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Measurement Of Apixaban, Dabigatran, Edoxaban And Rivaroxaban In Human Plasma Using Automated Online Solid-phase Extraction Combined With Ultra-performance Liquid Chromatography-tandem Mass Spectrometry And Its Comparison With Coagulation Assays.

J. Kuhn, T. Gripp, T. Flieder, A. Hammerschmidt, D. Hendig, I. Faust, C. Knabbe, I. Birschmann
Published 2018 · Chemistry, Medicine

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BACKGROUND Measurement of direct oral anticoagulants (DOACs) concentration in patient blood is essential in special clinical circumstances. METHODS We developed a fast, selective and sensitive method for simultaneous measurement of DOACs in human plasma consisting of an automated online solid-phase extraction method coupled with ultra-performance liquid chromatography electrospray ionization-tandem mass spectrometry (online SPE-UPLC-MS/MS). RESULTS The calibration curves of all DOACs were linear over the working range (apixaban: 0.25-760 μg/L, r > 0.99; dabigatran: 0.5-900 μg/L, r > 0.99; edoxaban: 0.6-800 μg/L, r > 0.99; rivaroxaban: 0.5-900 μg/L, r > 0.99). Limits of detection in the plasma matrix were < 0.2 μg/L, whereas the lower limits of quantification were < 0.6 μg/L for all DOACs. The intraassay and interassay CV for all DOACs were < 6%. Mean recoveries were between 61.4% and 91.6%. Method comparison between our online SPE-UPLC-MS/MS assay and commercially available functional based coagulation assays using patient samples showed a high degree of correlation for all investigated DOACs. CONCLUSIONS We developed and validated the first online SPE-UPLC-MS/MS method for fast, sensitive, specific, and reliable measurement of the new generation of DOACs and compared this method with commercial available coagulation assays.
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