Semin Thromb Hemost 2015; 41(02): 228-236
DOI: 10.1055/s-0035-1544158
Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.

Patients' Serum and Urine as Easily Accessible Samples for the Measurement of Non–Vitamin K Antagonist Oral Anticoagulants

Job Harenberg
1   Clinical Pharmacology, Medical Faculty Mannheim, Ruprecht-Karls University Heidelberg, Mannheim, Germany
,
Shanshan Du
1   Clinical Pharmacology, Medical Faculty Mannheim, Ruprecht-Karls University Heidelberg, Mannheim, Germany
,
Sandra Krämer
1   Clinical Pharmacology, Medical Faculty Mannheim, Ruprecht-Karls University Heidelberg, Mannheim, Germany
,
Christel Weiss
2   Biometry and Statistics, Medical Faculty Mannheim, Ruprecht-Karls University Heidelberg, Mannheim, Germany
,
Roland Krämer
3   Institute Inorganic Chemistry, University of Heidelberg, Heidelberg, Germany
,
Martin Wehling
1   Clinical Pharmacology, Medical Faculty Mannheim, Ruprecht-Karls University Heidelberg, Mannheim, Germany
› Author Affiliations
Further Information

Publication History

Publication Date:
15 February 2015 (online)

Abstract

Measurement of the anticoagulant effect of non–vitamin K antagonist oral anticoagulants (NOAC) may be desirable, in particular in patients with acute medical conditions. Useful methods should give results rapidly within minutes, should be easy to perform, specific, and sensitive. Using plasma samples, chromogenic assays can be made to be specific for the two types of NOAC (factor Xa and thrombin inhibitors), and also hemoclot and ecarin clotting time specific for dabigatran. If plasma samples anticoagulated with sodium citrate are not available, blood samples anticoagulated with ethylene diamine tetraacetic acid or serum samples may be regarded as alternatives for the determination of NOAC. At present, dabigatran cannot be determined from serum samples because it may be consumed during the clotting process to obtain serum. NOAC can be determined in urine samples due to their renal elimination. Quantitative methods are preferable to qualitative methods, although the latter may be advantageous in some situations, being developed as point-of-care tests for oral factor Xa and thrombin inhibitors. In these tests, the presence and absence of NOAC in urine can be identified with the naked eye after a few minutes and these tests are highly specific and sensitive. New assays such as a semiquantitative determination in urine samples and measurement using other sample matrices are currently under development.

