CN102838692A - Extraction method for heparin sodium - Google Patents
Extraction method for heparin sodium Download PDFInfo
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- CN102838692A CN102838692A CN 201110172251 CN201110172251A CN102838692A CN 102838692 A CN102838692 A CN 102838692A CN 201110172251 CN201110172251 CN 201110172251 CN 201110172251 A CN201110172251 A CN 201110172251A CN 102838692 A CN102838692 A CN 102838692A
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- heparin sodium
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Abstract
The present discloses a heparin sodium extraction method, wherein a purpose of the present invention is to solve a problem of a poor absorption effect of the resin in the conventional enzymolysis process. In the prior, grease can be decomposed in the enzymolysis process, a large pore size resin for heparin sodium production is easily clogged by the grease, such that an absorption effect is reduced, and a yield is reduced. With the present invention, a resin and enzymolysis liquid separation method is adopted, the enzymolysis liquid in the enzymolysis tank is subjected to constant temperature heating, and circulation is performed in a circulation device, such that the grease is not close to the resin so as not to clog the resin, the resin absorption and the yield are increased, resin pollution is avoided, and resin repair treatment difficulty is reduced.
Description
Technical field
The inventive method relates to be a kind of be the heparin sodium extracting technique of material with the pig intestinal mucosa, method has improved the prior art level through biological enzymolysis technology, chromatographic technique, thereby has improved a kind of process for extracting of heparin sodium output.
Background technology
Cardiovascular and cerebrovascular diseases is human No.1 disease killer.The Along with people's standard of living improves overnutrition, the deterioration of global environment, rhythm of life quickening, the aging population aggravation that brings; Cause the M & M of global cardiovascular and cerebrovascular diseases to increase just year by year, heparin appear as the miracle that numerous cardiovascular and cerebrovascular diseases patients have created life.Heparin is the maximum anticoagulation medicine of the most effective and clinical in the world consumption at present, is mainly used in cardiovascular and cerebrovascular diseases and hemodialysis, and wherein, it is unique effective specific medicament in hemodialysis.Clinical application and research show that heparin also has other multiple biological activity and clinical applications except that having blood coagulation resisting function, comprise effects such as reducing blood lipid, anti-middle film smooth muscle cell (SMC) hyperplasia, promotion fibrinolysis.In addition; Low molecular heparin is the one big type of antithrombotic medicine that further is processed into as raw material by the heparin bulk drug; Have clinical medicine purposes more widely, become the choice drug of acute phlebothrombosis of treatment and acute coronary artery syndrome diseases such as (stenocardia, myocardial infarctions).Heparin is the most complicated compound of known molecular structures up to now in the world, in a short time can't artificial chemosynthesis, and the heparin that only derives from pig intestinal mucosa at present can be used in clinical treatment.The raw material of heparin bulk drug is the heparin bullion; Its extraction can only be derived from the mucous membrane of small intestine of healthy live pig; Owing to contain impurity such as a large amount of impurity albumen, impurity nucleic acid, mikrobe; Need through physics and chemical extraction sepn process, orientation is obtained the complete heparin of natural structure group, thereby processes the heparin bulk drug.The heparin sodium bulk drug is the unique effective constituent of standard heparin preparation and the production starting point of Low molecular heparin raw material; Heparin preparations only is used for clinical according to the drug administration by injection mode at present; This makes the heparin bulk drug need very high purity, can guarantee the drug safety of preparation.
Report and adopt ultrasound-assisted enzymolysis and the salt process of separating can improve yield, as " Heilongjiang Bayi Agricultural Reclamation University, Song Da Wei Diplomarbeit; " high efficiency extraction of heparin and separating and purifying technology research ", through discovering, UW can help cell rupture really through the High Voltage cavitation effect; discharge more heparin, be free in the enzymolysis solution, but this method adopts the UW energy consumption huge; and hardware drops into high, and unsuitable volume production is not seen the industriallization report at present.
The research tradition method finds that all there is a problem in the method that adopts the resin absorption mode to extract heparin sodium, after resin drops into adsorption tanks; Not all resin all is fully used, and when the resin whip attachment, enzymolysis solution forms vortex in jar; The bottom resin density is greater than top, and exactly the bottom heparin concentration is low, and top concentration is high; And contamination precipitation is arranged at the bottom; Swim in adsorption tanks internal upper part resin in addition and be prone to be stopped up by the lubricant component in the enzymolysis solution, the resin absorption ability after the obstruction descends greatly, these problems affect resin absorption; The present invention be directed to the problems referred to above and adopt chromatography method to solve this difficult problem, do not find in heparin sodium crude is produced, to adopt the relevant report of chromatography method at present.
Summary of the invention
The process for extracting that the purpose of this invention is to provide a kind of heparin sodium, its cost is low, yield is high, energy consumption is low, pollution is little.
