Novel Rp-HPLC Method Development and Validation for the Simultaneous Estimation of Saxagliptin and Glimepiride

Abstract

Objective: The objective of the present study was to develop, and validate a novel reverse phase high performance liquid chromatographic (RP-HPLC) method, for simultaneous determination of saxagliptin (SAXA) and glimepiride (GLIM), in bulk mixtures, and in tablets.Methods: Determination of the drugs, SAXA and GLIM, was carried out employing ODS C18 column (250 mm X 4.6 mm i. d, 5 µm particle size), with diode array detector at λmax of 230 nm. The mobile phase employed for the current study, composed of two solvents, i.e., A (acetonitrile), and B (0.1 % w/v sodium di-hydrogen orthophosphate buffer, pH 3.8 adjusted with orthophosphoric acid). The mobile phase was pumped at a flow rate of 0.75 ml/min in the gradient mode. The validation study with respect to specificity, linearity, precision, accuracy, robustness, limit of detection (LOD), and limit of quantification (LOQ), was carried out employing the ICH Guidelines.Results: The developed method was selective and linear for both the drugs, i.e., between 15.63 µg/ml and 250.00 µg/ml for SAXA, and 7.81 µg/ml and 125.00 µg/ml for GLIM, with a correlation coefficient (R2) 0.9977 and 0.9982, for SAXA, and GLIM, respectively. The % recovery obtained was 102.98±0.14% for SAXA, and 101.84±1.96% for GLIM. The LOD and LOQ values for SAXA were obtained to be 1.30 µg/ml, and 3.94 µg/ml, respectively, while for GLIM, LOD was 0.82 µg/ml and LOQ was 2.48 µg/ml. The method also exhibits good robustness for different chromatographic conditions like wavelength, flow rate, mobile phase and injection volume.Conclusion: The method was successfully employed, for the quantification of SAXA and GLIM, in the quality control of in-house developed tablets, and can be applied for the industrial use.