1. Determination of dabigatran, rivaroxaban and apixaban by ultra-performance liquid chromatography - tandem mass spectrometry (UPLC-MS/MS) and coagulation assays for therapy monitoring of novel direct oral anticoagulants
- Author
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Jhpm Jack Emmen, E Ellen Schmitz, K Koen Boonen, van de Dh Daan Kerkhof, van der F Graaf, Mwm Schellings, van den Dja Heuvel, van Jlj Joost Dongen, Luc Brunsveld, Biomedical Engineering, Institute for Complex Molecular Systems, Soft Tissue Biomech. & Tissue Eng., Macromolecular and Organic Chemistry, Applied Physics and Science Education, and Chemical Biology
- Subjects
Quality Control ,Accuracy and precision ,Pyridones ,Morpholines ,Administration, Oral ,Thiophenes ,Thrombin time ,Pharmacology ,Tandem mass spectrometry ,Dabigatran ,Rivaroxaban ,Liquid chromatography–mass spectrometry ,Reference Values ,Tandem Mass Spectrometry ,medicine ,Humans ,Apixaban ,Blood Coagulation ,Chromatography, High Pressure Liquid ,Detection limit ,medicine.diagnostic_test ,Mass spectrometry ,Chemistry ,Anticoagulants ,Reproducibility of Results ,Hematology ,Drug monitoring ,Calibration ,beta-Alanine ,Pyrazoles ,Benzimidazoles ,Blood Coagulation Tests ,medicine.drug ,Factor Xa Inhibitors - Abstract
Background Three novel direct oral anticoagulants (DOACs) have recently been registered by the Food and Drug Administration and European Medicines Agency Commission: dabigatran, rivaroxaban, and apixaban. To quantify DOACs in plasma, various dedicated coagulation assays have been developed. Objective To develop and validate a reference ultra-performance liquid chromatography – tandem mass spectrometry (UPLC-MS/MS) method and to evaluate the analytical performance of several coagulation assays for quantification of dabigatran, rivaroxaban, and apixaban. Methods The developed UPLC-MS/MS method was validated by determination of precision, accuracy, specificity, matrix effects, lower limits of detection, carry-over, recovery, stability, and robustness. The following coagulation assays were evaluated for accuracy and precision: laboratory-developed (LD) diluted thrombin time (dTT), Hemoclot dTT, Pefakit PiCT, ECA, Liquid anti-Xa, Biophen Heparin (LRT), and Biophen DiXal anti-Xa. Agreement between the various coagulation assays and UPLC-MS/MS was determined with random samples from patients using dabigatran or rivaroxaban. Results The UPLC-MS/MS method was shown to be accurate, precise, sensitive, stable, and robust. The dabigatran coagulation assay showing the best precision, accuracy and agreement with the UPLC-MS/MS method was the LD dTT test. For rivaroxaban, the anti-factor Xa assays were superior to the PiCT-Xa assay with regard to precision, accuracy, and agreement with the reference method. For apixaban, the Liquid anti-Xa assay was superior to the PiCT-Xa assay. Conclusions Statistically significant differences were observed between the various coagulation assays as compared with the UPLC-MS/MS reference method. It is currently unknown whether these differences are clinically relevant. When DOACs are quantified with coagulation assays, comparison with a reference method as part of proficiency testing is therefore pivotal.
- Published
- 2014