Published on February 17, 2014
Int. J. Pharm. Res. Sci., 2014, 02(1), 42-46. www.ijprsonline.com ISSN: 2348 –0882 ============================================================================== Anthelmintic Activity of Ethanolic Extract Of Cassia Occidentalis Linn. Sayyad RR1*, Kare RD1, Jagtap SM1, Katkar ST1, Kadam JH1. 1 Department of Pharmacology, SPM’s College of Pharmacy, Akluj, India. * correspondence author:Mrs. Sayyad R.R.,Assistant Professor,SPM’s College ofPharmacy, Akluj, India.,Email: firstname.lastname@example.org --------------------------------------------------------------------------------------------------------------------------------------Abstract: The present study was done with the aim to evaluate anthelmintic activity of ethanolic extract of Cassia occidentalis Linn using adult earthworm Pheritima posthuma. Various concentrations (25, 50, 75 mg/ml) of all extracts were tested and results were expressed in terms of time for paralysis and time for death of worms. Albendazole was used as a reference standard and gum acacia in saline as a control group. Dose dependent activity was observed in all extracts Cassia occidentalis Linn. Keywords: activity. Cassia occidentals; Anthelmintic INTRODUCTION: Helminth infections are among the most widespread infections in humans, distressing a huge population of the world. Although the majority of infections due to helminths are generally restricted to tropical regions and cause enormous hazard to health and contribute to the prevalence of undernourishment, anaemia, eosinophilia and pneumonia. Parasitic diseases cause ruthless morbidity affecting principally population in endemic areas. The gastro-intestinal helminthes becomes resistant to currently available anthelmintic drugs therefore there is a foremost problem in treatment of helminthes diseases. Hence there is an increasing demand towards natural anthelmintics [1, 2, 3]. Cassia occidentalis called as Kasmard in Sanskrit, Kasondi in Hindi and Coffee Senna in English belongs to family Caesalpiniaceae. It is an ayurvedic plant with huge medicinal importance. Leaves of C. occidentalis plant have ethnomedicinal importance like paste of leaves is externally applied on healing wounds, sores, itch, cutaneous diseases, bone fracture, fever, ringworm, skin diseases and throat infection. The parts of the plant used are roots, leaves and seeds. The plant is used for fever, menstrual problems, tuberculosis, diuretic anemic, liver complaints, and as a tonic for general weakness and illness. The plant is also used to cure sore eyes, hematuria, rheumatism, typhoid, asthma, and disorder of haemoglobin and is also reported to cure leprosy. An infusion of the plant bark is given by the folklore in diabetes. Previous pharmacological investigations showed that C. occidentalis leaf extracts have antibacterial [6, 7], antimalarial , antimutagenic [9, 10], antiplasmodial  and hepatoprotective  activity. Chemical constituents isolated from C. occidentalis including sennoside, anthraquinone glycoside,  fatty oils, flavonoid glycosides, galactomannan, polysaccharides, and tannins . The present study was perform to study the anthelmintic activity of ethanolic extract of Cassia 42
Int. J. Pharm. Res. Sci., 2014, 02(1), 42-46. www.ijprsonline.com ISSN: 2348 –0882 ============================================================================== occidentalis on Indian earth worms (Pheretima posthuma). MATERIAL AND METHOD: Procurement of plant material: The fully mature, fresh leaves of Cassia occidentalis was collected from Akluj city, Solapur district, Maharashtra. Identification and Authentication: The fresh leaves was identified and authenticated by Mr.D. N. Kutwal, Professor, Head of Department of Botany, S.M.M. Akluj and leaves was deposited in the Herbarium of the same department. Preparation of extract of leaves of Cassia occidentalis: The leaves were air dried at room temperature and converted into fine powder with using blender. 500 gm of dry fine powder was suspended in 1.5 liters of ethanol then stirred magnetically for 24 hours at room temperature. The extract was double filtered by using musline cloth and whatmann No. 1 filter paper. The filtrate was concentrated to dryness to obtain crude extract. The dried ethanolic extracts of Cassia occidentalis was stored in desiccators under controlled conditions till it used for experimental purpose. Experimental animals: Anthelmintic activity was evaluated on adult Indian Earthworms (Pheretima posthuma) due its anatomical and physiological resemblance with the intestinal round worm of human beings . The Indian adult earthworms Pheretima posthuma (Annelida) were collected from moist soil of the field and washed with normal water and saline