The present invention adopts following technical scheme, and its characteristic comprises the steps:
A adopts conventional enzymolysis process salt solution to make the chitterlings enzymolysis solution; B adopts conventional method heating enzymolysis solution by the proteolytic enzyme inactivation; C adopts the conventional method impurity screening; D adopts the tomography devices that has circulation device to adsorb; E adopts conventional method wash-out resin; F adopts conventional method deposition heparin sodium; G adopts conventional method purified heparin sodium; H adopts the conventional method drying to obtain heparin sodium at last.Method therefor of the present invention is to prepare two hang-ups i.e. " 1, can not fully decompose mucous membrane 2, fail fine absorption " in the process to heparin sodium; The present invention solves is 2, fail the technical barrier of fine absorption; Adopt the benefit of D method to be; Changing in the past, dynamic adsorption is that Static Adsorption, avidity are desirable, resin is difficult for possibility blocked, resin can fully contact, stop outside biological pollution with enzymolysis solution, closed environment can keep-up pressure, and normality (need accomplish under certain pressure by resin absorption; China is geographical vast; Height above sea level just differs, constant pressure and adjustable when adopting the D described method to guarantee to adsorb through discovering, it is 1.025MPa that discover method D absorption optimum pressure is gone back in research).
The present invention has the following advantages:
1. technology is simple, and is easy and simple to handle, and less investment changes not quite existing installation, is fit to large-scale industrial production.
2. the heparin sodium yield improves, steady quality.The heparin sodium purity of producing with present method can reach 105-120IU/mg, and the heparin sodium yield can reach 100,000,000 IU/1500-1700 root weight at 1200 grams-1400 gram pig intestinal mucosas, improves 20-30% than prior art.
3. reduce and pollute, reduce cost.The inventive method flows into water exhaust system and then the water source is caused chronic pollution in the time of can stopping resin at adsorption tanks at wash-out.
4. cut down the consumption of energy, reduce production cost.The inventive method adopts circulating water system, than traditional technology economize on electricity power 30%.
Embodiment
Embodiment 1
A kind of process for extracting of heparin sodium may further comprise the steps:
A enzymolysis salt is separated mixed extraction: adopt the traditional technology enzymic hydrolysis, use 100 health pig mucous membrane of small intestine to be raw material, add entry, proteolytic enzyme, sausage-casing salt, sodium hydroxide; Its ratio was respectively 1: 5: 0.03: 0.15: 0.1, drop into together in the retort, after stirring; Transfer PH to 8.0-9.0; Be warming up to 55-60 ℃, constant temperature stirs enzymolysis 3H, and pH value should keep PH8.0-9.0 in the enzymolysis process.
B proteolytic enzyme inactivation: enzymolysis solution is heated to 85-90 ℃ fast, be incubated 10-30 minute, begin to filter.
C filters: treat that the proteolytic enzyme inactivation finishes, begin to filter, filter top impurity with the 100-120 mesh filter screen, collect filtrating.
D absorption: after the enzymolysis solution cooling,, cycle through enzymolysis solution the chromatography column of having pre-installed resin through self-circulating device; Circulation is for closed circulation, completely cut off and the contacting of air, and the post height is 3 times of resin height; Circulation 8H; Keep 60 ℃ of temperature simultaneously, tensimeter shows 1.01MPa, and tachograph shows 3L/m.
The E wash-out: the resin after will adsorbing stirs wash-out 2-4H 2 times with 15%-19% sodium chloride solution (amount of resin 1 times), collects elutriant, and then with 20% sodium chloride solution (amount of resin 1 times) wash-out 1 time, collection elutriant.
F heparin sodium deposition: elutriant in the step e is merged, add 85%-95% ethanol and stir, reach the 45-50 degree, stop to stir, leave standstill 12H, top alcohol is reclaimed treat to use next time, collect bottom heparin sodium throw out up to the precipitated liquid alcohol concn.
G heparin sodium purifying: step F gained heparin sodium deposition with the dissolving of 3% sodium chloride solution, is filtered with the 200-300 mesh filter screen, remove impurity; Adding the stirring of 85%-95% ethanol, reach 45%-50% up to strength of solution, staticly settle 8H; Reclaim top alcohol; Collect bottom heparin sodium deposition, add the 85%-95% ethanol dehydration then, collect the heparin sodium deposition.
The H oven dry: as for air-dry in the air dry oven, when extremely half-dried, the adding smectite is dry, can get heparin sodium with the heparin sodium deposition of collecting.
Embodiment 2
The step of present embodiment has just been done change with embodiment 1 on the flow velocity of adsorption chromatography post and pressure, embodiment 2 changes into: tensimeter shows 1.02MPa, tachograph demonstration 5L/m.
Embodiment 3
The step of present embodiment has just been done change with embodiment 1 on the flow velocity of adsorption chromatography post and pressure, embodiment 3 changes into: tensimeter shows 1.025MPa, tachograph demonstration 10L/m.
The present invention can summarize with other specific form without prejudice to spirit of the present invention or principal character; Under the situation that does not break away from the above-mentioned subject area of the present invention; Various changes and replacement according to ordinary skill knowledge and customary means are worked it out all fall into protection domain of the present invention.
