HPLC and HPTLC methods for simultaneous quantification of Metformin hydrochloride, Vildagliptin and Dapagliflozin propanediol with comparative evaluation by greenness and whiteness assessment tools

Abstract

The combination of Vildagliptin, Dapagliflozin propanediol monohydrate and Metformin hydrochloride has been approved by Central Drug Standard Control Organization in March 2023 for conducting Phase III trial for treatment of diabetes mellitus. None of the chromatographic methods are reported for the proposed combination. In context to this, the proposed work aims to develop and validate two chromatographic methods, high HPLC and HPTLC for simultaneous estimation of the said combination followed by validation in accordance to ICH Q2(R2). For HPLC, linearity range were, 300–700, 30–70 and 3–7 µg/ml for Metformin hydrochloride, Vildagliptin and Dapagliflozin propanediol respectively. Good resolution was obtained with the final mobile phase, Acetonitrile: 10 mM potassium dihydrogen phosphate buffer pH 6.5 set with TEA (75:25 %v/v). The flow rate was 1 ml/min and detection wavelength was 214 nm. Rt of Metformin hydrochloride, Vildagliptin and Dapagliflozin propanediol was 2.262, 3.956 and 11.411 min respectively. For HPTLC the linearity range set was 1000–5000, 5000–9000 and 1000–5000 ng/band for Metformin hydrochloride, Vildagliptin and Dapagliflozin propanediol respectively. Separation of all drugs was observed using optimized mobile phase, Toluene: Ethyl acetate: 3% Ammonium acetate: Triethylamine(4: 4: 3: 0.1). R_f for Metformin hydrochloride, Vildagliptin and Dapagliflozin propanediol was 0.19, 0.48 and 0.61 respectively at 214 nm. % Relative Standard Deviation for validation parameters of both methods were found to be <2, which indicates that the methods were validated properly as per guideline. The proposed methods were specific, reliable, precise and can be applicable in routine analysis. Analytical Eco-Scale, Analytical GREEnness metric, and complex Green Analytical Procedure Index were the three methods used to assess the greenness. Furthermore, the quality (R), ecological effect (G), and economic feasibility (B) of the new technique were assessed by RGBfast tool for whiteness assessment and the applicability evaluated by the BAGI metric tool for performance verification.