Abstract

A rapid and sensitive high-performance liquid chromatography (HPLC) method was developed and validated in human plasma for simultaneous determination of rosuvastatin and candesartan, using atorvastatin as internal standard. Chromatographic separation was achieved on a Waters C18 column (250 × 4.6 mm, 5 µm). A response surface methodology based 3 2 full factorial design was employed to optimize critical chromatographic conditions viz. pH and composition of mobile phase for achieving good resolution between the desired analytes. The optimised chromatographic conditions consisted of mobile phase of sodium acetate buffer (5 mM) with 0.1% acetic acid (pH 3.5)-acetonitrile (30:70, v/v), which was pumped at a flow rate of 1 mL/min. The detection was conducted at 254 nm for all the analytes. The calibration curves were linear over the concentration ranges of 10–150 ng/ml for both rosuvastatin and candesartan. The overall data of precision and accuracy were in accordance with US-FDA guidelines for bioanalytical method validation. Mean % extraction recovery observed for both analytes was above 80% as well as reproducible and consistent. No significant matrix effect was observed while carryover effect was deemed insignificant. Stability studies showed that the samples are stable over a long period which covers from sample collection to final analysis. Hence, the proposed method could be applied for routine laboratory analysis of rosuvastatin and candesartan in human plasma samples, clinical trials and pharmacokinetic studies.