Antioxidant and free radical scavenging activity of leaf extracts of Cleome chelidonii Linn.

Abstract

The plant extract studied could be an answer to the people
seeking for better therapeutic agents from natural sources, which is
believed to be more efficient with little or no side effects when
compared to the commonly used chemotherapeutic agents. The
present study verified the traditional use of Cleome chelidonii Linn.
for human ailments and partly explained its use in herbal medicine as
rich source of phytochemicals with the presence of tannins, phenols,
glycoside, flavanoids and terpenoids. Thus, this plant can be utilized
as an alternative source for chemo-drugs. Further studies are needed
with this plant to isolate, characterize and elucidate the structure of
bioactive compounds of this plant for industrial drug formulation. The
phytochemical analysis and antioxidant potential of Cleome chelidonii
Linn. were positive in all aspects. This may be due to the presence of
various compounds present in Cleome chelidonii Linn. The present
study reveals the potential of the plant taken for the study and further
extensive studies with a particular area of focus in future may form a
basis for new drug discoveries.

Full text

IJIPBART (2015) Volume 2, Issue (3), pp: 228-236 ISSN: 2349-865X
OPEN ACCESS
International Journal of Innovation in Pharma
Biosciences and Research Technology (IJIPBART)
Original Research Article
www.refsynjournals.com 228
Antioxidant and free radical scavenging activity of leaf
extracts of Cleome chelidonii Linn.
V.Sumitha
1
, M.Gurulakshmi
1
*
1
Department of Chemistry, Madras Christian College (Autonomous), Tambaram East, Chennai – 600059, India.
ABSTRACT
INTRODUCTION
Cleomaceae is a small family of flowering plants in the order Brassicales, comprising more
than 300 species belonging to 9 genera of which Cleome is the largest genus with about 180 - 200
species of medicinal, ethnobotanical, ecological importance (Aparadh et al., 2012). Cleome chelidonii
Linn. is a traditional plant belonging to the family Cleomaceae. It is grown as perennials throughout
dry seasons. It has a pink or white colour flowers. It is generally known for the treatment of colic,
dysentery, headache, otitis, and rheumatism (Kirtikar and Basu, 1987). Cleome chelidonii have many
medicinal applications, often in rubifacient and counter irritant preparations (Reddy and Raju, 2001).
They are also used for rheumatism and even headache. It contains glucocapparin and glucocleomin.
Article received
August 08, 2015
Article accepted
August 15, 2015
Article published
September 30, 2015
*
Corresponding Author:
M. Gurulakshmi,
Department of Chemistry,
Madras Christian College
Tambaram, Chennai – 600059,
India.
gurulakshmi.m@gmail.com
The plant extract studied could be an answer to the people
seeking for better therapeutic agents from natural sources, which is
believed to be more efficient with little or no side effects when
compared to the commonly used chemotherapeutic agents. The
present study verified the traditional use of Cleome chelidonii Linn.
for human ailments and partly explained its use in herbal medicine as
rich source of phytochemicals with the presence of tannins, phenols,
glycoside, flavanoids and terpenoids. Thus, this plant can be utilized
as an alternative source for chemo-drugs. Further studies are needed
with this plant to isolate, characterize and elucidate the structure of
bioactive compounds of this plant for industrial drug formulation. The
phytochemical analysis and antioxidant potential of Cleome chelidonii
Linn. were positive in all aspects. This may be due to the presence of
various compounds present in Cleome chelidonii Linn. The present
study reveals the potential of the plant taken for the study and further
extensive studies with a particular area of focus in future may form a
basis for new drug discoveries.
Keywords: Cleome chelidonii Linn., Phytochemical extraction,
Alkaloids, Terpenoids, Radical scavenging

