Lakshmi Narasimhudu. C et al. / Journal of Pharmacy Research 2011,4(8),2705-2706 Research Article Available online through ISSN: 0974-6943 http://jprsolutions.info In vitro anthelmintic activity of Phyllanthus amarus Schum. &Thonn (Euphorbiaceae) Lakshmi Narasimhudu. C1, B. Naga Raju2, G.H. Philip2 and R.R.Venkata Raju1 Department of Botany1, Sri Krishnadevaraya University, Anantapur – 515 003. Department of Zoology2, Sri Krishnadevaraya University, Anantapur – 515 003. Received on: 17-05-2011; Revised on: 12-06-2011; Accepted on:16-07-2011 ABSTRACT Phyllanthus amarus Schum. & Thonn. (Euphorbiaceae), a potential medicinal plant distributed all over India. The plant is used for aphrodisiac, bruises, diuretic, dropsy, dysentery, genito-urinary problems, gonorrhoea, jaundice, menorrhagia, ringworm, sores, stomachic, swellings, wounds, and ulcers. In the present work the alcoholic and aqueous extracts of P. amarus were evaluated for its anthelmintic activity against adult earthworms (Phertima prosthuma). The activities of the extracts were compared with standard reference and distilled water as control. The activity of three concentrations (25, 50, and 75 mg/ml) of each extract were studied, which involved the determination of time of paralysis and time of death of the test worms. It was found both the extracts exhibited significant anthelmintic activity while maximum was recorded in ethanol extract.

Key words: Phyllanthus amarus, anthelmintic activity, Phertima prosthuma, Albendazole.

INTRODUCTION Helminthiasis is a world wide and one of the common diseases of all ages involved determination of time of paralysis and time of death of the worms. especially in third world countries. The parasitic disease causes severe morbid- Albendazole was included as standard reference and distilled water as control. ity by affecting population in endemic areas with major economic and social [8] [1] The anthelmintic assay was carried as per the method of Ajaiyeoba et al with consequences . Anthelminthis are those agents that expel parasitic worms minor modifications. The anthelmintic activity was evaluated on the adult (helminthes) from the body, by either stunning or killing them. Intestinal Indian earthworms, due to its anatomical and physiological resemblance with infections with worms can be more easily treated than those infections that the intestinal roundworm parasites of human beings [9, 10, 11]. Because of easy occur in other location in the body because the worms need to be killed by the [2] availability, earthworms have been used widely for the initial evaluation of drug and the drug need not be absorbed when administered by oral route . anthelmintic compounds in vitro [12, 13, 14]. Indian adult earthworms collected from moist soil and washed with normal saline to remove all faecal matter Phyllanthus amarus belonging to the family Euphorbiaceae, called Nela Usiri were used for the anthelmintic activity. The earthworms of 6-8 cm in length in Telugu (local language). It is reputed in systems of medicines as the various and 0.2- 0.3 cm in width were used for all experimental protocol. The earth parts of the plant has been used traditionally for many of the diseases like worms were divided into ten groups containing six worms in each group. All the aphrodisiac, bruises, diuretic, dropsy, dysentery, genito-urinary problems, gon- extracts and standard drug solution were freshly prepared in normal saline orrhoea, jaundice, menorrhagia, ringworm, sores, stomachic, swellings, wounds, [3, 4] before starting the experiments. Different extracts and standard drug solutions ulcers . The whole plant was used for anaemia, antipyretic, diabetes, di- were poured in different petri plates. All the earth worms were released into uretic, dropsy, fever, jaundice, skin disease, while leaves are used for aphrodi- 10ml of solution of Albendazole, ethanol and aqueous extracts of the leaves of siac, dysentery, dyspepsia, gonorrhoea, malaria, stomach ache, swellings, fruits P. amarus (25, 50 and 75 mg/ml) in distilled water. Observations were made on for bruises, genito-urinary problems, sores, ringworm, wounds, ulcers and roots the basis of the time taken to paralysis and death of worms. Time for paralysis for menorrhagia. The present study aimed to investigate an anthelmintic was noted when no movement of any sort could be observed except when the activity of crude extracts of leaves of P. amarus to confirm the claims of worms were shaken vigorously. Death was concluded when the worms lost their herbal practitioners. motility when dipped in warm water (50ºC) followed with fading away of their body colors. MATERIALS AND METHODS: Table 1: Anthelmintic activity of the leaf extracts of Phyllanthus amarus Plant material: The plant samples were collected from the moist areas and waste lands of Treatment Conc. Paralysis Time Death time Anantapur district. The plant specimen was identified with the help of local (mg/ml) (min.) (min.) and regional floras [5, 6]. The plant was shade dried and pulverized in mechanical Ee Pa 25 48 112 grinder and stored in an air tight container till further successive extraction. 50 45 76 75 29 52 Preparation of aqueous extract: Aq Pa 25 85 175 50 43 70 The dried powder (70 gm) was extracted with water. The drug was extracted for 75 32 55 72hr. and the filtered extract was dried on water bath [7]. Standard 25 62 119 50 45 85 75 30 70 Preparation of ethanolic extract: Control - - - The dried powder (70gm) extracted in a soxhlet apparatus using ethanol (95%), at a temperature range of 55°C to 65°C. The filtrate was evaporated to RESULT AND DISCUSSION: dryness at 40°C under reduced pressure in rotoflash evaporator [7]. Preliminary phytochemical analysis showed the presence of alkaloids, caro- tenoids, catecholic compounds, coumarins, flavonoids, glycosides, lignans, Investigation of anthelmintic activity phenols, proteins, polyoses, saponins and steroids like phytoconstituents in Ethanol and aqueous extracts of the leaves of P. amarus were investigated for the extracts of Phyllanthus amarus. Some of these phytoconstituents may be their anthelmintic activity against Phertima prosthuma. Various concentra- responsible to show a potent anthelmintic activity. From the observations tions (25, 50 and 75 mg/ml) of each extract were tested in the bioassay, which made all the extracts of leaves of P. amarus was found to show a potent anthelmintic activity when compared to the standard drug. Ethanolic extract *Corresponding author. of P. amarus at 75 mg/ml concentration shows paralysis at 29 min and death 52 min, whereas aqueous extract shows paralysis at 32 min and death 55 min R.R.Venkata Raju respectively against the earthworm. The reference drug Albendazole exhibited Department of Botany, the same at 30 min and 70 min respectively [Table1, Fig1]. Albendazole Sri Krishnadevaraya University, exhibits anthelmintic activity by blocking glucose uptake and depletion of glycogen stores in test parasite. The ethanolic extract of P. amarus not only Anantapur – 515 003. demonstrated paralysis, but also caused death of worms especially at higher concentration of 75 mg/ml in shorter time as compared to reference drug Albendazole.

