CN108456262A - A kind of preparation process of high purity heparin sodium - Google Patents
A kind of preparation process of high purity heparin sodium Download PDFInfo
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- CN108456262A CN108456262A CN201810202915.9A CN201810202915A CN108456262A CN 108456262 A CN108456262 A CN 108456262A CN 201810202915 A CN201810202915 A CN 201810202915A CN 108456262 A CN108456262 A CN 108456262A
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- Prior art keywords
- heparin sodium
- oxidation
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- hydrogen peroxide
- sodium
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
- C08B37/0063—Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
- C08B37/0075—Heparin; Heparan sulfate; Derivatives thereof, e.g. heparosan; Purification or extraction methods thereof
- C08B37/0081—Reaction with amino acids, peptides, or proteins
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
- C08B37/0063—Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
- C08B37/0075—Heparin; Heparan sulfate; Derivatives thereof, e.g. heparosan; Purification or extraction methods thereof
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- General Health & Medical Sciences (AREA)
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- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
Abstract
The present invention provides a kind of preparation processes of high purity heparin sodium, mainly be dehydrated and be dried in vacuo including the sodium chloride salt solution of 3.0% (w/v), the trypsin digestion of 0.1%~0.2% (w/v), hydrogen peroxide ozone efficient oxidation, absolute ethyl alcohol precipitation, acetone and etc., wherein hydrogen peroxide ozone efficient oxidation system can generate HO2 ‑, it is the inducing agent that free radical OH is generated, free radical OH will cause chain reaction once generating, accelerate oxidation rate, improve oxidation effectiveness.It is compared, has been saved at least 40% time with traditional multistage hydrogen peroxide oxidation using the technique purification heparin sodium described in this programme, potency improves 9U/mg, and the potency rate of recovery improves 8%, and heparin sodium purity obtained is high, and color and luster is good.
Description
Technical field
The present invention relates to heparin sodium preparation fields, and in particular to a kind of preparation process of high purity heparin sodium.
Background technology
Heparin sodium is the practical widest anticoagulation medicine of clinical medicine domain, it is by D-Glucose amine, N- acetyl Portugal
The mucopolysaccharide sulfuric ester that grapes glucosamine, D- glucuronic acids and L- iduronic acids alternately form, is primarily present in pluck
Mast cell granule in, mostly combined with protein and existed with Protein-Heparin complexes, mainly in mucous membrane of small intestine and lungs
Content highest.Heparin sodium is mainly used for blood coagulation resisting function, is one of the biochemical drug of currently the most important ones anticoagulation and antithrombotic,
But heparin sodium be raw material prepare medicament often through injection by way of be administered, so heparin sodium must have it is very high-purity
Degree just can guarantee medical safety.
The heparin sodium for preparing high-purity at present is purified on the basis of crude heparin sodium, and the prior art is usually adopted
The protein and nucleic acid in crude product are removed with hydrogen peroxide secondary oxidation, process temperature is up to 95 degrees Celsius, and PH is 1.5~2.0
Between, under conditions of such highly acidity and high-temperature, heparin sodium is easy inactivation, and titer of heparin sodium is caused to be lost, and aoxidizes
Time is up to 20 hours or more, and production efficiency is low, while hydrogen peroxide use is bigger, and product color is better, but potency is lower.
In addition, also there is the prior art to remove the protein nucleic acid in crude product using the method for flocculation sediment and membrane filtration, but it is acid
Protein precipitation is not easy to filter out, and leads to filter out that the time is often long, and filter efficiency is low, therefore the prior art is using utilization
The glass powder or diatomite of silicone hydroxyl are as filter aid, but the effect is unsatisfactory, while filter aid also has absorption to heparin sodium
Effect, can cause the rate of recovery to reduce.Therefore, it is necessary to a kind of high purity heparin sodium preparation processes, to solve prior art preparation
The technical issues of long time period, inefficiency, titer of heparin sodium and the rate of recovery are lowered.
Invention content
It is an object of the invention to：A kind of preparation process of high purity heparin sodium, for solving the prior art preparation time
The technical issues of long working efficiency is low, and titer of heparin sodium and the rate of recovery reduce.
