Chinese Medicine, 2011, 2, 47-52
doi:10.4236/cm.2011.22009 Published Online June 2011 (htt p://www.SciRP.org/journa l/cm)
In Vitro Studies on Sida cordifolia Linn for Anthelmintic
and Antioxidant Properties
Rajesh Singh Pawa*, Ankit Jain, Preeti Sharma, Pradeep Kumar Chaurasiya, Pradeep Kumar Singour
Pharmacognosy and Phytochemistry Division, Facul t y of Pharmacy,
VNS Group of Educations, VNS Campus, Vidya Vihar, India
Received March 1, 2011; revised March 16, 2011; accepted April 22, 2011
Abstract
The present study was undertaken to evaluate in-vitro antioxidant and anthelmintic activity of ethanolic and
aqueous extract from whole plant Sida cordifolia Linn (Malvaceae). The antioxidant activities are evaluated
by various antioxidant assays like α, α-Diphenyl-β-picrylhydrazyl (DPPH) free radical scavenging, total re-
ducing power, nitric oxide scavenging and hydrogen peroxide scavenging. The various antioxidant activities
were compared to standard antioxidan ts such as asc orbic acid . The anti oxidant acti vity o f ethanolic ex tract is
almost quantitativ ely equivalent to that of the standards used , ascorbic acid. The further anth elmintic activity
of whole plant is investigated through using Indian earthworm (Pheretima posthuma) showed that it is one of
the most importan t local medicinal plants both fo r ritual and et hnomedical p ractices. Various concentrations
of ethanol and aqueous extract (10, 20, 30, 40 mg/ml) of whole plant of Sida cordifolia Linn were tested in
the bioassay, which involve determination of time of paralysis of the worms. Albendazole was included as
reference sta ndard. The most activity was o bserved with aqueou s extract as compared to sta ndard drug. The
results from the above studies indicate that plant Sida cordifolia Linn. possesses potent antioxidant and an-
thelmintic activity.
Keywords: Albendazole, Ant helmintic, Antioxidant, Ascorbic Acid, DPPH, Pheretima posthuma, Sida
cordifolia Linn
1. Introduction
Sida cordifolia Linn. (Malvaceae) syn. Country Mallow
is a small, erect, downy shrub [1]. The leaves of the plant
are cord ate-oblong or ovate-oblong and fruits with a pair
of awns on each carpel. Roots of the plant which consti-
tute a drug are 5 - 15 cm long with few lateral roots of
smaller size. It is almost odorless with slightly bitter taste
[2]. It grows well through the plains of India, especially,
in damp climates. The shrub grows up to 0.75 - 1.5 me-
ters in height. The leaves are 2.5 - 7 cm long and 2.5 - 5
cm broad, with 7 - 9 veins. They are heart shaped, serrate
and truncate. The flowers are small, yellow or white in
color, solitary and axillaries. The fruits are moong -sized,
6 - 8 mm in diameter [3]. Bark is considered as cooling.
It is useful in blood, throat, urinary system related
troubles, piles, phthisis, insanity etc [4]. Franco et al.
tested that rather than being a stimulant, Sida cordifolia
actually acts as a depressant and decreases CNS activity
[5]. It acts as a weight loss product is through its hypo g-
lycemic (blood sugar lowering) activity [6]. Fumaric acid
isolated from S. cordifolia was reported to be hepatopro-
tective [7]. The roots of Sida cordifolia have been re-
ported to possess astringent, diuretic and ton ic properties.
The drug has also demonstrated antibacterial, antiplaque
and antifungal activities. Also it contains phytoconstitu-
ents such as pseudoephedrine, vaccine, asparagin, ephe-
drine, vascicinone, vaccine, vascinol. The present study
was undertaken to validate its anthelmintic and antioxi-
dant activity.
2. Materials and Methods
2.1. Plant Material
The whole plant of Sida cordifolia was identified, and
collected in the month of October 2009 from the forest of
Pachmarhi region, district Hoshangabad (Madhya Pra-
desh). The plant was authenticated by a Mr. S Rao, at
Department of crop and herbal physiology college of

R. S. PAWAR ET AL.
48
agriculture, Jawaharlal Nehru Krishi Vishwavidhyala,
Jabalpur (Voucher specimen No./HD/CHPY825).
