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Thromb Haemost 1974; 31(03): 439-451
DOI: 10.1055/s-0038-1649184
Original Article
Schattauer GmbH

The Rivanol Method for the Quantitative Determination of Antithrombin III in Plasma

Robert H. Yue
1   Cardiovascular Research, Institute of Rehabilitation Medicine, New York University Medical Center, New York, New York 10016
,
Toby Starr
1   Cardiovascular Research, Institute of Rehabilitation Medicine, New York University Medical Center, New York, New York 10016
,
Menard M. Gertler
1   Cardiovascular Research, Institute of Rehabilitation Medicine, New York University Medical Center, New York, New York 10016
› Author Affiliations
Further Information

Publication History

Received 30 December 1973

accepted 20 February 1974

Publication Date:
29 June 2018 (online)

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Summary

A simple quantitative method for the determination of antithrombin III in plasma is described. It is based on the preferential precipitation of plasma antithrombin activities by a chemical compound 2,5-diamino-7-ethoxyacridin lactate (rivanol). When the incubation time of plasma with rivanol took place within 0.5 and 2 hours under specifically controlled experimental conditions, rivanol precipitated α2-macro-globulin selectively and quantitatively without any effect on antithrombin III. In the presence of rivanol, the inhibition of thrombin by plasma antithrombin III followed a second-order reaction with a rate constant of 80 1/M/sec. In this assay procedure, only the initial thrombin activity and the 30 minutes residual thrombin activity are required to evaluate the amount of antithrombin III present in the plasma quantitatively. “With proper routine laboratory care, this assay system was found to be accurate and reproducible.

 

  • References

  • 1 Abildgaard U. Purification of Two Progressive Antithrombin of Human Plasma. Scandinavian Journal of Clinical and Laboratory Investigation 1967; 19: 190
    CrossrefPubMedSearch in Google Scholar
  • 2 Abildgaard U. Binding of Thrombin to Antithrombin III. Scandinavian Journal of Clinical and Laboratory Investigation 1969; 24: 23
    CrossrefPubMedSearch in Google Scholar
  • 3 Abildgaard U, Egeberg O. Thrombin Inhibitory Activity of Fractions Obtained by Gel Filtration of Antithrombin III Deficient Plasma. Scandinavian Journal of Haematology 1968; 5: 155
    PubMedSearch in Google Scholar
  • 4 Abildgaard U, Gravern K, Godal H. C. Assay of Progressive Antithrombin in Plasma. Thrombosis et Diathesis Haemorrhagica 1970; 24: 224
    PubMedSearch in Google Scholar
  • 5 Bertho A, Grassmann W. Laboratory Methods of Biochemistry. New York: MacMillan & Co., Ltd.,; 1938
    Search in Google Scholar
  • 6 Biggs R, Denson K. W. E, Akman N, Borrett R, Hadden M. British Journal of Haematology 1970; 19: 283
    CrossrefPubMedSearch in Google Scholar
  • 7 Dombrose F. A, Seegers W. H, Sedensky J. A. Antithrombin: Inhibition of Thrombin and Autoprothrombin C (F — Xa) as a Mutual Depletion System. Thrombosis et Diathesis Haemorrhagica 1971; 26: 103
    PubMedSearch in Google Scholar
  • 8 Heimburger N, Haupt H, Schwick H. G. Proteinase inhibitor of human plasma. In Fritz H, Tschesche H. (ed.) Proceedings of the International Research Conference on Proteinase Inhibitors, Munich, November 4—61970. 1971: 1
    Search in Google Scholar
  • 9 Heimburger N, Trobisch H. Blood Coagulation and Fibrinolysis. Angewandte Chemie International Edition 1971; 10: 85
    PubMedSearch in Google Scholar
  • 10 Marciniak E, Tsukamura S. Two Progressive Inhibitors of Factor Xa in Human Blood. British Journal of Haematology 1972; 22: 341
    CrossrefPubMedSearch in Google Scholar
  • 11 Quick A. J. The Physiology and Pathology of Hemostasis . Philadelphia.: Lea and Febiger; 1951
    Search in Google Scholar
  • 12 Seegers W. H. Antithrombin as a Proteinase Inhibitor. Annals of New York Academy of Science 1968; 146: 593
    CrossrefPubMedSearch in Google Scholar
  • 13 Seegers W. H, Marciniak E. Inhibition of Autoprothrombin C Activity with Plasma. Nature 1962; 193: 1188
    PubMedSearch in Google Scholar
  • 14 Seegers W. H, Smith H. P. Antithrombin Activity of Plasma: Quantitative Interrelationship. Proceedings of the Society of Experimental Biology, New York 1943; 52: 159
    PubMedSearch in Google Scholar
  • 15 Seegers W. H, Cole E. R, Harmison C. R, Monkhouse F. C. Neutralization of Autoprothrombin C Activity with Antithrombin. Canadian Journal of Biology 1964; 42: 359
    PubMedSearch in Google Scholar
  • 16 Steinbuch M, Beatrix C, Josso F. A2-macroglobulin as a Progressive Antithrombin. Nature 1967; 216: 500
    CrossrefPubMedSearch in Google Scholar
  • 17 Steinbuch M, Quentin M, Pejaudier L. Specific Technique for the Isolation of Human A2-macroglobulin. Nature 1965; 205: 1227
    CrossrefPubMedSearch in Google Scholar
  • 18 Yin E. T, Wessler S, Butler J. V. Plasma Heparin: A Unique, Practical, Submicrogram-Sensitive Assay. Journal of Laboratory and Clinical Medicine 1973; 81: 298
    PubMedSearch in Google Scholar
  • 19 Yin E. T, Wessler S, Stoll P. J. Rabbit Plasma Inhibitor of the Activated Species of Blood Coagulation Factor X. Purification and Some Properties. Journal of Biological Chemistry 1971; a 246: 3694
    PubMedSearch in Google Scholar
  • 20 Yin E. T, Wessler S, Stoll P. J. Biological Properties of the Natural Occurring Plasma Inhibitor to Activated Factor X. Journal of Biological Chemistry 1971; b 246: 3703
    PubMedSearch in Google Scholar
  • 21 Yin E. T, Wessler S, Stoll P. J. Identity of Plasma-Activated Factor X Inhibitor with Antithrombin III and Heparin Cofactor. Journal of Biological Chemistry 1971; c 246: 3712
    PubMedSearch in Google Scholar
  • 22 Yue R. H, Starr T, Gertler M. M. Antithrombin III Levels in Ischemic Thrombotic Vascular Diseases. Proceedings of the III Congress of the International Society of Thrombosis and Haemostasis, August 22–261972. Washington, D. C.: 1972
    Search in Google Scholar
  • 23 Yue R. H, Starr T, Gertler M. M. The Rivanol Method for the Quantitative Determination of Antithrombin III in Plasma. Circulation 1973; a 48 Supplement 77
    PubMedSearch in Google Scholar
  • 24 Yue R. H, Starr T, Gertler M. M. Quantitative Determination of Total Antithrombin III in Plasma. Thrombosis et Diathesis Haemorrhagica 1973; b 30: 84
    PubMedSearch in Google Scholar