CN103804526A - Method for purifying crude product of heparin sodium - Google Patents
Method for purifying crude product of heparin sodium Download PDFInfo
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- CN103804526A CN103804526A CN201310631229.0A CN201310631229A CN103804526A CN 103804526 A CN103804526 A CN 103804526A CN 201310631229 A CN201310631229 A CN 201310631229A CN 103804526 A CN103804526 A CN 103804526A
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- heparin sodium
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Abstract
The invention discloses a method for purifying a crude product of heparin sodium. The method adopts the small intestinal mucosa of a healthy pig as a main raw material and obtains a crude product of heparin sodium by the steps of dissociation (enzymolysis), filtration, protein enzymolysis, protein removal and precipitation, dehydration and drying. The method has the advantages of short production cycle, low production cost and high yield, does not generate toxic substances, and is good for environmental protection.
Description
Technical field
The present invention relates to biological technical field, relate in particular to the extracting method of heparin sodium, is a kind of take pig intestinal mucosa as raw material, by biological enzymolysis technology, thus the method for purification heparin sodium.
Background technology
Heparin sodium is mucopolysaccharide sulfuric acid ester anticoagulant, it is the sodium salt of the CSSO3 extracted in the intestinal mucosa by pig or ox, belong to mucopolysaccharide material, because it has strong anticoagulation, it is the first medicine of the thrombotic diseases such as control deep-vein thrombosis formation, along with deepening continuously of research, it is found that heparin sodium not only has anti-freezing, antithrombotic to form and adjust the effect of blood fat, also have anti-inflammatory, antianaphylaxis, the various biological function such as antiviral, anticancer.
At present, in general the extraction of heparin sodium crude is all that animal small intestine is carried out to scraping, and the mucus obtaining after scraping is first carried out enzymolysis, filter, then adsorb wash-out with heparin sodium is resin dedicated, then through alcohol precipitation, after dehydration, gains are being dried.It is low that this traditional technology is being in yield, fluctuating yield, and loss is large, and discharging of waste liquid concentration is high, with serious pollution defect.
Summary of the invention
For the deficiency existing in existing production technique, the object of the present invention is to provide a kind of method of purification of heparin sodium crude, be a kind of technique of simplifying, while improving yield, energy-saving, be the novel method that object is developed, and this method be conducive to suitability for industrialized production.
Technical solution of the present invention is that this preparation method comprises the following steps:
1, (enzymolysis) dissociates: by mucous membrane of small intestine liquid (solid-to-liquid ratio is about 1:9) the suction retort of the health pig taking, adjust PH9.0-10.0 by industrial soda, add the sodium-chlor of 3%-5% (weight ratio), add organized enzyme by 0.5% of raw material weight, after mixing, be warming up to 45-55 ℃, constant temperature stirs, the 1-3 hour that dissociates, dissociation process keeps PH constant, is warming up to 83-91 ℃, insulation 10-15 minute, then cooling immediately.
2, filter: cooling fluid suction filtration machine is filtered 2 times, successively utilize the millipore filtration of 2 μ m and 0.22 μ m to filter, to remove impurity, obtain filtered liquid.
3, enzymolysis protein matter: filtrate is regulated to PH7.5-9.5, then be warming up to 40-55 ℃, add organized enzyme by 0.5% of raw material weight, keep 40-55 ℃ of temperature to stir 4-7 hour, in enzymolysis process, control the pH value of reaction system, then be warming up to 90-95 ℃ by anxious enzymolysis solution, maintain 10-15 minute, then cooling immediately.
4, except albumen: the heparin sodium in filtrate filtered with isolating protein and concentrated with micro-filtration membrane concentration device or distillation or resin, obtaining concentrated solution.
5, precipitation dehydrates: the isopyknic ethanol that is 85-90% by concentration joins in above-mentioned concentrated solution, the postprecipitation 10-12 hour that stirs, then removes supernatant liquor, collecting precipitation, add again isopyknic dehydrated alcohol dehydration, after dehydration, be drying to obtain heparin sodium crude.
Advantage of the present invention is: in the present invention for regulating the bronsted lowry acids and bases bronsted lowry of pH value to be respectively hydrochloric acid and sodium hydrate solid.
Compared with prior art, advantage of the present invention is: (1), rejected the troublesome operation that uses resin absorption in traditional technology, technique is simple.(2), adopt filtering with microporous membrane, can effectively remove the impurity in heparin sodium.
Embodiment
Embodiment 1
(1) (enzymolysis) dissociates: in mucous membrane of small intestine liquid 1kg (solid-to-liquid ratio is about 1:9) the suction retort of the health pig taking, adjust PH9.5 by industrial soda, add the sodium-chlor of 40g, add organized enzyme 5g, after mixing, be warming up to 48 ℃, constant temperature stirs, dissociate 2 hours, dissociation process keeps PH constant, is warming up to 85 ℃, be incubated 12 minutes, then cooling immediately.
(2) filter: cooling fluid suction filtration machine is filtered 2 times, successively utilize the millipore filtration of 2 μ m and 0.22 μ m to filter, to remove impurity, obtain filtered liquid.
(3) enzymolysis protein matter: filtrate is regulated to PH8.0, be then warming up to 48 ℃, add organized enzyme 5g, keep 48 ℃ of temperature to stir 6 hours, in enzymolysis process, control the pH value of reaction system, be then warming up to 92 ℃ by anxious enzymolysis solution, maintain 12 minutes, then cooling immediately.
(4) except albumen: the heparin sodium in filtrate filtered with isolating protein and concentrated with micro-filtration membrane concentration device or distillation or resin, obtaining concentrated solution.