Journal of Pharmacy Research Vol.4.Issue 8. August 2011 2705-2706 Lakshmi Narasimhudu. C et al. / Journal of Pharmacy Research 2011,4(8),2705-2706 REFERENCE: 1. Tagbota S, Townson S, Antiparasitic properties of medicinal and other naturally occurring products, Adv Parasitol, 50, 2001, 199-205. 2. Senthil Velan S, Suresh V, Senthil Kumar N, Aldrin Bright J, Karthikeyan P, In vitro anthelmintic activity of Samanea saman (Merr) bark, Journal of Pharmacy Research, 3 (12), 2010, 2977-2978. 3. Kirtikar KR and Basu BD, Indian Medicinal Plant, 2nd ed, Vol – III, Lalit Mohan Basu Published, Allahabad, India, 1981, page 2225. 4. Jain SK, Dictionary of Indian Folk Medicine and Ethnobotany, Deep Publica- tions, New Delhi, India, 1991, page 142. 5. Pullaiah T & Ali Moulali D, Flora of Andhra Pradesh, 1st ed, Vol - II, Scientific Publishers Jodhpur, 1977, page 694. 6. Gamble JS, Flora of Presidency of Madras, Vol – II, Botanical Survey of India, Calcutta, 1921, Page 1048. 7. Bibhilesh B. Mendhe, Umesh Nema, Piyush Gupta and Bhushan R. Gandhare, Evaluation of Anthelmintic activity of leaf extracts of Butea Monosperma, Inter- national Journal of Pharmaceutical Sciences and Research, 1(3), 2010, 69-72. Time for paralysis and death (min) 8. Ajaiyeoba EO, Onocha PA, Olarenwaju OT, Invitro anthelmintic properties of Buchholzia coriaceae and Gynandropsis gynandra extract, Pharm Biol, 39, 2001, 217-20. 9. Suresh V, Arunachalam G, Senthil Kumar N, In vitro anthelmintic activity of Nyctanthes arbor-tristis Linn bark. Journal of Pharmacy Research, 4(1), 2011, Extracts 283-284. 10. Vidyarthi RD, A text book of Zoology, 14th ed, New Delhi, S. Chand and Co; 1967. Ee Pa: Ethanol extract of P. amarus, Aq Pa: Aqueous extract of P. amarus, Conc.: Concentration 11. Chatterjee KD, Parasitology, Protozoology and Helminthology, 6th ed, Calcutta: Fig 1: Anthelmintic activity of various extracts of Phyllanthus amarus In Guha Ray Sree Saraswaty Press Ltd, 1967. 12. Sollmann T, Anthelmintics: Their efficiency as tested on earthworms, J Pharmacol. CONCLUSION: Exp. Ther, 12, 1918, 120-70. The traditional use of the leaves of Phyllanthus amarus as an anthelmintic 13. Das GK, Suresh P, Kar DM, Ganpaty S, Panda SB, Evaluation of Evolvulus was confirmed as the leaf extracts displayed activity against the worms used in alsinoids Linn. For anthelmintic and antimicrobial activity, J Nat Rem, 2, 2002, the study. Further it would be interesting to isolate the responsible 182-5. 14. Shivkar YM, Kumar VL, Anthelmintic activity of latex of Calotropis procera, phytoconstituents which are responsible for the anthelmintic activity and the Pharma Biol, 41, 2003, 263-5. mechanism of action, which is being attempted in the laboratory. Source of support: Nil, Conflict of interest: None Declared

Journal of Pharmacy Research Vol.4.Issue 8. August 2011 2705-2706