Development and validation of HPLC method for the determination of Candesartan in human plasma.

ABSTRACT

Candesartan (CAN), an ARB-blocker, antihypertensive, was analyzed in human plasma by a simple, accurate and precise RP-HPLC (reverse phase-High performance liquid chromatography assay method which was then validated for its accuracy, specificity and precision. The mobile phase has a constitution of acetone, diethylamine and distilled water, while Phosphoric acid was used to adjust the pH to 2.5±0.1. This mobile phase was run at 1.1ml/min and the fluorescence wavelength was set to 392 nm. A C-18 HPLC, column particle size (5 µm) Mediterranean Sea ® L x 1.D. 25cm x 4.6 mm (Supelcosil) , with auto sampler injection volume of 30µl ,an internal standard Valsartan was utilized for chromatographic detection. Candesartan took a retention time of 6±0.5 minutes. This method was validated by the parameters of selectivity, accuracy, precision, repeatability, reproducibility, recovery, linearity and stability. Candesartan's calibration curves were found to be linear in the range of 200ng/ml to 3.125ng/ml and the coefficient of determination (r2) was found to be 0.99. Analytical recovery obtained was above 88%. Hence, this method has been found to be useful for determining Candesartan in plasma.