The Spectrophotometry Determination of Thrombin Activity in Plasma^[*]

Summary

A method for the determination of thrombin activity, which is based on the spectrophotometry determination of the amount of fibrinogen clotted under standard conditions, was developed. The method was applied to the determination of the thrombin activity curve (TAC) of plasma and characteristic forms of the TAC were observed. The new method was applied to the preparation of a stable, platelet-poor defibrinogenated plasma, which was useful as a standard medium for making comparisons of normal and abnormal plasmas.

• References

• 1 Biggs R, MacFarlane R. G. Human Blood Coagulation and Its Disorders. Chap. 1. F. A. Davis Co.; Philadelphia: 1962
• 2 Seegers W. H. Thrombin and Prothrombin Activation. Ann. N. Y. Acad. Sci. 75: 182 1958;
• 3 Astrup T. The Haemostatic Balance. Thrombos. Diathes. haemorrh. (Stuttg.) 2: 347 1958;
• 4 Alexander B. Blood Coagulation and Thrombotic Disease. Circulation 25: 872 1962;
• 5 Kazal L. A. Coagulation Proteins. In: The Serum Proteins and Dysproteinemias. Sunderman W. F, and Sunderman Jr. W. F. Eds. J. B. Lippincott; Philadelphia: 1964
• 6 Biggs R, Douglas A. S. The Thromboplastin Generation Test. J. clin. Path. 6: 23 1953;
• 7 Ollendorff P. Defects in and Variability of the Thromboplastic System in Horse Plasma. Thrombos. Diathes. haemorrh. (Stuttg.) 4: 45 1959;
• 8 Ehrenpreis S, Scheraga H. A. Observations on the Analysis for Thrombin and the Inactiva-tion of Fibrin Monomer. J. biol. Chem. 227: 1043 1957;
• 9 Johnson J. F, Seegers W. H. Preparation, Purification and Assay of Thrombin. 120 In: The Coagulation of Blood. Methods of Study. Tocantins L. M. Ed. Grune and Stratton; New York: 1955
• 10 Kazal L. A, Amsel S, Miller O. P, Tocantins L. M. The Preparation and Some Properties of Fibrinogen Precipitated from Human Plasma by Glycine. Proc. Soc. exp. Biol. 113: 989 1963;
• 11 Kazal L. A, Miller O. P, Grannis G. F, Tocantins L. M. The Preparation of Human Fibrinogen from Cohn Fraction I. In: Blood Coagulation, Hemorrhage and Thrombosis – Methods of Study. Tocantins L. M, and Kazal L. A. eds. Grune and Stratton; 1964
• 12 Scheraga H. A, Laskowski Jr. M. The Fibrinogen-Fibrin Conversion. Advanc. Protein Chem. 12: 1-131 1957;
• 13 Waugh D. F, Livingstone B. J. Clotting Time and Reaction Velocity in the Interaction of Bovine Fibrinogen and Thrombin. Science 113: 121 1951;
• 14 Cocking E. C, Yemm E. W. Estimation of Amino Acids by Ninhydrin. Biochem. J. 58: XII 1954;
• 15 Gomori G. A Modification of the Colorimetrie Phosphorus Determination. J. Lab. clin. Med. 27: 955 1941; -2
• 16 Kazal L. A, Grannis G. F, Miller O. P. Studies of a Circulating Antithromboplastin. (In preparation.)
• 17 Pitney N. R, Dacie J. V. A Simple Method of Studying the Generation of Thrombin in Recalcified Plasma. J. clin. Path. 6: 9 1953;
• 18 Biggs R, MacFarlane R. G. Human Blood Coagulation and Its Disorders. 401 F. A. Davis, Co.; Philadelphia: 1962
• 19 Kazal L. A, Grannis G. F, Tocantins L. M. The Reactions of Thrombin with Fibrinogen and Plasma Antithrombin. Thrombos. Diathes. haemorrh. (Stuttg.) 13: 343 1965;