Technology for normal-temperature salt-free extraction of heparin sodium

An extraction process, the technology of heparin sodium, which is applied in the field of extracting heparin sodium, can solve the problems of difficult and harmless treatment of high-salt wastewater, damage to the use value of water bodies, and increase of production costs, so as to reduce wastewater treatment costs, high production costs, and save The effect of heat

CN103724456AInactive Publication Date: 2014-04-16SHENZHEN LEVEKING BIOLOGY ENG
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Publication Date
2014-04-16
Estimated Expiration
Not applicable · inactive patent
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Abstract

The invention discloses a technology for normal-temperature salt-free extraction of heparin sodium. The technology comprises the following steps of breaking intestinal mucosa, carrying out enzymolysis by a protease-lipase-nuclease compound enzyme preparation under the conditions of a normal temperature and no salt, then carrying out clear liquid centrifugal collection, adsorbing heparin sodium in the centrifugate by appropriate ion exchange resin, carrying out elution on the filtrate by high-concentration salt, precipitating the heparin sodium in the eluent by alcohol and carrying out drying to obtain a heparin sodium crude product. The technology can save a large amount of heat energy in enzymolysis and adsorption, reduce energy consumption and reduce a production cost. The technology realizes heparin sodium enzymolysis and adsorption under the condition of no salt so that the salinity of the discharged waste water is reduced, a waste water treatment cost is reduced and environmental benefits are improved.
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Description

technical field

[0001] The invention relates to the technical field of biopharmaceutical production, in particular to a process for extracting heparin sodium from pig small intestine under normal temperature and salt-free conditions. Background technique

[0002] Heparin sodium is an acidic mucopolysaccharide produced and secreted by mast cells of animal connective tissue. Medical heparin sodium is mainly used for anticoagulation. The crude extract of heparin sodium cannot be directly used as a clinical drug because it contains more impurities and the sample is not pure. Since the quality of the crude heparin sodium determines the quality of the high-quality heparin sodium to a certain extent, the preparation of the crude heparin sodium is very important. The traditional extraction methods of crude heparin sodium are as follows:

[0003] 1. Extraction by salting-out method: the earliest extraction of heparin sodium from pig small intestine was mainly by salting-out extrac...

Examples

Embodiment 1

[0022] 1) Homogenate: Dilute the fresh small intestinal mucosa with 5-6 kg of water for each small intestine, then homogenate or grind to obtain the homogenate of the small intestinal mucosa;

[0023] 2) Enzymolysis: get 200 liters of homogenate, adjust the pH of the homogenate to 8-9, then add 100 grams of compound enzyme preparation, and stir for 4 hours at room temperature; the compound enzyme preparation is protease: lipase: nucleic acid A mixture of enzymes mixed in a mass ratio of 10:1.5:0.4;

[0024] 3) Impurity removal: adopt continuous centrifugation or pressure filtration to discard solid impurities to obtain centrate;

[0025] 4) Adsorption: adjust the pH of the centrifuge to about 8.5, add 6g / L of FPA98 resin (based on the volume of the centrifuge, w / v), and collect the resin after stirring and adsorbing at 28°C to 30°C for 6 hours ;

[0026] 5) Cleaning and elution: wash the resin with water, and then use 5% brine to wash away some impurities. After cleaning, us...

Embodiment 2

[0029] 1) Homogenate: Dilute the fresh small intestinal mucosa with 5-6 kg of water for each small intestine, then homogenate or grind to obtain the homogenate of the small intestinal mucosa;

[0030] 2) Enzymolysis: get 400 liters of homogenate, adjust the pH of the homogenate to 8-9, then add 280 grams of compound enzyme preparation, and stir for 3 hours at room temperature; the compound enzyme preparation is protease: lipase: nucleic acid A mixture of enzymes mixed in a mass ratio of 10:1.7:0.5;

[0031] 3) Impurity removal: adopt continuous centrifugation or pressure filtration to discard solid impurities to obtain centrate;

[0032] 4) Adsorption: adjust the pH of the centrifuge to about 8.5, add 6g / L of FPA98 resin (based on the volume of the centrifuge, w / v), and collect the resin after stirring and adsorbing at 28°C to 30°C for 6 hours ;

[0033] 5) Cleaning and elution: wash the resin with water, and then use 5% brine to wash away some impurities. After cleaning, us...

Embodiment 3

[0036] 1) Homogenate: Dilute the fresh small intestinal mucosa with 5-6 kg of water for each small intestine, then homogenate or grind to obtain the homogenate of the small intestinal mucosa;

[0037] 2) Enzymolysis: Get 600 liters of homogenate, adjust the pH of the homogenate to 8-9, then add 600 grams of compound enzyme preparation, and stir for 2 hours at room temperature; the compound enzyme preparation is protease: nuclease: fat A mixture of enzymes mixed in a mass ratio of 10:2:0.6;

[0038] 3) Impurity removal: adopt continuous centrifugation or pressure filtration to discard solid impurities to obtain centrate;

[0039] 4) Adsorption: adjust the pH of the centrifuge to about 8.5, add 6g / L of FPA98 resin (based on the volume of the centrifuge, w / v), and collect the resin after stirring and adsorbing at 28°C to 30°C for 6 hours ;

[0040]5) Cleaning and elution: wash the resin with water, and then use 5% brine to wash away some impurities. After cleaning, use 23% to 2...