Volume : 2, Issue : 5, MAY 2016

METHOD DEVELOPMENT AND VALIDATION FOR SIMULTANEOUS QUANTIFICATION OF OLEANOLIC ACID AND ?-SITOSTEROL FROM ACHYRANTHES ASPERA AND FROM MARKETED FORMULATION BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

Mansee M. Pathak, Vikas V. Vaidya, Deepal D. Ghadge

Abstract

Objective: To develop simple, accurate, precise and reproducible High Performance Liquid Chromatographic (HPLC) method for simultaneous quantification of Oleanolic acid and β-sitosterol in the whole plant extract of Achyranthes aspera.
Methodology: The methanolic extract of the plant powder of A. aspera was separated on HPLC column (Phenomenox, Luna® 5 μm C18, LC Column 150 x 2 mm) 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 10-300 μg/mL for β-sitosterol and 20-400 μg/mL for Oleanolic acid respectively. 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 two components in a commercially available polyherbal formulation “Cystone” containing A.aspera extract.
Conclusion: A precise, accurate and reproducible HPLC method is developed and validated for simultaneous quantification of two bio-active components from A.aspera. The method can be used as quality control tool in standardization of raw materials and marketed formulations.

Keywords

HPLC, Oleanolic acid, ?-sitosterol.

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