I J P B C S Research Article - International Journal of...
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International Journal of Pharmaceutical
Biological and Chemical Sciences
International Journal of Pharmaceutical, Biological and Chemical Sciences (IJPBCS)
| JAN-MAR 2014 | VOLUME 3 | ISSUE 1 | 06-17 www.ijpbcs.net or www.ijpbcs.com
Research Article
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e6
ANTIOXIDANT AND FREE RADICAL SCAVENGING ACTIVITY OF ANNULAR
AND SEASONAL AYURVEDIC MEDICINAL PLANTS ELEPHANTOPUS SCABER L.
AND BIOPHYTUM SENSITIVUM DC.
A. Subramanian1, S. Santhosh Kumar
2*, S. K. Suja
3, M. Sudarshan
4 and A. Chakraborty
5
1Department of Biochemistry, Raja Serafoji Government College, Thanjavur-613005, Tamil Nadu, India
*2Department of Physics, Avvaiyar Government College for Women, Karaikal – 609 602,
U.T of Puducherry, India 3 Department of Chemistry, Lady Doak College, Tallakulam, Madurai - 625002, Tamilnadu, India
4Trace Element Sciences Laboratory, UGC-DAE Consortium for Scientific Research, Kolkata - 700 098, India
5Radiation Biology Laboratory, UGC-DAE Consortium for Scientific Research, Kolkata 700 098, India
*Corresponding Author Email: [email protected]
INTRODUCTION
Medicinal plants are widely used in the ancient system
of medicines such as Ayurveda, Siddha, Unani, and in
Tibetan medicine. 3/4th
of the population in developing
countries relies on traditional medicine, according to the
world health organisation (WHO) that too in the form of
crude extracts, decoction, arishta, lehya, surna, and even
in tablet form. Millions of rural households use
medicinal plants in a self help mode. In recent years
secondary plant metabolites phytochemical,
pharmacological activities have been extensively
investigated as a source of curative agent[1]. Recently
there has been an upsurge of interest in the therapeutic
potentials of plants, antioxidants in reducing free radical
induced tissue injury, although several butylated
hydroxy toluene (BHT), are commercially available, but
are quite unsafe and their toxicity is a problem of
concern. Hence strong restrictions have been placed on
their application and which is to be substituted with
naturally occurring antioxidants.
Many plant species have been investigated in the search
for novel antioxidants [2-5] but generally there is still a
demand to find more information concerning the
antioxidant potential of plant species as they are safe and
also bioactive. The antioxidant detoxifies the reactive
oxygen species (ROS), a potentially reactive derivative
of oxygen, present in the body. However excess
production of ROS or inadequate antioxidant defense
causes oxidative damage to various bio-molecules
including DNA, which is a critical etiological factor
implicated in several chronic diseases like, diabetes
mellitus, cancer, arthritis and also in ageing process[6].
Therefore, considerable attention has been directed, now
a days, towards the identification of plants with
antioxidant ability.
Natural antioxidants especially phenolics and flavanoids
from tea, wine, fruits, vegetables and spices are already
exploited commercially either as antioxidant additives or
as nutritional supplements [7]. Gulcin[8] reported the
antioxidant and radical scavenging activity of black
pepper seeds from both water extract and ethanol extract.
Due to the bioactive nature and safe to use of naturally
occurring antioxidants, many studies have been
conducted in recent years, and most recently, Vinay at al.
[6] have studied the antioxidant activity of commonly
available plants and found Hibiscus is comparatively
ABSTRACT:
Traditionally used Ayurvedic medicinal plants, Elephantopus scaber and Biophytum sensitivum are investigated for their
antioxidant potency. Preliminary phytochemical analysis of pet-ether fraction of these plant leaves show the presence of
alkaloids, tannins, saponins, flavanoids and phenolic compounds. Both the pet-ether and ethanol fractions exhibited
antioxidant activity significantly. The DPPH radical scavenging and the nitric oxide scavenging activity are high for ethanol
fraction than pet-ether and ascorbic acid. The pet-ether fraction of B.sensitivum showed comparatively good reducing power
even at low concentration and shows higher potentiality in antioxidant activity than other extracts, which is in good
coincidence with the phytochemical constituents. The results validate the potential usage of these plants in the traditional
Ayurvedic system of medicine, to a greater extent.