The technical solution adopted by the present invention is as follows：
A kind of preparation process of high purity heparin sodium, includes the following steps：
(1) proportionally every kilogram of crude heparin sodium is added in 8~10L purified waters and is dissolved, concentration 3.0% is added
(w/v) sodium chloride is stirred 45~60 minutes and is filtered, removes the contamination precipitation of precipitation, obtain crude product filtered fluid；
(2) trypsase of 0.1%~0.2% (w/v) is added in the crude product filtered fluid obtained in step (1), adjusts
PH values stir 1.5~2 hours, the foreign protein in heparin sodium is degraded to small-molecular peptides by trypsase, simultaneously to 7.5~8.5
Make heparin sodium molecule dissociate from the compound of heparin sodium-protein to come, obtains enzymolysis liquid；
(3) pH value adjustment for the enzymolysis liquid that step (2) obtains will be passed through to 1.5~2.0, after stirring 30~45 minutes, adopted
The small-molecular peptides that removal zymoprotein and protein are degraded are centrifuged with high-speed low temperature, it is molten to obtain the few heparin sodium of protein content
Liquid；
(4) hydrogen peroxide of 1.5%~2.0% (w/v) is added in the heparin sodium molten night obtained by step (3), and
It by pH value adjustment to 8.5~9.5, stirs 3~4 hours, the colors in oxidation removal heparin sodium molten night, then high speed
Low-temperature centrifugation obtains heparin sodium oxidation solution；
(5) absolute ethyl alcohol is added in the heparin sodium oxidation solution obtained by step (4), by alcoholic strength according to volume basis
Number is adjusted to 55~65 degree, and the sodium chloride of 1.0%~1.5% (w/v) is added in the solution, is stirred 1~1.5 hour and is filtered,
Obtain heparin sodium sediment；
(6) acetone is added in the heparin sodium sediment obtained by step (5) to be dehydrated, and by being dried in vacuo,
High purity heparin sodium is made.When preparation process using the present invention purifies heparin sodium, compared with conventional multi-level oxidation technology, potency
9U/mg is improved, the potency rate of recovery improves 8%.
Preferably, the hydrogen peroxide of 1.0%~1.5% (w/v) is added in the step (4) heparin sodium molten night, and will
PH values are adjusted to 8.5~9.5, then pass to ozonation aerated 2.5~3 hours, form the efficient oxidation body of hydrogen peroxide-ozone
It is colors and residual protein in oxidation removal heparin sodium molten night, then high-speed low temperature centrifuges to obtain heparin sodium oxidation
Liquid.Hydrogen peroxide-ozone efficient oxidation system can generate HO2 -, it is the inducing agent that free radical OH is generated, free radical OH is once
It generates, chain reaction will be caused, accelerate oxidation rate, improve oxidation effectiveness.
Compared to the prior art, the beneficial effects of the invention are as follows：
1. the mode that this programme uses trypsin digestion and level-one hydrogen peroxide oxidation removes albumen in heparin sodium
Matter and pigment impurity are compared with traditional multistage hydrogen peroxide oxidation, have saved at least 40% time, improved
Working efficiency.In addition, during heparin sodium purifies, the method for using high-speed low temperature centrifugation solves the prior art and adopts
With membrane filtration acidic protein precipitation is difficult, time-consuming and ineffective technical problem.Preparation process using the present invention
When purifying heparin sodium, compared with conventional multi-level oxidation technology, potency improves 9U/mg, and the potency rate of recovery improves 8%, is made
Heparin sodium purity it is high, color and luster is good.
2. HO can be generated using hydrogen peroxide-ozone efficient oxidation system2 -, it is the inducing agent that free radical OH is generated, from
By base OH once generating, chain reaction will be caused, accelerate oxidation rate, improve oxidation effectiveness.It is smelly using hydrogen peroxide-
The efficient oxidation system of oxygen decolourizes to pigment impurity and is removed residual protein, than simple hydrogen peroxide oxidation ratio
Get up, reduce the content of hydrogen peroxide, further improves the recycling potency of heparin sodium, while ozone is in addition to having sterilization to disappear
Outside malicious, decolouring and deodorizing function, also there is the advantage that can be aoxidized hardly degraded organic substance and improve flocculating effect etc., and smelly
Oxygen will not remain or generate secondary pollution, can further shorten the reaction time, improve working efficiency.
Specific implementation mode
All features disclosed in this specification can be with any other than mutually exclusive feature and/or step
Mode combines.