solution to remove soil and fecal matter. The Earth worms of 4-8 cm in length and 0.2- 0.3 cm in width were used for all experimental parameters. Drugs and Chemicals used: Albendazole (Glasko Smith Kline) was used as reference standard purchased from local medical shop, Akluj. Chemicals ethanol (95% V/V) (S.D fine chemicals, Mumbai). Preparation of test sample: Samples for experiments were prepared by dissolving extract to obtain a stock solution of 100 mg/ml, from the stock solution, different working dilutions were prepared to get concentration range of 25, 50 and 75 mg/ml of ethanolic extracts (ECO). For present study Albendazole taken as standard drug. The concentration of standard drug was prepared in 1% gum acacia to give 20 mg/ml concentration. Evaluation of Anthelmintic Activity: The evaluation of anthelmentic activity was followed by earlier reported method . Three different concentrations (as given earlier) were prepared and the group of six earthworms which having equal size were released into 10 ml of sample with desired concentration in petridish. Observations were made for the time taken to cause paralysis and death of the individual worms. Mean time for the paralysis in any sort could be observed, except when the worm was shaken vigorously. The worms neither moved when shaken vigorously nor when dipped in warm water (500C). Paralysis is assumed to occur when they do not revive even in saline solution. Potency is inversely proportional to time taken for paralysis and / or death of parasite. Statistical analysis: The results were expressed as mean ± SEM and statistically analyzed by ANOVA followed by Dunnett’s test, with level of significance set at p<0.05. 3. Results and Discussion: In the present work, ethanolic extracts of Cassia occidentalis leaves was used to evaluate anthelmintic activity against Indian earthworms Pheritima posthuma, Each extract containing 25, 43
Int. J. Pharm. Res. Sci., 2014, 02(1), 42-46. www.ijprsonline.com ISSN: 2348 –0882 ============================================================================== 50, and 75 mg/ml produced dose dependent paralysis ranging from loss of motility to loss of response to external stimuli, which eventually progressed to death. As shown in table no.1 Table 1: Anthelmintic activity of ethanolic Extracts of Cassia occidentalis leaves on Indian Earthworms (Pheretima posthuma). Time taken Time taken for for death paralysis in minute in minute (Mean ± (Mean ± SEM) SEM) Name of Grou p Name of extract & Concentratio n Group -I Gum acacia in saline (1%) --- --- Group -II Albendazole (20mg/ml) 34.7±0.55 39.0±0.52 Group -III ECO (25mg/ml) 62.2±1.70* * 70.5±0.81* * Group -IV ECO (50mg/ml) 52.5±0.56* * 64.3±2.40* * Group -V ECO (75mg/ml) 30.0±0.37* * 49.3±0.76* * Values are expressed as mean ± SEM. (n=6), ANOVA followed by Dunnett’s test, * p<0.05, **p<0.01 when ECO 25, ECO 50, ECO 75 groups were compared with Standard(Group-II). ECO- Ethanolic extract of Cassia occidentalis. The results of the current investigation indicate that the ethanolic extracts of Cassia occidentalis is the most potent and requires less time to the paralysis and death of the worms. Extracts showed a concentration depended anthelmintic property. Chemical constituents isolated from C. occidentalis including sennoside, anthraquinone glycoside,  fatty oils, flavonoid glycosides, galactomannan, polysaccharides, and tannins . Polyphenolic compounds shown anthelmentic activity and some synthetic phenolic anthelmentics are shown to interfere with energy generation in helmintic parasites by uncoupling oxidative phosphorylation . Tannins possess antiparasitic activity . Reported anthelmentic activity of tannin that they can bind to free protein in GIT of host animal or glycoprotein on the cuticle of the parasites and may cause death . It is possible that tannin contained in the ethanolic extract of Cassia occidentalis leaves may produce similar effect. From the observation ethanolic extracts of leaves of Cassia occidentalis was showed anthelmintic activity. The function of the anthelmintic drugs, like albendazole, is known to cause paralysis of the worms so that they are expelled in the feaces of man and animals. The extracts not only demonstrated this property, but they also caused death of the worms. Synthetic phenolic anthelmintics, like niclosamide, interfere with the energy generation in the helminth parasites by uncoupling the oxidative phosphorylation. Another possible mechanism of action is that they bind to free proteins in the gastrointestinal tract of the host animal or to glycoprotein on the cuticle of the parasite and by this cause death. Tannins have also been shown to produce anthelmintic activities . From the observation ethanolic extracts of leaves of Cassia occidentalis was showed anthelmintic activity. After a brief stimulant effect, earthworm lost their motility of exposure to crude extract of leaves Cassia occidentalis. Ethanolic extract of leaves Cassia occidentalis exhibited anthelmintic activity in dose dependent manner giving short time of paralysis minutes, and time of death in minutes respectively with 75 mg/ml. Therefore potency of drug was found to be inversely proportional to the time taken for 44
