CN106519077A - Preparation process of high-potency heparin sodium - Google Patents
Preparation process of high-potency heparin sodium Download PDFInfo
- Publication number
- CN106519077A CN106519077A CN201611010799.8A CN201611010799A CN106519077A CN 106519077 A CN106519077 A CN 106519077A CN 201611010799 A CN201611010799 A CN 201611010799A CN 106519077 A CN106519077 A CN 106519077A
- Authority
- CN
- China
- Prior art keywords
- resin
- heparin sodium
- heparin
- precipitate
- added
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- 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
-
- 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
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Materials Engineering (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Organic Chemistry (AREA)
- Sustainable Development (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention relates to a preparation process of high-potency heparin sodium. The preparation process comprises raw material treatment, enzymolysis extraction, flocculation treatment, resin adsorption, resin washing, heparin elution, alcohol precipitation and refinement. The preparation process has the advantages as follows: a heparin sodium enzymatic hydrolysate is subjected to flocculation treatment, protein, nucleic acid, mycelia and other macromolecular substances in the enzymatic hydrolysate are flocculated and precipitated, a foundation is laid for the following separation process, and the difficulty of following impurity removal is reduced; heparin elution is performed after resin washing, impurities in heparin sodium are reduced, and impurities in an eluent are relatively reduced; therefore, complete hydrolysis can be realized with the method, and the purity and yield of the product can be increased accordingly.
Description
Technical field
The invention belongs to pharmaceutical formulating art, more particularly to a kind of high-potency heparin sodium preparation technology.
Background technology
Heparin sodium is a kind of mucopolysaccharide sulfuric acid ester anticoagulant, is mainly used in treating acute infarct, can mitigate shakiness
Sizing angina pectoriss, antithrombus formation and blood fat reducing, the effect of cholesterol reducing.People are found that liver in liver first within 1916
Element, then, finds that the material is widely existed in animal liver, lung and mucous membrane of small intestine again.Due to China's pig aboundresources, small intestinal
Mucosa is cheap, therefore, people are often with pig small intestine as raw material production heparin sodium.
At present, traditional heparin process for producing sodium includes that Feedstock treating, enzymolysis and extraction, ion-exchange absorption echo refined, should
Technique presence hydrolysis is incomplete, product purity is low and the low shortcoming of yield.
The content of the invention
The technical problem to be solved in the present invention be to provide one kind can hydrolyze completely, product purity it is high
And the high-potency heparin sodium preparation technology of high income.
For solving above-mentioned technical problem, the technical scheme is that：A kind of high-potency heparin sodium preparation technology, its innovation
Put and be：The production technology is comprised the following steps：
（1）Feedstock treating：Fresh Intestinum Sus domestica clear water is carefully cleaned after the inside and outside dirt of removal and external skin fat, is rubbed into
Meat paste shape；
（2）Enzymolysis and extraction：To step（1）In add 7～10 times of water in the meat paste shape that obtains, add 2%～3% Trypsin
Enzyme, plus 30%～40% NaOH adjusted between pH to 8～9, and 3%～5% NaCl after 40～60min of stirring reaction, is added, is put
In reaction pot, extraction time is 10～15min, after terminating, filters, obtains enzymolysis solution；
（3）Flocculation treatment：3%～5% chitosan flocculant is added in above-mentioned enzymolysis solution, with the speed of 100～120r/min
Stirring 20～30min of mixed liquor, after being stored at room temperature 4～6h, sends into centrifuge and filters, obtain the supernatant containing heparin sodium
Liquid；
（4）Resin absorption：Supernatant is adsorbed using strong-base anion-exchange resin, the consumption of resin accounts for the mass ratio of mother solution and is
0.8%～1.2%, 8～9h of stirring and adsorbing, then collect strong-base anion-exchange resin, discard waste liquid；
（5）Resin is washed：First washed with tap water and warm water, then handed over 4～5 degree of aqueous salt solu-tion strong alkalinity anion
30～40min of resin is changed, to remove resin surface impurity；
（6）Heparin eluting：With resin on the 5%～8% of 3 times of volumes sodium chloride solution eluting, elution time is 3～4h, then with 2
Sodium chloride solution 3～the 5h of eluting of the 3%～5% of times volume, eluent will merge twice；
（7）Precipitate with ethanol：By above-mentioned eluent, the 80%～90% of 2 times of volumes ethanol is added, stir 20～30min, then place of settling
20～24h of reason, collects precipitate, obtains heparin sodium crude after being dried；
（8）It is refined：By heparin sodium crude add 10 times of volumes pure water dissolving, then with 30%～40% NaOH adjust pH to 9～
Between 10, sucting reaction tank is filtered to remove impurity with frame filter press, then is adjusted between pH to 6～7 with 30%～40% NaOH,
80%～90% ethanol is slow added into, is stirred, precipitate 8～9h；Collect multiple precipitate, centrifuge dehydration, then 70～80
It is dried more than 20 hours at a temperature of DEG C, you can obtain finished product.