Embodiment 1
A kind of preparation process of high purity heparin sodium, includes the following steps：
(1) proportionally every kilogram of crude heparin sodium is added in 8~10L purified waters and is dissolved, concentration 3.0% is added
(w/v) sodium chloride is stirred 45~60 minutes and is filtered, removes the contamination precipitation of precipitation, obtain crude product filtered fluid；
(2) trypsase of 0.1%~0.2% (w/v) is added in the crude product filtered fluid obtained in step (1), adjusts
PH values stir 1.5~2 hours, the foreign protein in heparin sodium is degraded to small-molecular peptides by trypsase, simultaneously to 7.5~8.5
Make heparin sodium molecule dissociate from the compound of heparin sodium-protein to come, obtains enzymolysis liquid；
(3) pH value adjustment for the enzymolysis liquid that step (2) obtains will be passed through to 1.5~2.0, after stirring 30~45 minutes, adopted
The small-molecular peptides that removal zymoprotein and protein are degraded are centrifuged with high-speed low temperature, it is molten to obtain the few heparin sodium of protein content
Liquid；
(4) hydrogen peroxide of 1.5%~2.0% (w/v) is added in the heparin sodium molten night obtained by step (3), and
It by pH value adjustment to 8.5~9.5, stirs 3~4 hours, the colors in oxidation removal heparin sodium molten night, then high speed
Low-temperature centrifugation obtains heparin sodium oxidation solution；
(5) absolute ethyl alcohol is added in the heparin sodium oxidation solution obtained by step (4), by alcoholic strength according to volume basis
Number is adjusted to 55~65 degree, and the sodium chloride of 1.0%~1.5% (w/v) is added in the solution, is stirred 1~1.5 hour and is filtered,
Obtain heparin sodium sediment；
(6) acetone is added in the heparin sodium sediment obtained by step (5) to be dehydrated, and by being dried in vacuo,
High purity heparin sodium is made.
Technique of the present invention uses trypsin digestion and the mode of level-one hydrogen peroxide oxidation removes heparin sodium
In protein and pigment impurity, compared with traditional multistage hydrogen peroxide oxidation, saved at least 40% when
Between, it improves work efficiency.In addition, during heparin sodium purifies, the method for using high-speed low temperature centrifugation solves existing
There is technology using membrane filtration acidic protein precipitation difficulty, time-consuming and ineffective technical problem.It is using the present invention
When preparation process purifies heparin sodium, compared with conventional multi-level oxidation technology, potency improves 9U/mg, and the potency rate of recovery improves
8%.
Embodiment 2
Embodiment 2 is theed improvement is that the improvement of embodiment 1：In step (4) described in embodiment 1, in heparin sodium
The hydrogen peroxide of 1.0%~1.5% (w/v) is added in molten night, and by pH value adjustment to 8.5~9.5, then passes to ozonation aerated
2.5~3 hours, form the efficient oxidation system of hydrogen peroxide-ozone, colors in oxidation removal heparin sodium molten night and
Residual protein, then high-speed low temperature centrifuge to obtain heparin sodium oxidation solution.
It is decolourized and is removed to pigment impurity residual protein using the efficient oxidation system of hydrogen peroxide-ozone, than
Simple hydrogen peroxide oxidation is compared, and the content of hydrogen peroxide is reduced, and further improves the recycling effect of heparin sodium
Valence, while ozone, other than having the function of sterilizing, decolouring and deodorizing, hardly degraded organic substance can be aoxidized and improve wadding by also having
The advantage of solidifying effect etc., and ozone will not remain or generate secondary pollution.
Hydrogen peroxide-ozone efficient oxidation system can generate HO2 -, it is the inducing agent that free radical OH is generated, free radical
OH will cause chain reaction once generating, and accelerate oxidation rate, improve oxidation effectiveness.Therefore with conventional oxidation system ratio
Get up, hydrogen peroxide-ozone efficient oxidation system can further shorten the reaction time, improve working efficiency.
Above-described embodiment is merely illustrative of the technical solution of the present invention, rather than its limitations；Although with reference to above-described embodiment
Invention is explained in detail, it will be understood by those of ordinary skill in the art that：It still can be to aforementioned each implementation
Technical solution recorded in example is modified or equivalent replacement of some of the technical features；And these modification or
It replaces, the spirit and scope for various embodiments of the present invention technical solution that it does not separate the essence of the corresponding technical solution.
Claims (2)
1. a kind of preparation process of high purity heparin sodium, which is characterized in that include the following steps：
(1) proportionally every kilogram of crude heparin sodium is added in 8~10L purified waters and is dissolved, 3.0% (w/ of concentration is added
V) sodium chloride is stirred 45~60 minutes and is filtered, removes the contamination precipitation of precipitation, obtain crude product filtered fluid；
(2) trypsase of 0.1%~0.2% (w/v) is added in the crude product filtered fluid obtained in step (1), adjusts pH value
To 7.5~8.5, stir 1.5~2 hours, the foreign protein in heparin sodium is degraded to small-molecular peptides by trypsase, while making heparin
Sodium molecule is dissociateed from the compound of heparin sodium-protein to be come, and enzymolysis liquid is obtained；
(3) pH value adjustment for the enzymolysis liquid that step (2) obtains will be passed through to 1.5~2.0, after stirring 30~45 minutes, using height
The small-molecular peptides that fast low-temperature centrifugation removal zymoprotein and protein are degraded, obtain the few heparin sodium aqua of protein content；
(4) it is added the hydrogen peroxide of 1.5%~2.0% (w/v) in by step (3) obtained heparin sodium molten night, and by PH
Value is adjusted to 8.5~9.5, stirs 3~4 hours, the colors in oxidation removal heparin sodium molten night, then high-speed low temperature from
Gains in depth of comprehension are to heparin sodium oxidation solution；
(5) absolute ethyl alcohol is added in the heparin sodium oxidation solution obtained by step (4), by alcoholic strength according to percentage by volume tune
It saves to 55~65 degree, and the sodium chloride of 1.0%~1.5% (w/v) is added in the solution, stir 1~1.5 hour and filter, obtain
Heparin sodium sediment；
(6) acetone is added in the heparin sodium sediment obtained by step (5) to be dehydrated, and by vacuum drying, is made
High purity heparin sodium.