KEYWORDS: Ayurvedic medicinal plant, phytochemical, antioxidant activity, DPPH, E.scaber, B.sensitivum.
S. Santhosh Kumar *
et al; ANTIOXIDANT AND FREE RADICAL SCAVENGING ACTIVITY OF ANNULAR ……………
International Journal of Pharmaceutical, Biological and Chemical Sciences (IJPBCS) | JAN-MAR 2014 | VOLUME 3 | ISSUE 1 | 06-17| www.ijpbcs.net
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rich in flavanoids, which is used in many ways by the
south Indian people, such as, oil, food supplement,
gulkand, lehya, shampoo, etc. Hence it is a necessity
now to identify potentially antioxidant plants, available
in common.
The oxidative stress, defined as “the imbalance between
oxidants and antioxidants in favour of the oxidants
potentially leading to damage” has been suggested to be
the cause of aging [9] and various diseases in humans
[10]. In modern western medicine, the balance between
antioxidation and oxidation is believed to be a critical
concept maintaining a healthy biological system [10-14].
Such concept exists in China since 2000 years, called
yin-yang (antioxidant-oxidation process). It was found
that yin-tonic herbs are mainly having flavanoids[15],
which are phenolic compounds with strong antioxidant
activity. Hence increasing the intake of foods rich in
antioxidant compounds, (e.g., polyphenols, cartenoids)
will enhance human health. As a consequence many
plants have been identified as having potentially active
antioxidant [10, 16, 17].
The majority of the antioxidant activity is due to the
flavones, isoflovones, flavonoids, anthocynin, coumarin
lignans, catechins and isocatechins [18]. Besides,
phenolic compounds and flavonoids are also
widely distributed in plants having multiple biological
effects, including antioxidant, free radical scavenging
abilities, anti-inflammatory, anti-carcinogenic
etc.[19,20].
The current focus is towards natural antioxidants, and it
is interesting to investigate the antioxidant properties of
plants, especially, which are in traditional medicinal
practice. Hence we have identified two important plants
such as Elephatopus scaber Linn, and Biophytum
sensitivum Dc., which are used in Ayurvedic practice for
several ailments.
Eventhogh many plants are having reported to be good
antioxidants the availability of plants is still a big
question. Even though the Biophytum sensitivum is
having good antioxidant and free radical scavenging
capacity, this plant is available only for around three to
four months after rainy season. And hence we selected
an annual/perennial plant Elephatopus scaber for such
study in comparison with Biophytum sensitivum.
Numerous applications of E. scaber in folk medicine as
well as the bioactivities shown by various extracts and
respective compounds of this plant are discussed earlier
[21,22]. Bioactivity screening of the extracts and their
compounds had shown that E. scaber possess wound
healing, anti-venom, anti-microbial, anti-inflammatory,
anti-diabetic, cytotoxic and anti-tumour activities.
The sacred plant B. sensitivum is one among the
Dashapushpam in the traditional culture of Kerala, is a
potent medicinal plant. It is having the radio-protective
activity, immune modulatory action, cardio protective,
antitumor activity, wound healing activity, etc. [23-25].
MATERIALS AND METHODS
The Plant Elephatopus scaber Linn, collected from the
southernmost part of India, the Kanyakumari District of
TamilNadu at coordinates 8°18'8"N 77°10'34"E, and
Biophytum sensitivum Dc, collected from Mahe region at
coordinates 11°42′4″N 75°32′12″E, located in Kerala
under U.T. of Puducherry, India, are washed well in pure
water and shadow dried. The dried leaves are separated
and powdered using mortar and pestle. The pet-ether
extract of the sample was obtained by adding the plant
powder with pet-ether in the ratio of 1gm for 10ml and
extracting for 48 hours using Soxhlet apparatus. The
solvent is concentrated at temperature below 40oC and
the resulting extracts are used for determination of
flavanoids, phenols and free radical scavenging activity.
