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Method for preparing heparin sodium by combining enzymolysis and salt decomposition

A technology of joint preparation and heparin sodium, which is applied in the field of biochemical industry, can solve the problems such as the inability to release heparin sodium, achieve the effect of solving the problem of poisoning and high product purity

Inactive Publication Date: 2012-08-22
叶青理
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0011] The technical problem to be solved by the present invention is to provide a method for the joint preparation of heparin sodium by salt hydrolysis and enzymatic hydrolysis, which can solve the problem that simple salt hydrolysis and simple enzymatic hydrolysis cannot completely release heparin sodium from intestinal mucosal cells , using the combined process of salt hydrolysis and enzymatic hydrolysis to completely dissociate heparin from intestinal mucosal cells, thereby completely extracting

Method used

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  • Method for preparing heparin sodium by combining enzymolysis and salt decomposition
  • Method for preparing heparin sodium by combining enzymolysis and salt decomposition

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Experimental program
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Embodiment 1

[0027] The specific steps of the present invention are as follows: before the test, the used utensils, reagents and consumables are autoclaved. Specific steps are as follows:

[0028] The purpose of the present invention is to solve the problem that simple salt hydrolysis and simple enzymatic hydrolysis cannot completely release heparin sodium from intestinal mucosal cells, that is, the combined process of salt hydrolysis and enzymatic hydrolysis can completely dissociate heparin from intestinal mucosal cells, thereby completely Extraction problem. And found more objective production parameters for industrial scale production.

[0029] The present invention realizes by following scheme:

[0030] 1) Dissociation: According to the weight ratio of water: pig small intestine mucosa is 4~5: 1, add water to the pig small intestine mucosa, and add salt to make the liquid salinity 5 degrees, add NaOH to adjust the pH value to 9, stir and heat up to 55°C, after 30 minutes of heat pr...

Embodiment 2

[0042] Pump out 400 pairs of pig small intestinal mucosa and water to account for 90% of the space in the reaction tank of 2000 liters, add 120 kilograms of salt, add alkali to adjust the pH value to 9, be warming up to 55 ℃, after insulated for 30 minutes, add alkaline protease ( 2709 type) 1000 grams, keep warm for 2 hours, then continue to heat up to 85°C, filter and cool to about 55°C.

[0043] (2) Add 8000 grams of basic resin Type 98 to the liquid at 55° C., stir and absorb for 8 hours, and filter to wash the resin.

[0044] (3) Add the same amount of 15°C saline as the resin, and perform elution twice, each time for 1 hour, and mix the above-mentioned elution water together.

[0045] (4) Add industrial alcohol to the upper eluent to make the alcohol concentration 45 degrees, let it stand for 24 hours, filter the lower sediment with polyester cloth, and dry it in a hot water bath at 65-75 ° C to obtain a purity of 90 The above activity units / mg heparin sodium product.

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Abstract

The invention relates to a method for preparing heparin sodium by combining enzymolysis and salt decomposition, comprises the following steps: 1) dissociation: according to the weight ratio of 4-5:1 between water and chatterling mucosa, adding the water into the chatterling mucosa, and adding salt to adjust the solution salinity to be 5; adding alkali to adjust the pH value to be 9, and stirring and heating to 55 DEG C; carrying out heat preservation for 30min, and then adding alkali protease 2709 into a reaction tank according to the weight ratio of 2000-3000:1 between the chatterling mucosaand the alkali protease 2709; and carrying out heat preservation for 2h, heating to 85 DEG C and filtering; and 2) absorbing, eluting, precipitating, filtering and drying filtrate obtained in the step 1), and finally obtaining the heparin sodium. By adopting the technique combining enzymolysis and salt decomposition, the method leads heparin to be thoroughly dissociated from the chatterling mucosa, thus completing full extraction. On the basis of the prior art, the method increases about one third of unit production value and controls the increased cost not more than 1% of the increased production value.

Description

technical field [0001] The invention belongs to the field of biochemical industry, and in particular relates to a method for preparing heparin sodium through salt hydrolysis and enzymatic hydrolysis. Background technique [0002] Heparin sodium is the sodium salt form of heparin. Heparin is widely present in animal body fluids and tissues. In animals, heparin generally exists in combination with proteins to form protein-polysaccharide complexes. The vast majority of heparin used in the market is extracted from the small intestinal mucosa and lungs of pigs and cattle. [0003] The structural formula of heparin is generally considered as follows: [0004] [0005] We have seen that heparin is a kind of heparin compound composed of multiple tetrasaccharide units. The molecular weight of highly active heparin extracted from pig intestinal mucosa and lung is generally between 3000-30000. [0006] Heparin has a wide range of uses, mainly for anticoagulant, antithrombotic, a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08B37/10C12S3/02
Inventor 叶青理
Owner 叶青理
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