2.2. Drugs and Chemicals
The following drugs and chemicals were used. Drug:
Albendazole (BANDY, Mankind Pharma Ltd, New Del-
hi). Chemicals: petroleum ether (40˚C - 60˚C) A.R. (PCL,
Pune), Ethanol A.R. (PCL, Pune), Dimethyl formamide
(DMF) A.R. (PCL, Pune), Saline water (Claris Lifesciences
Ltd., Ahmadabad), DPPH (Sigma Chemical Co.), Greiss
reagent (Sigma Chemical Co.), Ascorbic acid (Sigma
Chemical Co.) etc.
2.3. Animals
3 - 5 cm in length and 0.1 - 0. 2 cm in width Indian adult
earthworm (Pheretima posthuma) collected from moist
soil of Chambal fertilizer and chemical Ltd. MP Nagar,
star Arcade zone-I, Bhopal.
2.4. Preparation of Extracts
The plant was washed and dried in shade followed by
drying in hot air oven at low temperature then it was
coarsely powdered. Plant powder (60 gm) subjected to
successive solvent extraction in soxhlet apparatus using
various solvents viz. petroleum ether (40˚C - 60˚C),
ethanol and all the marc left was macerated with water.
The petroleum ether was used to defat the marc and dis-
carded. The ethanol and aqueous extracts were collected,
concentrated and dried in open air to give percentage
yield as 4.91% w/w and 26.24% w/w respectively [8].
2.5. Phytochemical Studies
Preliminary Phytochemical tests were performed as per
methods described by Ansari and Khandelwal [9,10].
Ethanolic and aqueous extracts tested to reveal the pres-
ence of diffe rent prim ary and sec ondary me taboli tes [ 11].
2.6. Anthelmintic Activity
The Invitro trial for anthelmintic activity of ethanol and
aqueous extract were conducted on earthworm Phereti-
ma posthuma as per previous method [12]. Ethanol and
aqueous extract of plant Sida cordifolia were dissolved in
minimum amount of DMF and the volume was adjusted to
10 ml with saline water. All drugs and extract solution
were freshly prepared before starting the experiment.
2.7. Description of Groups
In each group, 6 earthwo rms were released into 10 ml o f
desired formulations as follows: vehicle (5% DMF in
normal saline), Albendazole (10 - 40 mg/ml), and vari-
ous concentration (10 - 40 mg/ml) of ethanol extract or
aqueous extract of plant Sida cordifolia in normal saline
solution containing 5% DMF. Observations were made
for the time taken to paralysis and death of individual
worm. Paralysis is said to occur when the worm is not
able to move even in saline solution.
2.8. Antioxidant Estimation
The assessment of antioxidant activity was done through
various in-vitro assays. The free radical scavenging ac-
tivity of various concentrations of ethanol, aqueous ex-
tract of plant and ascorbic acid was measured in ter ms of
hydrogen donating and radical-scavenging ability using
the stable radical DPPH. Nitric oxide was generated from
sodium nitroprusside and measured by the Greiss reac-
tion. The activity was further confirmed by Reducing
power assay and Hydrogen Peroxide Radical Scavenging
Activity.
2.9. Free Radical Scavenging Activity
Antioxidant scavenging activity was studied using 1,
1-diphenyl-2-picrylhydrazyl free radical (DPPH) by the
method of Brand William W. Various concentrations of
test solutions in 0.1 ml were added to 0.9 ml of 0.1 mM
solution of DPPH in methanol. Methanol (0.1 ml) was
used as experimental control. After 30 min of incubation
at room temperature, the reduction in the number of free
radical was measured by reading the absorbance at 517
nm. Ascorbic acid was used as reference standard. The
scavenging activity of the samples corresponded to the
intensity of quenching DPPH. The percent inhibition was
calculated from the following equation: % Inhibition =
[(Absorbance of control – Absorbance of test sample)/
Absorbance control] × 100.
2.10. Reducing Power Assay
The reducing power of plant extracts were determined by
the method of Oyaizu. The capacity of extract to reduce
the ferric-ferricyanide complex to the ferrous-ferricya-
nide complex of Prussian blue was determined by re-
cording the absorbance at 700 nm after incubation. For
this purpose, different concentrations of aqueous and
ethanolic plant extract in 1 ml of distilled water were
mixed with pho sphate buffer (2.5 ml, 0. 2 M, pH 6.6) and
potassium ferricyanide (2.5 ml, 1%). The mixture was
incubated at 50˚C for 20 minutes Aliquots (2.5 ml) of
Trichloroacetic acid (TCA, 10%) were added to the
mixture. The 2.5 ml of solution was mixed with distilled
water (2.5 ml) and FeCl3 (0.5 ml, 0.1%). The absorbance

R. S. PAWAR ET AL.
49
was measured at 700 nm by spectrophotometer. In-
creased absorbance of the reaction mixture indicates in-
creased reducing ability.
