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Method for detecting potency of heparin or heparin with low molecular weight through a coagulometer

A technology of low molecular weight heparin and blood coagulation instrument, which is applied in the detection field of heparin (salt) or low molecular weight heparin (salt) anti-Xa titer by blood coagulation instrument, and can solve the problem of large error, high cost and chromogenic substrate method. Cumbersome and other problems, to achieve the effect of fast speed, low cost, and improved experimental accuracy

Inactive Publication Date: 2015-12-30
山东万邦赛诺康生化制药股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is: chromogenic substrate method is loaded down with trivial details, error is big, cost is high

Method used

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  • Method for detecting potency of heparin or heparin with low molecular weight through a coagulometer
  • Method for detecting potency of heparin or heparin with low molecular weight through a coagulometer
  • Method for detecting potency of heparin or heparin with low molecular weight through a coagulometer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] (1) Configuration of standard series solutions

[0034] 1.1 Tris-NaCl buffer solution (PH7.4)

[0035] Take 6.08g of trishydroxymethylaminomethane and 8.77g of sodium chloride, add 500ml of water to dissolve, add 10g of bovine serum albumin, adjust the pH value to 7.4 with hydrochloric acid, and add water to dilute to 1000ml.

[0036] 1.2 Tris-EDTA disodium buffer solution (PH8.4)

[0037] Take 5.12g of sodium chloride, 3.03g of trishydroxymethylaminomethane and 1.4g of disodium edetate, add 250ml of water to dissolve, adjust the pH value to 8.4 with hydrochloric acid, and dilute to 500ml with water.

[0038] 1.3 Antithrombin (ATIII) solution

[0039] Make a solution containing 1 IU per 1 ml of water.

[0040] 1.4 Chromogenic substrate S-2765 (or other FXa-specific chromogenic substrate) solution

[0041] Make a 0.003mol / L solution with water, and dilute to 0.0005mol / L with Tris-EDTA disodium buffer solution (PH8.4) before use.

[0042] 1.5 Factor Xa (FXa) solution...

Embodiment 2

[0061] The concentrations of the standards and other reagents in the assay can be varied as long as the absorbance at several points is linearly related to the logarithm of the concentration. Use the first sample with a concentration of 0.2IU / ml to draw a standard curve (the addition of FXa is 60ul and the dilution ratio is 0.8).

[0062] The experimental data are as follows:

[0063]

[0064] Draw a logarithmic curve with this data, such as figure 2 , the standard curve regression equation is: y=-0.9717x-0.5464, the linear correlation coefficient R 2 =0.9998 (x is logarithm of concentration, y is absorbance).

[0065] Take the 0.2IU / ml sample as the sample to be tested, measure the absorbance after the reaction, and calculate the actual concentration by using the concentration logarithm-absorbance standard curve. Three parallel experiments were done at the same concentration (0.2IU / ml), and the results were as follows:

[0066] Absorbance 0.134 0.132 0.13...

Embodiment 3

[0069] The potency determination of the sample to be tested is heparin sodium: take the sample to be tested and estimate the potency, dilute and detect according to the low molecular weight heparin sodium assay method in Example 1, as long as the absorbance is within the linear range, the obtained linear equation can be used to calculate the potency. As the dilution ratio 0.8 times of embodiment 1 and embodiment 2 becomes 0.5 times, the results can be obtained as follows:

[0070]

[0071] Draw a logarithmic curve with this data, such as image 3 , the standard curve regression equation is: y=-0.7236x-0.3544, the linear correlation coefficient R 2 =0.9822 (x is the logarithm of the concentration, y is the absorbance).

[0072] That is to say, increasing the dilution ratio can make the standard curve cover a larger range and further save reagents.

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Abstract

The invention relates to a detection method for detecting the anti-Xa potency of heparin (salt) or heparin (salt) with the low molecular weight through a coagulometer. The method comprises the following steps of 1 standard serial solution preparing; 2 standard curve drawing; 3 sample processing and detecting; 4 calculating. According to the method, four kinds of standard solution concentrations are achieved through automatic diluting performed with the coagulometer, personal errors are few, all the operation steps are automatically completed, the man-made influences of the time, the temperature and stirring are completely eradicated, and the experiment precision is high; reaction of the used reagents is changed from semimicro reaction to micro reaction, and therefore the expensive reagents are saved; a linear equation method is adopted and is intuitional.

Description

technical field [0001] The present invention relates to a detection method of heparin (salt) or low molecular weight heparin (salt) potency, in particular to a detection method for measuring heparin (salt) or low molecular weight heparin (salt) anti-Xa potency with a hemagglutination meter. Background technique [0002] Heparin (salt) or low molecular weight heparin (salt) is an anticoagulant and antithrombotic drug. Heparin (salt) or low molecular weight heparin (salt) can isolate antithrombotic activity, that is, anti-Xa potency. Xa activity in human plasma is closely related to thrombosis, that is to say, high factor Xa activity will lead to clinical thrombosis, so heparin (salt) or low molecular weight heparin (salt) anti-Xa is often used clinically. potency activity. [0003] Now both Chinese Pharmacopoeia and European Pharmacopoeia adopt the chromogenic substrate method to detect the anti-Xa potency of heparin (salt) or low molecular weight heparin (salt). This metho...

Claims

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Application Information

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IPC IPC(8): G01N21/31G01N1/28
Inventor 郑会武丛义国刘清凉
Owner 山东万邦赛诺康生化制药股份有限公司
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