9

Antioxidant Activity of Combined Extract of Some Medicinal

Plants of Indian Origin
1

H. Ali1 and S. Dixit2

Department of Chemistry, Research scholar, MANIT, Bhopal

2
Department of Chemistry, HOD, MANIT, Bhopal
Email: sana_soni26@yahoo.co.in, savitadixit1@yahoo.com

Abstract
Azdirachta indica, Tinospora cordifolia, Ocimum sactum, Triticum aestivum, Aloe barbandesis are Indian
medicinal plants and has several medicinal properties. Antioxidants are essential substances which possess
the ability to protect the body from damaged caused by free radical induced oxidative stress. A variety of free
radical scavenging antioxidants exist with in the body which many of them derived from nutritional sources
like food, vegetables. In this study the antioxidant activity of acetone and methanol (30:70) combine extract
of selected plant materials, traditionally used by Indian population was evaluated against 2,2-diphenyl-1-picrylhydrazyl radical. This study suggests the possible mechanism of antioxidant activity due to the presence of
flavonoids in each plant extract.

Introduction
About 80% of the world population depends absolutely on plants for their health and healing. Whereas
in the developed world, confidence on surgery and
pharmaceutical medicine is more usual but in the recent years, more and more people are complementing
their treatment with natural supplements [1]. Furthermore, motivation of people towards herbs are increasing due to their concern about the side effect of drugs,
those are prepared from synthetic materials. The
people want to concern their own health rather than
merely submitting themselves to impersonal health
care system. Many botanical and some common dietary supplements are good sources of antioxidants
compounds [2]. Plant materials containing phenolic
constituents are increasingly of interest as they retard
oxidative degradation of lipids and thereby improving quality and nutritional value of food [3, 4]. The
importance of the antioxidant constituents of plant
material is the maintained of health and protection
from cancer [5]. They are very important substances
that possess the ability to protect the body from damage caused by free radical induced oxidative stress [6,
7]. Several researches on the phenolic constituent and
antioxidant activities in various plants have been con-

ducted [8]. In the meantime, Galvez etal. [9] pointed

out that there was a correlation between antioxidant
capacity and phenolic content.
Recently, natural have received much attention as
source of biological active substances including antioxidants. Numerous studies have been carried out on
some plants, vegetables, fruits because they are rich
sources of antioxidants such as vitamin C, vitamin A,
vitamin E, carotenoids, polyphenolic compounds and
flavonoids [10] which prevent free radical damage, reducing risk of various diseases. Thus the consumption
of antioxidants from these resources is beneficial in
preventing disease. The search for newer antioxidant
compounds, especially of plant origin has ever since
increased.

Materials and Methods

Collection of plant material

Fresh leaves of Azadirachta indica, Ocimum sactum, Aloe barbandesis, Stem of Tinospora cordifolia,
Whole grass of Triticum aestivum were collected from
their proper origin.

M.M. Srivastava, L.D. Khemani, S. Srivastava, Chemistry of Phytopotentials: Health, Energy and Environmental Perspectives, DOI:10.1007/9783-64223394-4_9, Springer-Verlag Berlin Heidelberg 2012

43

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Section A Health Perspectives

Extraction of plant material

The plant materials (leaves of Azadirachta indica,

Ocimum sactum, Aloe barbandesis, stem of Tinospora
cordifolia, Whole grass of Triticum aestivum were airdried at room temperature (26C) for 2 weeks, after
which it was grinded to a uniform powder. The mixture of acetone and methanol (30:70) extract were
prepared by using 75g each of the dry powdered
plant materials in Soxhlet apparatus at 40C for 48 h.
The extract were filtered after 48 h, the extracts were
concentrated using a rotary evaporator with the water bath set at 40C. The percentage yield of extracts
ranged from 717%w/w. The extract of each plant
material were mixed in equal proportion and make a
single sample.

methanol and DPPH were used as the standard control. Thirty minutes later, the absorbance was measured at 517nm. The absorbance of DPPH solution
decreases when kept in contact with antioxidant test
sample and free radical scavenging activity is inversely proportional to the absorbance of DPPH solution [11, 12]. Percent inhibition of DPPH free radical
scavenging activity was calculated using the following formula.
DPPH Scavenged (%) = ( Acont Atest) / Acont X 100
Where Acont is the absorbance of the control reaction.
Atest is the absorbance in the presence of the sample
of the extract.

