A novel stability indicating Rp-Uplc-Dad method for determination of metformin and empagliflozin in bulk and tablet dosage form

Abstract

The objective of the present work was to develop and validate a novel stability indicating RP-UPLC-DAD method for the simultaneous analysis of Metformin and Empagliflozin in bulk and tablet dosage form. This new RP-UPLC method has superior in technology to formal Reverse phase-HPLC with retention time, solvent utilization, resolution and less cost. The peak area separation was accomplished on a Waters model UPLC system equipped with PDA detector and autosampler. A volume of 5 µL of sample and standard were injected into the column and all analytes were separated by using the mobile phase contains mixture 0.1% ortho phosphoric acid buffer (the pH was adjusted to 3.4 with 0.1 N NaOH) and methanol in the ratio 40:60% v/v at a flow rate of 0.25ml/ min through C18 BEH(Ethylene Bridged Hybrid) UPLC (100mm x 2.1mm,1.8µm) at 350C column temperature and the detector wavelength was set at 254 nm. The system suitable parameters such as tailing factor, resolution and plate count of two drugs were 1.16, 1.37;3.47;2314.34 and 4723 respectively. Retention time and peak area and of Metformin and Empagliflozin were found to be 0.882 & 3.471, 4887835 & 163463 respectively. Regression equation shows an r value (correlation coefficient) of greater than 0.999 for Metformin and Empagliflozin. Percent recovery of Metformin and Empagliflozin was found to be 99.92%-100.12% & 100.12%-100.56% respectively .Both Metformin and Empagliflozin were subjected to stress conditions such as acidic, basic, oxidative, thermal and photo degradation but substantial degradation was observed in acid studies. The newly developed RP-UPLC-DAD chromatographic method was validated with regard of system suitability, linearity, robustness, accuracy,, LOD,precision and LOQ.