A simple, specific, sensitive and rapid reverse phase high performance liquid chromatographic (HPLC) method for the determination of irbesartan (IRB) in small volumes of rat plasma was developed and validated. Biological sample preparation involved simple extraction with organic solvent, followed by dilution with mobile phase to eliminate any chromatographic solvent effects. IRB was quantitated on a C8 column (4.6 mm i.d. × 300 mm length), using a mobile phase composed of methanol: acetonitrile (70:30 %v/v) which was delivered at a flow rate of 1.0 mL/min. The method was proven to be linear over a plasma concentration range of 10 -1000 ng/mL with a mean correlation coefficient of 0.9913. The intra-day and inter-day precision (coefficient of variation) were in the range of 1.9% to 4.3% and 2.7% to 5.4%, respectively. The intra-day accuracy (relative error) was in the range of 2.2% to 5.1% and the inter-day accuracy were in the range of 3.4% to 5.8%. The mean recovery of IRB from rat plasma was found to be 96.93%. The limit of detection (LOD) and the limit of quantification (LOQ) of the developed method were determined to be 1 ng/mL and 10 ng/mL respectively. The developed method was successfully applied to quantitatively assess the pharmacokinetics of IRB in wister rats following a single oral dose (6.75 mg/kg). The developed method was established as a rapid analytical tool in a pharmacokinetic study as it required short retention time, high precision, sensitivity and small volumes of plasma for analysis.
Loading....