(5) precipitation dehydrates: the isopyknic ethanol that is 90% by concentration joins in above-mentioned concentrated solution, stir postprecipitation 12 hours, then removes supernatant liquor, collecting precipitation, add again isopyknic dehydrated alcohol dehydration, after dehydration, be drying to obtain heparin sodium crude 9g.
Claims (2)
1. a method of purification for heparin sodium crude, is characterized in that comprising the following steps:
(1) (enzymolysis) dissociates: by mucous membrane of small intestine liquid (solid-to-liquid ratio is about 1:9) the suction retort of the health pig taking, adjust PH9.0-10.0 by industrial soda, add the sodium-chlor of 3%-5% (weight ratio), add organized enzyme by 0.5% of raw material weight, after mixing, be warming up to 45-55 ℃, constant temperature stirs, the 1-3 hour that dissociates, dissociation process keeps PH constant, is warming up to 83-91 ℃, insulation 10-15 minute, then cooling immediately.
(2) filter: cooling fluid suction filtration machine is filtered 2 times, successively utilize the millipore filtration of 2 μ m and 0.22 μ m to filter, to remove impurity, obtain filtered liquid.
(3) enzymolysis protein matter: filtrate is regulated to PH7.5-9.5, then be warming up to 40-55 ℃, add organized enzyme by 0.5% of raw material weight, keep 40-55 ℃ of temperature to stir 4-7 hour, in enzymolysis process, control the pH value of reaction system, then be warming up to 90-95 ℃ by anxious enzymolysis solution, maintain 10-15 minute, then cooling immediately.
(4) except albumen: the heparin sodium in filtrate filtered with isolating protein and concentrated with micro-filtration membrane concentration device or distillation or resin, obtaining concentrated solution.
(5) precipitation dehydrates: the isopyknic ethanol that is 85%-90% by concentration joins in above-mentioned concentrated solution, the postprecipitation 10-12 hour that stirs, then removes supernatant liquor, collecting precipitation, add again isopyknic dehydrated alcohol dehydration, after dehydration, be drying to obtain heparin sodium crude.
2. method according to claim 1, is characterized in that: in (1) step, add the organized enzyme of raw material weight 0.5%, to strengthen the dissociation effect of heparin sodium.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201310631229.0A CN103804526B (en) | 2013-11-26 | 2013-11-26 | A kind of method of purification of heparin sodium crude |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201310631229.0A CN103804526B (en) | 2013-11-26 | 2013-11-26 | A kind of method of purification of heparin sodium crude |
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| CN103804526A true CN103804526A (en) | 2014-05-21 |
| CN103804526B CN103804526B (en) | 2016-08-17 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105884935A (en) * | 2014-10-17 | 2016-08-24 | 北京海吉星医疗科技有限公司 | Method for purifying heparin sodium |
| CN111560087A (en) * | 2020-06-28 | 2020-08-21 | 揭阳市润达肠衣有限公司 | Purification method of high-quality heparin sodium |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109467620A (en) * | 2017-09-08 | 2019-03-15 | 山阳县恒瑞肉制品有限公司 | A kind of method of one step of enzyme process combination membrane technology preparation refined heparin sodium |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3451996A (en) * | 1968-02-12 | 1969-06-24 | Thompson Farms Co | Method for the preparation of heparin |
| CN1837244A (en) * | 2005-03-22 | 2006-09-27 | 孙剑鸣 | Process for the production of heparin sodium crude |
| CN101544999A (en) * | 2009-04-10 | 2009-09-30 | 湖北五瑞生物工程有限公司 | Method for producing and purifying high purity and low molecular weight sodium heparin |
| CN102993336A (en) * | 2011-09-14 | 2013-03-27 | 浦江亚太肠衣有限公司 | Crude heparin sodium purification technology |
-
2013
- 2013-11-26 CN CN201310631229.0A patent/CN103804526B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3451996A (en) * | 1968-02-12 | 1969-06-24 | Thompson Farms Co | Method for the preparation of heparin |
| CN1837244A (en) * | 2005-03-22 | 2006-09-27 | 孙剑鸣 | Process for the production of heparin sodium crude |
| CN101544999A (en) * | 2009-04-10 | 2009-09-30 | 湖北五瑞生物工程有限公司 | Method for producing and purifying high purity and low molecular weight sodium heparin |
| CN102993336A (en) * | 2011-09-14 | 2013-03-27 | 浦江亚太肠衣有限公司 | Crude heparin sodium purification technology |
Non-Patent Citations (1)
| Title |
|---|
| 徐俊涛: "酶解法制备肝素钠关键工艺技术研究", 《中国优秀硕士学位论文全文数据库 工程科技Ⅰ辑》, no. 4, 15 April 2013 (2013-04-15) * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105884935A (en) * | 2014-10-17 | 2016-08-24 | 北京海吉星医疗科技有限公司 | Method for purifying heparin sodium |
| CN111560087A (en) * | 2020-06-28 | 2020-08-21 | 揭阳市润达肠衣有限公司 | Purification method of high-quality heparin sodium |
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| Publication number | Publication date |
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| CN103804526B (en) | 2016-08-17 |
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Address after: 266100 Zhuzhou Road, Laoshan District, Shandong, No. 97, No. Patentee after: Qingdao Jiulong biological medicine group Co., Ltd. Address before: 266100 Zhuzhou Road, Laoshan District, Shandong, No. 97, No. Patentee before: Qingdao Jiulong Bio-Pharmaceutical Co., Ltd. |
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