Highlights
- •ROTEM® clotting times correlated well with dabigatran plasma concentrations.
- •Ecarin chromogenic assay and diluted thrombin time correlated with concentrations.
- •Coagulability similar in dabigatran-spiked samples and those from treated patients.
- •Storage for one year at −80 °C changed neither dabigatran levels nor coagulability.
Abstract
Background
Issues with laboratory measurement of dabigatran include: 1. Do coagulation assays
reflect dabigatran plasma concentrations? 2. Do samples from patients treated with
dabigatran have the same coagulability as dabigatran-spiked samples from healthy volunteers?
3. What is the long-term stability of dabigatran after storage at −80 °C? This study
aims to evaluate these questions.
Materials and methods
Ecarin chromogenic assay (ECA), a laboratory-developed diluted thrombin time (LD-dTT),
prothrombin time (PT) and activated partial thromboplastin time (APTT) and ROTEM®
were used to measure dabigatran anticoagulant activity and liquid chromatography-tandem
mass spectrometry (LC-MS/MS) to measure dabigatran plasma concentrations. ROTEM® (EXTEM,
INTEM, FIBTEM) was performed in whole blood and the other assays in platelet poor
plasma (PPP), both in samples spiked with dabigatran (0, 25, 50, 100, 250, 500 and
1000 ng/mL) from healthy donors and in ex vivo samples from patients treated with dabigatran etexilate. Citrated PPP samples were
frozen and stored at −80 °C, 1, 3, 6 and 12 months until analysis.
Results
EXTEM and FIBTEM clotting time (CT), ECA and LD-dTT correlate well with dabigatran
plasma concentrations. With the exception of few ROTEM® parameters, there were no
differences between spiked and patient samples. Samples were stable for at least 12 months
at −80 °C.
Conclusions
EXTEM and FIBTEM CT, ECA and LD-dTT are suitable for measuring the effect of dabigatran
in treated patients. In general, results from spiked plasma samples are similar to
those of patient samples. Storage of dabigatran plasma samples for up to 12 months
does not influence measured levels.
Abbreviations:
APTT (activated partial thromboplastin time), APTT-Cepha (APTT using Cephascreen® (Diagnostica Stago) reagent), APTT-PSL (APTT using Pathromtin®SL (Siemens Healthcare) reagent), ANOVA (analysis of variance), CAT (calibrated automated thrombinography), CT (clotting time), DMSO (dimethyl sulfoxide), DTI (direct thrombin inhibitor), ECA (ecarin chromogenic assay), ETP (endogenous thrombin potential), EXTEM (thromboelastometry testing the extrinsic haemostatic system), HCl (hydrochloric acid), INTEM (thromboelastometry testing the intrinsic haemostatic system), FIBTEM (thromboelastometry testing the fibrin part of clot formation), LC-MS/MS (liquid chromatography-tandem mass spectrometry), LD-dTT (a laboratory developed diluted thrombin time), MCF (maximum clot firmness), MRM (multiple reaction monitoring), NOAC (non-vitamin K antagonist oral anticoagulants), PT (prothrombin time), PT-INN (PT using Dade® Innovin® (Siemens Healthcare) reagent), PT-Owren (PT using Owrens (Medirox) reagent), ROTEM (rotational thromboelastometry), SE (standard error), SEE (standard error of the estimates), TF (tissue factor), TQD (tandem quadrupole), UPLC (ultra-performance liquid chromatography), VKA (vitamin K-antagonist)Keywords
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Article info
Publication history
Published online: February 17, 2018
Accepted:
February 16,
2018
Received in revised form:
February 8,
2018
Received:
December 13,
2017
Identification
Copyright
© 2018 Elsevier Ltd. All rights reserved.