Phytochemical screening
The Phytochemical studies are carried out using the
freshly prepared pet-ether extracts of these plant leaves
to detect the presence of stereoids, terpenoids, tannins,
flavonoids, saponins, glycosides, amino acids etc., using
standard procedures[26-30].
Test for Flavonoid
Alkaline reagent test: Extracts are treated with few drops
of sodium hydroxide solution. Formation of intense
yellow colour, which becomes colourless on addition of
dilute acid, indicates the presence of flavonoids.
Flavanoids are found to be useful in disease resistance
[31]. They are hydroxylated phenolic and they have been
found to be antimicrobial substances against wide array
of microorganisms in vitro. Their activity is probably
due to their ability to complex with extracellular and
soluble proteins and to complex with bacterial cell wall
[32]. They also are effective antioxidant and show strong
anticancer activities [33-35].
Test for carbohydrates
Molisch’s test: Filtrates are treated with 2 drops of
alcoholic α- naphthol solution in a test tube. Formation
of the violet ring at the junction indicates the presence of
carbohydrates.
Systematical reduction of the amount of dietary
carbohydrates could suppress, or at least delay the
emergence of cancer, and that proliferation of already
existing tumor cells could be slowed down[36], but the
cell surface carbohydrates are acting as tumor
suppressors.
S. Santhosh Kumar *
et al; ANTIOXIDANT AND FREE RADICAL SCAVENGING ACTIVITY OF ANNULAR ……………
International Journal of Pharmaceutical, Biological and Chemical Sciences (IJPBCS) | JAN-MAR 2014 | VOLUME 3 | ISSUE 1 | 06-17| www.ijpbcs.net
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Test for alkaloids
Dragendroff’s test:
Filtrates are treated with Dragendorffs’ reagent (solution
of potassium bismuth iodide). Formation of red
precipitate indicates the presence of alkaloids.
Alkaloids are known to play some metabolic roles and
control development in living system [37]. These have
protective role in human and animal and it is used in
medicine especially the steroidal alkaloids which
constitute most of the valuable drugs. One of their
biological properties is cytotoxicity [38]. Alkaloids also
have analgesic [39, 40], antispasmodic and antibacterial
[41, 42] properties.
Test for saponin
Froth test: Extract aere diluted with distilled water to
20ml and this was shaken in a graduated cylinder for 15
minutes. Formation of 1cm layer of foam indicates the
presence of saponins.
Saponins, which are known to produce inhibitory effect
on inflammation [43] and has the property of
precipitating and coagulating red blood cells. Crude
saponins are having inhibition of glucose transport.
Glycosides are known to lower the blood pressure [44].
Steroids have been reported to have antibacterial
properties [45] and are important especially due to their
relationship with compounds such as sex hormones [46].
Plant sterols could act as a natural preventive dietary
product [47]. It helps in the lowering of plasma
cholesterol and LDL (low-density lipoprotein)
cholesterol. Hence, it will assist in reducing drastically
the morbidity and mortality caused by cardiovascular
disease.
Plant extracts containing alkaloids, flavanoids, phenolic
and/or terpenes showing active trypanocidal activity
[48]. As per Sofowora [26], these compounds are potent
bioactive compounds that could be used for therapeutic
purpose or which are precursors for the synthesis of
useful drugs. The phenolic compounds are one of the
largest and most ubiquitous groups of plant metabolites
[49]. They possess biological properties such as anti-
apoptosis, anti-aging, anti-carcinogen, anti-
inflammation, anti-atherosclerosis, cardiovascular
protection and improvement of endothelial function, as
well as inhibition of angiogenesis and cell proliferation
activities [18]. The phenols are reported to inhibit alpha-
amylace, sucrose, as well as the action of sodium
glucose-transporter 1 (SGLUT-1) of the intestinal brush
border. The polyphenol compound has antimicrobial and
antifungal effect and has been used in disinfections.