DPPH scavenging activity

1mg of extract powder were dissolved in 1ml of 90%

methanol solution to obtain 1000g/ml sample solutions were series diluted in to concentration ranging
from 4001000g/ml (i.e. 400, 500, 600, 700, 800,
900 and 1000g/ml). 200 M solution of DPPH in
methanol was prepared and 1.5ml of this solution was
added to 1.5ml of methanol extract solution at different concentrations (4001000g/ml). Mixture of

% in h ib itio n

Determination of antioxidant activity

100
90
80
70
60
50
40
30
20
10
0

y = 0.0716x + 22.306
R = 0.8224

Se rie s1
L in e a r (Se rie s1 )

500
1000
Concentration l/m l

1500

Figure 1: Inhibition of DPPH by the combine extract Azadirachta indica, Ocimum sactum, Aloe barbandesis, Tinospora
cordifolia, Triticum aestivum

Table 1: Characteristics of the used medicinal plants

Extract yield
%w/w

Part used

Family name

Scientific name

Chemical constituent

17.24

Leaves

Meliaceae

Azadirachta indica

Reducing sugar, Terpenoids, Flavonoids, Saponins, Alkaloids, Cardiac

glycosides

16.09

Leaves

Lamicae

Ocimum sactum

Flavonoids, Tannins

10.08

Branch

Asphodelaceae

Aloe barbandesis

Flavonoids, Tannins, Alkaloids

9.26

Stem

Menispermaceae

Tinospora cordifolia

Reducing sugar, Flavonoids,

Saponins, Alkaloids, Cardiac
Glycosides, Tannins

7.94

Grass

Poaceae

Triticum aestivum

Flavonoids, Tannins, Alkaloids

9 Antioxidant Activity of Combined Extract of Some Medicinal Plants of Indian Origin

Scavenging of hydrogen peroxide

Sample with different concentration (i.e. 400, 500,

600, 700, 800, 900 and 1000g/ml) were added to
0.1M phosphate buffer solution (pH 7.4, 3.4ml) respectively and mixed with 43mM H2O2 solution
(0.6ml). After 10 minutes, the reaction mixture absorbance was determined at 230nm. The reaction mixture without sample was used as the blank [13].
The % inhibition activity = ( Acont Atest) / Acont X 100
Where Acont is the absorbance of the control reaction.
Atest is the absorbance in the presence of the sample
of the extract.
90

y = 0.0682x + 15.688
R = 0.9866

80

% in h ib itio n

70
60
50

S e rie s1

40

L in e a r (S e rie s1 )

30
20
10
0
0

500

1000
Concentration g/m l

1500

Figure 2: Inhibition of Hydrogen peroxide by the combine extract of Azadirachta indica, Ocimum sactum, Aloe barbandesis, Tinospora cordifolia, Triticum aestivum

Results and Discussion

The percentage yield of Azadirachta indica, Ocimum
sactum, Aloe barbandesis, Tinospora cordifolia, Triticum aestivum was found to be 717%w/w. Preliminary phytochemical screening of the extracts of individual plant showed the presence of flavonoids.
DPPH assay is based on the measurement of scavenging ability of antioxidant towards the stable DPPH
radical. DPPH is relatively stable nitrogen centered
free radical that can accept an electron or hydrogen
radical to become a stable diamagnetic molecule.
DPPH radicals react with suitable reducing agent as a
result of which electron become paired off forming the
corresponding hydrazine. The solution therefore loses
color stoichiometrically at 517nm [14]. From result it
may be postulated that the combine extract have hydrogen donor thus scavenge free radical DPPH.

45

H2O2 is a weak oxidizing agent and can inactivate

a few enzymes directly, usually by oxidation of essential thiol (SH) groups. Hydrogen peroxide can cross
cell membrane rapidly. Once inside the cell, H2O2 can
probably react with Fe2+ and possibly Cu2+ to form hydroxyl radical and this may be the origin of many of
its toxic effects. It is therefore biologically advantageous for cell to control the amount of hydrogen peroxide that is allowed to accumulate. The decomposition of H2O2 by the combine extract may result from
its antioxidant activity.
Results obtained in the present study indicate that
the combine extract of Azadirachta indica, Ocimum
sactum, Aloe barbandesis, Tinospora cordifolia, Triticum aestivum showed the maximum antioxidant activity with IC50 386.7g/ml in DPPH and 504.7g/ml
in hydrogen peroxide.

Conclusion
Results obtained in the present study indicate that the
combine extract inhibit free radical scavenging activity. The overall antioxidant activity of extract might
be attributed to its polyphenolic content and other
phytochemical constituents. It could be a source of
natural oxidant that could have greater importance as
therapeutic agent in preventing various diseases.

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Section A Health Perspectives

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