Development and validation of stability indicating reverse Phase high performance liquid chromatography method with Photodiode array detection for the simultaneous estimation of hypoglycemic agents, Dapagliflozin and Metformin

Abstract

The aim of the present study was to develop and validate a stability indicating reverse phase high performance liquid chromatographic (RP-HPLC) method for simultaneous quantitative analysis of dapagliflozin and metformin in bulk and tablets. Dapagliflozin, metformin and stress degradation products were eluted on the Supelco C8, 250 mm x 4.6 mm, 5 μm analytical column with a mobile phase consisting of 0.1 M dipotassium hydrogen phosphate, acetonitrile and methanol (60:30:10 v/v/v), pumped at 1.2 mL/min flow rate. The column temperature was set 30°C and 10 μL of the samples were injected. Detection was done at 285 nm. According to ICH guidelines, the developed RP-HPLC method was validated. The retention times of dapagliflozin and metformin were 2.847 min and 3.804 min, respectively. Linearity was observed in the concentration range of 2-6 μg/mL (R² = 0.9999) for dapagliflozin and 200-600 μg/mL (R² = 0.9998) for metformin. The limit of detection for dapagliflozin and metformin were found to be 0.004 μg/mL and 0.272 μg/mL, respectively. The percent recovery and percent relative standard deviation for the selected drugs were in the range of 99.00%-99.82% and 0.098%-0.291%, respectively. Non interference of peaks from stress degradation products in acidic, alkaline, oxidative, thermal and photolytic conditions demonstrated the stability indicating power of the method. The proposed RP-HPLC method can be used as a stability indicating method for the assay of dapagliflozin and metformin simultaneously in bulk and tablet dosage form.