Determination of antioxidant activity
The pet-ether fractions of the two plants, E. scaber and
B. sensitivum, and the ethanolic fractions of B.
sensitivum were screened for free radical scavenging
activity against DPPH, reducing power and Nitric oxide
radical at different concentrations. All determinations
were performed in triplicate. The percentage inhibition
of the DPPH radical by the sample was calculated as
follows [2, 50],
Absorbance of Control- Absorbance of Sample
% DPPH. Scavenging = 100 x --------------------------------------------------------------
Absorbance of Control
Where Absorbance of Control is the absorbance in absence of standards or extracts and Absorbance of Sample is the
absorbance in presence of standards or extracts.
The DPPH radicals are widely used to investigate the
scavenging activity of some natural compounds. 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 method[51].
The diluted working solutions of the test extracts are
prepared in pet-ether. Ascorbic acid is used as standard
in 100-500 µg/ml solution. 0.002% of DPPH was
prepared in pet-ether and 1ml of this solution was mixed
with 1ml of sample solution and standard solution
separately. These solution mixtures were kept in dark for
30 min and optical density was measured at 517 nm
using Amile photo calorimeter.
In the DPPH assay, the antioxidants were able to reduce
the stable radical DPPH to the yellow coloured diphenyl-
picrylhydrazine, resulting a colour change from purple to
yellow. The absorbance decreased when the DPPH was
scavenged by an antioxidant through donation of
hydrogen to form a stable DPPH molecule. In the radical
form, the molecule had an absorbance at 517nm which
disappeared after acceptance of an electron or hydrogen
radical from an antioxidant compound to become a
stable diamagnetic molecule [52]. This method is based
on the reduction of alcoholic DPPH solution in the
presence of a hydrogenating antioxidant due to the
formation of the non- radical form (DPPH-H) in the
reaction [53]. The experiment was repeated for three
times. Ascorbic acid was used as standard control. IC50
values denote the concentration of sample, which is
required to scavenge 50% of DPPH free radicals.
S. Santhosh Kumar *
et al; ANTIOXIDANT AND FREE RADICAL SCAVENGING ACTIVITY OF ANNULAR ……………
International Journal of Pharmaceutical, Biological and Chemical Sciences (IJPBCS) | JAN-MAR 2014 | VOLUME 3 | ISSUE 1 | 06-17| www.ijpbcs.net
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Reducing power determination Yen and Chen [54] method was followed to determine
the reducing power. Different amounts of extracts (100-
500 μg/ ml) in water were mixed with phosphate buffer
(2.5 ml, 0.2 M, pH 6.6) and potassium ferricyanide
[K3Fe (CN) 6] (2.5 ml, 1%). The mixture was incubated
at 50oC for 20 min. and the reaction was stopped by
adding 2.5 ml of trichloroacetic acid (10%) and then it
was centrifuged at 3000 rpm for 10 min. 2.5ml of the
upper layer solution was mixed with equal quantity of
distilled water and 0.5ml of FeCl3, and the absorbance
was measured at 700nm[55]. Increased absorbance of the
reaction mixture indicated increased reducing power.
Ascorbic acid is used as positive control.
Nitric oxide-scavenging activity This procedure is based on the principle that, sodium
nitro-prusside in aqueous solution at physiological pH
spontaneously generates nitric oxide which interacts with
oxygen to produce nitrite ions that can be estimated
using Griess reagent. Scavengers of nitric oxide compete
with oxygen, leading to reduced production of nitrite
ions. For the experiment, sodium nitroprusside (10 mM),
in phosphate-buffered saline, was mixed with different
concentrations of each extracts dissolved in water and
incubated at room temperature for 60 minutes. After the
incubation period, 0.5 ml of Griess reagent was added.
The absorbance of the chromophore formed was read at
540nm [55]. Ascorbic acid is used as positive control.
RESULTS Phytochemical analysis
The phytochemical constituents of E.scaber and
B.sensitivum are analysed using the pet-ether fraction of
the samples. Many studies have been conducted in
E.scaber leaf [56] with benzene, chloroform, and
methanol extracts, but pet-ether fraction shows the
presence of alkaloids, tannins, saponins, flavanoids,
phenolic copmpounds and carbohydrates. The
B.sensitivum also shows the presence of such compounds
(Table.1)
In vitro antioxidant activity
DPPH- radical scavenging activity
DPPH is a stable nitrogen-centered free radical, the color
of which changes to yellow upon reduction by either the
process of hydrogen- or electron- donation. Substances
which are able to perform this reaction can be considered
as antioxidants and therefore radical scavengers [57]. It
is found that the radical-scavenging activities of all the
extracts increased with increasing concentration (Fig.1).
