DEVELOPMENT AND VALIDATION OF HPLC METHOD FOR SIMULTANEOUS DETERMINATION OF BIO-ACTIVE MARKERS COROSOLIC ACID, ASIATIC ACID AND β-SITOSTEROL FROM LEAVES OF LAGERSTROEMIA SPECIOSA LINN. AND FROM MARKETED FORMULATION

摘要

Objective: To develop simple, accurate and reproducible High Performance Liquid Chromatographic (HPLC) method for simultaneous quantification of Corosolic acid, Asiatic acid and β-sitosterol in the leaf extract of L.speciosa. Methodology: The methanolic extract of the leaf powder of L.speciosa was separated on HPLC column (HyPurity C18, 100mm X 2.1mm, 5μ column) with gradient mixture of HPLC grade Acetonitrile and Water as mobile phase. The separated components were detected using Photo Diode Array (PDA) detector at 210 nm. Results: Linear response was found in the concentration range of 0.5-600 μg/mL for Asiatic acid, 1.0-600 μg/mL for Corosolic acid and 2.0-500 μg/mL for β-sitosterol respecticvely. The relative standard deviation for inter-day and intra-day precision was found to be <2%. The validated method was successfully applied for quantification of the three components in a commercially available dietary supplement "Banaba" containing L.speciosa leaf extract. Conclusion: A precise, accurate and reproducible HPLC method is developed for simultaneous quantification of three bio-active components from L.speciosa. The method can be used as quality control tool in standardization of raw materials and marketed formulations