2.11. Nitric Oxide Scavenging Activity
Nitric oxide scavenging activity was measured by the
spectrometry metho d of Madan MP. Sodium nitropruside
(5 mmol) in phosphate-buffered saline was mixed w ith a
control without the test compound, but with an equiva-
lent amount of methanol. Test solution of aqueous and
ethanolic extracts at different concentrations were dis-
solved in methanol and incubated at 25˚C for 30 min.
After 30 min, 1.5 ml of the incubated solution was re-
moved and diluted with 1.5 ml of Griess reagent (1%
Sulphanilamide, 2%) Phosphoric acid, and 0.1% Naph-
thyl ethylenediamine dihydrochloride). The absorbance
of the chromophore formed during the diazolization of
the nitrite with sulphanilamide and the subsequent
coupling with Naphthyl ethylenediamine dihydrochloride
was mea s ured at 546 nm.
2.12. Hydrogen Peroxide Radical Scavenging
Activity
The plant extract radical scavenging activity against hy-
drogen peroxide was determined using the method of
Ruch et al. Samples of aqueous and ethanolic of different
concentration were added to 0.1 M phosphate buffer so-
lution (pH 7.4, 3.4 ml), respectively, and mixed with 43
mM hydrogen peroxide solution (0.6 ml). After 10 min,
the reaction mixture absorbance was determined at 230
nm. The reaction mixture without sample was used as the
blank. Ascorbic acid was used as a reference compound.
The percentage inhibition activity was calculated [13].
3. Results and Discussion
The work presented here deals with preliminary phyto-
chemical study, Anthelmintic and Antioxidant estimation
of ethanolic and aqueous extract of whole plant Sida
cordifolia Lin n .
3.1. Phytochemical Studies
Tests for ethanol and aqueous extracts of whole plant
Sida cordifolia showed presence of alkaloids, resins,
steroids, proteins and flavonoids (Table 1).
3.2. Anthelmintic Activity
The results showed that aqueous extract of Sida cordifo-
lia plant took least time to cause paralysis and death of
the earthworms (Table 2). But the anthelmintic activity
Table 1. Preliminary phytochemical tests for ethanol and
aqueous extracts.
Chemical tests Phytochemicals Ethanol
extract Aqueous extract
Mayer’s Test Alkaloids + +
Killer Killani Test Glycosides _ _
Vanillin- HCl Test Tannins and phe-
nolic compounds _ _
Color detection
with ferric chloride Resins + +
Lead acetate test Flavonoids + +
Libermann-Bur
chard Test
Steroids + +
Ninhydrin Test A mino-acids + _
Biuret Test Proteins + +
Molisch’s Test Carbohydrate + +
Solubility test Fats and oils _ _
+= Present, - = Absent
Table 2. Invitro anthelmentic activity of Sida cordifolia.
Test
samples Concentration mg/ml
Time taken
for paralysis
(minutes)
Time taken for
death (mintues)
Albendazole
10
17.26 ± 0.437*
26.35 ± 1.026*
20
10.00 ± 0.25*
15.33 ± 0.31*
30
4.37 ± 0.15*
18.37 ± 0.121*
40
3.75 ± 0.67*
8.51 ± 1.231*
Aqueous
extract
10 19.18 ± 1.38* 30.05 ± 1.547*
20
13.38 ± 3.12**
21.58 ± 1.225*
30
6.35 ± 0.85*
20.00 ± 4.24**
40
05.35 ± 0.69*
10.23 ± 1.025*
Ethanolic
extract
10 27.00 ± 0.895* 34.28 ± 0.49*
20
10.00 ± 0.617*
23.45 ± 0.89*
30
8.27 ± 0.327*
15.38 ± 1.026*
40
5.52 ± 0.246*
18.00 ± 4.05**
Control earthwor ms (Pheretima posthuma) were alive up to 24 hours of the
experiment; All values represent mean _+ SEM; values are significant dif-
ferent f r o m reference s t an d ard (albendazole) wh er e, *p < 0.01; **p < 0.05 v/s
Albendazol e, SEM = Standa r d Error Mean
is found to be present in both aqueous as well as ethanol
extracts of Sida cordifolia plant [14].