Among the three extracts of two plants, ethanol extract
of E. scaber showed the best activity, i.e., IC50 = 517.56
g/ml. IC50 for DPPH radical scavenging activity is
reported in figures plotted separately (Figs. 2-5).
Reducing power Fe (III) reduction is often used as an indicator of electron
donating activity, which is an important mechanism of
phenolic antioxidant action [58]. In the reducing power
assay, the presence of antioxidants in the samples would
result in the reducing of Fe3+
to Fe2+
by donating an
electron. Increasing absorbance at 700 nm indicates an
increase in reductive ability. Fig. 6 shows dose-response
curves for the reducing powers of the samples. It is
found that the reducing powers of all the samples
increased with their concentrations. Ethanol extract of
B.sensitivum and pet-ether extract of E. scaber show
good reducing power at lower concentrations compared
to ascorbic acid.
Nitric oxide-scavenging activity The % inhibition was increased with increasing
concentration of the extracts (Fig.7). The ethanol extract
of B.sensitivum showed better reducing power than other
extracts. However, activity of ascorbic acid is very more
pronounced than that of the samples at the concentration
>300g/ml. In addition to reactive oxygen species, nitric
oxide is also implicated in inflammation, cancer and
other pathological conditions [58].
Antioxidants scavenge for free radicals and consequently
are a very special group of nutritional supplements. The
free radicals are all along present in the body, due to the
occurrences of biochemical process, but it is harmful if it
exceeds the normal level. Antioxidants are substances
with free radical chain breaking properties. Among the
numerous antioxidants available, flavonoids are known
to inhibit lipid peroxidation to scavenge free radicals and
active oxygen to chelate iron ion and to inactive
lipoxygenase [59].
Table.1 Phytochemical constituents of E.scaber, and B.sensitivum.
Compound E.Scaber B.Sensitivum
Benzene* Chloroform* Methanol* Pet-ether Pet-ether
Alkaloids -- -- -- + +
Tannins -- + + + +
Saponins -- -- -- + +
Flavonoids -- -- + + +
Phenolic
compound -- -- + + +
Carbohydrate -- -- -- + +
* From Ref.[56].
S. Santhosh Kumar *
et al; ANTIOXIDANT AND FREE RADICAL SCAVENGING ACTIVITY OF ANNULAR ……………
International Journal of Pharmaceutical, Biological and Chemical Sciences (IJPBCS) | JAN-MAR 2014 | VOLUME 3 | ISSUE 1 | 06-17| www.ijpbcs.net
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Fig.1. DPPH radical scavenging activity of the pet-ether extract of E.scaber leaves
and pet-ether & ethanol extract of leaves compared with standard, Ascorbic acid.
Fig.2. IC50 value of ethanol extract of B.sensitivum.
The IC50 value marked in figure is obtained through linear fitting of the curve.
100 200 300 400 500
0
10
20
30
40
50
60
70
80
90
100
% in
hib
itio
n
Concentration (gm)
B. Sensitivum
E. Scaber
B. Sensitivum (Ethanol)
Ascorbic acid
100 200 300 400 500
0
10
20
30
40
50Linear Regression
Y = A + B * X
Parameter Value Error
-----------------------------------------
A -4.849 4.37839
B 0.10605 0.0132
-----------------------------------------
R SD N P
-----------------------------------------
0.97754 4.17463 5 0.00403
-----------------------------------------
IC50
=517.560
% in
hib
itio
n
Concentration(gm/ml)
S. Santhosh Kumar *
et al; ANTIOXIDANT AND FREE RADICAL SCAVENGING ACTIVITY OF ANNULAR ……………
International Journal of Pharmaceutical, Biological and Chemical Sciences (IJPBCS) | JAN-MAR 2014 | VOLUME 3 | ISSUE 1 | 06-17| www.ijpbcs.net
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Fig.3. IC50 value of pet-ether extract of B.sensitivum.
