SU1429025A1 - Method of analysing heparin in blood - Google Patents

Abstract

The invention relates to medicine, intended for the early detection of disorders of the blood anticoagulant system associated with a change in the size of the reserve intracellular heparin, which is able to penetrate into the bloodstream in patients with various pathologies. The purpose of the invention is to improve the accuracy of the method by detecting the reserve of heparin in the blood cells. To do this, a sample of venous blood is treated with Lactobacterin at a concentration of not less than 0.33 mg / ml, the plasma is separated by incubation and centrifugation. Then the thrombin solution is added and the plasma coagulation time is measured in comparison with the control. The time difference indicates the amount of free heparin in the blood. S (l

Description

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The invention relates to medicine and can be used for the early detection of disorders of the anticoagulant system of the meek, associated with the generation of the size of the reserve intracellular heparin, which can be divided into the bloodstream in patients with various pathologies (cardiovascular system, respiratory system, kidney, etc.),

The purpose of the invention is to improve the accuracy of the method by detecting a reserve of heparin in blood cells.

The method is carried out as follows.

i Into two siliconized tubes with Orz ml of sodium citrate solution (0.11 M), draw 3.0 ml of venous blood freely flowing out of the needle and mix thoroughly by inverting the tube three times with a covered polyethylene stopper. Add 0.3 ml to the first tube: lactobacterin at a concentration of 10 mg / ml in the second 0.3 ml of physiological solution and the tubes are thoroughly mixed. The tubes are placed us. 30 min in

; thermostat at J / S. After incubation: the tubes with blood are centrifuged at; 1000 rev / kcn for 10 min. In the half-I samples of plasma and the second and second test tubes, the content of heparin i is determined using the C -1 p and 3 method, and this is measured in a test tube; the test tube is measured on I “05 ml. Of l ziological rastkl plasma.

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 c: 1: 1 bath at, after 30 s pribsvlut ml of thrombin solution (production of Kaunas pre-ti bgteriologicheskie drugs at the Research Institute of Epidemiology, epidemiology, microbiology and gkgkeny) p. co-center 5 DG1 mg / ml and mark the clotting. and the plasma. Thrombin solution in such concentration should be folded. About ml Healthy person’s diluted in 0.05 ml of physiological saline in 30 seconds. In another test tube, measure 0 ml; OfOS ml of toluidine blue solution at a concentration of 1 mg / ml. The tube is placed in a water bath. After 30 seconds, a ml of thrombin working solution is added and the time is checked for 3 MI NB ani mp. about

The difference in coagulation time in the first and second agguration products

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tags indicates quantity. blood heparin,

In the blood plasma of the first tube, to which lactobacterin was added, the magnitude of the total blood heparin involved in the regulation of the hemostasis anticoagulant system is determined, and in the blood plasma of the second tube (without lactobacterian), the amount of free heparin in the blood. The difference in the amount of heparin in the blood plasma of the first and second tubes will be the reserve intracellular heparin.

Heparin was determined in two ways: known (S.E. Sirmoi) and proposed.

At the same time, paracoagulant tests were made: 3-naphthol sample by the method of Cummine N., Lyons R.A. (1948), ztalonova sample by the method of F. F. A, Tallis G.L. (1969), a common fibrinogen according to R.A.Rutberg (1961), a thromboelastogram was recorded on the device Hemocoagulograf. SHCGM 4.

Example 1. The blood of a healthy woman was examined for 25 years. Plasma heparin, determined by a known method, was 4 s, while plasma obtained from blood was subjected to lactobacterin action, 7.4 s. Consequently, the total blood heparin value, which determines the state of the anticoagulant system, is 7.4 s, the amount of free heparin is 4c, and the content of the reserve intracellular heparin is 7.4-4 3.4 s. Paracoagulation samples were negative, the total fibrinogen value was 2.7 g / l, the time R of the thromboelastogram was 6 minutes and 38 seconds.

Example 2. A 63-year-old patient's blood was investigated with a diagnosis of ischemic heart disease, stable angina pectoris, functional class 2, H st. The magnitude of heparin plasma. Not by the door-to-door effect of lactobacterin was 4.2 s, and the plasma obtained from blood subjected to the action of lactobacterin was 11 s. Consequently, the value of the total blood heparin involved in the regulation of the state of backlash is 11 seconds, free blood heparin, and the reserve intracellular 11-4, 8 seconds. The ethanol sample (one and: -; paracoagulation samples) was positive by 2 pluses, the value of total fibrinogen was 4.8 g / l, and the time R of the thromboelastogram was 4 min. 45 s.

From the above examples it can be seen that in comparison with a healthy person, a patient (Example 2) showed activation of blood coagulation: a positive result of a ethanol test.

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This method allows the activation of the anticoagulation system to be established by increasing the synthesis of intracellular heparin.

Claims (1)

Invention Formula The method of determining heparin in blood high concentration of total fibrin Qi by separation of plasma, incubated gene, shortened R thromboelastogram time. However, the largest free heparin studied by a known method is difficult to note at 37 ° C, adding a solution of thrombine with the subsequent measurement of the plasma coagulation time in comparison with a control of 5 o tl and h 0 u and c the change in the anticoagulation state | 5 so that, with a chain, the accuracy the blood of the patient: the content of heparin in the patient did not differ from that of a healthy person (example 1). Assessment of the patient’s anticoagulant system according to prev20 The pathway due to the detection of a reserve of heparin in blood cells, before plasma separation, blood is treated with lactobacterin in a concentration of at least 0.33 mg / mp. it at 37 ° C, adding a solution of thrombin, followed by measuring the time and plasma coagulation in comparison with a control of 5 o tl and h and The pathway due to the detection of a reserve of heparin in blood cells, before plasma separation, blood is treated with lactobacterin in a concentration of at least 0.33 mg / mp.