CN102212149B - Preparation process for extracting crude sodium heparin from pig lungs - Google Patents
Preparation process for extracting crude sodium heparin from pig lungs Download PDFInfo
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- CN102212149B CN102212149B CN2011101611386A CN201110161138A CN102212149B CN 102212149 B CN102212149 B CN 102212149B CN 2011101611386 A CN2011101611386 A CN 2011101611386A CN 201110161138 A CN201110161138 A CN 201110161138A CN 102212149 B CN102212149 B CN 102212149B
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- pig lung
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- heparin sodium
- enzymolysis
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Abstract
The invention relates to a preparation process for extracting crude sodium heparin from pig lungs, which comprises the following steps: mincing fresh pig lungs into slurry, carrying out heat insulation and enzymolysis to obtain an enzymolysis liquid, filtering the enzymolysis liquid to collect filtrate, carrying out ion exchange adsorption treatment on the filtrate, washing and eluting resin, precipitating sodium heparin, and drying to obtain crude sodium heparin. The enzymolysis method employed by the invention can thoroughly dissolve pig lung slurry and effectively improve and stabilize the quality of the product and the yield of sodium heparin; and simultaneously, the obtained crude sodium heparin has low impurity content and high titer.
Description
Technical field
The present invention relates to a kind of preparation technology who utilizes animal viscera to extract heparin sodium, especially a kind of preparation technology who utilizes the pig lung to extract heparin sodium crude.
Background technology
Heparin sodium is a kind of grape amine glycan that extensively is present in animal organ's (as: mucous membrane of small intestine of ox, sheep, liver, pancreas and lung tissue etc.); The same with most of mucopolysaccharides in vivo as the mixture existence of protein bound, the anticoagulating active of this mixture increases along with proteinic removal.Heparin is received and the verivate medicine is classical anti-freezing, antithrombotic reagent, is widely used in treatment nephropathy patient's oozing of blood clinically, and Acute Myocardial Infarction disease prevents thrombosis; Cerebrovascular disease, tetter is removed uremic and partner treatment fulminant epidemic meningitis septicemia that ephrosis forms; Ephritis etc., simultaneously, heparin sodium also has better action at aspects such as reducing blood-fat and immunity; And along with it is understood in depth gradually, its medical use enlarges day by day, and supply falls short of demand in market.
At present, China utilizes chitterlings usually, extracts fresh intestines mucosa liquid; Use traditional salt to separate the explained hereafter heparin sodium; This preparation method since the production cycle long, hydrolysis not exclusively, not thorough, the heparin sodium purity that makes is low, yield is low, it is low to tire; Be difficult to carry out the production of scale property, and can not satisfy the heavy demand in market.
Summary of the invention
The objective of the invention is to overcome the weak point of prior art; A kind of product purity height, few, the high preparation technology who utilizes pig lung extraction heparin sodium crude that tires of impurity are provided, and this process using enzyme enzymolysis process extracts heparin sodium crude from the pig lung that starting material are easy to get.
The present invention realizes that the technical scheme of purpose is following:
A kind of preparation technology who utilizes the pig lung to extract heparin sodium crude, step is following:
(1) broken slurry: the pig lung is rubbed the pulp thing of regrinding, under fully stirring, add the water of weight ratio 1: 1-2, mixing gets pig lung slurries;
(2) enzymolysis: the pH value of pig lung slurries adjustment slurries is 8-10, adds the heparin sodium specific enzyme of the 1-1.5% of pig lung slurry weight, stirs, and is warming up to 45-60 ℃; After insulation reaction 1.5-2.5 hour, keeping the pH value of reaction solution is 8-10, adds the 4-5% sodium-chlor of enzymolysis weight; Stir, be incubated 0.5-1.5 hour, continue to be warmed up to 90-100 ℃; Kept 10-10 minute, cooling obtains pig lung enzymolysis solution;
(3) filter, adsorb: filter pig lung enzymolysis solution, remove surperficial oil layer, filtrating is warmed up to 40-55 ℃ again; Whipped state adds the macroporous adsorbent resin of the 2-4% of filtrate volume down; Stir and leave standstill filtration after 3-5 hour, remove upper strata liquid, collect the resin that is adsorbed with heparin sodium;
(4) wash-out: the water flushing filter of resin elder generation is done, and carries out gradient elution with salt solution again, filters dried resin at last; The filtrating that merges gradient elution gets elutriant;
(5) deposition obtains bullion: after isopyknic 95% ethanol sedimentation of adding spends the night in elutriant, remove supernatant liquid, throw out is a heparin sodium crude, and final drying stores.
