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A technology of heparin sodium and pig small intestine, applied in the field of biochemical industry, can solve the problem of not being able to release heparin sodium, and achieve the effect of solving the problem of poisoning and high product purity
Inactive Publication Date: 2019-11-12
山阳县恒瑞肉制品有限公司
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[0011] The technical problem to be solved by the present invention is to provide a method for preparing heparin sodium, which can solve the problem that simple salt hydrolysis and simple enzymatic hydrolysis cannot completely release heparin sodium from intestinal mucosal cells. The combined process of dissociation completely dissociates heparin from intestinal mucosal cells, thereby completely extracting
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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] (1) Pump 400 pairs of pig small intestinal mucosa and water to occupy 90% of the space in a 2000-liter reaction tank, add 120 kg of salt, add alkali to adjust the pH value to 9, raise the temperature to 55 ° C, keep warm for 30 minutes, and then add alkali Protease (2709 type) 1000 grams, heat preservation 2 hours, then continue to heat up to 85 ℃, filter and cool to about 55 ℃.
[0043] (2) Add 8,000 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 twice as much saline at 15°C as the resin, and elute twice, each time for 1 hour, and mix the above eluents 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 more than 90 Activity units / mg heparin sodium product.
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Abstract
The invention relates to a method used for preparing heparin sodium. The method comprises following steps: 1, dissociation is carried out, wherein water is added into pig small intestinal mucosa at awater / pig small intestinal mucosa weight ratio of 4-5:1, a salt is added to adjust solution salinity to be 5 DEG, an alkali is added to adjust pH value to 9, heating to 55 DEG C is carried out with stirring, after 30min of thermal insulation treatment, alkaline protease is introduced into a reaction tank at a weight ratio of pig small intestinal mucosa : alkaline protease 2709 of 2000-3000:1, thermal insulation treatment is carried out for 2h, the temperature is increased to 85 DEG C, and filtering is carried out; and 2, a filtrate obtained in step 1 is subjected to adsorption, elution, deposition filtering, and drying to obtain heparin sodium. According to the method, salt hydrolysis and enzymatic hydrolysis combined technology is adopted to realize complete dissociation of heparin from intestine mucous membrane cells, so that complete extraction is realized, based on the prior art, unit yield is increased by about 1 / 3, and cost is increased by less than 1% of the increased output 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. 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, anti-cell aging, anti-tumor metastasis, anti-virus...
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