Method Development and Validation for Simultaneous Estimation of Glimepiride and Simvastatin by using Reversed Phase High-performance Liquid Chromatography

Abstract

An analytical method was developed using reverse phase high performance liquid chromatograph (RP-HPLC) system and a C-18 reverse-phase column (Nucleodur C18, 250mm × 4.6mm i.d.,5μ) for simultaneous estimation of simvastatin and glimepiride. Acetonitrile and potassium dihydrogen phosphate buffer pH 4 (75:25, v/v) were used as mobile phase. The flow rate was 1 mL min−1 and the chromatogram of both drugs was detected at wavelength of 232 nm. Method was validated as per ICH Q2 (R1) guidelines. The retention times of glimepiride and simvastatin was found to be 4.726 min and 9.829 min, respectively. Both drugs have shown linearity over the concentration range 2–10μg/mL with r2 of 0.997 for GLM and 0.998 for SIM. The mean percentage recovery of both the drugs was found within 98–102% at all the levels which indicated that the method was accurate. The percentage relative standard deviation was found less than 2% which indicated that method was satisfactorily precised. The LOD and LOQ were found to be 0.24 and 0.73 for simvastatin and 0.32 and 0.96 for glimepiride. The method was found to be robust as there was no significant change in response with variation in pH, flow rate and mobile phase composition. It was concluded that the developed method has passed all the validation tests and can be successfully applied to estimate the presence of both the drugs in bulk as well as in pharmaceutical formulations.