Gurulakshmi et al IJIPBART (2015) Volume 2, Issue (3), pp: 257-265
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Free radicals are continuously produced in the human body, as they are essential for energy
supply, detoxification, chemical signaling and immune function (Gulcin, 2005). Free radicals can
initiate the oxidation of biomolecules, such as protein, lipid, amino acids and DNA which will lead to
cell injury and can induce numerous diseases (Hsu, 2003). This activity is due to the ability of
antioxidants to reduce oxidative stress by neutralizing or scavenging reactive species by hydrogen
donation (Erkan, 2008). Studies have shown that phenolics play an important preventive role in the
development of cancer, heart diseases and ageing related diseases (Larsomn, 1988). Oxidative
metabolism is an essential process for the survival of living things, drugs, and foodstuff and yet
causes formation of free radicals (Antolovich et al., 2002, Pourmorad et al., 2006). There are several
coupled reactions or antioxidant interactions between them, which is also an important pathway for
antioxidant protection in humans (Sami, 1995).
Antioxidants are molecules capable of preventing the oxidation of other molecules (Ames,
1993) thereby; protecting the body from damage caused by free radical induced oxidative stress.
Enzymatic antioxidants include superoxide dismutase, catalase and glutathione peroxidase (Mates et
al., 1999). The non-enzymatic antioxidants include ascorbic acid (vitamin C), α-tocopherol (vitamin
E), carotenoids, thiol antioxidants, flavonoids and metallonin (McCall and Frei, 1999). Flavonoids
have played a major role in successful medical treatments of ancient times (Cook and Samman, 1996,
Di Carlo et al., 1999, Cushnie and Lamb, 2005). In addition, they also inhibit lipid peroxidation,
platelet aggregation as well as the activity of enzyme systems including cyclooxygenase and
lipoxygenase (Awad et al., 2001). Flavonoids exert these effects as antioxidants, free radical
scavengers and chelators of divalent cations (Afanas’ev et al., 1989, Hollman et al., 1999) and are
reported to have unique cardioprotective effects (Rajadurai and Prince, 2007). In an earlier study, rats
fed a flavonoid-rich diet are reported to exhibit reduced myocardial post-ischemic damage (Heim et
al., 2002). The ultimate aim of the study is to reveal the antioxidant and free radical scavenging
vitality of Cleome chelidonii Linn. leaves as the leaves are widely used for external applications.
METHODS
Collection of Source
The specimens of Cleome chelidonii Linn. were collected from karuneelam village at
Kanchipuram district.
Cleome chelidonii Linn. Leaf Powder Preparation
The leaves of Cleome chelidonii Linn. were dried at 45⁰C for 24 hours and the dried leaves
were grinded and the moisture content was determined. 24g of Cleome chelidonii Linn. leaf powder
was measured using analytical weighing balance (Shimadzu, Japan).
Extraction
The leaf powder was extracted with four different solvents ie., ethanol, methanol, aqueous
and acetone by cold extraction.

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Phytochemical Screening
The different solvent extracts of
(Harborne, 1983; Sazada et al.,
2003)
Quantitat
ive Estimation of Total Alkaloid
The total alkaloid content was gravimetrically measured adopting the method Harborne (1973)
using the whole leaf powder.
Estimation of total Terpenoid
The total terpenoid content of the acetone, ethanol, methanol and aque
spectrophotometrically using Linalool as standard (Ghorai
Antioxidant Property of Extracts
Hydrogen Peroxide Scavenging Capacity
The ability of the
Cleome chelidonii
determined according to the method of Ruch
Hydroxyl Radical Scavenging Activity
The scavenging activity of methanol extract of
was measured according to the method of Arad
Reducing Power Assay
The antioxidants present in the sample reduce the oxidant probe and the respective product
interacted with some colouring agents to form a coloured complex with the method of
(2012).
RESULTS AND DISCUSSION
The leaves of
Cleome chelidonii
by cold extraction method. Diffe
extracts
. The extract were analysed for phytochemical screening to identify
bioactive compounds (Figure 2 & 3).
Figure 1.
Fresh leaves and powder of
These tests showed the presence of various phytochemical constituents like Terpenoids,
Saponins, Tannins, Sugar, Phenolics, Flavon
IJIPBAR
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The different solvent extracts of
C. chelidonii
leaf was screened for the phytochemical substances
2003)
ive Estimation of Total Alkaloid
The total alkaloid content was gravimetrically measured adopting the method Harborne (1973)
The total terpenoid content of the acetone, ethanol, methanol and aque
ous extract was evaluated
spectrophotometrically using Linalool as standard (Ghorai
et al., 2012)
Antioxidant Property of Extracts
Hydrogen Peroxide Scavenging Capacity
Cleome chelidonii
Linn.
extracts to scavenge hydrogen peroxide was
determined according to the method of Ruch
et al. (1989).
Hydroxyl Radical Scavenging Activity
The scavenging activity of methanol extract of
Cleome chelidonii
Linn.
was measured according to the method of Arad
et al. (1992).
The antioxidants present in the sample reduce the oxidant probe and the respective product
interacted with some colouring agents to form a coloured complex with the method of
RESULTS AND DISCUSSION
Cleome chelidonii
Linn.
were dried and powdered (Figure 1) and then extracted
by cold extraction method. Diffe
rent solvents of extracts like ethanol, methanol, a
. The extract were analysed for phytochemical screening to identify
the presence of different
bioactive compounds (Figure 2 & 3).
Fresh leaves and powder of
Cleome chelidonii
Linn.
These tests showed the presence of various phytochemical constituents like Terpenoids,
Saponins, Tannins, Sugar, Phenolics, Flavon
oids, Cardiac glycosides, Anthroquinones, Reducing
T (2015) Volume 2, Issue (3), pp: 2
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230
leaf was screened for the phytochemical substances
The total alkaloid content was gravimetrically measured adopting the method Harborne (1973)
ous extract was evaluated
extracts to scavenge hydrogen peroxide was
Linn.
on hydroxyl radical
The antioxidants present in the sample reduce the oxidant probe and the respective product
interacted with some colouring agents to form a coloured complex with the method of
Islam et al.
were dried and powdered (Figure 1) and then extracted
rent solvents of extracts like ethanol, methanol, a
queous and acetone
the presence of different
Linn.
These tests showed the presence of various phytochemical constituents like Terpenoids,
oids, Cardiac glycosides, Anthroquinones, Reducing

