CN110218269A - A kind of heparin process for producing sodium of short-term crude product - Google Patents
A kind of heparin process for producing sodium of short-term crude product Download PDFInfo
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- CN110218269A CN110218269A CN201910555803.6A CN201910555803A CN110218269A CN 110218269 A CN110218269 A CN 110218269A CN 201910555803 A CN201910555803 A CN 201910555803A CN 110218269 A CN110218269 A CN 110218269A
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- water
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- heparin
- crude product
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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
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
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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
Abstract
The invention discloses a kind of heparin process for producing sodium of short-term crude product, it is characterized in that, the production technology includes the following steps: pre-treatment step: selection ox lung and chitterlings stir into mud after mixing according to 1:3.5 ratio first, and the water containing 2% sodium chloride solution will be filled in ox lung after stirring and chitterlings, after aqueous solution is stood 3 hours, aqueous solution cooling is condensed into ice cube；Extraction step: will be ice melting by 60 DEG C of warm water, the NaOH solution allotment aqueous solution of pH9.0~9.2 of 110mol/L is deployed, and after ice cube melts completely, stand cooling；Absorption phase: being heated up to 50 DEG C for mixed solution, the aqueous solution of 110mol/LNaOH solution tune pH8.0~9.0 is added and resin that addition is handled well, weight resin is the 5% of mixed solution, 1h is stood after stirring 6h, with 80 mesh nylon net filters after having stood；Elution stage: resin is first rinsed with water and is filtered dry, then carries out gradient elution with salt water, is finally filtered dry resin；The filtrate for merging gradient elution, obtains eluent.
Description
Technical field
The present invention relates to a kind of production technologies, and in particular to a kind of heparin process for producing sodium of short-term crude product.
Background technique
Heparin sodium is be widely present in animal organ (such as: ox, the mucous membrane of small intestine of sheep, liver, pancreas and lung tissue) one
The compound that kind grape amine glycan is combined as protein in vivo as most of mucopolysaccharides exists, this compound
Anticoagulating active increases with the removal of protein.Heparin sodium and its derivative drug are anticoagulant, the antithrombotic reagent of classics,
It is clinically widely used in the oozing of blood for the treatment of Patients With Kidney Diseases, acute myocardial infarction disease prevents thrombosis, cranial vascular disease, skin
Skin disease, the uremic and partner treatment fulminant epidemic meningitis septicemia that removing nephrosis is formed, ephritis etc., meanwhile, heparin sodium is in drop blood
Rouge and it is immune etc. also have preferable effect, and with gradually understanding in depth to it, medical application is growing, market
Supply falls short of demand.
Heparin is Mike's human relations in 1916 when studying blood coagulation, the natural bioactive mucopolysaccharide object found from the liver of dog
Matter.After more than ten years, European and American developed countries just find and extract from ox lung, still later discovery pig, content in small sheep intestines mucous membrane again
It is extremely abundant, and start to carry out medical clinical research.Heparin is formally included in United States Pharmacopeia within 1940, and is widely used in anticoagulant
Blood prevents thrombosis, treatment cardiovascular and cerebrovascular disease etc..China started to introduce production the seventies, had nearly 40 year calendar so far
Shi Liao, existing market supply mainly extracted from pig intestinal mucosa or animal lungs crude heparin sodium (heparin and mostly
Number mucopolysaccharide is the same, the presence mostly in the form of being combined into compound with protein in vivo).During the extraction process due to heparin solution
From incomplete, so that there is always a certain number of protein in crude product heparin, pharmacy or outlet cannot be directly used to, it is therefore desirable to
Purification is further purified.Heparin sodium after purification as the generally attention by countries in the world the world of medicine of anticoagulation material medicine,
It is simultaneously also one of the main biochemical product that China's export is earned foreign exchange.
