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Human Journals
Research Article
August 2015 Vol.:4, Issue:1
© All rights are reserved by Chintala Shivakoti et al.
Evaluation of
In Vitro
Antioxidant Activity of
Sateria verticillata
Leaves by Using DPPH Scavenging Assay
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Keywords: Sateria verticillata, poaceae, DPPH assay,
antioxidant activity
ABSTRACT
Drugs from the plant origin are easily available, less
expensive, safe, efficient and rarely have side effects. The
compounds such as alkaloids, tannins, flavanoids and phenols
have a major role in preventing a number of chronic diseases
by a definite physiological action on the human body like anti-
inflammatory, anti-thrombotic, anti-oxidant, hepatoprotective
and anticarcinogenic activities. The aim of present study was
to evaluate the undertaken to analyze the antioxidant activity
of various extract of Sateria verticillata plant. Standard
methods were adopted to assess antioxidant activity of the
plant materials. The extent of radical scavenging was
determined by calculated IC50 value. The results revealed that
aqueous, ethyl acetate and ethanol extracts showed good
antioxidant activity when compared to Ascorbic acid standard.
Chintala Shivakoti1*, O.Ishwarya2, Alluri Ramesh3,
D.Rakesh goud4
Vishnu Institute of Pharmaceutical Education and
Research, Vishnupur, Narsapur, Medak, India.
Department of Chemistry, Vishnu Institute of
Pharmaceutical Education and Research,
Telangana, India.
Submitted: 11 August 2015
Accepted: 16 August 2015
Published: 25 August 2015
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Citation: Chintala Shivakoti et al. Ijppr.Human, 2015; Vol. 4 (1): 230-234.
231
INTRODUCTION
Oxygen is an essential element for life to perform biological functions such as catabolic and
anabolic process of fats, proteins and carbohydrates in order to generate energy for growth
and other activities of the cell. Although oxygen is not dangerous by itself, but it is involved
in the generation of various kinds of "reactive oxygen species" (ROS). ROS can interact with
biomolecules and ultimately lead to free radical chain reactions. Free radical chain reactions
are produced in the mitochondrial respiratory chain, liver mixed function oxidase, xanthine
oxidase activity, atmospheric pollutants and for transition metal catalysts, drugs and
xenobiotics [1, 2]. ROS attacks the unsaturated fatty acids present in the biomembranes
resulting in membrane lipid peroxidation, a decrease in membrane fluidity, loss of enzymes
and receptor activity and damage to membrane protein leading to cell inactivation [3],
mutation leading to cancers [4]. ROS also leads to pathological conditions such as ischemia,
anemia, asthma, arthritis, inflammation, neurodegenration, Parkinson’s diseases, mongolism,
ageing process and dementia. Antioxidants are used in the treatment of diseases caused by
ROS. Antioxidants are composed of a group of compounds and enzymes potent enough to
scavenge free radicals before they cause tissue damage [5]. Vitamin E, vitamin C,
carotenoids, natural flavanoids etc. are the natural antioxidants produced in the body while
others must be sequestered from the diet or through supplementation. Most antioxidants were
found in citrus and dried fruits, cruciferous vegetables, garlic, onions, carrots, tomatoes, sweet
potatoes, sesame and olive oil. There are thousands of naturally occurring and synthetic
antioxidants known; these antioxidants belong to different classes of compounds and may
cause some side effects [6]. Plant secondary metabolites such as phenolic compounds,
carotenoid, ascorbic acid, thiols and tocopherols have shown antioxidant activity that includes
scavenging free radical species, inhibiting the production of reactive species, inhibiting the
production of reactive species resulting from normal cell metabolism. Thereby prevent the
damage to lipids, proteins, nucleic acids and subsequent cellular damage and death [7].
Sateria verticillata (family: poacae) commonly known as bristly foxtail and hoocked bristle
grass. Parts of the plant are also being used for many disorders like rheumatism, psoriasis and
chronic eruptions.
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Citation: Chintala Shivakoti et al. Ijppr.Human, 2015; Vol. 4 (1): 230-234.
232
MATERIALS AND METHODS
Collection and authentication
Sateria verticillata were collected from Narsapur, Medak District and authenticated by D.
Venkaeswara Rao, Deputy Director, A. P. Forest Academy, Dullapally, Hyderabad, Ranga
Reddy Dist.
Processing of plant materials
Each part of the plant was washed under running water to make it free from dust and foreign
particles. The plant parts were powdered and kept in air tight container before analysis.
Preparation of Extracts
Powdered plant material (200 gm) was extracted with water, ethanol, and ethyl acetate using
cold maceration method. All the extracts were filtered with a muslin cloth and the filtrate was
concentrated in vacuum evaporator. Dried extracts were used for further studies [8].