It is an advantage of the current invention that：The present invention carries out flocculation treatment to heparin sodium enzymolysis solution, make protein in enzymolysis solution,
The macromolecular substances such as nucleic acid, mycelium produce flocculation, precipitate, are that subsequent separation process lays the foundation, and reduce follow-up remove impurity
Difficulty；In the present invention after resin washing, then to heparin eluting, and then reduce the impurity in heparin sodium so that eluting
Impurity in liquid is also relative to be reduced；Thus, can be hydrolyzed completely by the inventive method, and cause to produce
Product purity and yield are also correspondingly improved.
Specific embodiment
The following examples can make professional and technical personnel that the present invention is more fully understood, but not therefore by this
It is bright be limited in described scope of embodiments among.
Embodiment 1
The high-potency heparin sodium preparation technology of the present embodiment, comprises the steps：
（1）Feedstock treating：Fresh Intestinum Sus domestica clear water is carefully cleaned after the inside and outside dirt of removal and external skin fat, is rubbed into
Meat paste shape；
（2）Enzymolysis and extraction：To step（1）In add 7 times of water in the meat paste shape that obtains, add 3% trypsin, plus 40%
NaOH adjust pH to 9, after stirring reaction 40min, add 5% NaCl, be placed in reaction pot, extraction time is 10min,
After end, filter, obtain enzymolysis solution；
（3）Flocculation treatment：5% chitosan flocculant is added in above-mentioned enzymolysis solution, with the speed stirring mixed liquor of 120r/min
20min, after being stored at room temperature 4h, sends into centrifuge and filters, obtain the supernatant containing heparin sodium；
（4）Resin absorption：Supernatant is adsorbed using strong-base anion-exchange resin, the consumption of resin accounts for the mass ratio of mother solution and is
1.2%, then stirring and adsorbing 8h collects strong-base anion-exchange resin, discards waste liquid；
（5）Resin is washed：First washed with tap water and warm water, then exchanged with 5 degree of aqueous salt solu-tion strong alkalinity anion
Resin 30min, to remove resin surface impurity；
（6）Heparin eluting：With resin on the 8% of 3 times of volumes sodium chloride solution eluting, elution time is 3h, then with 2 times of volumes
5% sodium chloride solution eluting 3h, will twice eluent merge；
（7）Precipitate with ethanol：By above-mentioned eluent, the 90% of 2 times of volumes ethanol is added, stir 20min, then precipitation process 20h, received
Collection precipitate, obtains heparin sodium crude after being dried；
（8）It is refined：Heparin sodium crude being added into the pure water dissolving of 10 times of volumes, then pH to 9 being adjusted with 40% NaOH, suction is anti-
Tank being answered, impurity being filtered to remove with frame filter press, then adjust pH to 7 with 40% NaOH, be slow added into 90% ethanol, stirring is equal
It is even, precipitate 8h；Multiple precipitate, centrifuge dehydration is collected, then is dried more than 20 hours at a temperature of 80 DEG C, you can obtain finished product.
After testing, the heparin sodium product potency ＞ 200U/mg for obtaining.
