Sarah Bos

53 Anticoagulant activity of edoxaban in patients with cirrhosis 4 Additional exclusion criteria for the control group included the presence of known liver disease and a history of clinically relevant bleeding complications. The study protocol was approved by the local medical ethical committee (METc 2016/226), and was registered at Netherlands Trial Register (NTR6397). Written informed consent was obtained from each subject before inclusion in the study. Plasma Samples Blood samples were drawn by venipuncture twice at day 1 (baseline and 2 hrs after the first dose) and once on day 3 and day 7 at peak level, two hours after ingestion of edoxaban. Specifically, 18 ml of blood in tubes containing 3.2% sodium citrate was drawn at the following time points: The blood was processed to platelet-poor plasma (PPP) by double centrifugation at 2000 g and 10,000 g respectively for 10 min. Plasma was stored at -80 ◦ C until use. Thrombin generation assay The thrombin generation assay (TGA) was performed in platelet-poor plasma (PPP) with the fluorimetric method described by Hemker, Calibrated Automated Thrombography® (CAT).(19) Coagulation was activated using commercially available reagents containing recombinant tissue factor (TF, final concentration 5 pM), phospholipids (final concentration 4 µM), in the presence of soluble thrombomodulin (TM, the concentration of which is not revealed by the manufacturer). Reagents were purchased from Thrombinoscope BV, Maastricht, the Netherlands, and thrombin generation experiments were executed following protocols provided by Thrombinoscope. The endogenous thrombin potential (ETP; which represents the total enzymatic work performed by thrombin during the time that it was active) was calculated from the thrombin generation curve using the Thrombinoscope software, and the percentual decrease in ETP following edoxaban administration was calculated. Conventional coagulation assays The prothrombin time (PT), activated partial thrombin time (APTT), and levels of prothrombin (FII), fibrinogen (FI) and D-dimer were assessed on an automated coagulation analyzer (ACL 300 TOP) with reagents (Recombiplastin 2G for PT, Hemosil