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Method for detecting plasmin titer by bubble rising process

A plasmin and titer technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of irregular shape of the dissolved circle, affecting plasmin, brittleness, etc. Overcome the effect of long reaction time and easy control

Active Publication Date: 2015-04-01
BEIJING SAISHENG PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] 1. The measured value is easily affected by factors such as plate uniformity and constant temperature incubation time
[0007] 2. The plate making operation is troublesome, and the thickness and composition of the plate are not easy to be uniform
[0009] 4. The reaction time is long, which is not convenient for production process control
[0010] 5. The shape of the melting ring is sometimes irregular, and the error of detecting the diameter of the melting ring is large
[0012] 1. After the hydrolysis reaction of this method, the remaining fibrin is washed many times. During the washing process, the glue block is easily broken and lost, which affects the test results.
[0013] 2. The operation is cumbersome and time-consuming, which is not convenient for production process control
[0014] 3. Fibrinogen is derived from animal blood, with complex components and uncertain impurities. Impurities can inhibit the titer of plasmin to varying degrees, and even different batches of fibrinogen will affect the titer of plasmin.

Method used

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  • Method for detecting plasmin titer by bubble rising process
  • Method for detecting plasmin titer by bubble rising process
  • Method for detecting plasmin titer by bubble rising process

Examples

Experimental program
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Embodiment 1

[0030] The test materials used in the present invention, the plasmin standard product is self-made, and all the other materials are commercially available products unless otherwise specified.

[0031] Preparation of reagents:

[0032] Tris buffer solution with pH 7.6: Take 6.05 grams of Tris, add 500 ml of water to dissolve, add 360 ml of 0.1 mol / L hydrochloric acid solution, mix well, adjust the pH to 7.6, add water to 1000 ml.

[0033] Thrombin solution with 2.5 BP units / ml: take thrombin, add the above-mentioned tris buffer solution with pH 7.6 to dissolve and dilute to a solution containing 2.5 BP units per 1 ml. .

[0034] 2.5 mg / ml fibrinogen solution: take fibrinogen, add the above-mentioned tris buffer with pH 7.6 to dissolve and dilute to a solution containing about 2.5 mg of coagulable protein per 1 ml.

[0035] Preparation of the standard solution: take the plasmin standard, add the above-mentioned tris buffer solution of pH 7.6 and dilute it into solutions of 50...

Embodiment 2

[0039] The test materials used in the present invention, the plasmin standard product is self-made, and all the other materials are commercially available products unless otherwise specified.

[0040] Preparation of reagents:

[0041] Tris buffer solution with pH 7.6: Take 6.05 grams of Tris, add 500 ml of water to dissolve, add 360 ml of 0.1 mol / L hydrochloric acid solution, mix well, adjust the pH to 7.6, add water to 1000 ml.

[0042] Thrombin solution with 3BP units / ml: take thrombin, add the above-mentioned tris buffer solution of pH 7.6 to dissolve and dilute to a solution containing 3BP units per 1 ml. .

[0043] 3 mg / ml fibrinogen solution: take fibrinogen, add the above-mentioned tris buffer with pH 7.6 to dissolve and dilute to a solution containing about 3 mg of coagulable protein per 1 ml.

[0044] Preparation of the standard solution: take the plasmin standard, add the above-mentioned tris buffer solution of pH 7.6 and dilute it into solutions of 50, 100, 150, ...

Embodiment 3

[0048] The test materials used in the present invention, the plasmin standard product is self-made, and all the other materials are commercially available products unless otherwise specified.

[0049] Preparation of reagents:

[0050] Tris buffer solution with pH 7.8: take an appropriate amount of Tris, add 500 ml of water to dissolve, add an appropriate amount of 0.1 mol / L hydrochloric acid solution, mix well, adjust the pH to 7.8, add water to 1000 ml.

[0051] Thrombin solution with 8BP units / ml: Take thrombin, add the above-mentioned tris buffer solution of pH 7.8 to dissolve and dilute to a solution containing 8BP units per 1 ml. .

[0052] 8 mg / ml fibrinogen solution: take fibrinogen, add the above-mentioned tris buffer with pH 7.8 to dissolve and dilute to a solution containing about 8 mg of coagulable protein per 1 ml.

[0053] Preparation of the standard solution: take the plasmin standard, add the above-mentioned tris buffer solution of pH 7.8 and dilute it into so...

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Abstract

The invention relates to the field of pharmaceutical technology and particularly relates to a method for detecting plasmin titer by bubble rising process. The method includes using tris(hydroxymethyl)aminomethane buffered solution with pH 6-9 as solvent, and using 1-10BP / ml thrombin solution and 1-10mg / ml fibrinogen solution to prepare substrate. Standard plasmin solution is made by diluting tris(hydroxymethyl)aminomethane buffered solution with pH 6-9 into the standard solution containing 10-1000 units per milliliter, and standard regression curves are prepared by the method. A sample is made by diluting tris(hydroxymethyl)aminomethane buffered solution with pH 6-9 into sample solution containing about 10-1000 units per milliliter, estimation is performed by the same method, and titer of the sample is calculated according to the standard curves. By the method, interference of different sources of fibrinogens on estimation of plasmin titer is eliminated, inspecting operation is simple and fast, estimation results are accurate and reliable, key techniques of plasmin quality control are achieved, and accordingly quality control in plasmin industrialization process is accurate, stable and feasible.

Description

technical field [0001] The invention relates to the technical field of pharmacy, in particular to a method for detecting the titer of plasmin by a bubble rising method. Background technique [0002] Plasmin proteolytic enzyme is used for peripheral vascular diseases such as cerebral infarction, hypercoagulation state and thromboangiitis. This product acts on fibrinogen and fibrin (thrombosis precursor protein), degrading it into small molecular soluble fragments, which are easy to decompose and clear from the blood circulation, thereby producing defibrinogen effect; this product promotes the activation of tissue plasminogen The substance (t-PA) is released by endothelial cells and enhances its activity, so it has antithrombotic function; this product can reduce platelet aggregation and blood viscosity; this product also has the function of reducing myocardial oxygen consumption and improving microcirculation. This product is intravenously injected into the human body, and t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/34
Inventor 彭兴华马骉吴丹蒲丽萍
Owner BEIJING SAISHENG PHARMA
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