3.3. Antioxidant Activity
3.3.1. DPPH Radical Scavenging Activity
The result of DPPH radical scavenging activity of the
ethanolic and aqueous extract of Sida cordifolia with
IC50 (% Inhibition) is shown in Figure 1. The IC50 value
of ethanolic and aqueous extract of Sida cordifolia was
found to be 15 µg/ml and 16 µg/ml respectively where as
the IC50 value of standard (Ascorbic acid) was found to
be 8 µg /ml. The results showed a significant (p < 0.01)
decrease in the concentration of DPPH radical due to the
scavenging ability of both extracts as compared to stan-
dard (ascorbic acid). Thus, it can be concluded that both

R. S. PAWAR ET AL.
50
Figure 1. Invitro DPPH radical scavenging activity of ascorbic acid and various extracts of Sida cordifolia.
Figure 2. Invitro reducing power assay of ascorbic acid and various extracts of Sida cordifolia.
extracts of Sida cordifolia have antioxidant activity. But
ethanolic extract was more significant as compared to
aqueous extract [15].
3.3.2. Reducing Power Assay Method
The Reducing ability of ethanolic and aqueous extract of
Sida cordifolia and ascorbic acid is shown in Figure 2.
The IC50 value of ethanolic and aqueous extract of Sida
cordifolia was found to be 16 µg/ml and 22 µg/ml re-
spectively where as the IC50 Value of Ascorbic acid
standard was found to be 11 µg/ml. Both the extracts
show significant (p < 0.01) reducing property when
compared with standard (ascorbic acid). Thus, it can be
concluded that ethanolic extract show more potent re-
ducing capacity as compared to aqueous extract [16].
3.3.3. Nitric Oxide Radical Scavenging Activity
The nitric oxide scavenging activity of ethanolic and
aqueous extract of Sida cordifolia and ascorbic acid
shown in Figure 3 illustrates the % inhibition of nitric
oxide generation by ethanolic and aqueous extract of
Sida cordifolia. Ascorbic acid was used as a reference.
The IC50 value of ethanolic and aqueous extracts were
found to be 112 µg/ml and 117 µg/ml, respectively,
whereas the IC50 value of ascorbic acid was found to be
85 µg/ml. The results indicate significant (p < 0.01) de-
crease in the concentration of nitric oxide radical due to
the scavenging ability of both ethanolic and aqueous
extract as compared to standard (ascorbic acid). Both the
extracts show significant scavenging activity but etha-
nolic extract showed more significant activity as com-
pared to aqueous extract [17].
3.3.4. Hydr oge n Pe roxi de Rad ic al Scave nging Ac tivity
Hydrogen peroxide scavenging activity of ethanolic and
aqueous extract of Sida cordifolia and ascorbic acid are
shown in Figure 4. The IC50 values for ethanolic and
aqueous extracts were 183 µg/ml, 190 µg/ml whereas

R. S. PAWAR ET AL.
51
Figure 3. Invitro Nitric oxide scavenging activity of ascorbic acid and various extracts of Sida cordifolia.
Figure 4. Invitro Hydrogen peroxide free radical scavenging activity of ascorbic acid and various ex-
tracts of Sida cordifolia.
170 µg/ml was the value of ascorbic acid. It showed sig-
nificant scavenging activity of hydroxyl radical generat-
ed from H2O2 system [18]. The results indicate that
ethanolic and aqueous extract possess significant anti-
oxidant activity (p < 0.01). Comparison of hydroxyl rad-
ical activity of both ethanolic and aqueous extract with
ascorbic acid showed ethanolic extract more significantly
active than aqueous extract [19].
4. Conclusions
The present pharmacological studies on plant Sida cor-
difolia for anthelmintic and antioxidant activity reveal
that the both extracts, i.e., ethanolic as well as aqueous
show the anthelmintic activity. For antioxidant activity
ethanolic extract was more significant as compared to
aqueous extract due to the presence of various phyto-
constituents such as alkaloid (asparagin, ephedrine, vas-
cicinone, vascinol, pseudoephedrine), flavonoids (5,7-
dihydroxy-3-isoprenyl flavone and 5-hydroxy-3-iso-
prenyl flavone) and phenolic compounds [20,21]. Thus,
the whole plant of Sida cordifolia can be used as folklore
medicine for the treatment of both causes.
5. Acknowledgements
The authors are tha nkful to Group dire ctor of VNS Institute
of Pharmacy and Research Centre for financial support,
providing necessary facilities, chemicals and other needful
environment for successful completion of this research.
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