The subtractive process of heparin mainly carries out removing protein purification and decoloration to crude heparin sodium, reaches National Pharmacopeia standard
It is required that.In China, the purification of heparin mostly uses two that potassium permanganate and hydrogen peroxide two-step penetration method or hydrogen peroxide are added by several times
Secondary oxidizing process production technology；Also there is document announcement simultaneously, the record of oxidizing process purification heparin is combined using enzymatic hydrolysis.But it is above-mentioned several
Kind technique all Shortcomings: there are manganese dioxide to suck heparin for the first potassium permanganate and hydrogen peroxide two-step penetration method, makes liver
Plain loss of activity is big, and the rate of recovery is low；The disadvantages of filtration difficulty, the production cycle is long, and color is bad, of poor quality；Second of peroxidating
The Two-step anodization technique that oxygen time is added is generally used at present because product color is preferable.But the technique is primarily present secondary
Hydrogen oxide processing damages heparin structure, it is difficult to obtain the deficiency of the fine work heparin product of more efficient valence；The third enzymatic hydrolysis
It in conjunction with oxidizing process technique, is not widely adopted though having been reported that, exists simultaneously enzymatic hydrolysis condition, oxidizing condition and R. concomitans biology
The deficiency of the technological designs optimal selection problems such as product extraction, separation, purification technique.
Currently, China usually utilizes chitterlings, fresh intestinal mucosa liquid is extracted, produces heparin with traditional salt solution technique
Sodium, for this preparation method since the production cycle is long, hydrolysis is incomplete, is not thorough, and heparin sodium purity obtained is low, yield is low, effect
Valence is low, is difficult to carry out Produce on a large scale, and be not able to satisfy the wilderness demand in market.
Summary of the invention
Long the technical problem to be solved by the present invention is to the production cycle, hydrolysis is incomplete, is not thorough, and heparin sodium obtained is pure
Spend that low, yield is low, potency is low, and it is an object of the present invention to provide a kind of heparin process for producing sodium of short-term crude product, solves the problem above-mentioned.
The present invention is achieved through the following technical solutions:
A kind of heparin process for producing sodium of short-term crude product, which is characterized in that the production technology includes the following steps: pre- place
Reason step: selection ox lung and chitterlings stir into mud after mixing according to 1:3.5 ratio first, and will ox lung and chitterlings after stirring
Aqueous solution cooling after aqueous solution is stood 3 hours, is condensed into ice cube by interior water of the filling containing 2% sodium chloride solution；Extract step
It is rapid: will be ice melting by 60 DEG C of warm water, the NaOH solution allotment aqueous solution of pH9.0~9.2 of 110mol/L is deployed, and to ice cube
After melting completely, cooling is stood；Absorption phase: mixed solution is heated up to 50 DEG C, 110mol/LNaOH solution tune pH8.0 is added
The resin that~9.0 aqueous solution and addition are handled well, weight resin are the 5% of mixed solution, stand 1h after stirring 6h, have stood
Afterwards with 80 mesh nylon net filters；Elution stage: resin is first rinsed with water and is filtered dry, then carries out gradient elution with salt water, is finally filtered dry
Resin；The filtrate for merging gradient elution, obtains eluent；Precipitating obtains crude product: 95% isometric ethyl alcohol being added in eluent
After precipitates overnight, supernatant liquor is removed, sediment is heparin sodium crude, last stored dry.By by fresh pluck
Mud is stirred into, makes it possible to and quickly takes out mucous membrane liquid from internal organ, and the water that sodium chloride solution is added can save
It is preceding first to carry out preliminary chemical reaction.
Further, molten containing 2% sodium chloride in stirring ox lung and chitterlings filling simultaneously in the pre-treatment step
The water of liquid is divided into and adds water three times, and second is carried out after first time plus water spacer 3 minutes and adds water, divides in second plus water spacer 5
Third time is carried out after clock to add water and stir.In three times plus water can allow ox lung and chitterlings mixed raw material sufficiently to react with sodium chloride,
Further, the water containing 2% sodium chloride solution filled in three times, weight and ox lung and chitterlings mixture
Weight ratio be 1:4.Sodium chloride solution is needed when being sufficiently mixed with mixed raw material, and providing sufficient moisture can tie by liquid
Ice, and liquid level and liquid floor height difference are higher, are conducive to the precipitating of the purees of ox lung and chitterlings raw material, can freeze
Raw material transport is preferably carried out afterwards.
Further, in the extraction step after ice melting, after mixed solution restores room temperature, mixed solution is stood
12 hours, enter next step after sediment sufficiently precipitates in solution to be mixed.Mixed solution is stood, convenient after ice melting
Liquid sufficiently separated with sediment.