 
  • References

  • 1 Hylek EM, Ko D, Cove CL. Gaps in translation from trials to practice: non-vitamin K antagonist oral anticoagulants (NOACs) for stroke prevention in atrial fibrillation. Thromb Haemost 2014; 111 (5) 783-788
  • 2 Harenberg J, Wehling M. Future of anticoagulant therapy. Cardiovasc Ther 2011; 29 (5) 291-300
  • 3 Antovic JP, Skeppholm M, Eintrei J , et al. Evaluation of coagulation assays versus LC-MS/MS for determinations of dabigatran concentrations in plasma. Eur J Clin Pharmacol 2013; 69 (11) 1875-1881
  • 4 Douxfils J, Dogné JM, Mullier F , et al. Comparison of calibrated dilute thrombin time and aPTT tests with LC-MS/MS for the therapeutic monitoring of patients treated with dabigatran etexilate. Thromb Haemost 2013; 110 (3) 543-549
  • 5 Harenberg J, Kraemer S, Du S , et al. Determination of direct oral anticoagulants from human serum samples. Semin Thromb Hemost 2014; 40 (1) 129-134
  • 6 Gous T, Couchman L, Patel JP, Paradzai C, Arya R, Flanagan RJ. Measurement of the direct oral anticoagulants apixaban, dabigatran, edoxaban, and rivaroxaban in human plasma using turbulent flow liquid chromatography with high-resolution mass spectrometry. Ther Drug Monit 2014; 36 (5) 597-605
  • 7 Schmitz EM, Boonen K, van den Heuvel DJ , et al. 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. J Thromb Haemost 2014; 12 (10) 1636-1646
  • 8 Delavenne X, Moracchini J, Laporte S, Mismetti P, Basset T. UPLC MS/MS assay for routine quantification of dabigatran - a direct thrombin inhibitor - in human plasma. J Pharm Biomed Anal 2012; 58: 152-156
  • 9 Harenberg J, Du S, Krämer S , et al. Novel methods for assessing oral direct factor Xa and thrombin inhibitors: use of point-of-care testing and urine samples. Semin Thromb Hemost 2013; 39 (1) 66-71
  • 10 Harenberg J, Du S, Weiss C, Krämer R, Hoppensteadt D, Walenga J ; working party: methods to determine apixaban of the Subcommittee on Control of Anticoagulation of the International Society of Thrombosis and Haemostasis. Report of the Subcommittee on Control of Anticoagulation on the determination of the anticoagulant effects of apixaban: communication from the SSC of the ISTH. J Thromb Haemost 2014; 12 (5) 801-804
  • 11 Hauel NH, Nar H, Priepke H, Ries U, Stassen JM, Wienen W. Structure-based design of novel potent nonpeptide thrombin inhibitors. J Med Chem 2002; 45 (9) 1757-1766
  • 12 Stangier J, Stähle H, Rathgen K, Roth W, Reseski K, Körnicke T. Pharmacokinetics and pharmacodynamics of dabigatran etexilate, an oral direct thrombin inhibitor, with coadministration of digoxin. J Clin Pharmacol 2012; 52 (2) 243-250
  • 13 Du S, Weiss C, Giese C , et al. Determination of dabigatran in plasma, serum and urine samples: comparison of six methods. Clin Chem Lab Med . In press
  • 14 Harenberg J, Giese C, Marx S, Krämer R. Determination of dabigatran in human plasma samples. Semin Thromb Hemost 2012; 38 (1) 16-22
  • 15 Nowak G. The ecarin clotting time, a universal method to quantify direct thrombin inhibitors. Pathophysiol Haemost Thromb 2003; 33 (4) 173-183
  • 16 Fenyvesi T, Jörg I, Weiss C, Harenberg J. Effects of lepirudin, argatroban and melagatran and additional influence of phenprocoumon on ecarin clotting time. Thromb Res 2003; 111 (1-2) 89-94
  • 17 Stangier J, Feuring M. Using the HEMOCLOT direct thrombin inhibitor assay to determine plasma concentrations of dabigatran. Blood Coagul Fibrinolysis 2012; 23 (2) 138-143
  • 18 Harenberg J, Du S, Krämer S, Krämer R, Wehling M, Weiss C. Measurement of non-vitamin K antagonist oral anticoagulants in patient plasma using Heptest-STAT coagulation method (epub ahead of print). Ther Drug Monit 2014;
  • 19 Harenberg J, Giese C, Hagedorn A, Traeger I, Fenyvesi T. Determination of antithrombin-dependent factor Xa inhibitors by prothrombin-induced clotting time. Semin Thromb Hemost 2007; 33 (5) 503-507
  • 20 Calatzis A, Peetz D, Haas S, Spannagl M, Rudin K, Wilmer M. Prothrombinase-induced clotting time assay for determination of the anticoagulant effects of unfractionated and low-molecular-weight heparins, fondaparinux, and thrombin inhibitors. Am J Clin Pathol 2008; 130 (3) 446-454
  • 21 Samama MM, Martinoli JL, LeFlem L , et al. Assessment of laboratory assays to measure rivaroxaban—an oral, direct factor Xa inhibitor. Thromb Haemost 2010; 103 (4) 815-825
  • 22 Tripodi A, Chantarangkul V, Guinet C, Samama MM. The International Normalized Ratio calibrated for rivaroxaban has the potential to normalize prothrombin time results for rivaroxaban-treated patients: results of an in vitro study. J Thromb Haemost 2011; 9 (1) 226-228
  • 23 Van Blerk M, Bailleul E, Chatelain B , et al. Influence of dabigatran and rivaroxaban on routine coagulation assays. A nationwide Belgian survey. Thromb Haemost 2014; 112 (6)
  • 24 Hapgood G, Butler J, Malan E, Chunilal S, Tran H. The effect of dabigatran on the activated partial thromboplastin time and thrombin time as determined by the Hemoclot thrombin inhibitor assay in patient plasma samples. Thromb Haemost 2013; 110 (2) 308-315
