RP-HPLC Method Development and Validation for Simultaneous Estimation of Berberine and Ursolic Acid in Formulation

Medicines used in ayurveda are typically made of many herbs or are polyherbals. Standardization of these medicines can be carried out by quantitative estimation of two or more of its markers simultaneously. Berberine and ursolic acid are two active markers which are present in an Ayurvedic formulation, Giloy Tulsi tablets. In the present research work, the Reverse Phase High Performance Liquid Chromatography (RP-HPLC) method was developed and validated for the simultaneous quantitative determination of berberine and ursolic acid from the formulation. With an RP Hemochrom C18 column, chromatographic separation was accomplished at 292 nm with eluent composed of methanol: acetonitrile (4:1, v/v) at a flow rate of 1.0 mL/min. The retention time of berberine and ursolic acid was obtained to be 1.5 ± 0.02 and 3.2 ± 0.02 min, respectively. A simple, rapid, accurate, specific, and robust analytical method was developed to estimate berberine and ursolic acid. This validated method can be used for routine quality control of ayurvedic formulations or herbal formulations having plants which contain berberine and ursolic acid.