Embodiment 2
The high-potency heparin sodium preparation technology of the present embodiment, comprises the steps：
（1）Feedstock treating：Fresh Intestinum Sus domestica clear water is carefully cleaned after the inside and outside dirt of removal and external skin fat, is rubbed into
Meat paste shape；
（2）Enzymolysis and extraction：To step（1）In add 10 times of water in the meat paste shape that obtains, add 2% trypsin, plus
30% NaOH adjusts pH to 8, after stirring reaction 60min, adds 3% NaCl, is placed in reaction pot, and extraction time is
15min, after terminating, filters, obtains enzymolysis solution；
（3）Flocculation treatment：3% chitosan flocculant is added in above-mentioned enzymolysis solution, with the speed stirring mixed liquor of 100r/min
Min, after being stored at room temperature 6h, sends into centrifuge and filters, obtain the supernatant containing heparin sodium；
（4）Resin absorption：Supernatant is adsorbed using strong-base anion-exchange resin, the consumption of resin accounts for the mass ratio of mother solution and is
0.8%, then stirring and adsorbing 9h collects strong-base anion-exchange resin, discards waste liquid；
（5）Resin is washed：First washed with tap water and warm water, then exchanged with 4 degree of aqueous salt solu-tion strong alkalinity anion
Resin 40min, to remove resin surface impurity；
（6）Heparin eluting：With resin on the 5% of 3 times of volumes sodium chloride solution eluting, elution time is 4h, then with 2 times of volumes
3% sodium chloride solution eluting 5h, will twice eluent merge；
（7）Precipitate with ethanol：By above-mentioned eluent, the 80% of 2 times of volumes ethanol is added, stir 30min, then precipitation process 24h, received
Collection precipitate, obtains heparin sodium crude after being dried；
（8）It is refined：Heparin sodium crude being added into the pure water dissolving of 10 times of volumes, then pH to 9 being adjusted with 30% NaOH, suction is anti-
Tank being answered, impurity being filtered to remove with frame filter press, then adjust pH to 6 with 30% NaOH, be slow added into 80% ethanol, stirring is equal
It is even, precipitate 9h；Multiple precipitate, centrifuge dehydration is collected, then is dried more than 20 hours at a temperature of 70 DEG C, you can obtain finished product.
After testing, the heparin sodium product potency ＞ 200U/mg for obtaining.
Embodiment 3
The high-potency heparin sodium preparation technology of the present embodiment, comprises the steps：
（1）Feedstock treating：Fresh Intestinum Sus domestica clear water is carefully cleaned after the inside and outside dirt of removal and external skin fat, is rubbed into
Meat paste shape；
（2）Enzymolysis and extraction：To step（1）In add 9 times of water in the meat paste shape that obtains, add 2.5% trypsin, plus
35% NaOH adjusts pH to 8.5, after stirring reaction 50min, adds 4% NaCl, is placed in reaction pot, and extraction time is
13min, after terminating, filters, obtains enzymolysis solution；
（3）Flocculation treatment：4% chitosan flocculant is added in above-mentioned enzymolysis solution, with the speed stirring mixed liquor of 110r/min
25min, after being stored at room temperature 5h, sends into centrifuge and filters, obtain the supernatant containing heparin sodium；
（4）Resin absorption：Supernatant is adsorbed using strong-base anion-exchange resin, the consumption of resin accounts for the mass ratio of mother solution and is
1.0%, then stirring and adsorbing 8.5h collects strong-base anion-exchange resin, discards waste liquid；
（5）Resin is washed：First washed with tap water and warm water, then handed over 4.5 degree of aqueous salt solu-tion strong alkalinity anion
Resin 35min is changed, to remove resin surface impurity；
（6）Heparin eluting：With resin on the 7% of 3 times of volumes sodium chloride solution eluting, elution time is 3.5h, then with 2 times of bodies
The sodium chloride solution eluting 4h of long-pending 4%, eluent will merge twice；
（7）Precipitate with ethanol：By above-mentioned eluent, the 85% of 2 times of volumes ethanol is added, stir 25min, then precipitation process 22h, received
Collection precipitate, obtains heparin sodium crude after being dried；
（8）It is refined：By heparin sodium crude add 10 times of volumes pure water dissolving, then with 35% NaOH adjust pH to 9.5, suction
Retort, is filtered to remove impurity with frame filter press, then adjusts pH to 6.5 with 35% NaOH, is slow added into 85% ethanol, stirs
Mix uniform, precipitate 8.5h；Multiple precipitate, centrifuge dehydration is collected, then is dried more than 20 hours at a temperature of 75 DEG C, you can obtained into
Product.
After testing, the heparin sodium product potency ＞ 200U/mg for obtaining.