Int. J. Pharm. Res. Sci., 2014, 02(1), 42-46. www.ijprsonline.com ISSN: 2348 –0882 ============================================================================== paralysis / death of worms. The higher concentrations of ethanolic extract produced paralytic effect much earlier and necrotic spots were observed externally on the worms, with higher concentrations. The effect of each ethanolic extract was compared with Albendazole as standard drug (20 mg/ml). In conclusion, the traditional claim of leaf of Cassia occidentalis as an anthelmintic have been confirmed as the extracts displayed activity against the worms used in the study. Further studies are required to isolate and reveal the active compound contained in the crude extracts of Cassia occidentalis and to establish the mechanism of action. Conclusion: Using the Pheretima Posthuma as the animal models, we have shown that (ethanolic extract) crude leaves powder of Cassia occidentalis has potential to act against helminthiasis. Moreover, the extent of anthelmintic effect of the leaves powder is comparable to that of standard drug, Albendazole being used against helminthiasis, in general. This observation unambiguously suggests that the leaves powder of Cassia occidentalis must contain lead compounds that may provide profound implications on designing de novo anthelmintic drugs. Acknowledgement: The authors wish to express thanks to Mr. D. N. Kutwal, Professor, Head of Department of Botany, S.M.M. Akluj for identification and authentication of plant. References:  Bundy DA. Trans Royal Soc, Trop Med Hyg, 8, 1994, 259-261.  Tagbota S, Townson S. Adv Parasitol, 50,2001, 199-205.  Sondhi SM, Shahu R, Magan Archana. Indian Drugs, 31,7,1994, 317-320.  Krithikar KR, Basu BD. Cassia occidentalis, Indian Medicinal Plants, II edition, 1999,860.  The Wealth of India. A dictionary of Indian Raw Material and Industrial Products. New Delhi, Council of Scientific and Industrial Research, 1998, 350.  Jain SC, Sharma RA, Jain R, Mittal C. Antimicrobial screening of Cassia occidentalis L in vivo and in vitro, Phytotherapy Research, 121,1998, 200-204.  Saganuwan AS, Gulumbe ML. Evaluation of in vitro antimicrobial activities and phytochemical constituents of Cassia occidentalis. Animal Research International, 3, 2006, 566-569. Tona L, Ngimbi NP, Tsakala M, Mesia K, et al. Antimalarial activity of 20 crude extracts from nine African medicinal plants used in Kinshasa Congo. Journal of Ethnopharmacology, 681,1999, 193-203. Jafri MA, Subhani MJ, Javed K, Singh S. Hepatoprotective activity of leaves of Cassia occidentalis against paracetamol and ethyl alcohol intoxification in rats, Journal of Ethnopharmacology, 661, 1999, 355-61. Sharma N, Trikha P, Athar M, Raisuddin S. In vitro inhibition of carcinogen induced mutagenicity by Cassia occidentalis and Emblica officinalis. Drug and Chemical Toxicology, 23, 2000, 477-84. Tona L, Cimanga RK, Mesia K, Musuamba CT, et al. In vitro antiplasmodial activity of extracts and fractions from seven medicinal plants used in the Democratic Republic of Congo. Journal of Ethnopharmacology, 93,2004, 27-32. Yadav JP, Arya V, Yadav S, Panghal M, et al. Cassia occidentalis: A review on its ethnobotany, phytochemical and pharmacological process. Fitoterapia, 2009, 9-8.  Yadav JP, Arya V, Yadav S, Panghal M, et al. Fitoterapia, 81,4, 2010, 223-230.  Lal J, Gupta PC. Experientia,30,1974, 850-1. 45
Int. J. Pharm. Res. Sci., 2014, 02(1), 42-46. www.ijprsonline.com ISSN: 2348 –0882 ============================================================================== Vidyarthi RD. A Text book of Zoology,14th Edition, S. Chand and Co., New Delhi, 1967, 45.  Bhusan M, Pawar, Nayana D, Int. J. Pharma Tech Res,2,2,2010, 1483-1487.  Nirmal SA. J. Sci. Technol. 29, 3, 2007, 755757.  Wei Li. Afr. J. Agri. Res.4,9, 2009,898-902.  Trapti R, Vijay B. Komal M. Aswar PB. Asian J.Res.Chem. 2,2,2009, 181-182.  Athnasiadou S, Kyriazakis I, Jackson F, Coop RL. Direct anthelmintic effects of condensed tannins towards different gastrointestinal nematodes of sheep: In vitro and in vivo studies, Veterinary Parasitol, 99, 2001, 205-19.  Martin RJ. Mode of action of anthelmintic drugs. Veterinary Journal,154,1997, 11-34. 46
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