Abstract

PHYTOCHEMICAL SCREENING AND SEPARATION OF BIOACTIVE COMPOUNDS FROM AZADIRACHTA INDICA LEAVES BY TLC AND COLUMN CHROMATOGRAPHIC TECHNIQUES

T. Anju*, A. Divya, V. Sri Varsha, R. Nandini, K. Prasanna Bhavani, Y. Bhavani, V. Varma and T. Satyanarayana

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Abstract

Many medicinal plants were studied for their phytochemical contents. Azadirachta indica is one of these plants. This study, primarily aimed to carry out a preliminary phytochemical screening to detect the major classes of bioactive compounds presented in Azadirachta indica dry leaves. Many solvents (Methanol, Ethanol, Ethyl acetate, Acetone, Acetic acid and Distilled water) were used to determine the best solvent that can be used for extraction and to perform Thin layer chromatography profiling of all sequential extracts. TLC was carried out in Silica gel plates using mobile phase protocol, Chloroform:Ethanol:Methanol (1:1:1) and using six mentioned solvents. Phytochemical screening of six extracts showed the presence of Alkaloids, glycosides, flavonoids, tannins, phenols, saponins and steroids in all solvent extracts. However, the concentration of these compounds is more with Methanol extract (+++). As well as thin layer chromatographic studies of the Azadirachta indica leaf extract (Methanol, Ethanol, Ethyl acetate, Acetone, Acetic acid and Distilled water) constituted different coloured phytochemical compounds with different Rf values. Methanol extract is the best which shows seven bands with Rf less than 1 for 24 hrs. After seven days among all the extracts, methanol extract was found to contain the maximum number of bioactive compounds which showed seven bands with Rf less than 1. Further Column chromatographic studies of methanolic extract using the solvent system Chloroform:ethanol:methanol, shows the better separation with methanol solvent. From these studies it was concluded that the methanol extract for Azadirachta indica leaves contain a higher content of bioactive compounds and methanol as a solvent shows effective separation of bioactive compounds, Which can be used for further research on this plant.

Keywords: Azadirachta indica, Thin layer chromatography, column chromatography, Methanolic extract.