Novel RP-HPLC Method For Simultaneous Estimation of Andrographolide And Berberinein Polyherbal Formulation

A novel,simple, rapid, isocratic reverse phase high-performance liquid chromatographic method was developed for simultaneous estimation of andrographolide and berberine. The analysis was carried on Prontosil C18 (250 ×4.6 mm,5μ)column, with mobile phase consisting of acetonitrile and potassium dihydrogen phosphate buffer (pH 2.2 adjusted with ortho phosphoric acid) in the ratio of 30:70. The flow rate was set at 1.0ml/minand effluents were monitored at 235nm.The retention time of andrographolide and berberine werefound to be 10.08±0.2 and 12.47±0.2 minrespectively.Linear responses were obtained in concentration ranges of 5-50µg/ml, with R2 of 0.9989 and 0.9993 for andrographolide and berberine respectively. As per ICH guidelines, this method is accurate, simple, rapid and sensitive, hence can also be used in the evaluation of other formulations containing andrographolide and berberine.