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A B S T R A C T
RGUHS Journal of Pharmaceutical Sciences
Evaluation of Analgesic and Antipyretic Activity of Tridax Procumbens Leaves
Extract
1 1 1 2 1
Narayan Singh Patel , Deepak Kumar Jain* , Hemant Nagar , Arti Patel and H.S.Chandel
*1
Truba Institute of Pharmacy, Karond Gandhi Nagar Bypass Road, Bhopal, M.P – 462038, India
2
Radharaman College of Pharmacy, Ratibad Bhopal, M.P – 462036, India
This study was designed to evaluate the analgesic and antipyretic activity of leaves extract of Tridax procumbens Linn. (ethanolic and
ethyl acetate). Analgesic activity was investigated against Acetic acid and sodium chloride induced Writhing test in rats. TPEE at
300mg/kg (P.O.) shows better analgesic activity than TPEAE while both extracts at 200mg/kg (P.O.) have very low analgesic activity.
Ethanolic and Ethyl acetate extracts of Tridax procumbens were evaluated for antipyretic activity using hyperpyrexia induced in rabbits
by Typhoid-Paratyphoid A, B vaccine. Like Paracetamol 100 mg/kg (I.V.) Tridax procumbens showed significant reduction in elevated
body temperature at 300 mg/kg (P.O.) .The analgesic and antipyretic activity of Tridax procumbens leaves could be at least in part due to
the presence of flavonoids and other polyphenols in the extracts.
Keywords: Analgesic, Antipyretic activity, Tridax procumbens.
INTRODUCTION
Tridax procumbens L. (Compositae) is a hispid, procumbent
herb, found as a weed throughout India. Tridax procumbens is a
perennial plant, 15–40cm high, often rooting at the nodes,
1
sometimes developing a woody base . Its common names
include coat buttons and Tridax daisy in English, cadillo
chisaca in Spanish, herbe caille in French, Jayanti Veda in
Sanskrit, ghamra in Hindi, Bishalya karani in Oriya, Dagadi
pala in Marathi, Gaddi Chemanthi in Telugu,Thata poodu in
2
Tamil, and kotobukigiku in Japanese . The poultice of whole
plant is used as an anti-inflammatory remedy by the tribal
3
healers . The previous pharmacological investigations on
4,5
Tridax procumbens reported acute anti-inflammatory
6 7 8,9
Immunomodulatory wound healing , hepatoprotective
10 11
anti-leishmanial anti-hyperglycemic and hair growth
12 13 14.
promoting activities anti-diabetic Antimicrobial The
Phyotochemical investigation reports the isolation of lipid
constituents, sterols, flavonoids, polysaccharide, and bergenin
15, 16
derivatives from T. procumbens . The leaves are reportedly
used to treat bronchial catarrh dysentry and diarrhoea and as
a hair restorative. In southern Orissa a paste prepared from
the whole plant is taken orally to relieve diarrhoea. A fine
paste of the leaves is applied externally to reduce swelling of
hemorrhoids. The whole plant and seed being used to treat a
variety of ailments the leaves are cooked and eaten as a
17
vegetable .
RGUHS Journal of Pharmaceutical Sciences
Received: 8/7/2011, Modified: 11/9/2011, Accepted: 25/11/2011
226
Original Research Article
By considering the ethnopharmacology and research articles
on Tridax procumbens and being an anti-inflammatory herb due
4, 5
to presence of flavanoinds , so that it may have activity
against Pain and Pyrexia; hence aim of present study is to
evaluate the in Vivo Analgesic and Antipyretic activity of
Tridax procumbens leaves extracts by Writhing test and
Hyperpyrexia Induced by Typhoid-Paratyphoid A, B Vaccine
in rats and rabbits respectively.
MATERIAL AND METHODS
The plant was collected in the month of September-October
2010 from the fields of village Mau, Dist Narsinghpur (M.P).
