K.G.Lalitha et al.

/ Journal of Pharmacy Research 2011,4(3),639-640

Research Article
ISSN: 0974-6943 Available online through
www.jpronline.info
Anti-inflammatory activity of barks of Evodia lunu-ankenda (Geartn) Merr
K.G.Lalitha1*, R.Sathish 1, T.Regupathi1, K. Natarajan1, P.Kalaiselvi 2, T.Venkatachalam2
1
Department of Pharmaceutical Chemistry, Ultra College of Pharmacy, Madurai – 625 020, Tamil Nadu, India.
2
Department of Pharmaceutical Chemistry, Annai JKK Sampoorani Ammal College of Pharmacy, B. Komarapalayam - 638 183, Tamilnadu, India.
Received on: 14-10-2010; Revised on: 14-12-2010; Accepted on:08-02-2011
ABSTRACT
The aim was to evaluate the anti-inflammatory activity of Evodia lunu-ankenda ethyl acetate extract. The ethyl acetate fraction of E. lunu-ankenda bark extract was prepared and administered
orally to rats at 125, 250 and 500 mg/kg p.o. doses. The anti-inflammatory activity of E. lunu-ankenda was determined by carrageenan-induced paw edema and cotton pellet granuloma models.
Oral administration of E. lunu-ankenda extract reduced inflammation significantly (P<0.01) in the carageenan paw edema and the cotton pellet granuloma models. The results of the study
validates the traditional use of E. lunu-ankenda in the treatment for inflammatory disease.

Key words: Evodia lunu-ankenda; Anti-inflammatory activity; Carrageenan; Cotton pellet granuloma.

INTRODUCTION
Evodia lunu-ankenda (Geartn) Merr. (Rutaceae) is an endemic plant found in Western Ghats of p.o.), Group II was treated with indomethacin (10 mg/kg,p.o.), Groups III, IV and V were
India. In folk medicine, the juice of the leaves and barks are prescribed for fever. [1] Dictamine, treated with ELA at doses 125, 250 and 500 mg/kg p.o. respectively.
Evolitrine, kokusaginine, N-methyl-4-methoxy -2-quinolone, marmesin has been isolated
from bark and seeds of E. lunu-ankenda. [2] Although E. lunu-ankenda is frequently used to treat Carrageenan-induced rat paw edema
fever and inflammation, its potency has not been pharmacologically examined. Inflammation Acute inflammation was caused by injecting 0.1 ml of 1 % (w/v) carrageenan in saline into the
is the complex biological response of vascular tissues to harmful stimuli including pathogens sub-plantar region of the right hind paw of each rat. The paw volume was measured
irritants or damaged cells. Currently available steroidal as well as non-steroidal anti-inflamma- plethysmometrically at 0 h and 3 h after the carrageenan injection. Edema was expressed as
tory agents are associated with inherent problems. There is continuous search especially from mean increase in paw volume relative to control animals. The percentage inhibition of edema
natural sources for alternative agents. Large number of herbal extracts as well as products being was calculated by the following equation: % inhibition of edema= 100 (1-Vt/Vc), where Vc is
employed in the treatment of inflammatory disorders by natural healers. In this study, we the edema volume in the control group and Vt is the edema volume in tested groups. A single
tested the ethyl acetate fraction of E. lunu-ankenda barks for preventing inflammation of dose of drug pretreatment was given 1 h before the injection of carrageenan.[6]
carrageenan-induced edema and cotton pellet granuloma in rat models.
Cotton pellet granuloma
MATERIALS AND METHODS In the cotton pellets-induced granuloma, the rats were anaesthetized with pentobarbitone (30
mg/kg) and after removing the fur on the back, a single midline incision was made on the dorsal
Preparation of extract and phytochemical screening surface. Cotton pellets weighing 10±1mg, sterilized and then implanted in both axillae and
Barks of the plant were collected from the Kolli kodu, Trivandrum, India, in December 2008 groin region of each rat. The standard indomethacin and ELA were administered daily in the
and authenticated by Dr P.Danial, Scientist, Botanical Survey of India, Coimbatore, India. morning for 7 days. On 8th day the animals were anaesthetized and the pellets together with the
The barks (500 g) were dried at room temperature, protected from dust and sunlight. They were granuloma tissues were carefully removed and made free from extraneous tissues. The wet
then pulverized manually and extracted first with petroleum ether (60-80oC) using a Soxhlet pellets were weighed and then dried in an oven at 60°C for 24 h to constant weight, after that
apparatus and subsequently with ethyl acetate. The ethyl acetate extract was concentrated in the dried pellets were weighed again. Increment in the dry weight of the pellets was taken as a
vacuo (12 g), designated as ELA. ELA was administered as a suspension (1 % w/v Tween 80) measure of granuloma formation.[7]
to experimental animals. Chemical tests were carried out with ELA using standard procedures
to identify the phyto constituents. [3, 4] Statistical analysis
All values were expressed as mean ± SEM. The data were statistically analyzed by one-way
Drug and chemicals ANOVA followed by Dunnett’s test. P values < 0.05 were considered significant.
Indomethacin (Ranbaxy Laboratories Ltd, Mumbai) was used as the standard anti-inflamma-
tory agent. Petroleum ether 60-800C and ethanol were procured from Merck India( Ltd), RESULTS
Carageenan was purchased from Sigma- Aldrich, St Louis, MO, USA. Phytochemical screening of ELA revealed the presence of phenolic compounds, phenyl propanoid,
flavonoids , chromenes, alkaloids and sterols. Oral administration of ELA up to 5000 mg/kg
Experimental animals did not produce any toxic effects in mice. No mortality was observed and ELA was found to be
Male Swiss albino mice (25-30 g) and male Wistar albino rats (150-200 g) were used as safe at the given doses. The results showed significant (P < 0.01) inhibition of rat hind paw
experimental models. They were housed in standard polypropylene cages and kept under edema and cotton pellet granuloma in a dose-dependent manner when compared to the
ambient temperature (25±2o C), relative humidity (45-55 %) and in a 12 h light-dark cycle. The respective control group. The ELA displayed marked anti-inflammatory effects at a dose of 500
animals were received standard pellet diet and water ad libitum ( M/s Hindustan Lever mg/kg ; the percentage inhibition of carrageenan-induced paw edema was 54 % after 3 h of drug
Limited, Bangalore, India). They were divided randomly into five groups of six animals in treatment. The standard indomethacin 10 mg/kg produced 65 % inhibition. In the granuloma
each for both models. The experiments were carried out during the light cycle. The institu- assay, ELA at 500 mg/kg reduced the wet and dry weights by 59 % and 57 % respectively,
tional animal ethical committee clearance was obtained for carrying out the experiment. Indomethacin (10 mg/kg) produced 65 and 67% reductions in wet and dry weights respectively
(Table 1).
Acute oral toxicity studies
The OECD guidelines 425 were followed for the acute oral toxicity study for fixing the dose. Table 1: Effect of ELA on carrageenan-induced rat paw edema and cotton pellet granu-
The dose level up to 5000 mg/kg of ELA did not produce any mortality on oral administration. loma in rats
So 125, 250 and 500 mg/kg doses were fixed for the study. [5]
Treatment Dose Increase in % Granuloma % Granuloma %
group (mg/kg) paw volume Inhibition weight inhibition weight inhibition
Anti-inflammatory activity (ml) (wet)(mg) (wet) (dry) (mg) (dry)
The anti-inflammatory activity of ELA was evaluated using the carrageenan-induced rat hind
paw edema method and the cotton pellet granuloma method. In both the models the rats were I 1% Tween 80 1ml/kg 0.52±0.02 - 226.0±1.20 - 56.70±1.45 -
II Indomethacin 10 0.18±0.02a 65 80.20±1.14a 65 18.60±1.45a 67
divided as follows, Group I served as control and received vechile (1% Tween 80, 1ml/kg III ELA 125 0.40±0.03a 23 140.21±1.44a 37 37.00±1.15a 35
IV ELA 250 0.31±0.04a 40 118.73±1.16a 46 32.00±1.15a 44
*Corresponding author. V ELA 500 0.24±0.01a 54 92.04± 1.10a 59 24.33±1.76a 57