Further, the resin used in the absorption phase is D-254 macroporous absorbent resin.Using macroporous absorbent resin
It is adsorbed, the heparin sodium floated in aqueous solution can be absorbed into resin, heparin sodium is separated with water.
Further, the drying is to be dried under reduced pressure, temperature 45 C~55 DEG C, and vacuum degree is -0.1~-0.09Mpa, is done
40~60 hours dry time.
Compared with prior art, the present invention having the following advantages and benefits:
1, a kind of heparin process for producing sodium of short-term crude product of the present invention rubs fresh ox lung, chitterlings at slurries, slurries
Heat preservation enzymatic hydrolysis, enzymolysis liquid are collected by filtration filtrate, carry out ion exchange adsorption processing, the washing of resin and elution, heparin to filtrate
The precipitating of sodium, dry heparin sodium crude, the enzyme solution that the present invention uses more thoroughly can comprehensively dissolve pig lung slurries, effectively
The quality and heparin sodium yield with stable prod are improved, while the crude product impurity obtained is few, potency is high；
2, a kind of heparin process for producing sodium of short-term crude product of the present invention can be convenient for transporting after taking cooling to freeze,
And after this lysate rehydration, subsequent production is re-started by what precipitating can be convenient.
Specific embodiment
To make the objectives, technical solutions, and advantages of the present invention clearer, below with reference to embodiment, the present invention is made
Further to be described in detail, exemplary embodiment of the invention and its explanation for explaining only the invention, are not intended as to this
The restriction of invention.
Embodiment one
A kind of heparin process for producing sodium of short-term crude product of the present invention, which is characterized in that the production technology includes following step
Rapid: pre-treatment step: selection ox lung and chitterlings stir into mud after mixing according to 1:3.5 ratio first, and will ox lung after stirring
With water of the filling containing 2% sodium chloride solution in chitterlings, after aqueous solution is stood 3 hours, aqueous solution cooling is condensed into ice
Block；Extraction step: will be ice melting by 60 DEG C of warm water, NaOH solution allotment pH9.0~9.2 for deploying 110mol/L are water-soluble
Liquid, and after ice cube melts completely, stand cooling；Absorption phase: mixed solution is heated up to 50 DEG C, 110mol/LNaOH is added
The resin that the aqueous solutions of solution tune pH8.0~9.0 and addition are handled well, weight resin are the 5% of mixed solution, are stirred quiet after 6h
1h is set, with 80 mesh nylon net filters after having stood；Elution stage: resin is first rinsed with water and is filtered dry, then carries out gradient with salt water and wash
It is de-, finally it is filtered dry resin；The filtrate for merging gradient elution, obtains eluent；Precipitating obtains crude product: being added in eluent isometric
95% ethanol precipitation overnight after, remove supernatant liquor, sediment is heparin sodium crude, last stored dry.By will be fresh
Pluck stir into mud, make it possible to and quickly take out mucous membrane liquid from internal organ, and the water of sodium chloride solution is added
Preliminary chemical reaction can be first carried out before preservation.
The entire process flow of production technology of entire crude heparin sodium at present is by animal lung homogenizing into slurries, slurries heat preservation
Enzymatic hydrolysis, filtrate is collected by filtration in enzymolysis liquid, carries out ion exchange adsorption processing to filtrate, the washing and elution of resin, clarification filtration,
It is finally dry that heparin sodium crude, this mode are the most common process flows in crude heparin sodium manufacture, slurries are being made
When, slurries are kept the temperature and are digested, just bigger with present specification difference, present specification chooses ox lung and chitterlings by weight first
Amount ratio is mixed, its crude product heparin extracted amount of the more single internal organ of this mixed internal organ can be higher, maximum difference
Point is that sodium chloride solution can be first added in this mixed slurries, is then frozen by cooling method by solution, this icing
Low-temperature condition can allow the pause of the chemical reaction in solution, and can be convenient and transported, ice cube is then allowed by heating
Rehydration, slurries in the case where being sufficiently stirred, with diluted sodium hydroxide solution fine-tune its pH value be 9 when, after mixing evenly, slowly heat up
To 40 DEG C, continue to stir, and keep the pH value 9~9.2 of slurries, is digested 4 hours at 37 DEG C -40 DEG C of temperature；Then heat to 47
DEG C -50 DEG C, plasm PH value 8-9 is kept, continues enzymatic hydrolysis 5 hours to obtain enzymolysis liquid.In entirely heat preservation enzymolysis process, such as enzymolysis liquid
When pH value check is declined, just should carefully it be adjusted with dilute sodium hydroxide in time.
