Abstract

ANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR SIMULTANEOUS ESTIMATION OF DAPAGLIFLOZIN AND HYDROCHLOROTHIAZIDE IN SYNTHETIC MIXTURE

Hetal Rathva*, Mitali Dalwadi and Dr. Umesh Upadhyay

ABSTRACT

In the present study UV-spectroscopy and RP-HPLC method were developed validated for the simultaneous analysis of Dapagliflozin and Hydrochlorothiazide in synthetic mixture. UV spectroscopy were developed using First Order Derivative Spectroscopy method. The zero-crossing point (ZCP) was obtained at 277.00 nm and 241.20 nm for estimation of DAPA and HCT respectively, RP-HPLC method was developed using C18 (250mm x 4.6mm, 5µm) column as a stationary phase and Acetonitrile: Methanol: Phosphate buffer pH 3.5 with ortho phosphoric acid (30:35:35 % v/v/v) as mobile phase. Flow rate was 1ml/ min at 223 nm. The methods were validated in terms of linearity, accuracy (% Recovery), precision (inter day, intraday and reproducibility) and robustness. Both the methods were linear (R2 = 0.9994- 0.9955) and accurate (% recovery was 98.89 - 102.33% for DAPA and 98.08-103.62% for HCT for UV method and 98.63 – 102.10% for DAPA and 98.30 – 101.35% for HCT for RP-HPLC method). The method was also found precise (% RSD< 2%) and robust. Any one of the validated methods can be used for the analysis of Dapagliflozin and Hydrochlorothiazide synthetic mixture. The result of analysis was validated according to ICH Q2 R1 Guidelines. So The method was found to be accurate, precise, specific, selective, repeatable, robust and reproducible.

Keywords: UV spectroscopy, First Derivative, Spectra, HPLC, Chromatography, Stationary Phase, Mobile Phase, Development, Validation Parameters.