Preparation of standard solution
Required quantity of Ascorbic acid was dissolved in ethanol to give the concentration of 20,
30, 40, 50 μg/ml.
Preparation of test solution
Stock solutions of samples were prepared by dissolving 10 mg of dried plant extract in 10 ml
of ethanol to give concentration of 1 mg/ml. Then prepared sample concentrations of 20, 30,
40, 50 μg/ml.
Preparation of DPPH solution
3.9 mg of DPPH was dissolved in 3.0 ml ethanol; it was protected from light by covering the
test tubes with aluminum foil.
In-vitro antioxidant assay
A great number of in vitro methods have been developed to measure the efficiency of natural
antioxidants either as pure compounds or as plant extracts. α,α-diphenyl-6 picrylhydrazyl
radical Scavenging assay (DPPH), Ferric reducing antioxidant power (FRAP), Nitric oxide
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Citation: Chintala Shivakoti et al. Ijppr.Human, 2015; Vol. 4 (1): 230-234.
233
Scavenging assay, Superoxide anion radical Scavenging assay, ABTS radical Scavenging
assay, hydroxyl radical Scavenging assay, are the in vitro antioxidant assay methods used to
assess the antioxidant activity of the leaves extracts of Sateria verticillata. The absorption
maximum of a stable DPPH radical in method was at 517 nm. The decrease in absorption of
DPPH radical caused by antioxidants, because of the reaction between antioxidant molecule
and radical progresses, results in the Scavenging of the radical by hydrogen donation.
Statistical analysis
All data were expressed as mean±SD. Statistical analysis was performed by One-way
ANOVA using Origin version 6.0 software and p<0.05 and p<0.001 was considered as
statistically significant.
RESULTS AND DISCUSSION
These diverse groups of compounds have potential of natural antioxidant and have ability to
act as both efficient radical scavengers. The antioxidant activity of phenols is due to their
redox properties, hydrogen donors and singlet oxygen quenchers [13]. The antioxidative
characteristics might be attributed to the presence of phytochemical such as flavanoids and
other phenolic compounds. Polyphenols have been known to show medicinal activity as well
as exhibiting physiological activity. The compounds such as flavanoids; which contain
hydroxyls are responsible for the radical scavenging activity in plant [10, 9, 2,]. The reducing
capacity of a compound may be used as a significant indicator of its potential antioxidant
activity [11]. Reducing power is to the measure of the reductive ability of antioxidant and it is
judged by the transformation of Fe3+ to Fe2+ in the presence of extracts [12]. The reduction
power of aqueous and other extracts was summarized in Table1. The data showed that
reducing power of the extracts increased with increased concentration of extracts. The extracts
showed potent ferric reducing power.
The result of DPPH scavenging activity assay in this study indicates the ethyl acetate extract
was potentially active. The aqueous and other extracts produced more or less similar DPPH
anion scavenging power of 44.36±2.09% ethyl acetate & 40.12±5.36% ethanol extract at 50
μg/ml concentration with 92.648±30.68 μg/ml of IC50 for ethyl acetate extract & 106.15
±25.33 μg/ml of IC50 value for ethanol extract and 63.99 ±25.24 μg/ml for Ascorbic acid
(Table 1). The scavenging activity of ethyl acetate soluble fractions compared with the
standard drug ascorbic acid suggests that the plant phytochemicals are a potent scavenger of
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Citation: Chintala Shivakoti et al. Ijppr.Human, 2015; Vol. 4 (1): 230-234.
234
free radicals. However further study aimed at characterization of active constituents
responsible for antioxidant activity. Overall, the ethyl acetate extract of Sateria verticiilata
Linn leaves have most potent antioxidant activity.
Table 1. In vitro free radical scavenging effect of Sateria verticillata leaves by DPPH
method:
Extracts
Conc/ percentage Scavenging
IC50 g/ml
20 mg/ml
30 mg/ml
40 mg/ml
50 mg/ml
Aqueous
31.23±4.26**
33.32±4.81*
34.62±3.04*
37.73±4.46*
84.54±4.62
Ethanol
21.33±2.08**
25.08±1.00*
30±4.35**
44.36±2.09**
92.648±30.368
Ethyl acetate
13.40±4.01**
22.48±2.04**
28.35±8.95**
29.25±9.47**
106.158±25.332
Standard
4.95±6.85**
12.77±6.96**
27.96±1.41*
27.72±3.77**
63.997±25.244
Significant at *p=<0.05; **p=<0.001
CONCLUSION
On the basis of the results it is concluded that the extracts contain higher quantities of
phenolic compounds, which exhibit antioxidant and free radical scavenging activity. It also
chelates iron and possesses reducing power. In vitro assay systems confirm Sateria
verticillata leaves as natural antioxidants. Further in vivo assessment also needed to confirm
the antioxidant nature of Sateria verticillata leaves.
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