And said step (3) macroporous adsorbent resin is the D-254 resin.
And said step (3) is filtered with 60 order nylon cloths.
And said gradient elution is: use 1-2 times of salinity of resin volume to be 4-5 degree Beaume sodium-chlor wash-out 1 hour, filter is done; Continue to use 0.5-0.9 times of salinity of resin volume to stir wash-out 45 minutes～70 minutes, the filter dried resin as 4-5 degree Beaume sodium chloride solution; Use 2-3 times of salinity of resin volume to stir wash-out 2-3 hour again as 23-24 degree Beaume sodium chloride solution; The filter dried resin continues to use 1-2 times of salinity to be 23-24 degree Beaume sodium chloride solution wash-out 3-4 hour, filters dried resin.
And said stored dry step is: heparin sodium crude places container to drain, and puts into moisture eliminator then, regularly replaces siccative.
Advantage of the present invention and beneficial effect are:
1, the present invention is rubbed into slurries, slurries insulation enzymolysis, enzymolysis solution with the fresh pig lung and is filtered and collect filtrating, filtrating carried out IX absorption is handled, the washing of resin and the deposition of wash-out, heparin sodium, the dry heparin sodium crude that gets; The enzyme solution that the present invention adopts can more thoroughly dissolve pig lung slurries all sidedly; The quality and the heparin sodium yield of effective raising and stable prod, few, the height of tiring of bullion impurity that obtains simultaneously.
2, preparation technology of the present invention is reasonable in design, and no waste gas, waste residue, waste water are discharged contaminate environment, and the operation production cost is low, is suitable for suitability for industrialized production.
3, nontoxic, harmless, the biological environmental production of explained hereafter of the present invention, pig lung raw material sources are easy to get, and cost is low, is fit to large-scale industrial production.
Embodiment
Through specific embodiment the present invention is made further detailed description below, following examples are descriptive, are not determinate, can not limit protection scope of the present invention with this.
The heparin sodium that the heparin sodium specific enzyme of using in the present embodiment all adopts permanent magnificent road, Nanning east biotechnology Ltd to produce extracts specific enzyme.
A kind of preparation technology who utilizes the pig lung to extract heparin sodium crude, step is following:
(1) broken slurry: at first the fresh pig lung is carefully cleaned with clear water, remove inside and outside dirt and outer cortical fat after, with its rubbing pulp thing of regrinding, get 1000g pig lung soup compound, adding 1000g water mixing gets pig lung slurries under fully stirring;
(2) enzymolysis: pig lung slurries are under fully stirring, and using the pH value of sodium hydroxide solution adjustment slurries is 8.5 o'clock, adds 10g heparin sodium specific enzyme, stirs; Be warming up to 53 ℃, insulation reaction is after 2 hours, and keeping the pH value of reaction solution is 8.5, adds 90g sodium-chlor; Stir, be incubated 1 hour, continue to be warmed up to 98 ℃; Kept 15 minutes, naturally cooling obtains pig lung enzymolysis solution;
(3) filter, adsorb: behind the pig lung enzymolysis solution naturally cooling, filter with 60 order nylon cloths, and skim surperficial oil layer; Filtrating is warmed up to 48 ℃ again, and whipped state adds 60g D-254 resin down, stirs and leaves standstill filtration after 4 hours; Remove upper strata liquid with the U trap suction, collect the resin that is adsorbed with heparin sodium;
(4) wash-out: the water flushing filter of resin elder generation is done, and using the 70g salinity again is 4.5 degree Beaume sodium-chlor wash-outs 1 hour, and filter is done; Continuing to use 0.7 times of salinity of resin volume is that 4.5 degree Beaume sodium chloride solutions stirred wash-out 45 minutes, the filter dried resin; Using the 240g salinity again is that 23 degree Beaume sodium chloride solutions stirred wash-out 2 hours; The filter dried resin continues to use the 240g salinity to be 23-24 degree Beaume sodium chloride solution wash-out 3 hours, filters dried resin; Merge above-mentioned filtrate elutriant;
(5) deposition obtains bullion: after 95% ethanol sedimentation of adding 3000g spends the night in elutriant, remove supernatant liquid with U trap, throw out is a heparin sodium crude;
(6) stored dry: suction filtration to the heparin sodium throw out of doing places B to drain; Put into moisture eliminator then, regularly replace till siccative such as Vanadium Pentoxide in FLAKES no longer absorb water, smash into particulate state to pieces; Promptly get heparin sodium crude 2.0g, it is tired and is every milligram 90 unit.
