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A kind of thrombin time determination reagent and preparation method thereof

A technology of thrombin time and bovine thrombin, which is applied in the field of thrombin time measurement reagents and its preparation, can solve the problems of increasing the use and control of raw materials, lowering the glass transition temperature of the mixed solution, increasing the viscosity of the mixed solution, etc., and reducing the dosage , reduce catalysis and improve accuracy

Active Publication Date: 2016-07-20
QINGDAO GUGAO BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Polyols or polysaccharides can improve the stability of thrombin, but excessive addition of such substances will increase the viscosity of the mixture, thereby reducing the accuracy of the TT value measured by optical methods
The addition of excess chlorate will reduce the glass transition temperature of the mixed solution while adjusting the ionic strength of the solution.
Sodium azide has good antibacterial properties (Lichstein, H.C.; Soule, M.H.Studies of the effect of sodium azide on microbic growth and respiration: I. The action of sodium azide on microbic growth". Journal of Bacteriology47(3):221–230), but its toxicity is relatively high. Clinical studies have shown that inhalation of azide Sodium can cause nasal and eye irritation, headache, dizziness, weakness, bronchitis, lower blood pressure and other symptoms. At the same time, under the condition of impact, sodium azide is easy to detonate. Therefore, the use of raw materials in the production process is increased. and control difficulty

Method used

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  • A kind of thrombin time determination reagent and preparation method thereof
  • A kind of thrombin time determination reagent and preparation method thereof
  • A kind of thrombin time determination reagent and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Weigh 6.2g Tris and 2.5g sodium chloride, add them into a glass bottle with a volume of 1.5L, add deionized water to 500mL, stir to dissolve, adjust the pH to 7.3 with dilute hydrochloric acid, add 2g bovine serum albumin, 1.4g chloride Calcium chloride, 2g of polyethylene glycol, 15g of mannitol, 10g of maltose, and 0.5g of sodium azide were thoroughly mixed, and the volume was adjusted to 1L, and bovine thrombin was added to make the bovine thrombin concentration 5U / ml. After stirring and mixing, the reagent for measuring thrombin time is obtained.

Embodiment 2

[0018] Weigh 6.2g Tris and 2.5g sodium chloride, add them into a glass bottle with a volume of 1.5L, add deionized water to 500mL, stir to dissolve, adjust the pH to 7.3 with dilute hydrochloric acid, add 1.5g bovine serum albumin, 1.4g Calcium chloride, 2g polyethylene glycol, 15g mannitol, 10g maltose, 0.5g sodium azide, 10g valine, after thorough mixing, set the volume to 1L, add bovine thrombin to make the concentration of bovine thrombin It is 5U / ml. After stirring and mixing, the thrombin time determination reagent is obtained.

Embodiment 3

[0020] Weigh 6.2g Tris and 2.5g sodium chloride, add them into a glass bottle with a volume of 1.5L, add deionized water to 500mL, stir to dissolve, adjust the pH to 7.3 with dilute hydrochloric acid, add 1.5g bovine serum albumin, 1.4g Calcium chloride, 2g polyethylene glycol, 15g mannitol, 10g maltose, 10g valine, 2mM coumaric acid, after thorough mixing, set the volume to 1L, add bovine thrombin to make the bovine thrombin concentration 5U / ml, after stirring and mixing, the thrombin time determination reagent is obtained.

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Abstract

The invention relates to a thrombin time measuring reagent which is prepared from bovine thrombin and a stabilizer. The stabilizer is prepared from valine and cumaric acid, wherein the adding amount of the valine ranges from 0.5% to 2.5% by the mass volume ratio, and the adding amount of the cumaric acid ranges from 0.1 mM to 4 mM. The preparing process of the thrombin time measuring reagent comprises the steps that a Tris buffer solution of 50 mM is prepared, pH is regulated to be neutral, the stabilizer is added into the buffer solution, the bovine thrombin is added after even mixing is conducted, and the thrombin time measuring reagent is obtained after even mixing. According to the thrombin time measuring reagent, the valine and the cumaric acid are selected to replace common triazacyclononane to serve as the stabilizer, the production and use processes of the thrombin time measuring reagent are safer, and the stability of the reagent is higher.

Description

technical field [0001] The invention relates to a coagulation reagent and a preparation method thereof, in particular to a thrombin time measurement reagent and a preparation method thereof. Background technique [0002] Thrombin time (TT) is used to detect coagulation, anticoagulation and fibrinolysis system functions, and is one of the commonly used clinical detection items. The main component of TT determination reagent is thrombin, as a biological macromolecule, thrombin is easily inactivated. In order to avoid denaturation of thrombin, it is necessary to add stabilizing components in the reagent. At present, most of the existing TT assay reagents on the market use polyols or polysaccharides or hydrochloride as a stabilizer for thrombin, and add sodium azide as an antibacterial agent or stabilizer at the same time, including patents US4696812, WO01 / 23536. Polyols or polysaccharides can improve the stability of thrombin, but excessive addition of such substances will ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/86
CPCG01N33/86
Inventor 胡小伟黄翔宇王华
Owner QINGDAO GUGAO BIOTECH CO LTD
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