High-Performance Thin Layer Chromatography Method for Identification and Quantification of Oleanolic Acid in the Roots of Achyranthes aspera Linn

Abstract

Aim: A correlation exists between biological activities and presence of secondary metabolites in plants. The present research work is carried out to develop and validate a chromatographic method using HPTLC for identification and quantification of Oleanolic Acid from the roots of Achyranthes aspera Linn. Method: The External Standard Method is used for quantification analysis in present study. It assures accuracy and precision in quantitative analysis and is especially suited for HPTLC studies. Chromatography was performed on aluminium-backed silica gel 60F254 HPTLC plates of 250 μm thickness with solvents toluene: ethyl acetate: methanol: acetone 14:4:1:1 (v/v/v/v) as the mobile phase. Derivatisation was carried out with Anisaldehyde sulphuric acid, scanned, and quantified at 540 nm. Results: The concentration of Oleanolic Acid in roots of the plant is found to be 1.277ng/ μg. The statistical analysis proved that the developed method is suitable and specific. Conclusion: This is a sensitive, specific and reproducible HPTLC method for the quantification of Oleanolic Acid from roots of Achyranthes aspera Linn.