It was authenticated by Prof. A.B.Tiwari, Head of
Department of Crop and Herbal Physiology, Jawaharlal
Nehru Krishi Vishwavidyalaya, Jabalpur (M.P.) and the
specimen voucher no. assigened was DCHP/1610/
Sep10/JNKV.
The leaves were dried under shade and pulverized to coarse
powder which was passed through sieve No.20 to maintain
uniformity and used for the further study like screening of
leaf, powder and crude extract to determine the leaf
characters, physiochemical analysis such as Loss on Drying,
Total ash value, Foaming Index, Water soluble ash value and
Acid insoluble ash value while the crude extract was tested for
several phytochemical test for different phytoconstituents
such as Alkaloids by Dragendorff's test and Mayer's test,
Glycosides by Legal's test and Baljet's test ,Carbohydrate by
Benedict's test and Molisch's test, Steroids by Salkowski test
and Liebermann-Burchard test, Proteins by Biuret test,
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Flavonoids by Shinoda test and Alkaline reagent test also
18, 19
tested for Saponins and Tannins.
Animals care and handling
The experiments were carried out on albino rabbits of 5-8
months of either sex weighing between 1.5 to 1.6 kg and
Wistar albino rats of either sex of weight 150 ±20 grams.
They were provided from Truba Institute of Pharmacy,
Bhopal and were acclimatized to the standard laboratory
conditions in well cross ventilated animal house at
temperature 25±2°C relative humidity 44 –56% and light
and dark cycles of 10 and 14 hours respectively for 1 week
before and during the experiments. Rabbits were kept in mesh
bottom iron cages to avoid caprophagy habit they fed with
cauliflower, cabbage, carrot and water for 10 days before the
experiment while Rats were fed with standard diet (Golden
Feeds, Delhi) and water ad libitum. The experiments were
approved by the Institutional Animal Ethics Committee and
a s p e r C P C S E A g u i d e l i n e s ( a p p r o v a l n o .
1196/a/08/CPCSEA).
Drugs and chemicals
Diclofenac sodium provided by Novartis pharmaceuticals
Pvt. Ltd, Paracetabmol provided by Lupin Pharmaceuticals
Pvt. Ltd., Typhoid-Paratyphoid A, B vaccine (Sanofi pasture)
was purchased and all other chemicals utilized were of
analytical grade.
Acute toxicity studies
The LD 50 was calculated by “Staire case” method. The LD50
was determined in rats and mice by oral and intraperitonea1
route. The initial dosing was 2000 mg/kg p.o. and 800 mg/kg
4
i.p. in both the species .
Preparation of extract dose
2% acacia suspension was prepared by suspending 2 gram of
accurately weighed acacia powder in 100 ml of 0.9% saline.
20 ml of vehicle was taken separately to which 2 gram of dried
extract was added and sonicated, this produce suspension of
100 mg/ml strength. Both TPEE and TPEAE suspension
were prepared in such manner.
Animal grouping
Patel et al Evaluation of Analgesic and Antipyretic Activity of Tridax Procumbens Leaves Extract.
For Analgesic activity, rats were divided into six groups and
each group contains six animals. Group I (Negative control,
vehicle treated), Group II (Standard, Diclofenac sodium)
10mg/kg treated), Group III (TPEE 200mg/kg treated),
Group IV (TPEE 300mg/kg treated), Group V
(TPEAE200mg/kg treated), Group VI (TPEAE 300mg/kg
treated).
For Antipyretic activity, rabbits were divided into four groups
227
and each group contains four animals. Group I (Negative
control, vehicle treated), Group II (Standard, Paracetamol
100mg/kg treated), Group III (TPEE 300mg/kg treated),
Group IV (TPEAE 300mg/kg treated).
