Quantitative determination of amlodipine in serum by liquid chromatography with atmospheric pressure chemical ionization tandem mass spectrometry.

Abstract

A sensitive and specific liquid chromatographic method coupled with tandem mass spectrometry was developed for the quantification of amlodipine in human and rat serum, which is a dihydropyridine derivative with calcium antagonist activity. An atmospheric pressure chemical ionization interface was used as the ion source and the analysis was performed in the selected reactive monitoring (SRM) mode. Deuterated amlodipine was used as the internal standard, and serum samples were treated with diethyl ether extraction prior to analysis. Serum levels in the range 0.014-7.2 ng ml-1 were measured accurately by this method, and the lower limit of quantitation (LOQ) was 0.014 ng ml-1 using 1 ml of human serum. The accuracy was within 7% of the expected values. The intra-assay precision was less than 3% and the inter-assay precision was less than 6%. The method was applied to a pharmacokinetic study of amlodipine in rats, in which the measurable range was 0.14-72 ng ml-1 using 0.1 ml of serum because of a limitation on the sample volume.