Quantification of peramivir in rat plasma by liquid chromatography/tandem mass spectrometry employing precolumn derivatization and its application in pharmacokinetics / 军事医学科学院院刊

ABSTRACT

Objective:To develop a liquid chromatography/tandem mass spectrometry employing precolumn derivatization method for determination of peramivir in rat plasma.Methods: The sample preparation consisted of a protein precipitation extraction followed by derivatization with hydrochloric acid (10 mol/L) methanol (10∶90, v/v) and determined by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). The positive ion SRM mode was performed and the precursor-to-the-product ion transitions of m/z 343→284 and 299→152 were used to measure the derivative of peramivir and Ro 64-0802 (I.S.). The chromatographic separation was achieved using a Zorbax RX-C8 (2.1 mm × 150 mm × 5 μm) analytical column with an isocratic mobile phase composed of acetonitrile-water-formic acid (30∶70∶0.1, v/v/v, 0.2 ml/min). Results:The method was linear over a concentration range of 10 to 10 000 ng/ml. The LLOQ was 10 ng/ml. The average inter-day/intra-day precision values ranged from 5.0% to 7.1%, respectively, while the accuracy values ranged from 89.9% to 106.1%.Conclusion: In this method, retention time is greatly improved, the matrix effects are decreased by chemical derivatization. This method has been successfully applied to the preclinical and clinical research of peramivir.