Development And Validation Of RP-HPLC Method For Simultaneous Quantification Of Three Anti-Diabetic Agents In Pharmaceutical Formulation

The simultaneous determination of sitagliptin, metformin, and glimepiride in pharmaceutical preparation hasbeen accomplished through the development and validation of a novel, straightforward, quick, and sensitiveHPLC approach. Using an isocratic mobile phase made of 0.01M phosphate buffer: acetonitrile (40:60),separation was accomplished on a 250mm x 4.6mm, 5µm particle, C18 column. The pH of the mobile phasewas set at 5.6 with 50% orthophosphoric acid. A flow rate of 1 mL/min was used for the analysis. All chemicalswere detected using UV absorbance at 230 nm, and the analytes were eluted in less than ten minutes.According to ICH criteria, the approach was validated. Over the concentration range of 10-70 µg mL-1 for MET,5-35 µg mL-1 for SITA, and 1-30 µg mL-1 for GLI beyond, the linearity, accuracy, and precision of the methodwere satisfactory to good. The lower detection limit was found to be 0.67, 0.36, and 0.96µg mL-1 for, Metformin,Sitagliptin, and Glimepiride individually. The developed method is successfully used for the routine analysis offormulations for quality control and stability valuation.