Liquid chromatographic method for simultaneous quantitation of clopidogrel, aspirin and atorvastatin in rat plasma and its application to the pharmacokinetic study

Abstract

A simple and robust analytical reversed-phase high-performance liquid chromatography method was developed and validated for simultaneous chromatographic elution of three cardiovascular drugs, namely clopidogrel, aspirin (ASP) and atorvastatin. The method was developed in rat plasma and dosage formulation with high-quality chromatographic separation between the drug peaks by using a stainless steel analytical column thermo beta-basic, C18 (25 3 0.46 cm, 5 mm). The system was operated at 258C using a mobile phase consisting of acetonitrile and phosphate buffer (pH 3.0) in the gradient ratio at a flow rate of 1 mL min21 with ultraviolet detection monitored at 232 nm. The parametric statistics, i.e., correlation coefficient of 0.999, was assessed for all the drugs having linearity over the tested concentration range (10-10,000 ng mL21) in rat plasma using an unweighted calibration curve. The accuracy of samples for six replicate measurements at lower limit of quantitation level was within limit. The method was applicable for the quality control of the mentioned drugs in raw material, bulk drug and pharmaceutical formulations as well as in pharmacokinetic studies.