It should be understood by those skilled in the art that, the present invention is not restricted to the described embodiments, above-described embodiment and explanation
Merely illustrating the principles of the invention described in book, without departing from the spirit and scope of the present invention, the present invention also has
Various changes and modifications, these changes and improvements are both fallen within scope of the claimed invention.The claimed scope of the invention
By appending claims and its equivalent thereof.
Claims (1)
1. a kind of high-potency heparin sodium preparation technology, it is characterised in that：The production technology is comprised the following steps：
（1）Feedstock treating：Fresh Intestinum Sus domestica clear water is carefully cleaned after the inside and outside dirt of removal and external skin fat, is rubbed into
Meat paste shape；
（2）Enzymolysis and extraction：To step（1）In add 7～10 times of water in the meat paste shape that obtains, add 2%～3% Trypsin
Enzyme, plus 30%～40% NaOH adjusted between pH to 8～9, and 3%～5% NaCl after 40～60min of stirring reaction, is added, is put
In reaction pot, extraction time is 10～15min, after terminating, filters, obtains enzymolysis solution；
（3）Flocculation treatment：3%～5% chitosan flocculant is added in above-mentioned enzymolysis solution, with the speed of 100～120r/min
Stirring 20～30min of mixed liquor, after being stored at room temperature 4～6h, sends into centrifuge and filters, obtain the supernatant containing heparin sodium
Liquid；
（4）Resin absorption：Supernatant is adsorbed using strong-base anion-exchange resin, the consumption of resin accounts for the mass ratio of mother solution and is
0.8%～1.2%, 8～9h of stirring and adsorbing, then collect strong-base anion-exchange resin, discard waste liquid；
（5）Resin is washed：First washed with tap water and warm water, then handed over 4～5 degree of aqueous salt solu-tion strong alkalinity anion
30～40min of resin is changed, to remove resin surface impurity；
（6）Heparin eluting：With resin on the 5%～8% of 3 times of volumes sodium chloride solution eluting, elution time is 3～4h, then with 2
Sodium chloride solution 3～the 5h of eluting of the 3%～5% of times volume, eluent will merge twice；
（7）Precipitate with ethanol：By above-mentioned eluent, the 80%～90% of 2 times of volumes ethanol is added, stir 20～30min, then place of settling
20～24h of reason, collects precipitate, obtains heparin sodium crude after being dried；
（8）It is refined：By heparin sodium crude add 10 times of volumes pure water dissolving, then with 30%～40% NaOH adjust pH to 9～
Between 10, sucting reaction tank is filtered to remove impurity with frame filter press, then is adjusted between pH to 6～7 with 30%～40% NaOH,
80%～90% ethanol is slow added into, is stirred, precipitate 8～9h；Collect multiple precipitate, centrifuge dehydration, then 70～80
It is dried more than 20 hours at a temperature of DEG C, you can obtain finished product.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611010799.8A CN106519077A (en) | 2016-11-17 | 2016-11-17 | Preparation process of high-potency heparin sodium |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611010799.8A CN106519077A (en) | 2016-11-17 | 2016-11-17 | Preparation process of high-potency heparin sodium |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106519077A true CN106519077A (en) | 2017-03-22 |
Family
ID=58352265
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611010799.8A Pending CN106519077A (en) | 2016-11-17 | 2016-11-17 | Preparation process of high-potency heparin sodium |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106519077A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107011464A (en) * | 2017-05-31 | 2017-08-04 | 广元市海天实业有限责任公司 | A kind of efficient crude heparin sodium production technology |
| CN107098989A (en) * | 2017-04-27 | 2017-08-29 | 甘肃省金羚集团药业有限公司 | A kind of preparation method of liquaemin |
| CN116655827A (en) * | 2023-06-12 | 2023-08-29 | 江苏千牧生物科技股份有限公司 | Extraction process for heparin sodium preparation |
| CN117362476A (en) * | 2023-08-10 | 2024-01-09 | 浩泰健康济盛(吉林)生物科技有限公司 | Method for preparing refined heparin by using bovine lung and corresponding refined heparin |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1876687A (en) * | 2006-06-27 | 2006-12-13 | 胡世辉 | Heparin sodium production process |