Dr. K.G.Lalitha DISCUSSION

Prof & Head The Evodia lunu-ankenda ethyl acetate extract was evaluated for its anti-inflammatory activity
Department of Pharmaceutical Chemistry, Ultra College in acute and sub-chronic models. Carrageenan-induced paw edema is the standard experimen-
of Pharmacy, Madurai – 20, Tamil Nadu, India. tal model for acute inflammation. Carrageenan is the phlogistic agent of choice for testing anti
Tel: + 91-9894893301 inflammatory drugs as it is not known to be antigenic and is devoid of apparent systemic effects.
E-mail:kg.lalitha@gmail.com
Moreover the experimental model exhibits high degree of reproducibility. The development of
Journal of Pharmacy Research Vol.4.Issue 3. March 2011 639-640
 K.G.Lalitha et al. / Journal of Pharmacy Research 2011,4(3),639-640
edema has been described as biphasic. [8] The first phase (1h) is mediated through the release Research, New Delhi; 1952.
of serotonin and histamine and the second phase (over 1 h) is mediated by prostaglandins, 2. Mohan PS, Ramesh M, Shanmugam P. Studies on the Indian rutaceae. Chemical investigation of
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cyclooxygenase products. Continuity between the two phases is provided by kinins. [9-11] ELA 3. Evan WC. Trease and Evans Pharmacognosy,15 ed, W.B. Saunders, New York; 2002.
showed a significant (P<0.01) anti-inflammatory effect, which may be due to inhibition of the 4. Odebedy O. Sofowora Phytochemical screening of Nigerian medicinal plants. Lloydia 41, 1978,
mediators of inflammation induced by the phlogogenic stimuli. The cotton pellet induction of 41-234.
5. OECD Guidelines 425 for the testing of chemicals adopted 23.03.2006. Acute oral toxicity – up and
granuloma is a model for the proliferative phase of inflammation which develops over a period down procedure (UDP).
of several days. Chronic inflammation occurs by means of the development of proliferative 6. Winter CA, Risely EA, Nussm GW. Carrageenin-induced edema in hind paws of the rat as an assay
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Source of support: Nil, Conflict of interest: None Declared

Journal of Pharmacy Research Vol.4.Issue 3. March 2011 639-640