Greener Liquid Chromatography for Simultaneous Determination of Rosiglitazone and Glimepiride in Rat Sera and its Application to a Pharmacokinetic Study

In this research, greener chromatography employing a more eco-friendly microemulsion mobile phase was utilized for the analysis of rosiglitazone (ROS) and glimepiride (GLM) in rat sera. ROS, GLM, and darfenacin (an internal standard) were detected using UV wavelength of 237 nm. HPLC analysis by using a monolithic column was performed by directly injecting the sample after appropriate dilution with the mobile phase. The chromatographic behavior of these compounds was studied to demonstrate their chromatographic efficiency, retention, and peak symmetry. Regression analyses showed excellent linearity (r = 0.999) over concentrations ranging from 100 to 5000 ng mL-1for both ROS and GLM. The developed method by using rat sera had limits of detection of 40.0 ng mL-1for both drugs. The assay was statistically validated for linearity, accuracy, precision and specificity according to ICH guidelines. The intra- and inter-assay precisions were satisfactory and the accuracy of the method was proved. This assay was successfully applied in a pharmacokinetic study.