Abstract
To establish a rapid and sensitive ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for the determination of rivaroxaban, apixaban and edoxaban in rat plasma. The analytes and the internal standard (diazepam) were separated on an Acquity UPLC BEH C18 chromatography column (2.1mm×50mm, 1.7μm) using gradient elution with a mobile phase of acetonitrile and 0.1% formic acid in water at a flow rate of 0.4mL/min. The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring mode to monitor the precursor-to-product ion transitions of m/z 436.1→145.1 for rivaroxaban, m/z 460.0→443.1 for apixaban, m/z 548.2→366.1 for edoxaban and m/z 285.2→193.1 for diazepam (IS) using a positive electrospray ionization interface. The method was validated over a concentration range of 1.0-200ng/mL for rivaroxaban, 1.0-100ng/mL for apixaban and 1.0-500ng/mL for edoxaban. Total time for each chromatograph was 3.5min. The intra- and inter-day precision and accuracy of the quality control samples at low, medium, and high concentration levels exhibited relative standard deviations <10.5% and the accuracy values ranged from -9.9 to 11.3%. The method was successfully applied to a pharmacokinetic study of rivaroxaban, apixaban and edoxaban in rats.
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