Abstract
The present paper describes a novel, isocratic, simple, precise, accurate, and robust reversed-phase high-performance liquid chromatographic (RP-HPLC) method development and validation for simultaneous quantitative estimation of berberine and aloe-emodin in polyherbal formulation. The proposed chromatographic estimation was carried out isocratically using Cosmosil C18 (250 × 4.6mm) SH 5.0μm column, the mobile phase was a mixture of 0.05 M potassium dihydrogen phosphate buffer (pH adjusted to 2.5 using ortho-phosphoric acid): acetonitrile in a ratio of 60: 40 v/v with a flow rate of 1 mL/min and column oven adjusted to 30°Cwith injection volume 10μL. The ultraviolet (UV) detection was carried out at 225 nm. The retention time of berberine and aloe-emodin was found to 4.78±0.2 min and 15.49±0.2 min respectively. Calibration curves were linear over the tested concentration range of 4 to 20μg/mL. The present study reveals that this novel isocratic HPLC method is well validated, reliable and can be used for routine analysis of berberine and aloe-emodin in polyherbal formulations containing these phytoconstituents as one of the ingredients.
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