Approaches Development and Validation of RP-HPLC Method for Estimation of Glimepiride in Rat Plasma-Application to Pharmacokinetic Studies

Abstract

A simple and sensitive method was developed for estimation of Glimepiride (GLIM) in
rat Plasma by reverse phase high performance liquid chromatography (RP-HPLC). The
drug samples were extracted by liquid-liquid extraction with 300 µL of acetonitrile and
5 mL of diethyl ether. Chromatographic separation was achieved on C18 column using
methanol: water (85:15 v/v) as mobile phase at a flow rate of 1ml/min and UV detection
at 230 nm. The retention time of GLIM was found to be 2.5 min. The developed method
was validated for accuracy, precision, linearity and recovery. Linearity studies were
found to be acceptable over the range of 100 – 6000 ng/mL. The method was
successfully applied for the analysis of rat plasma sample for application in
pharmacokinetic study, bioavailability.