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

Abstract

A simple and sensitive method was developed for simultaneous estimation of Glimepiride (GLIM) and Sildenafil citrate (SIL) 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 1 ml/min and UV detection at 230 nm. The retention time of GLIM and SIL was found to be 2.5 and 4.0 min respectively with total run time of 7 min. The developed method was validated for accuracy, precision, linearity and recovery. The method was linear and found to be acceptable over the range of 100–12 000 ng/ml. The method was successfully applied for the analysis of rat plasma sample for application to pharmacokinetic.

Key words

glimepiride - sildenafil citrate - RP-HPLC - rat plasma

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