  • 25 Samama MM, Contant G, Spiro TE , et al. Laboratory assessment of rivaroxaban: a review. Thromb J 2013; 11 (1) 11
  • 26 Suzuki S, Otsuka T, Sagara K , et al. Dabigatran in clinical practice for atrial fibrillation with special reference to activated partial thromboplastin time. Circ J 2012; 76 (3) 755-757
  • 27 Harenberg J, Krämer R, Giese C, Marx S, Weiss C, Wehling M. Determination of rivaroxaban by different factor Xa specific chromogenic substrate assays: reduction of interassay variability. J Thromb Thrombolysis 2011; 32 (3) 267-271
  • 28 Emanuele RM, Racanelli A, Fareed J. Pharmacokinetics of heparins differing in mean molecular weight using a Xa amidolytic and Heptest clotting method. Ther Drug Monit 1988; 10 (2) 153-159
  • 29 Yin ET, Wessler S, Butler JV. Plasma heparin: a unique, practical, submicrogram-sensitive assay. J Lab Clin Med 1973; 81 (2) 298-310
  • 30 Hoffmann U, Harenberg J, Bauer K , et al. Bioequivalence of subcutaneous and intravenous body-weight-independent high-dose low-molecular-weight heparin Certoparin on anti-Xa, Heptest, and tissue factor pathway inhibitor activity in volunteers. Blood Coagul Fibrinolysis 2002; 13 (4) 289-296
  • 31 Jeske WP, Walenga JM, Samama MM, Hoppensteadt D, Mayuga M, Fareed J. Functionality of fondaparinux (pentasaccharide) depends on clinical antithrombin levels. Blood Coagul Fibrinolysis 2011; 22 (3) 206-210
  • 32 Harenberg J, Jeschek M, Acker M, Malsch R, Huhle G, Heene DL. Effects of low-molecular-weight dermatan sulfate on coagulation, fibrinolysis and tissue factor pathway inhibitor in healthy volunteers. Blood Coagul Fibrinolysis 1996; 7 (1) 49-56
  • 33 Kubitza D, Becka M, Mück W, Krätzschmar J. Pharmacodynamics and pharmacokinetics during the transition from warfarin to rivaroxaban: a randomized study in healthy subjects. Br J Clin Pharmacol 2014; 78 (2) 353-363
  • 34 Mueck W, Lensing AW, Agnelli G, Decousus H, Prandoni P, Misselwitz F. Rivaroxaban: population pharmacokinetic analyses in patients treated for acute deep-vein thrombosis and exposure simulations in patients with atrial fibrillation treated for stroke prevention. Clin Pharmacokinet 2011; 50 (10) 675-686
  • 35 Miyares MA, Davis K. Newer oral anticoagulants: a review of laboratory monitoring options and reversal agents in the hemorrhagic patient. Am J Health Syst Pharm 2012; 69 (17) 1473-1484
  • 36 Dempfle CE, Zharkowa U, Elmas E, Ahmad-Nejad P, Neumaier M, Borggrefe M. Heptest-STAT, a new assay for monitoring of low-molecular-weight heparins, is not influenced by pregnancy-related changes of blood plasma. Thromb Haemost 2009; 102 (5) 1001-1006
  • 37 Kluft C, Meijer P, Kret R, Burggraaf J. Preincubation in the Prothrombinase-induced Clotting Time test (PiCT) is necessary for in vitro evaluation of fondaparinux and to be avoided for the reversible, direct factor Xa inhibitor, rivaroxaban. Int J Lab Hematol 2013; 35 (4) 379-384
  • 38 Lippi G, Favaloro EJ. Recent guidelines and recommendations for laboratory assessment of the direct oral anticoagulants (DOACs): is there consensus?. Clin Chem Lab Med 2014;
  • 39 Rosén S. Chromogenic methods in coagulation diagnostics. Hamostaseologie 2005; 25 (3) 259-266
  • 40 Douxfils J, Mullier F, Loosen C, Chatelain C, Chatelain B, Dogné JM. Assessment of the impact of rivaroxaban on coagulation assays: laboratory recommendations for the monitoring of rivaroxaban and review of the literature. Thromb Res 2012; 130 (6) 956-966
  • 41 Du S, Krämer S, Giese C, Weiss C, Wehling M, Krämer R, Harenberg J Chromogenic assays for measurement of rivaroxaban from EDTA anticoagulated plasma samples (epub ahead of print) Thromb Haemost 2015; 113 (5)
  • 42 Mani V, Wang S, Inci F, De Libero G, Singhal A, Demirci U. Emerging technologies for monitoring drug-resistant tuberculosis at the point-of-care. Adv Drug Deliv Rev 2014; 78C: 105-117
  • 43 Lippi G, Salvagno GL, Montagnana M, Lima-Oliveira G, Guidi GC, Favaloro EJ. Quality standards for sample collection in coagulation testing. Semin Thromb Hemost 2012; 38 (6) 565-575
  • 44 Harenberg J, Krämer S, Du S , et al. Measurement of rivaroxaban and apixaban in serum samples of patients. Eur J Clin Invest 2014; 44 (8) 743-752
  • 45 Stangier J, Rathgen K, Stähle H, Mazur D. Influence of renal impairment on the pharmacokinetics and pharmacodynamics of oral dabigatran etexilate: an open-label, parallel-group, single-centre study. Clin Pharmacokinet 2010; 49 (4) 259-268
  • 46 Harenberg J, Krämer S, Du S, Weiss C, Krämer R. Concept of a point of care test to detect new oral anticoagulants in urine samples. Thromb J 2013; 11 (1) 15
  • 47 Cuker A, Siegal DM, Crowther MA, Garcia DA. Laboratory measurement of the anticoagulant activity of the non-vitamin K oral anticoagulants. J Am Coll Cardiol 2014; 64 (11) 1128-1139
  • 48 Baglin T, Hillarp A, Tripodi A, Elalamy I, Buller H, Ageno W. Measuring Oral Direct Inhibitors (ODIs) of thrombin and factor Xa: A recommendation from the Subcommittee on Control of Anticoagulation of the Scientific and Standardisation Committee of the International Society on Thrombosis and Haemostasis. J Thromb Haemost 2013; 11 (4) 756-760