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Method for fast separating fast-moving heparin and dermatan sulfate

A dermatan sulfate, fast-moving technology, applied in the field of separation of mucopolysaccharide components in heparin anticoagulant drugs, can solve problems such as long electrophoresis time and band dispersion, and achieve the effect of reducing electrophoresis voltage and prolonging electrophoresis time.

Inactive Publication Date: 2016-04-06
DONGYING TIANDONG PHARM CO LTD
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Problems solved by technology

[0007] The present invention aims at the problems of long electrophoresis time, dispersed bands and two electrophoresis in existing agarose gel electrophoresis separation of mucopolysaccharides, and provides a method for rapidly separating fast-moving heparin and dermatan sulfate

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  • Method for fast separating fast-moving heparin and dermatan sulfate
  • Method for fast separating fast-moving heparin and dermatan sulfate

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[0050] Preparation of agarose gel: The specification of the electrophoresis gel is 100×70, the concentration is 0.6%, the buffer solution is 0.04M barium acetate (PH5.8), the thickness of the electrophoresis gel is 3mm, the agarose is melted in the buffer solution and cooled to 60 At about ℃, pour it into the glue tank and wait for it to solidify before use;

[0051] Electrophoresis: put the solidified gel in the electrophoresis tank, use 0.04M barium acetate (pH5.8) as the buffer, load 10 micrograms of sample on each gel well, and slowly add a small amount to the top of the buffer after loading the sample Petroleum ether, put the whole set of electrophoresis equipment in a 4°C cold room, 120V, electrophoresis for 2 hours;

[0052] Staining and decolorization: After electrophoresis, take out the gel plate, soak it in Cetavlon solution for 3 hours, cover it with dry filter paper after taking it out, dehydrate it several times with careful pressure; use toluidine blue solution, ...

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Abstract

The invention discloses a method for fast separating fast-moving heparin and dermatan sulfate. The method includes the steps that agarose gel is prepared with a barium acetate solution with the concentration of 0.04 M as a buffering solution, sampling is performed on each sample application hole, the gel is placed in the 0.04 M barium acetate buffering solution, a small amount of petroleum ether is added into the upper layer of the buffering liquid so that a gas-liquid interface can be sealed; a whole electrophoresis tank system is placed in a cold chamber for electrophoresis, and slight pressing and dewatering are performed after electrophoresis ends; 0.2% of toluidine blue is dissolved in a solvent formed by ethanol, water and acetic acid, a staining solution is prepared, gel is suspended in the staining solution, and then washing is performed with a solution formed by mixing ethanol, water and acetic acid. According to the method, due to the fact that one time of agarose gel electrophoresis is performed in the 0.04 M barium acetate buffering solution, cooling is performed through electrophoresis liquid, electrophoresis voltage is reduced, electrophoresis time is prolonged, and the gas-liquid interface is sealed through petroleum ether, and the fast-moving heparin and dermatan sulfate can be effectively separated.

Description

technical field [0001] The invention belongs to the field of separation of mucopolysaccharide components in heparin anticoagulant drugs in the biomedicine industry, and in particular relates to a method for rapidly separating fast-moving heparin and dermatan sulfate. Background technique [0002] Mucopolysaccharide compounds are an important class of biological macromolecules, which have a variety of heavy [0003] The important biological functions and pharmacological activities include: anticoagulant, antitumor, hypolipidemic, antithrombotic, etc. The representative types of mucopolysaccharides include heparin, heparan sulfate, dermatan sulfate, chondroitin sulfate, etc., each of which has different biopharmacological effects. [0004] The structure of mucopolysaccharides is similar, and they are all sulfated and acetylated sugar amino-uronic acid disaccharide units in different degrees. [0005] Repeated long-chain structure, each mucopolysaccharide is composed of molec...

Claims

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Application Information

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IPC IPC(8): C08B37/10C08B37/08
CPCC08B37/0069C08B37/0075
Inventor 张在忠郭林李荣陈少鹏李玮涛
Owner DONGYING TIANDONG PHARM CO LTD
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