Login to MyKarger

New to MyKarger? Click here to sign up.

Login with Facebook

Forgot Password? Reset your password

Authors, Editors, Reviewers

For Manuscript Submission, Check or Review Login please go to Submission Websites List.

Submission Websites List

Institutional Login (Shibboleth)

For the academic login, please select your country in the dropdown list. You will be redirected to verify your credentials.

Table of Contents
Vol. 33, No. 4, 2003
Issue release date: July – August
Section title: Original Paper
Pathophysiol Haemost Thromb 2003;33:202–205

Ecarin Chromogenic Assay – A New Method for Quantitative Determination of Direct Thrombin Inhibitors Like Hirudin

Lange U.a · Nowak G.b · Bucha E.a
aHaemoSys GmbH and bResearch Unit ‘Pharmacological Hemostaseology’, Medical Faculty, Friedrich Schiller University, Jena, Germany
email Corresponding Author

Dr. Ute Lange

HaemoSys GmbH

Winzerlaer Strasse 2

DE–07745 Jena (Germany)

Tel. +49 36 41 50 83 21, Fax +49 36 41 50 83 01, E-Mail u.lange@haemosys.de


  1. Tripodi A, Chantarangkul V, Arbini AA, Moia M, Mannucci PM: Effects of hirudin on activated partial thromboplastin time determined with ten different reagents. Thromb Haemost 1993;70:286–288.
  2. Nurmohamed MT, Berckmans RJ, Morrien-Salomons WM, Berends F, Hommes DW, Rijnierse JJ, Sturk A: Monitoring anticoagulant therapy by activated partial thromboplastin time: Hirudin assessment. An evaluation of native blood and plasma assays. Thromb Haemost 1994;72:685–692.
  3. Cullberg M, Eriksson UG, Larsson M, Karlsson MO: Population modelling of the effect of inogatran, a thrombin inhibitor, on ex vivo coagulation time (APTT) in healthy subjects and in patients with coronary artery disease. Br J Clin Pharmacol 2001;51:71–79.
  4. Despotis GJ, Hogue CW, Saleem R, Bigham M, Skubas N, Apostolidou I, Qayum A, Joist JH: The relationship between hirudin and activated clotting time: Implications for patients with heparin-induced thrombocytopenia undergoing cardiac surgery. Anesth Analg 2001;93:28–32.
  5. Reid TJ 3rd, Alving BM: A quantitative thrombin time for determining levels of hirudin and hirulog. Thromb Haemost 1993;70:608–616.
  6. Spinner S, Stoffler G, Fink E: Quantitative enzyme-linked immunosorbent assay (ELISA) for hirudin. J Immunol Methods 1986;87:79–83.
  7. Spinner S, Scheffauer F, Maschler R, Stoffler G: A hirudin catching ELISA for quantitating the anticoagulant in biological fluids. Thromb Res 1988;51:617–625.
  8. Iyer L, Adam M, Amiral J, Fareed J, Bermes E Jr: Development and validation of two enzyme-linked immunosorbent assay (ELISA) methods for recombinant hirudin. Semin Thromb Hemost 1995;21:184–192.
  9. Griessbach U, Stürzebecher J, Markwardt F: Assay of hirudin in plasma using a chromogenic thrombin substrate. Thromb Res 1985;37:347–350.
  10. Stürzebecher J: Methods for determination of hirudin. Semin Thromb Hemost 1991;17:99–102.
  11. Spannagl M, Bichler J, Birg A, Lill H, Schramm W: Development of a chromogenic substrate assay for the determination of hirudin in plasma. Blood Coagul Fibrinolysis 1991;2:121–127.
  12. Groetsch H, Damm D, Ben Youssef R, Haertel D: Comparison of two different methods for the determination of rDNA-hirudin in plasma samples: HPLC vs a chromogenic thrombin substrate. Thromb Res 1991;64:273–277.
  13. Hafner G, Fickenscher K, Friesen HJ, Rupprecht HJ, Konheiser U, Ehrenthal W, Lotz J, Prellwitz W: Evaluation of an automated chromogenic substrate assay for the rapid determination of hirudin in plasma. Thromb Res 1995;77:165–173.
  14. Calatzis A, Spannagl M, Gempeler-Messina P, Kolde HJ, Schramm W, Haas S: The prothrombinase-induced clotting test: A new technique for the monitoring of anticoagulants. Haemostasis 2000;30(suppl 2):172–174.
    External Resources
  15. Nowak G, Bucha E: A new method for the therapeutic monitoring of hirudin. Thromb Haemost 1993;69:306–310.
    External Resources
  16. Nowak G: Monitoring of the action of antithrombin agents by ecarin clotting time; in Pifarré R (ed): New Anticoagulants for the Cardiovascular Patient. Philadelphia, Hanley & Belfus, 1997, pp 539–550.
  17. Nowak G: Clinical monitoring of hirudin and direct thrombin inhibitors. Semin Thromb Hemost 2001;27:537–541.
  18. Hafner G, Roser M, Nauck M: Methods for the monitoring of direct thrombin inhibitors. Semin Thromb Hemost 2002;28:425–430.
  19. Ratnoff OD, Menzie C: A new method for the determination of fibrinogen in small samples of plasma. J Lab Clin Med 1954;37:316–320.
  20. Nowak G, Bucha E: Quantitative determination of hirudin in blood and body fluids. Semin Thromb Hemost 1996;22:197–202.
  21. Morita T, Iwanaga S, Suzuki T: The mechanism of action of bovine prothrombin by an activator isolated from Echis carinatus venom and characterization of the new active intermediates. J Biochem (Tokyo) 1976;79:1089–1108.
  22. Rhee MJ, Morris S, Kosow DP: Role of meizothrombin and meizothrombin-(des F1) in the conversion of prothrombin to thrombin by the Echis carinatus venom coagulant. Biochemistry 1982;21:3437–3443.
  23. Cote HC, Stevens WK, Bajzar L, Banfield DK, Nesheim ME, MacGillivray RT: Characterization of a stable form of human meizothrombin derived from recombinant prothrombin (R155A, R271A, and R284A). J Biol Chem 1994;269:11374–11380.
  24. Schoen P, Lindhout T: The in situ inhibition of prothrombinase-formed human alpha-thrombin and meizothrombin (des F1) by antithrombin III and heparin. J Biol Chem 1987;262:11268–11274.
  25. Lindhoff-Last E, Piechottka GP, Rabe F, Bauersachs R: Hirudin determination in plasma can be strongly influenced by the prothrombin level. Thromb Res 2000;100:55–60.
  26. Lange U, Wiesenburg A, Olschewski A, Nowak G, Bucha E: Quantitative determination of direct thrombin inhibitors using the ecarin chromogenic assay (ECA) – both POCT and automated method. Ann Hematol 2003;82 (suppl 1):S53.
  27. Kathiresan S, Shiomura J, Jang IK: Argatroban. J Thromb Thrombolysis 2002;13:41–47.
  28. Fenyvesi T, Jörg I, Harenberg J: Monitoring of anticoagulant effects of direct thrombin inhibitors. Semin Thromb Hemost 2002;28:361–368.