| CN102952204A (en) * | 2012-10-09 | 2013-03-06 | 江苏联众肠衣有限公司 | Novel production technique of heparin sodium |
| CN103848929A (en) * | 2014-03-21 | 2014-06-11 | 广元市申达实业有限公司 | Process for high-efficiently extracting sodium heparin |
| CN104725531A (en) * | 2013-12-24 | 2015-06-24 | 安徽宝迪肉类食品有限公司 | Production process of effectively extracting heparin sodium |
-
2016
- 2016-11-17 CN CN201611010799.8A patent/CN106519077A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1876687A (en) * | 2006-06-27 | 2006-12-13 | 胡世辉 | Heparin sodium production process |
| CN102952204A (en) * | 2012-10-09 | 2013-03-06 | 江苏联众肠衣有限公司 | Novel production technique of heparin sodium |
| CN104725531A (en) * | 2013-12-24 | 2015-06-24 | 安徽宝迪肉类食品有限公司 | Production process of effectively extracting heparin sodium |
| CN103848929A (en) * | 2014-03-21 | 2014-06-11 | 广元市申达实业有限公司 | Process for high-efficiently extracting sodium heparin |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107098989A (en) * | 2017-04-27 | 2017-08-29 | 甘肃省金羚集团药业有限公司 | A kind of preparation method of liquaemin |
| CN107011464A (en) * | 2017-05-31 | 2017-08-04 | 广元市海天实业有限责任公司 | A kind of efficient crude heparin sodium production technology |
| CN116655827A (en) * | 2023-06-12 | 2023-08-29 | 江苏千牧生物科技股份有限公司 | Extraction process for heparin sodium preparation |
| CN117362476A (en) * | 2023-08-10 | 2024-01-09 | 浩泰健康济盛(吉林)生物科技有限公司 | Method for preparing refined heparin by using bovine lung and corresponding refined heparin |
| CN117362476B (en) * | 2023-08-10 | 2024-05-17 | 浩泰健康济盛(吉林)生物科技有限公司 | Method for preparing refined heparin by using bovine lung and corresponding refined heparin |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106519077A (en) | Preparation process of high-potency heparin sodium | |
| CN106397630B (en) | A method of Sodium Hyaluronate is extracted using membrane separation technique | |
| CN102952204A (en) | Novel production technique of heparin sodium | |
| CN104788510A (en) | Method for extracting glucosamine from self-fermentation liquid | |
| CN109438532B (en) | Method for extracting D-glucosamine | |
| CN106831894B (en) | A kind of method of deacetylation Coupling Adsorption separation D-Glucosamine Hydrochloride | |
| CN1137998C (en) | Zymolysis process of producing heparin sodium | |
| CN101704905A (en) | Method for extracting hyaluronic acid | |
| CN109265498A (en) | A kind of method that integrated poly sialic acid separating-purifying prepares N-acetyl-neuraminate | |
| CN1111171C (en) | Heparin and its preparing process | |
| CN107936129A (en) | A kind of preparation process of complete series molecular weight tremella polysaccharides | |
| CN103848929B (en) | A kind of high efficiency extraction technique of heparin sodium | |
| CN103130904A (en) | High-valued utilization method for patinopecten yessoensis offal | |
| CN106496363A (en) | A kind of efficient preparation technology of heparin sodium | |
| CN103724456B (en) | The Technology for normal-temperature salt-free extraction of heparin sodium | |
| CN104530143A (en) | Method for preparing soybean whey oligosaccharide through coupling of heating, flocculating, air floating and ultra-filtering | |
| CN107641161A (en) | A kind of optimal reparation technology of casing accessory substance liquaemin | |
| CN114907498A (en) | Process for improving adsorption of heparin sodium in enzymolysis liquid by resin | |
| CN113045610B (en) | Method for extracting glucosamine from N-acetylglucosamine fermentation liquor | |
| CN208594226U (en) | A kind of animal heparin sodium extraction collection device | |
| CN113980090A (en) | Method for recovering biological source protein by using biological medicine fermentation fungus residues | |
| CN1042703A (en) | Water diacolation-ion exchange method is extracted the levodopa novel process | |
| CN113354698A (en) | Method for preparing N-acetylglucosamine fermentation clear liquid by using composite flocculant | |
| CN103214512A (en) | Technology for preparing high-purity phytic acid with membrane technology by taking defatted rice bran as raw material | |
| CN109517092A (en) | The heparin process for producing sodium of pig intestinal mucosa |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20170322 |
|
| RJ01 | Rejection of invention patent application after publication |