METHOD DEVELOPMENT AND VALIDATION FOR THE SIMULTANEOUS ESTIMATION OF DAPAGLIFLOZIN, VILDAGLIPTIN, AND METFORMIN IN BULK AND PHARMACEUTICAL DOSAGE FORM BY RP-HPLC

In the present study, a reverse phase high performance liquid chromatography method was developed and validated for the simultaneous estimation of Dapagliflozin, Vildagliptin and Metformin in pharmaceutical formulations. To assess the effect of method parameters on chromatographic separation of the Dapagliflozin, Vildagliptin and Metformin statistically designed experiments were performed by varying different method parameters such as buffer concentration, pH of mobile phase, flow rate, and column temperature. The separation was performed on Prontosil ODS C18 Column (250 x 4.6 mm and 5μm) at room temperature using Methanol: acetonitrile and pH 5.6 phosphate buffer in the ratio of 55:25:20 (v/v) in isocratic condition at a flow rate of 1.0 mL/min. The detection was performed by a photo diode array (PDA) detector at 223 nm with total run time of 15 min. Calibration curves were linear in the concentration range of 2.0-12.0 μg/mL for Dapagliflozin, 20-120 μg/mL for Vildagliptin and 100-600μg/mL for Metformin. The LOD was noticed to be 0.15 μg/mL, 1.5 μg/mL and 7.5 μg/mL whereas the LOQ was calculated as 0.5 μg/mL, 5 μg/mL and 25 μg/mL respectively for Dapagliflozin, Vildagliptin and Metformin proves the sensitivity of the developed method. The %RSD was below 2 which confirms the ruggedness and robustness of the method. The % assay in formulation analysis was found to be 98.90, 98.40 and 99.67 for Dapagliflozin, Vildagliptin and Metformin respectively. Hence, the method was used for the routine analysis of Dapagliflozin, Vildagliptin and Metformin in bulk drug as well as in pharmaceutical formulations.