2. a kind of preparation process of high purity heparin sodium according to claim 1, it is characterised in that：In the step (4)
The hydrogen peroxide of 1.0%~1.5% (w/v) is added in heparin sodium molten night, and by pH value adjustment to 8.5~9.5, then passes to smelly
Oxygen is aerated 2.5~3 hours, forms the efficient oxidation system of hydrogen peroxide-ozone, the pigment in oxidation removal heparin sodium molten night
Substance and residual protein, then high-speed low temperature centrifuge to obtain heparin sodium oxidation solution.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810202915.9A CN108456262A (en) | 2018-03-13 | 2018-03-13 | A kind of preparation process of high purity heparin sodium |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201810202915.9A CN108456262A (en) | 2018-03-13 | 2018-03-13 | A kind of preparation process of high purity heparin sodium |
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| CN108456262A true CN108456262A (en) | 2018-08-28 |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109485750A (en) * | 2018-12-12 | 2019-03-19 | 大英县添峰生物制品有限公司 | A kind of heparin sodium low-temperature vacuum drying technology |
| CN109762079A (en) * | 2019-01-15 | 2019-05-17 | 湖北亿诺瑞生物制药有限公司 | A method of the separating-purifying Sulodexide bulk pharmaceutical chemicals from heparin byproduct |
| CN111057165A (en) * | 2020-01-13 | 2020-04-24 | 苏州健飞肠衣有限公司 | Preparation method of high-purity heparin sodium |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101831008A (en) * | 2009-03-11 | 2010-09-15 | 四川茂森生物科技有限公司 | New production process for refining crude heparin sodium |
| CN102225973A (en) * | 2011-06-22 | 2011-10-26 | 郓城绅联生物科技有限公司 | Production method for refined heparin sodium |
| CN105399867A (en) * | 2015-12-07 | 2016-03-16 | 青岛九龙生物医药有限公司 | Preparation method for heparin sodium |
| CN106084090A (en) * | 2016-06-30 | 2016-11-09 | 郭舒洋 | A kind of preparation method of refined heparin sodium |
-
2018
- 2018-03-13 CN CN201810202915.9A patent/CN108456262A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101831008A (en) * | 2009-03-11 | 2010-09-15 | 四川茂森生物科技有限公司 | New production process for refining crude heparin sodium |
| CN102225973A (en) * | 2011-06-22 | 2011-10-26 | 郓城绅联生物科技有限公司 | Production method for refined heparin sodium |
| CN105399867A (en) * | 2015-12-07 | 2016-03-16 | 青岛九龙生物医药有限公司 | Preparation method for heparin sodium |
| CN106084090A (en) * | 2016-06-30 | 2016-11-09 | 郭舒洋 | A kind of preparation method of refined heparin sodium |
Non-Patent Citations (1)
| Title |
|---|
| 朱灵峰: "《水与废水处理新技术》", 31 August 2007, 西安地图出版社 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109485750A (en) * | 2018-12-12 | 2019-03-19 | 大英县添峰生物制品有限公司 | A kind of heparin sodium low-temperature vacuum drying technology |
| CN109762079A (en) * | 2019-01-15 | 2019-05-17 | 湖北亿诺瑞生物制药有限公司 | A method of the separating-purifying Sulodexide bulk pharmaceutical chemicals from heparin byproduct |
| CN109762079B (en) * | 2019-01-15 | 2021-04-27 | 湖北亿诺瑞生物制药有限公司 | Method for separating and purifying sulodexide bulk drug from heparin by-product |
| CN111057165A (en) * | 2020-01-13 | 2020-04-24 | 苏州健飞肠衣有限公司 | Preparation method of high-purity heparin sodium |
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Application publication date: 20180828 |