The IC50 value obtained is marked in the figure.
Fig.4. IC50 value of pet-ether extract of E.scaber. Linear interpolation is used to obtain the IC50 value.
100 200 300 400 500
0
10
20
30
40
50
60
70Linear Regression
Y = A + B * X
Parameter Value Error
-------------------------------------------
A -7.457 5.24374
B 0.14727 0.01581
-------------------------------------------
R SD N P
-------------------------------------------
0.983 54.99971 5 0.00262
-------------------------------------------
IC50
=393.99
% in
hib
itio
n
concentration(gm/ml)
100 200 300 400 500
0
10
20
30
40
50
60
70
80Linear Regression
Y = A + B * X
Parameter Value Error
----------------------------------------
A -0.578 4.64138
B 0.14807 0.01399
----------------------------------------
R SD N P
----------------------------------------
0.98686 4.42538 5 0.0018
----------------------------------------
IC50
=339.316
% inh
ibitio
n
Concentration(gm/ml)
S. Santhosh Kumar *
et al; ANTIOXIDANT AND FREE RADICAL SCAVENGING ACTIVITY OF ANNULAR ……………
International Journal of Pharmaceutical, Biological and Chemical Sciences (IJPBCS) | JAN-MAR 2014 | VOLUME 3 | ISSUE 1 | 06-17| www.ijpbcs.net
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Fig.5. IC50 value of the standard, Ascorbic acid.
Fig.6. Reducing power of B.sensitivum and E.scaber. Ascorbic acid is used as control.
100 200 300 400 500
30
40
50
60
70
80
90
100Linear Regression
Y = A + B * X
Parameter Value Error
-------------------------------------
A 16.61072 3.37404
B 0.15592 0.01017
------------------------------------
R SD N P
------------------------------------
0.99368 3.21702 5 6.032E-4
------------------------------------
IC50
= 217.5836
%in
hib
itio
n
Concentration(g/ml)
100 200 300 400 500
0
100
200
300
400
500
% inh
ibitio
n
Concentration(g/ml)
B.sensitivum (pet.ether)
B.sensitivum (ethanol)
E.scaber
Ascorbic acid
S. Santhosh Kumar *
et al; ANTIOXIDANT AND FREE RADICAL SCAVENGING ACTIVITY OF ANNULAR ……………
International Journal of Pharmaceutical, Biological and Chemical Sciences (IJPBCS) | JAN-MAR 2014 | VOLUME 3 | ISSUE 1 | 06-17| www.ijpbcs.net
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Fig.7. Nitric oxide scavenging activity of pet-ether extract of E.scaber, pet-ether
& ethanol extract of B.sensitivum and standard (Ascorbic acid).
Fig.8. Comparision of IC50 vales of different extracts of the plants E.scaber and
B.sensitivum and ascorbic acid.
DISCUSSION
In the present paper we have evaluated the free radical
scavenging activity, of E.scaber and B.sensitivum. The
crude extracts of these plants showed antioxidant
activity, of which B.sensitivum Dc. showed a weak
value. It has been shown that the scavenging effects on
the DPPH radical increases sharply with the increasing
concentration of the samples and standards to a certain
100 200 300 400 500
0
20
40
60
80
100
120
140
160
180
200
B. Sensitivum
E. Scaber
B. Sensitivum (Ethanol)
Ascorbic acid
% in
hib
itio
n
Concentration (gm)/ml
E.scaber B.sens (P) B.sens(E) Ascorbic Acid
0
100
200
300
400
500
IC50
Samples
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et al; ANTIOXIDANT AND FREE RADICAL SCAVENGING ACTIVITY OF ANNULAR ……………
International Journal of Pharmaceutical, Biological and Chemical Sciences (IJPBCS) | JAN-MAR 2014 | VOLUME 3 | ISSUE 1 | 06-17| www.ijpbcs.net
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extent [60] and hence are said to be strongly dependent
on the phenols and antioxidant activity. The radical
scavenging activities of the selected plant extracts are
still less effective than the commercial available
synthetic like BHT, as the plant extracts are quite safe
and their toxicity is a not a problem of concern unlike
those of BHT, they could be exploited as antioxidant
additives or as nutritional supplements.
