VALIDATION OF STABILITY INDICATING HIGH PERFORMANCE LIQUID CHROMATOGRAPHIC METHOD FOR SIMULTANEOUS DETERMINATION OF ASSAY OF GLYBURIDE AND METFORMIN DRUGS IN THE PHARMACEUTICALS TABLET FORMULATIONS USING ATOVAQUONE AS A COMMON INTERNAL STANDARD

Shaikh Javed Shaikh Afzal* and Pathan Mohd Arif Ali Khan

ABSTRACT

Glyburide is an oral antihyperglycemic agent used for the treatment of non-insulin-dependent diabetes mellitus (NIDDM). Metformin is a biguanide antihyperglycemic agent used for treating non-insulin-dependent diabetes mellitus (NIDDM).Validation of stability indicating Simple, Specific, Precise, Accurate, Linear, Rugged, Robust High Performance Liquid Chromatographic method of analysis for simultaneous determination of assay of Glyburide and Metformin drugs in the pharmaceuticals Tablet formulations using Atovaquone as a common internal standard was performed. The assay was accomplished using a mixture of Ammonium Dihydrogen phosphate in water and methanol in the volume ratio of 30:70 v/v as mobile phase on an OYSTER ODS3, 150 mm x 4.6mm, 5μ as chromatographic column at a flow rate 1.000 mLmin-1 and with a uv detector at a wavelength 230nm. The temperature of auto injector and column oven was 100C and 300C receptively. The Injection volume of HPLC system kept as 30 μL. linearity of the analytical method was evaluated at concentration range of 0.5937 μg/ml to 15.0057 μg/ml for Glyburide and 100.0644 μg/ml to 3000.0089 μg/ml for Metformin respectively with Correlation coefficient (r) value more than 0.999.The LOD and LOQ was 0.1242 μg/ml and 0.3764 μg/mL for Glyburide and 21.4220 μg/ml and 64.9153 μg/mL for Metformin respectively. The retention time found to be 2.96 min for Glyburide, 7.14 min for Metformin and 4.24 min for internal standard. Specificity, Method Precision, System Precision, Ruggedness, Robustness, Recovery, Stability of analytical solution, Filter paper selection study, Stress testing(Force Degradation) at various conditions were performed as per the ICH (Q2) recommendations. All the results were found with in acceptance criteria.

Keywords: Glyburide, Metformin Hydrochloride, Atovaquone, High Performance Liquid Chromatographic, Force degradation studies, Assay.