions of the world. Some cooking methods
may be better than others when it comes to maintaining beneficial antioxidant levels. Depending on the
vegetable, cooking on a flat metal surface with no oil (griddling) and microwave cooking maintains the highest
antioxidant levels (12). It was recently demonstrated that thermal treatments can induce the formation of
compounds with new antioxidant properties. These compounds are called Millard Reaction Products (MRPs).
Thus, on thermal treatment, antioxidant properties can be maintained or even enhanced by the development of
MRPs. (13) The current research project aims at the comparative study of antioxidant activity of the monsoon
plant Chlerodendrum serratum for the three types of leaf samples viz. fresh, dried and cooked.

II.

OBJECTIVES:

Collection of the monsoon plant Chlerodendrum serratum from the vegetable vendor.
Sample preparation into three types- fresh, dried and cooked leaves
Extraction of all the samples by soxhlet apparatus
Analysing it by gas chromatography
Antioxidant activity exhibited by the three samples by DPPH method

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Comparing the antioxidant activity of all the three samples along with the commercial sample (bharangi
churna, taken as standard) and checking for the highest antioxidant activity amongst the four samples.

III.

MATERIALS AND METHODS:

The plant sample was collected from local markets of Thane.

IV.

METHODS:

A. Sample Preparation:
[1]. Soxhlet Extraction (14): 14.92g of each sample was used in the Soxhlet Apparatus (15), using 140mL of
ethanol. 7 cycles were carried out for each extraction.
[2]. Rotary Evaporation (16) (17): It was carried out to concentrate the samples by evaporation of alcohol and to
reduce their volumes.
B. Analysis:
[1].
Gas Chromatography-Mass Spectrometry (18) (19): The samples were sent to UNIPOS Environment
Pvt.
Ltd.
Laboratory
for
GC/MS.
The
results
obtained
are
as
follows (20)
OBSERVATIONS:
a.

Fresh Sample:
O
HO

O
O

OH

Pectolinarigenin
5, 7-Dihydroxy-6-methoxy-2-(4-methoxyphenyl) chromen-4-one

OH

HO

H
HO

OH

1-Phenyl-3-(2, 4, 6-trihydroxy) cyclohexyl-3-hydroxy-1-propene

OH

b.Dried sample:

HO

O
OH

Apigenin
(5, 7-Dihydroxy-2, (4-hydroxylphenyl)-4H-1-benzopyran-4-one

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The Comparative Study Of Antioxidant...

O
O

O
2

Dimer of cleroflavone
1.

The DPPH Assay(21)(22): The antioxidant activity of the plant extracts and the standard was assessed on the
basis of the radical scavenging effect of the stable 1,1-diphenyl-2-picryl-hydrazyl (DPPH) free radical
activity by modified methods. The diluted working solutions of the test extracts were prepared in ethanol.
Commercially available Bharangi powder was used as standard.
0.002% DPPH was prepared in ethanol.
1mL of this solution was mixed with 1mL of sample solutions and standard solution separately.
These solution mixtures were kept in dark for 30 minutes.
Optical density was measured at 517nm using Spectrophotometer(23)(24)
Formula for calculating the antioxidant activity:[(A1-A2)/A1]x100(25)
Where, A1=Absorbance of control
A2=Absorbance of sample

V.

RESULTS AND DISCUSSIONS:

RESULTS:
SAMPLE
1.
2.
3.
4.

Commercial
Fresh
Dried
Cooked

OPTICAL DENSITY (nm)

VI.

OPTICAL DENSITY(nm)
(SAMPLE)
0.08
0.72
1.19
0.15

ANTIOXIDANT
ACTIVITY (%)
61.90
50.34
20.66
74.13

DISCUSSIONS:

By observing the values displayed in the observation table, it can be concluded that the cooked sample
exhibited the highest antioxidant activity (74%), followed by the fresh sample (50%) and least activity
exhibited by the dried sample (20%). The cooked sample was blanched below 100C. Thus, there was no overheating. Its high result can be attributed to the following sequence of events; Because of the heat, cells get
ruptured exposing maximal cell content. The antioxidant moiety comes out and the heat also leads to the
formation of new antioxidant-like compounds. All this will happen maximally if cooking is done on the flat
metal surface with no oil.This does not hold well in the case of dried leaves, as they were first sun dried and
then oven dried, above 100C. Excessive heat resulted in loss of volatile components, thereby reducing its
radical scavenging power. The commercial sample showed 61% antioxidant activity, which were still less than
that of the cooked sample. Thus based on these results, it can be suggested that one can eat Bharangi, the leafy
vegetable under investigation, in the cooked form which still has the high antioxidant activity.

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The Comparative Study Of Antioxidant...

VII.

for public

ACKNOWLEDGEMENTS:

We thank our principal, Ms. Manju Nichani, and K.C. College for Science Honors Program and
Jigyaasa. We also thank the UNIPOS ENVIRONMENT PRIVATE LIMITED LABORATORY, Dr. Gandhale
and Dr. Sopan for the analytical study of our sample.

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