The results of the present study show that the extract of
E.scaber contains the highest amount of flavonoids and
alkaloids, which corresponds to higher nitric oxide
activity. Therefore the plant can be further studied for
novel antioxidant compounds, which is very well
evidenced by the present work. However , the chemical
constituents present in the extract, which are responsible
for this activity, need to be investigated, and it is obvious
that the constituents like alkaloids, tannins, saponins, and
flavanoids present in the extract may be responsible for
such activity. Several of such compounds are known to
possess potent antioxidant activity [17]. Some of these
constituents have already been isolated from these
plants. Hence, the observed antioxidant activity may be
due to the presence of any of these constituents. The
plants exhibited strong anticancer, hepato protective,
antiviral and several other activities which may be due to
its antioxidant activity. The crude pet ether extracts
merits further experiments in vivo. The DPPH
scavenging activity IC50 values (Fig.2-5) show a higher
inhibition of ethanol extract (IC50 = 517.560; Fig.2) of
B.sensitivum than its pet-ether extract (IC50 = 393.999;
Fig.3) which are still higher than the pet-ether extract
(IC50 = 339.316; Fig.4) of E.scaber. The standard,
ascorbic acid shows only IC50=217.5836 (Fig.5).
The order of DPPH radical scavenging activity is,
ethanol fraction of B.sensitivum, pet.ether fraction of
B.sensitivum and E.scaber and then ascorbic acid
(Fig.8). Ganga Rao et al.[61] extensively studied the
antioxidant and radical scavenging activity of E.scaber
using hydro alcohol, methanol, ethyl acetate and hexane
extracts. In this paper we have taken the pet.ether extract
of E.scaber for the study of radical scavenging activity.
The DPPH radical scavenging activity of this plant
shows a higher value at 50% inhibition than the other
extracts, compared to Ganga Rao et al.[61].
Increasing absorption at 700nm indicates an increase in
reductive ability. Fig.6 shows the reducing powers of the
extracts. It is found that the reducing power increased
with the increase of their concentrations. The pet-ether
extract had shown a good reducing power compared to
ascorbic acid, but at concentrations above 300g/ml, all
the fractions showed good reducing powers. And the
ethanol fraction of B.sensitivum showed a smooth
increase with concentration.
The nitric oxide scavenging capacity (Fig.7) also shows
IC50 value is higher for ethanol extract (IC50 = 681.361)
of B.sensitivum than its pet-ether extract (IC50 = 216.666)
followed by the pet-ether extract (IC50 = 158.207) of
E.scaber.
In very recent articles [23, 62] on B. sensitivum reports
only the usage of aqueous and methanolic extracts for its
study, but pet-ether and ethanol extract give additional
information about the chemical constituents of this plant
and hence possess higher antioxidant activity.
CONCLUSION
In conclusion, even though different extracts exhibited
different levels of antioxidant activities in different
models studied, ethanol and pet-ether extracts, in general
show a good result of antioxidant activity than any other
fraction and hence this may be the first report of these
plants on pet-ether and ethanol extract. The B.Sensitivum
can be replaced with E.scaber only for its phytochemical
constituents and its availability throughout the year . But
in the context of antioxidant and free radical scavenging
capacity ethanol and pet-ether extracts of B.Sensitivum
values higher than E.scaber. But further studies are
needed to identify the unknown antioxidant components
to establish their pharmacological properties.
ACKNOWLEDGEMENT
This research was partially supported by the UGC-DAE
Consortium for Scientific Research, Kolkata Center,
through a facility utilization project.
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*Corresponding author address:
S. Santhosh Kumar
Department of Physics,
Avvaiyar Government College for Women,
Karaikal – 609 602, U.T of Puducherry, India,
Telephone: 094864-59600
E-mail: [email protected]