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sugar, Starch, Emodols, Steroid, Vitamin C, Proteins, Free amino acid,
Alkaloids. Methanol and e
thanolic extracts shows the presence of majority of phytoconstituents
(Table 1).
Large quantity of compounds in
radical scavenger, which indicates that the extract has good potential as a source for natural
antioxidants to prevent free radical mediated oxidative damage. This plant can be used f
ailments as explained by its use
in herbal medicine as rich source of phytochemicals
of tannins, phenols, glycoside, flavanoids and terpenoids.
Figure 2.
Screening of phytochemicals in the
Table 1.
Phytochemical Screening
Sl. No. Substance
1. Terpenoid
2. Saponin
3. Tannin
4. Sugar
5. Phenolics
6. Flavonids
7. Cardiac glycoside
8. Anthraquinone
9. Reducing sugar
10. Starch
11. Emodol
12. Steriod
13. Vitamin c
14. Proteins
15. Free amino acid
16. Carbohydrate
17. Glycoside
18. Alkaloid
Note: + for positive; -
for negative
In the present study, there was a significant in
methanol extract, then acetone, ethanol and lesser amount in a
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sugar, Starch, Emodols, Steroid, Vitamin C, Proteins, Free amino acid,
Carbohydrate, Glycosi
thanolic extracts shows the presence of majority of phytoconstituents
Large quantity of compounds in
Cleome chelidonii Linn.
extract makes it a strong free
radical scavenger, which indicates that the extract has good potential as a source for natural
antioxidants to prevent free radical mediated oxidative damage. This plant can be used f
in herbal medicine as rich source of phytochemicals
of tannins, phenols, glycoside, flavanoids and terpenoids.
Screening of phytochemicals in the
Ethanol (A), Methanol (B),
A
Acetone (D) extract of leaves
Phytochemical Screening
of Extracts
Methanol Ethanol Acetone
+ + +
- + -
+ - -
- - -
+ + -
+ - +
+ - +
+ + -
- - -
- - -
- - -
- - +
- - -
- - -
+ + +
- + -
+ + -
+ + +
for negative
In the present study, there was a significant in
crease in terpenoids content found
methanol extract, then acetone, ethanol and lesser amount in a
q
ueous extract. The total alkaloid was
T (2015) Volume 2, Issue (3), pp: 2
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231
Carbohydrate, Glycosi
de and
thanolic extracts shows the presence of majority of phytoconstituents
extract makes it a strong free
radical scavenger, which indicates that the extract has good potential as a source for natural
antioxidants to prevent free radical mediated oxidative damage. This plant can be used f
or human
in herbal medicine as rich source of phytochemicals
with the presence
A
queous (C) and
Aqueous
+
+
-
-
-
+
+
-
-
-
-
+
-
-
+
-
+
+
crease in terpenoids content found
highest in
ueous extract. The total alkaloid was