Screening of Analgesic Activity in Rats
Acetic acid induced writhing test
Acetic acid 1%v/v 1ml/100gm was injected intraperitonialy
to animals of each group. Standard and test groups were
treated with diclofenac sodium (I.P.) 15 minute and ethanolic
extract, ethyl acetate extract (P.O.) 1 hour prior to the
administration of acetic acid respectively while control group
was treated with vehicle. Wriths were the stretching of the
abdomen with simultaneous stretching of at least one hind
limb. The total no. of wriths was recorded for 10 min starting
20, 2
10 min after injection of acetic acid.
NaCl induced writhing test
Sodium chloride 4%w/v at 1ml/kg concentration was
injected intraperitonialy to animals of each group. Standard
and test groups were treated with diclofenac sodium (I.P.) 15
minute and ethanolic extract, ethyl acetate extract (P.O.) 1
hour prior to the administration of Nacl respectively while
control group was treated with vehicle. Then they were
observed for a period of 10 min after 10 min of injection of
acetic acid and the number of writhes is recorded for each
20, 21
animal. The percent inhibition of wriths was calculated.
Screening of Antipyretic Activity in Rabbits
Typhoid-Paratyphoid A, B vaccine 0.1 ml was injected into
the marginal ear-vein of rabbits of each group. Standard and
test groups were treated with paracetamol (I.V.) 15 minute and
ethanolic extract, ethyl acetate extract (P.O.) 1 hour after the
administration of Typhoid-Paratyphoid A, B vaccine
respectively while control group was treated with vehicle. The
rectal temperature each animal was recorded 15 minute
before and at the interval of 30 minute after treatment using
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Telethermometer up to 4 hours
Statistical Analysis
The results wereexpressed as mean ±SEM. The results of the
present study were analyzed using one way ANOVA followed
by dunnett's t-test computed statically by using Graph pad
prism software (version 5.04).
RESULTS
Leaves were green, characteristic odour with slight bitter taste.
Leaves were of size 3-5 cm in length and 1-1.5 cm width.
There fracture was soft and rough texture. (Table 1). The
findings of physiochemical analysis were that the total ash
value was 6.6%, acid insoluble ash (2.5%), water soluble ash
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Table 1: Morphological characteristics of leaves of
Tridax procumbens.
S.No. Characters Observation
1 Color Green
2 Odour Characteristic
3 Taste Acridic
4 Size 3-5cm lenght,1-1.5cm width
5 Texture Rough
6 Fracture Soft
(1.14%), loss on drying was 0.1% and foaming index was less
than 20%. These parameters suggest that the plant part
contains nil moisture, have sufficient amount of organic
content. (Table No.2). On performing several tests for
Phyotochemical analysis on the ethanolic and ethyl acetate
extract it was observed that both the extract contains
appreciable amount of flavonoids and Phytosterols while
other Phytoconstituents such as alkaloids, tannins, resins,
proteins and carbohydrate were also present while they lack
glycosides. (Table No.3). The Analgesic activity of Tridax
Table 2: Physiochemical analysis of powdered leaves of
Tridax procumbens.
S.No. Parameters Observations (%)
1 Loss on drying 0.1
2 Foaming index 20
3 Total ash value 6.6
4 Acid insoluble ash value 2.5
5 Water soluble ash value 1.14
Patel et al Evaluation of Analgesic and Antipyretic Activity of Tridax Procumbens Leaves Extract.
procumbens leaves extract was due to the presence of active
Phytoconstituents which were determined in qualitative
analysis of crude extracts. On administration of extract
before I.P. administration of 1% v/v 1ml/100gm acetic acid
leads to inhibition of wriths (Table No. 4). The Analgesic
activity of Tridax procumbens leaves extract was due to the
presence of active Phytoconstituents which were determined
in qualitative analysis of crude extracts. On administration of
extract before I.P. administration of 4%w/v 1ml/kg sodium
chloride leads to inhibition of wriths (Table No. 5). The effect
of TPEE and TPEAE after I.V. injection of 0.1 ml of
Typhoid-Paratyphoid A, B vaccine into the marginal ear-vein
of rabbits leads to decrease in temperature measured by
telethermometer (Table No. 6)
Fig. 1: Effects of TPEE and TPEAE on Acetic acid induced
writhing response in rats CTR-Control, STD-Standard, TPEE-Tridax
procumbens ethanolic extract, TPEAE-Tridax procumbens ethyl
acetate extract (1-200mg/kg, 2-300mg/kg)
Table 3: Phyotochemical Analysis of Tridax procumbens TPEE and TPEAE.
