UPLC-MS/MS method for the simultaneous determination of imatinib, voriconazole and their metabolites concentrations in rat plasma

Abstract

In the present study, a simple ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method used to measure the plasma concentrations of imatinib, voriconazole and their metabolites (N-desmethyl imatinib and N-oxide voriconazole) in rats simultaneously making use of diazepam as the internal standard (IS) had been developed and validated. A simple protein precipitation by acetonitrile was employed for the sample preparation, then the analytes (imatinib, voriconazole and their metabolites) were eluted on an Acquity UPLC BEH C18 column (2.1 mm × 50 mm, 1.7 μm) using the mobile phase that made up by acetonitrile (A) and 0.1% formic acid in water (B). In positive ion mode, four analytes and IS were monitored by multiple reaction monitoring (MRM) as the following mass transition pairs: m/z 494.3→394.2 for imatinib, m/z 480.3→394.2 for N-desmethyl imatinib, m/z 350.1→281.1 for voriconazole, m/z 366.1→224.1 for N-oxide voriconazole, and m/z 285.0→154.0 for IS. This method exhibited a good linearity for each analyte. Inter-day and intra-day precision were determined with values of 0.3–14.8% and 2.6–14.8%, respectively; the accuracy values were from −12.5% to 10.2%. Finally, data of matrix effect, extraction recovery, and stability were all conformed to the bioanalytical method validation of acceptance criteria of FDA recommendations. This method is an efficient tool for simultaneous determination of the four analytes and has been successfully applied for pharmacokinetic study in rats.