Xây dựng và thẩm định phương pháp LC-MS/MS định lượng đồng thời amlodipin, perindopril và chất chuyển hóa có hoạt tính perindoprilat trong huyết tương người.

Abstract

A fast, simple, high throughput and specific method using ultra performance liquid chromatography tandem mass spectrometry was developed for simultaneous quantitation of amlodipine, perindopril and its active metabolite perindoprilat in human plasma. The Sciex Qtrap 6500+ UHPLCMS/MS was operated under the multi reaction-monitoring (MRM) mode using positive electrospray ionization technique. Amlodipine, perindopril, perindoprilat and internal standards were extracted from human plasma by protein precipitation technique using methanol. The samples were chromatographed on a C18 column (dimention 50 x 2,1 mm, particle 1,9 μm) with a gradient elution at a flow rate of 0.3 ml/min using 0.1 % formic acid in water – methanol. The calibration curves were found to be linear in the range 0.15 to 15 ng/ml, 1.25 to 125 ng/ml and 0.2 to 20 ng/ml for amlodipine, perindopril and perindoprilat, respectively, with mean correlation coefficient of > 0.99 for all analysis. The accuracy was within 85 % - 115.0 % and the precision was less than 15 %. Total run time was 4.5 min. This method can be used for BA-BE studies of amlodipine and perindopril combination preparations.