S.N Phytoconstituents Test Ethyl Acetate Ethanolic
extract(TPEAE) extract (TPEE)
1 Carbohydrates Benedict’s test +ve +ve
2 Proteins Biuret test +ve +ve
3 Flavonoids Shinoda ’ s Test +ve +ve
4 Alkaloids Mayer ’ s test, Dragendorff’s test +ve +ve
5 Glycosides Legal’s test, Baljet’s test -ve -ve
6 Saponins By shaking the extract in test tube +ve +ve
7 Tannins Ferric chloride test +ve +ve
8 Steroids Salkowski test +ve +ve
Table 4: Effects of TPEE and TPEAE on Acetic acid induced writhing response in rats.
Group No. Drug Dose Wriths % Inhibition of
(Mean±SEM)
wriths
I Vehicle 1 ml/100gm 14.17±0.40 0
II Diclofenac sodium 10 mg/kg 2.67±0.33 ** 81.2
III TP Ethanolic extract 200 mg/kg 10.2±0.93 * 28.0
IV TP Ethanolic extract 300 mg/kg 4.4±0.68 ** 68.2
V TP Ethyl acetate extract 200 mg/kg 12.32±0.86 13.0
IV TP Ethyl acetate extract 300 mg/kg 9.8±1.73 * 30.84
TP-Tridax procumbens, N = 6, Data expressed as mean ± S.E.M, One way ANOVA followed by Dunnett's
t-test Group I negative control, II standard, III, IV, V and VI test, All test and standard groups were
ompared with group I (negative control) ** P

DISCUSSION
Patel et al Evaluation of Analgesic and Antipyretic Activity of Tridax Procumbens Leaves Extract.
Table 5: Effects of TPEE and TPEAE on NaCl induced writhing response in rats.
Group No. Drug Dose Wriths % Inhibition of
(Mean±SEM)
wriths
I Vehicle 1 ml/100gm 16.3±0.21 0
II Diclofenac sodium 10 mg/kg 2.7±0.21** 83.4
III TP Ethanolic extract 200 mg/kg 13.2±0.42* 19.0
IV TP Ethanolic extract 300 mg/kg 4.7±0.51** 71.2
V TP Ethyl acetate extract 200 mg/kg 14.52±1.05 10.9
IV TP Ethyl acetate extract 300 mg/kg 12.58±1.12* 22.8
TP-Tridax procumbens N = 6, Data expressed as mean ± S.E.M One way ANOVA followed by Dunnett's
t-test Group I negative control, II positive control, III, IV, V and VI test. All test and standard groups were
compared with group I (negative control) ** P

as compared to acetic acid which were inhibited up to 71.2%
by 300mg/kg Tridax procumbens ethanolic extract and 22.8 %
by 300mg/kg Tridax procumbens ethyl acetate extract as
compared with the wriths of control group, whereas both
extract at 200mg/kg inhibits wriths by 19.0% and 10.9%
respectively. The 300mg/kg dose of Tridax procumbens
ethanolic extract shows significant (P

Patel et al Evaluation of Analgesic and Antipyretic Activity of Tridax Procumbens Leaves Extract.
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Address for Correspondence
Deepak Kumar Jain, Truba Institute of Pharmacy, Karond Gandhi Nagar Bypass
Road, Bhopal (M.P.)–462038 (India).
E-mail: jaindeepak2081@yahoo.com
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