CEEPC/IPM/CMSC - Abstrakt prezentace

(CEEPC/IPM/CMSC 2022 - FrP-40)
LC-MS/MS determination of direct oral anticoagulants in plasma of lupus positive patients

Barbora Piskláková 1,2 *, Jana Úlehlová 3,2, Eliška Ivanovová 1,2, Jana Procházková 3,2, Pavla Bradáčová 3,2, Aleš Kvasnička 1,2, David Friedecký 1,2, Luděk Slavík 3,2

  1. Laboratory of Inherited Metabolic Disorders, Department of Clinical Biochemistry,
  2. Palacký University Olomouc and University Hospital Olomouc
  3. Hemato-Oncology Clinic, Faculty of Medicine and Dentistry,


Direct oral anticoagulants (DOAC) are targeted inhibitors of coagulation factors that are commonly used for anticoagulation therapy. Functional assays used for their determination, are suspect of possible interference with antibodies present, especially in patients who have developed lupus anticoagulans (LA). An alternative option for the determination of DOAC is LC-MS, which should give correct results unencumbered by interferences. To evaluate the efficacy of DOAC treatment in lupus-positive patients, 31 plasma samples were investigated. All patient samples were spiked with three types of DOAC (dabigatran, rivaroxaban, and apixaban) at concentrations that significantly affected the screening test for LA and may mask the presence of LA. Determination of DOAC levels was performed both by routinely used functional assays and by our developed LC-MS/MS method. An UltiMate 3000 RS liquid chromatography system (Dionex, Sunnyvale, CA) coupled with a Triple Quad 6500 tandem quadrupole mass spectrometer (Sciex, Foster City, CA) was used for LC-MS/MS analysis. Separation was performed using a Luna Omega C18 polar column (Phenomenex, Torrance, CA) within 3 min. The results showed significant differences between these two approaches. When comparing them before and after DOAC binding, the fold change for the functional assays ranged from 1.4-1.7, whereas for LC-MS 62-183. This work suggests that the presence of LA-type antibodies substantially affects the determination of DOAC by functional assays and in this case the LC-MS/MS method should be used for their determination.

* Korespondující autor: barbora.pisklakova@upol.cz


This project was supported by MZ ČR – RVO (FNOl, 00098892), MZ ČR AZV NU20-08-00367, Doctoral Student Grant Competition UPOL DSGC-2021-0098.

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