Denomination of invention
A kind of process for extracting of heparin sodium
Summary
The invention discloses a kind of process for extracting of heparin sodium, may further comprise the steps:
A adopts conventional enzymolysis process salt solution to make the chitterlings enzymolysis solution; B adopts the conventional method heating let the proteolytic enzyme inactivation; C adopts the conventional method impurity screening; D adopts the tomography devices that has the constant temperature circulation device to adsorb; E adopts conventional method wash-out resin; F adopts conventional method deposition heparin sodium; G adopts conventional method purified heparin sodium; H adopts the conventional method drying to obtain heparin sodium.The inventive method heparin sodium yield improves, steady quality.The heparin sodium purity of producing with present method can reach 105-120IU/mg, and the heparin sodium yield can reach 100,000,000 IU/1500-1700 root weight at 1200 grams-1400 gram pig intestinal mucosas, improves 20-30% than prior art.
Claims (3)
1. the process for extracting of a heparin sodium, concrete steps are following:
A adopts conventional enzymolysis process salt solution to make the chitterlings enzymolysis solution; B adopts conventional method heating enzymolysis solution by the proteolytic enzyme inactivation; C adopts the conventional method impurity screening; D adopts the tomography devices that has the constant temperature circulation device to adsorb; E adopts conventional method wash-out resin; F adopts conventional method deposition heparin sodium; G adopts conventional method purified heparin sodium; H adopts the conventional method drying to obtain heparin sodium at last.
2. the process for extracting of a kind of heparin sodium according to claim 1 is characterized in that: the method for a kind of constant temperature that adopts among the said step D, circulation, sealing, chromatography.
3. the process for extracting of a kind of heparin sodium according to claim 2, its characteristic comprises the steps:
A enzymolysis salt is separated mixed extraction: adopt the traditional technology enzymic hydrolysis, use the health pig mucous membrane of small intestine to be raw material, add entry, proteolytic enzyme, sausage-casing salt, sodium hydroxide; Its ratio was respectively 1: 5: 0.03: 0.15: 0.1, drop into together in the retort, after stirring; Transfer PH to 8.0-9.0; Be warming up to 55-60 ℃, constant temperature stirs enzymolysis 3H, and pH value should keep PH8.0-9.0 in the enzymolysis process.
B proteolytic enzyme inactivation: enzymolysis solution is heated to 85-90 ℃ fast, be incubated 10-30 minute, begin to filter.
C filters: treat that the proteolytic enzyme inactivation finishes, begin to filter, filter top impurity with the 100-120 mesh filter screen, collect filtrating.
D absorption: after the enzymolysis solution cooling,, cycle through enzymolysis solution the chromatography column of having pre-installed resin through self-circulating device; Circulation is for closed circulation, completely cut off and the contacting of air, and the post height is 3 times of resin height; Circulation 8H keeps 60 ℃ of temperature, pressure 1.025MPa simultaneously.
The E wash-out: the resin after will adsorbing stirs wash-out 2-4H 2 times with 15%-19% sodium chloride solution (amount of resin 1 times), collects elutriant, and then with 20% sodium chloride solution (amount of resin 1 times) wash-out 1 time, collection elutriant.
F heparin sodium deposition: elutriant in the step e is merged, add 85%-95% ethanol and stir, reach the 45-50 degree, stop to stir, leave standstill 12H, top alcohol is reclaimed treat to use next time, collect bottom heparin sodium throw out up to the precipitated liquid alcohol concn.
G heparin sodium purifying: step F gained heparin sodium deposition with the dissolving of 3% sodium chloride solution, is filtered with the 200-300 mesh filter screen, remove impurity; Adding the stirring of 85%-95% ethanol, reach 45%-50% up to strength of solution, staticly settle 8H; Reclaim top alcohol; Collect bottom heparin sodium deposition, add the 85%-95% ethanol dehydration then, collect the heparin sodium deposition.
The H oven dry: as for air-dry in the air dry oven, when extremely half-dried, the adding smectite is dry, can get heparin sodium with the heparin sodium deposition of collecting.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110172251 CN102838692A (en) | 2011-06-24 | 2011-06-24 | Extraction method for heparin sodium |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110172251 CN102838692A (en) | 2011-06-24 | 2011-06-24 | Extraction method for heparin sodium |
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| CN102838692A true CN102838692A (en) | 2012-12-26 |
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| CN 201110172251 Pending CN102838692A (en) | 2011-06-24 | 2011-06-24 | Extraction method for heparin sodium |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104490793A (en) * | 2014-11-26 | 2015-04-08 | 山东辰中生物制药有限公司 | Method for improving heparin sodium thermostability |
| CN105801723A (en) * | 2016-04-12 | 2016-07-27 | 浙江大学 | Rapid purification method of marine sulfated polysaccharides |
-
2011
- 2011-06-24 CN CN 201110172251 patent/CN102838692A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104490793A (en) * | 2014-11-26 | 2015-04-08 | 山东辰中生物制药有限公司 | Method for improving heparin sodium thermostability |
| CN104490793B (en) * | 2014-11-26 | 2017-08-29 | 山东辰中生物制药有限公司 | The method for improving liquaemin heat endurance |
| CN105801723A (en) * | 2016-04-12 | 2016-07-27 | 浙江大学 | Rapid purification method of marine sulfated polysaccharides |
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Application publication date: 20121226 |