A green reversed phase HPLC and HPTLC methods and their validation for simultaneous estimation of remogliflozin, vildgliptin and metformin in fixed dose combination formulation

To estimate remogliflozin, vildagliptin and metformin in a new fixed dose combination two chromatographic methods were developed and validated. The initial method is a green RP- HPLC method which uses a (150 x 4.6) mm, 2.7 mm C18 column, Ascentis as stationary phase for chromatographic separation and mobile phase made up of acetonitrile (ACN) and phosphate buffer in a ratio of 35:65 by volume. A PDA detector was used for quantification at 220 nm. 1.5 mL/min is the flow rate. Linearity was observed at 62.5-375 mu g/mL, 6.25-37.5 mu g/mL and 12.5-75 mu g/mL for remogliflozin, vildagliptin and metformin respectively. A subsequent technique developed is HPTLC densitometric method which was also found as greener method for the separation and quantification of the three analytes in which pre-coated silica gel 60F254 aluminum plates were taken as stationary phase and the mobile phase contain phosphate buffer: ACN: glacial acetic acid in the ratio of 3: 7: 0.2 by volume. The densitometric measurements were achieved at a wavelength of 205 nm. Linearity was observed at concentrations 200-600 ng/spot, 20-60 ng/spot, 40-120 ng/spot for metformin, vildagliptin and remogliflozine respectively. Mean % recovery was 98.92-100.41% for remogliflozin, 99.92-100.33% for vildagliptin and 99.17-100.19% for metformin. The method was tested precise, specific and robust. [GRAPHICS] .