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Bioassay- guided fractionation and anti-fungal activity studies on pisonia grandis rInternational Journal of Pharmacognosy and Phytochemical Research 2013; 5(1); 1-3 Bioassay- Guided Fractionation and Anti-Fungal Activity Studies on *Shubashini K. Sripathi, Poongothai G.
Department of Chemistry, Avinashilingam University for Women, Coimbatore Tamilnadu, India.
Bioassay- guided fractionation of ethanol extract of leaves of Pisonia grandis was studied for its anti-fungal activity for
the microorganism Candida albicans, Aspergillus niger, Pencillium citrinum and Monascus purpureus by disc diffusion
method. The ethanol extract showed good anti-fungal activity for Monascus purpureus compared to standard
Keywords: Pisonia grandis, Nyctaginaceae, Monascus purpureus, Clotrimazole.
combined. A portion of these two layers are concentrated separately and subjected to anti-fungal study. The rest of distributed throughout India and is a widespread the CHCl3 layer was distilled completely; the residue was evergreen commonly grown lettuce tree1. Leaves, stem dissolved in 10% aqueous ethanol for further extraction and root of this species are extensively used by the tribals with pet-ether. The LLE with pet-ether is continued until in the preparation of several folk medicines. It has been the organic layer was colorless. Then the entire organic extensively used in Indian traditional medicine as an and aqueous layer was combined and distilled by vacuum antidiabetic, anti-inflammatory agent, and used in the and the residue was stored for anti-fungal study.
treatment of an algesia, ulcer, dysentery and snake bite2-7.
A column of silica gel (400 g) built in CHCl3 and was The plant has been studied by different workers with eluted with CHCl3, Chloroform- methanol mixtures of special reference to its pharmacological activity but no increasing polarity. The homogeneity of the fractions was isolation of phytochemicals has been reported8. Also no examined by TLC and similar fractions were combined report on the antifungal effects of Pisonia grandis exists.
and tested for anti-fungal activity (Table 2). Three This paper reports the anti-fungal effects of its extracts compounds isolated from the column were also tested for and with clotrimazole as reference drug. their anti-fungal activity.
Anti-Fungal assay : The anti-fungal activity was assayed MATERIALS AND METHODS
by Disk diffusion method. Sabouarud’s dextrose agar is Collection of plant material: The plant material (leaves) the most suitable medium because fungal growth is was collected during January- March 2009 in the local favored by a high sugar concentration and is relatively areas of Coimbatore, Tamilnadu, India. The identity of tolerant to acidity (pH 5.4) 9. Nutrient agar plates were plant material was confirmed by the taxonomist Dr.
prepared when the agar medium was amended with C.Kunhikannan, Scientist D, Biodiversity Division, complexes separately. Then the medium was warm and poured into petri plates. After solidification of the Coimbatore. The leaves were dried in shade and cut into medium, mycelia discs (6 mm dia) of the test fungi were small pieces and then used for phytochemical study.
inoculated at the centre of the plates. Diameter of the Preparation of leaf extract: Air dried pieces of leaves of inhibition zone for each fungus was measured after 48 hrs Pisonia grandis were extracted with 100% ethanol for 6 hour at reflux temperature. The extract was filtered; thefiltrate was evaporated to one tenth volume under reduced RESULTS AND DISCUSSION
pressure to get a greenish black pasty solid (sample A).
Table 1 shows the results of anti-fungal activity. The leaf Fractionation and column chromatography Procedure: A
ethanol extract and the various fractionated portion of the small portion of the sample A was set aside for testing its leaf extract showed varying degrees of inhibition against anti-fungal activity and the rest was macerated with equal all the fungal stains but only the ethanol soluble extract of volume of water and extracted with equal volumes of Pisonia grandis possesses maximum anti-fungal activity chloroform (CHCl3). The Liquid liquid extraction (LLE) with CHCl3 was continued until the CHCl3 layer was clotrimazole. Hence the ethanol extract of the leaves was colorless. Then the entire CHCl3 and aqueous layer were selected for column chromatographic analysis.
*Author for correspondence: Email: email@example.com Shubashini K. Sripathi, Poongothai G./ Bioassay- Guided Fractionation… Table 1 Anti-fungal screening result (Zone of inhibition in mm)Sample Concentrated pet-etherresidueConcentrated Table 2 Anti-fungal screening result for column fractions and for isolated compounds against Monascus purpureus Table 2 shows anti-fungal screening result for column Alloxan-induced diabetic rats. International journal fractions against Monascus purpureus and column of Applied Research in natural products. 2009; fractions 4, 5,6,7,8 and 11 showed a higher zone of inhibition than the standard Clotrimazole. The Minimum Radha R, Arokiyaraj S, Agastian P, Balaraju K, Inhibitory Concentration (MIC) values of these column Mohan Kumar R, Bula P. Phytochemical analysis fractions were also shown in Table 2. This study also and anti-inflammatory activity of Pisonia grandis revealed the potentially active fractions from which the R.Br. Journal of Biomedical and Pharmacology.
active principles would be isolated in the same Prabu D, Nappinnai M, Ponnudurai K, Prabhu K.
Evaluation of Wound-Healing Potential of Pisonia ACKNOWLEDGEMENTS
grandis R.Br: A Preclinical Study in Wistar Rats.
The authors thank the authorities of Avinashilingam The International Journal of Lower Extremity University for women, Coimbatore, India for providing necessary facilities and technical assistance. Authors are Anbalagan N, Rajinikanth KN, Kishore Gnanasam S, also grateful to The SAIF, Indian Institute of Technology Thomas Leonard J, Balakrishna K, Ramachandran S, Madras, India for the spectral data.
Sridhar SK. Analgesic, Anti-inflammatory andDiuretic Activities of Pisonia grandis. Journal of REFERENCE
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ȱɧɫɬɢɬɭɰɿɨɧɚɥɶɧɿ ɩɪɨɛɥɟɦɢ ɪɨɡɜɢɬɤɭ ɚɝɪɚɪɧɨʀ ɫɮɟɪɢ Privatdozent Dr. habil. VLADISLAV VALENTINOV, Schumpeter Fellow of the Volkswagen Foundation, Leibniz Institute of Agricultural Development in Central and Eastern Europe, Halle, Germany Third sector organizations in rural development: a transaction cost perspective1