Available online on www.ijppr.com International Journal of Pharmacognosy and Phytochemical Research 2013; 5(1); 60-65 ISSN: 0975-4873 *Author for correspondence: E-mail: sarithavedulla@gmail.com Research Article Isolation of Active Compound in Ficus Religiosa Linn *Saritha Vedulla, S. Angala Parameswari , C. Gopinath Department of Pharmaceutical analysis & Quality assurance, Annamacharya college of pharmacy, Rajampet ABSTRACT : Herbs have always been the principal form of medicine in India. Ficus religiosa (L.), commonly known as pepal belonging to the family Moraceae, is used traditionally as antiulcer, antibacterial, antidiabetic, in the treatment of gonorrhea and skin diseases. The plant was subjected to extraction and also fractionated with two various solvent like diethyl ether and n- butanol. The Phytochemical test, TLC and HPTLC reports shows presence of compound only in n- butanol fraction.So this fraction subjected to structural elucidation. The identified compound was stigmasterol. Keywords : Ficus religiosa leaf powder ,extraction, elucidation. INTRODUCTION Medicinal plants have curative properties due to the presence of various complex chemical substances of different composition, which are found as secondary plant metabolites in one or more parts of these plants. Ficus religiosa (L.) is a large perennial tree, glabrous when young, found throughout the plains of India upto 170m altitude in the Himalayas. The stem bark and leaves of F. religiosa are reported phytoconstituents of phenols, tannins, steroids, lanosterol, stigmasterol, lupen-3-one. The active constituent from the root bark F. religiosa was found to be β-sitosteryl-D-glucoside, The seeds contain phytosterolin, β-sitosterol, and its glycoside, albuminoids. The fruit of F. religiosa contained appreciable amounts of total phenolic contents, total flavonoid 1 . MATERIALS AND METHODS Plant material: The plant of Ficus religiosa (Linn) leaves were collected in the month of Jan 2009 in and around of Rajampet. These were authentificated by Dr.K.Madhava chetty, prof, dept.of botony,S.V.University, Thirupathi. Chemicals: Methano,10% acetic acid ,Diethyl ether ,n- butanol Instruments :Weighing balance ,Hot air oven ,Heating mantle ,HPTLC, IR spectrophotometer ,NMRspectrometer ,Mass spectrometer EXPERIMENTAL INVESTIGATION Procedure for extraction: Weighed accurately 50gms of fine leaves powder and 500ml of methanol was added. The crude drug was extracted through soxhlet apparatus for 72 hrs. Then filtered the solution and collected, to the residue of leaf powder again added 200ml of methanol, allowed the leaf powder to further maceration for 2days. Collected the filtrate and evaporated by using heating mantle until to get the dried powder. Fractionation of active constituent: Weighed 3gms of extract powder and dissolved with 10% of acetic acid and allowed to reflux for 1hr.The content was transferred to separating funnel. Then equal quantity of diethyl ether was added. Repeat the procedure for 2times then collect the bottom layer, it contains the extract. Added equal quantity of n-butanol, shaken it well and kept aside for 15min. Then collected the n-butanol fraction and evaporated to dryness. The residue was taken for further evaluation 2 . TLC PROCEDURE: Thin layer chromatography can be used to monitor the progress of a reaction, identify compounds present in a given mixture, and determine the purity of a substance. Silicagel G used as adsorbent and n- butanol: acetic acid : water in the ratio of 4:0.5:5 used as a mobile phase. Plate was prepared by pouring silica gel on glass plate and activated by heating at110 0 c for 30 min. Take the n-butanol fraction in methanol which was washed by dietylether , spotted on TLC. The spots are detected under long uv at 365 nm and Rf values are calculated 3 . High performance thin layer chromatography: HPTLC is characterized by efficient separation used either for identification or quantitation of chemical substances. Aluminium plates are normally used. Silica gel is the most widely used adsorbent. HPTLC plates are produced from 4.5 μm silica gel with an inert binder to form a 200μm layer.Plates exposed to high humidity are kept out to make them activated by placing in an oven at 110-120 0 C for 30 min prior to sample spotting. n-butanol :acetic acid :water in the ratio of 4:0.5:5 .Sample are solubilized with specified solvent of the extract.The sample volume normally applied on to the plate is around 0.2 μl. Streaking the sample on the plate results in better separation than spotting.In Ascending development the optimum separation distance is 20-25 mm with separation time of about 4 min. Detection of coloured substances or colourless substances absorbing in longer wave length UV region (365 nm) or substances with intrinsic fluorescence can be easily detected. Quantization was performed with photometric measurement of absorbed light or emitted fluorescence. In absorption densitometry, the spots in