ABSTRACT:
Medicinal plants play important roles in drug discovery with many pharmaceutical products which originated from plants. Leucas aspera Linn.commonly known as 'Pandharpheda' and is widely distributed throughout southern part of India,having many medicinal properties. The present work is about the development of Thin-layer chromatography-direct bioautography method for the identification of bioactive compounds from the Leucas aspera Linn, Thin-layer chromatography-direct bioautography method is very convenient for searching plant constituents with biological activity, sach as antibiotics. TLC-DB is a high throughput method enabling analyses of many samples in parallel and the comparison of their activity. Both screening and semi-quantitative analysis is possible. The phytochemicals were extracted and used to check the antibacterial activity against Pathogenic strain of Escherichia coli using agar cup diffusion methods. In order to identify the bioactive compound Thin-layer chromatography-direct bio-autography method was used. The active compound was transferred online to LC-MSMS using TLC-MS interface from developed TLC silica plate for finding out molecular weight. Different collisions into MSMS was used to predict the structure of the antibacterial compound i.e. B-sitosterol further a sensitive, simple and accurate HPTLC method has been established for identification and quantification of B- sitosterol as a antibacterial agent from Leucas aspera Linn. And again CAMAG TLC-MS interface was used for the confirmation of m/z values of B-sitosterol in standard as well as sample. The m/z value was found to be 397(M+H+ -H20) The method ie. Bioautography, HPTLC and TLC-MS together provides a rapid and secure method for detection, identification and quantification of biomarker B-sitosterol.