Analysis of carvedilol in human plasma using hydrophilic interaction liquid chromatography with tandem mass...

A rapid, sensitive and selective method for the determination of carvedilol in human plasma was developed using hydrophilic interaction liquid chromatography with tandem mass spectrometry (HILIC-MS/MS). Carvedilol and cisapride (internal standard) were extracted from human plasma with methyl tert-butyl ether at basic pH and analyzed on an Atlantis HILIC Silica column with the mobile phase of acetonitrile-ammonium formate (50 mM, pH 4.5) (90:10, v/v). The analytes were detected using an electrospray ionization tandem mass spectrometry in the multiple-reaction-monitoring mode. The standard curve was linear (r=0.9998) over the concentration range of 0.1-200 ng/ml. The lower limit of quantification for carvedilol was 0.1 ng/ml using 50 microl plasma sample. The coefficient of variation and relative error for intra- and inter-assay at four QC levels were 1.6-4.5% and -6.4 to 4.8%, respectively. The absolute and relative matrix effect for carvedilol and cisapride were practically absent. The extraction recoveries of carvedilol and cisapride were 81.6 and 85.2%, respectively. This method was successfully applied to the bioequivalence study of carvedilol in humans.