Cooling above-mentioned filtrate, carefully skims the grease lamella for floating on liquid level, and then control is warming up to 45 DEG C, stops adding
Heat, 80 grams of heparin adsorptions of addition are resin dedicated under stirring, to the effective component in absorption filtrate, at agitated absorption
Reason stood filtering after 8 hours.
Through ion exchange adsorption, treated that heparin adsorption is resin dedicated is rinsed with water, is filtered dry；Again with 70 grams of 4.5-5 Baumes
It is primary to spend NaCl, is filtered dry；Continue with 240 grams of 23-24 baume sodium chloride solutions, and 14 grams of sodium chloride are added；Then
Resin dedicated to the heparin adsorption washed to carry out elution action as eluent using sodium chloride concentrated solution, eluent salinity is
18-20 baume, temperature be 30-35 DEG C when, for the first time with 1500 grams elution 3.5 hours, for the second time washed with 1200 grams
De- liquid elutes 3 hours；After elution, it is filtered dry resin, eluent is merged, and is filtered with 80 mesh aperture nylon cloths, is received
Collect eluent.
The water containing 2% sodium chloride solution is filled simultaneously in stirring ox lung and chitterlings in the pre-treatment step, point
To add water three times, second is carried out after first time plus water spacer 3 minutes and adds water, carry out after five minutes in second plus water spacer
Third time adds water and stirs.In three times plus water can allow ox lung and chitterlings mixed raw material sufficiently to react with sodium chloride.
Embodiment two
The present embodiment optimizes on the basis of example 1, the water containing 2% sodium chloride solution filled in three times,
Its weight and the weight ratio of ox lung and chitterlings mixture are 1:4.Sodium chloride solution is needed when being sufficiently mixed with mixed raw material,
There is provided sufficient moisture can freeze by liquid, and liquid level and liquid floor height difference are higher, be conducive to ox lung and chitterlings are former
The precipitating of the purees of material can preferably carry out raw material transport after icing.
In the extraction step after ice melting, after mixed solution restores room temperature, mixed solution is stood 12 hours, to
Enter next step after sediment sufficiently precipitates in mixed solution.Mixed solution is stood, and the convenient liquid after ice melting fills
Divide and is separated with sediment.
The resin used in the absorption phase is D-254 macroporous absorbent resin.It is adsorbed using macroporous absorbent resin,
The heparin sodium floated in aqueous solution can be absorbed into resin, heparin sodium is separated with water.
The drying be dried under reduced pressure, temperature 45 C~55 DEG C, vacuum degree be -0.1~-0.09Mpa, drying time 40~
60 hours.It needs to grow very much in drying time, by way of slowly drying, can sufficiently evaporate the moisture in heparin sodium,
Finally obtain the crude heparin sodium of powdered drying.
Above-described specific embodiment has carried out further the purpose of the present invention, technical scheme and beneficial effects
It is described in detail, it should be understood that being not intended to limit the present invention the foregoing is merely a specific embodiment of the invention
Protection scope, all within the spirits and principles of the present invention, any modification, equivalent substitution, improvement and etc. done should all include
Within protection scope of the present invention.
