Determination of Glimepiride in Rat Serum by Rp-Hplc Method

Abstract

A simple and sensitive reverse phase high performance liquid chromatography (RP-HPLC) method was developed for the determination of glimepiride in rat serum. The assay involves one step liquid-liquid extraction with methanol. Gliclazide was used as an internal standard. Chromatographic separation was performed on a C 18 column using a mobile phase of methanol: 10 mM phosphate buffer (80:20 v/v) adjusted to pH 3.0 with orthophosphoric acid, at a flow rate of 1.0 ml/min and UV detection at 230 nm. The retention time of glimepiride and gliclazide was found to be 5.5 and 4.0 min and separation was complete in less than 10 min. The method was validated for linearity, accuracy and precision were found to be acceptable over the range of 0.5-500 µg/ml for glimepiride. The method was found suitable to analyse rat serum samples for application in pharmacokinetic, pharmacodynamic, bioavailability/bioequivalence studies.