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shown almost 2.19% in whole leaf powder of
showed that the terpinoids are more present in leaf of
Figure 3.
Determination of terpenoids in ac
ethanol extract of leaves of
Table 2. Total Terpenoids a
nd Alkaloids
Sl. No.
1
Aqueous
2
Methanol
3
Acetone
4
Ethanol
5
Total
Figure
Antioxidant assay
Present data showed that
scavenging and antioxidant activities. Drying process would generally result in the depletion of
naturally occurring antioxidants in raw materials from plants (Tomaino
show that sun
drying of green leafy
property and free radical scavenging activity (Oboh
possessed higher Hydroxyl radical scavenging activity, H
extracts, as methanol was able to denature polyphenol oxidases. Since methanol was an organic and
volatile solvent, it was more efficient in plant cell wall degradation.
19.21
0
20
40
60
Aqueous
Quantitative analysis of total
alkaloids and terpenoids
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shown almost 2.19% in whole leaf powder of
Cleome chelidonii Linn.
. (Table 3). These results
showed that the terpinoids are more present in leaf of
Cleome chelidonii Linn.
Determination of terpenoids in ac
etone, aqueous, methanol and
ethanol extract of leaves of
Cleome chelidonii Linn.
nd Alkaloids
Extract Result
Aqueous
19.21mg/g
Methanol
52.10 mg/g
Acetone
42.43 mg/g
Ethanol
30.24 mg/g
Total
Alkaloids(whole leaf) 2.19 %
Figure
4. Total Terpenoids and alkaloids content
Present data showed that
Cleome chelidonii Linn.
leaf extract exhibited strong free radical
scavenging and antioxidant activities. Drying process would generally result in the depletion of
naturally occurring antioxidants in raw materials from plants (Tomaino
et al.,
2004). Earlier reports
drying of green leafy
vegetables could cause a significant increase in the reducing
property and free radical scavenging activity (Oboh
et al., 2009).
Methanolic extracts of fresh samples
possessed higher Hydroxyl radical scavenging activity, H
2
O
2
and antioxidant activity than cool water
extracts, as methanol was able to denature polyphenol oxidases. Since methanol was an organic and
volatile solvent, it was more efficient in plant cell wall degradation.
52.1 42.43 30.24
Aqueous
Methanol
Acetone
Ethanol
Quantitative analysis of total
alkaloids and terpenoids
Aqueous
Methanol
Acetone
Ethanol
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. (Table 3). These results
etone, aqueous, methanol and
leaf extract exhibited strong free radical
scavenging and antioxidant activities. Drying process would generally result in the depletion of
2004). Earlier reports
vegetables could cause a significant increase in the reducing
Methanolic extracts of fresh samples
and antioxidant activity than cool water
extracts, as methanol was able to denature polyphenol oxidases. Since methanol was an organic and
Aqueous
Methanol
Acetone
Ethanol

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Figure 6.
Hydroxyl radical scavenging and Catalase activity
Figure
8
Figure
37.59
0
10
20
30
40
Catalase activity of
17.01
0
50
100
Hydroxyl radical scavenging activity of
0
500
1000
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Figure 5. Reducing power assay
Hydroxyl radical scavenging and Catalase activity
Figure 7. Catalase activity (%)
8
. Hydroxyl radical scavenging activity (%)
Figure
9. Reducing power assay (mcg/g)
37.59
29.93 37.22 29.11
Catalase activity of
Celome chelidonii Linn.
AQUEOUS
METHANOL
ACETONE
ETHANOL
17.01
65.95 55.15 53.6
Hydroxyl radical scavenging activity of
Celome
chelidonii Linn.
AQUEOUS
METHANOL
ACETONE
ETHANOL
470 600
200 150
Reduced power assay of Cleome
chelidonii linn
AQUEOUS
METHANOL
ACETONE
ETHANOL
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Hydroxyl radical scavenging and Catalase activity
AQUEOUS
METHANOL
ETHANOL

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The extracts were found to possess stronger radical scavenging activity as seen in reduced
power assay and hydroxyl radical scavenging activity assay, respectively (Figures 7, 8 & 9). Ethanol
and methanol were able to completely inactivate degradative enzymes present in fresh plant materials,
thus yielding higher than that in aqueous and acetone extracts. Antioxidant activity was more in the
methanol extract compared to acetone, ethanol and water extracts, respectively. Cleome chelidonii
Linn. possessed higher concentration of antioxidant activity in the methanol extract (Figure 9).
Whereas the free radical scavenging activities for ethanol, acetone and water extracts were lesser in
the Cleome chelidonii Linn. leaf extract.
CONCLUSION
In the present study, the findings revealed that the four extracts from leaf of Cleome
chelidonii Linn. contain phytochemical constituents like Terpenoids, Saponins, Tannins, Sugar,
Phenolics, Flavonoids, Cardiac glycosides, Anthroquinones, Reducing sugar, Starch, Emodols,
Steroid, Vitamin C, Proteins, Free amino acids, Carbohydrates, Glycosides and Alkaloids. Methanol,
alcohol, acetone and aqueous extracts showed significant antioxidant activity, Catalase activity, H
2
O
2
,
terpenoids and total alkaloid content, respectively. The phytochemical analysis and antioxidant
potential of Cleome chelidonii Linn. were positive in all aspects especially in methanol extract. This
may be due to the presence of various compounds present in Cleome chelidonii Linn. The present
study reveals the potential of the plant used and further extensive studies with a particular area of
focus in future may form the basis for new drug discoveries. Further studies and investigations should
be aimed at isolation and characterization of the active principles responsible for the reported
activities. This should include analysis of the active principles using Mass Spectrometry, Infra Red
Spectroscopy, Nuclear Magnetic Resonance spectroscopy, High Performance Thin Layer
Chromatography and confirmatory studies like mechanism of action at organic and inorganic levels in
animal models.
ACKNOWLEDGMENT
We thank M/S. Refsyn Biosciences Pvt. Ltd., Puducherry for their support in carrying out this
research work.
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Cite this article in press as Gurulakshmi et al. (2015) Antioxidant and free radical
scavenging activity of leaf extract of Cleome chelidonii Linn., IJIPBART, 2(03); 257-265.
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