Claims (6)
1. a kind of heparin process for producing sodium of short-term crude product, which is characterized in that the production technology includes the following steps:
Pre-treatment step: selection ox lung and chitterlings stir into mud after mixing according to 1:3.5 ratio first, and will ox lung after stirring
With water of the filling containing 2% sodium chloride solution in chitterlings, after aqueous solution is stood 3 hours, aqueous solution cooling is condensed into ice
Block；
Extraction step: will be ice melting by 60 DEG C of warm water, NaOH solution allotment pH9.0~9.2 for deploying 110mol/L are water-soluble
Liquid, and after ice cube melts completely, stand cooling；
Absorption phase: mixed solution is heated up to 50 DEG C, the aqueous solution of 110mol/LNaOH solution tune pH8.0~9.0 is added simultaneously
The resin handled well is added, weight resin is the 5% of mixed solution, 1h is stood after stirring 6h, with 80 mesh nylon wires after having stood
Filtering；
Elution stage: resin is first rinsed with water and is filtered dry, then carries out gradient elution with salt water, is finally filtered dry resin；Merge gradient to wash
De- filtrate, obtains eluent；
Precipitating obtains crude product: after 95% isometric ethanol precipitation is added in eluent overnight, removing supernatant liquor, sediment
For heparin sodium crude, last stored dry.
2. a kind of heparin process for producing sodium of short-term crude product according to claim 1, which is characterized in that the pretreatment step
Ox lung and chitterlings water of the filling containing 2% sodium chloride solution simultaneously are being stirred in rapid, is being divided into and adds water three times, add in first time
Second plus water are carried out after water spacer 3 minutes, are carried out third time after five minutes in second plus water spacer and are added water and stirred.
3. a kind of heparin process for producing sodium of short-term crude product according to claim 2, which is characterized in that fill in three times
The weight ratio of water containing 2% sodium chloride solution, weight and ox lung and chitterlings mixture is 1:4.
4. a kind of heparin process for producing sodium of short-term crude product according to claim 1, which is characterized in that the extraction step
In after ice melting, after mixed solution restores room temperature, mixed solution is stood 12 hours, sediment is abundant in solution to be mixed
Enter next step after precipitating.
5. a kind of heparin process for producing sodium of short-term crude product according to claim 1, which is characterized in that the absorption phase
The middle resin used is D-254 macroporous absorbent resin.
6. a kind of heparin process for producing sodium of short-term crude product according to claim 1, which is characterized in that the drying is to subtract
Pressing dry dry, temperature 45 C~55 DEG C, vacuum degree is -0.1~-0.09Mpa, drying time 40~60 hours.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201910555803.6A CN110218269A (en) | 2019-06-25 | 2019-06-25 | A kind of heparin process for producing sodium of short-term crude product |
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| CN201910555803.6A CN110218269A (en) | 2019-06-25 | 2019-06-25 | A kind of heparin process for producing sodium of short-term crude product |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2810588C1 (en) * | 2022-12-12 | 2023-12-27 | Общество с ограниченной ответственностью "РУССИНЕРГИЯ" | Method of obtaining heparin |
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| CN101891842A (en) * | 2010-07-16 | 2010-11-24 | 杭州龙扬生物科技有限公司 | Process for producing heparin sodium |
| CN103467624A (en) * | 2013-08-29 | 2013-12-25 | 武汉多宝微生物技术有限公司 | Extraction method of heparin sodium crude product |
| CN106967185A (en) * | 2017-03-22 | 2017-07-21 | 潢川县鹏升畜产品有限公司 | The method of comprehensive utilization of liquaemin is extracted from chitterlings |
| CN108314749A (en) * | 2018-03-08 | 2018-07-24 | 广元市海鹏生物科技有限公司 | A kind of method of driven Object Extraction heparin sodium |
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| CN1876687A (en) * | 2006-06-27 | 2006-12-13 | 胡世辉 | Heparin sodium production process |
| CN101891842A (en) * | 2010-07-16 | 2010-11-24 | 杭州龙扬生物科技有限公司 | Process for producing heparin sodium |
| CN103467624A (en) * | 2013-08-29 | 2013-12-25 | 武汉多宝微生物技术有限公司 | Extraction method of heparin sodium crude product |
| CN106967185A (en) * | 2017-03-22 | 2017-07-21 | 潢川县鹏升畜产品有限公司 | The method of comprehensive utilization of liquaemin is extracted from chitterlings |
| CN108314749A (en) * | 2018-03-08 | 2018-07-24 | 广元市海鹏生物科技有限公司 | A kind of method of driven Object Extraction heparin sodium |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2810588C1 (en) * | 2022-12-12 | 2023-12-27 | Общество с ограниченной ответственностью "РУССИНЕРГИЯ" | Method of obtaining heparin |
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Application publication date: 20190910 |