Objective: The objective of this study was to develop and validate a novel, simple, rapid, precise and accurate reversed-phase high performance liquid chromatographic (RP-HPLC) method for simultaneous quantitative estimation of berberine, quercetin, and piperine in Ayurvedic formulation.Methods: The chromatographic separation was achieved using a stationary phase C18 shim-pack (150 mm x 4.6 mm, 5µ) column and mobile phase consisted of acetonitrile: 0.04 M potassium dihydrogen phosphate buffer (pH 3.0 adjusted using orthophosphoric acid) in a ratio of 65:35 v/v, with a flow rate of 1 ml/min and UV detection at 255 nm.Results: The retention time of berberine, quercetin, and piperine were found to be 2.7, 3.0 and 6.3 min respectively. Linearity for berberine, quercetin, and piperine were found in the range of 12-28 µg/ml. All calibration curve showed good linear correlation coefficients (r2˃ 0.999) within the tested ranges. Mean percent recoveries for berberine, quercetin, and piperine were found to be within the acceptance limits (98-120%). The percent relative standard deviation (% RSD) for precision was found to be less than 2% which indicates method is precise.Conclusion: The developed method is novel, simple, precise, accurate and can be used for quantitative analysis and quality control of the raw material as well as other commercial formulations containing these three markers.

Objective: The objective of this study was to develop and validate a simple, rapid, accurate, and precise reversed-phase high-performance liquid chromatographic method (RP-HPLC) for simultaneous estimation of drotaverine hydrochloride, ethamsylate, and tranexamic acid in tablet dosage form.Methods: The chromatographic separation was achieved using stationary phase C18 shim-pack GIST (150 mm ×4.6 mm, 5 μ) column and mobile phase consists of methanol:potassium dihydrogen phosphate buffer (pH 3.0 adjusted using orthophosphoric acid) in a ratio of 30:70 v/v, with a flow rate of 1 ml/min and ultraviolet detection at 220 nm.Results: The retention time of tranexamic acid, ethamsylate, and drotaverine hydrochloride was found to be 3.6, 4.0, and 5.0 min, respectively. The developed method was validated for linearity, accuracy, precision, and robustness according to the international conference on harmonization guidelines. The method was found to be linear over the tested concentration range of 48–112 μg/ml for drotaverine hydrochloride, 150–350 μg/ml for ethamsylate, and 150–350 μg/ml for tranexamic acid. Mean percentage recoveries were found to be 99.59 for drotaverine hydrochloride, 99.27 for ethamsylate, and 99.71 for tranexamic acid. The correlation coefficient for all components was found to be more than 0.999.Conclusion: A novel RP-HPLC method was developed and validated for simultaneous for the estimation of drotaverine hydrochloride, ethamsylate, and tranexamic acid to their commercially available tablet dosage form.