Claims (5)
1. preparation technology who utilizes the pig lung to extract heparin sodium crude, it is characterized in that: step is following:
⑴ broken slurry: the pig lung is rubbed the pulp thing of regrinding, under fully stirring, add the water of weight ratio 1:1-2, mixing gets pig lung slurries;
⑵ enzymolysis: the pH value of adjustment pig lung slurries is 8-10, adds the heparin sodium specific enzyme of the 1-1.5% of pig lung slurry weight, stirs, and is warming up to 45-60 ℃; After insulation reaction 1.5-2.5 hour, keeping the pH value of reaction solution is 8-10, adds the 4-5% sodium-chlor of enzymolysis weight; Stir, be incubated 0.5-1.5 hour, continue to be warmed up to 90-100 ℃; Kept 10 minutes, cooling obtains pig lung enzymolysis solution;
The heparin sodium that described heparin sodium specific enzyme all adopts permanent magnificent road, Nanning east biotechnology Ltd to produce extracts specific enzyme;
⑶ filter, absorption: filter pig lung enzymolysis solution, remove surperficial oil layer, filtrating is warmed up to 40-55 ℃ again; Whipped state adds the macroporous adsorbent resin of the 2-4% of filtrate volume down; Stir and leave standstill filtration after 3-5 hour, remove upper strata liquid, collect the resin that is adsorbed with heparin sodium;
⑷ wash-out: the water flushing filter of resin elder generation is done, and carries out gradient elution with salt solution again, filters dried resin at last; The filtrating that merges gradient elution gets elutriant;
⑸ deposition obtains bullion: after isopyknic 95% ethanol sedimentation of adding spends the night in elutriant, remove supernatant liquid, throw out is a heparin sodium crude, and final drying stores.
2. the preparation technology who utilizes the pig lung to extract heparin sodium crude according to claim 1, it is characterized in that: said step ⑶ macroporous adsorbent resin is the D-254 resin.
3. the preparation technology who utilizes the pig lung to extract heparin sodium crude according to claim 1 is characterized in that: said step ⑶ filters in the pig lung enzymolysis solution and filters with 60 order nylon cloths.
4. the preparation technology who utilizes the pig lung to extract heparin sodium crude according to claim 1, it is characterized in that: said gradient elution is: uses 1-2 times of salinity of resin volume to be 4-5 degree Beaume sodium-chlor wash-out 1 hour, filter is dried; Continue to use 0.5-0.9 times of salinity of resin volume to stir wash-out 45 minutes～70 minutes, the filter dried resin as 4-5 degree Beaume sodium chloride solution; Use 2-3 times of salinity of resin volume to stir wash-out 2-3 hour again as 23-24 degree Beaume sodium chloride solution; The filter dried resin continues to use 1-2 times of salinity to be 23-24 degree Beaume sodium chloride solution wash-out 3-4 hour, filters dried resin.
5. the preparation technology who utilizes the pig lung to extract heparin sodium crude according to claim 1, it is characterized in that: said stored dry step is: heparin sodium crude places container to drain, and puts into moisture eliminator then, regularly replaces siccative.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN2011101611386A CN102212149B (en) | 2011-06-15 | 2011-06-15 | Preparation process for extracting crude sodium heparin from pig lungs |
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| CN2011101611386A CN102212149B (en) | 2011-06-15 | 2011-06-15 | Preparation process for extracting crude sodium heparin from pig lungs |
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| CN102212149A CN102212149A (en) | 2011-10-12 |
| CN102212149B true CN102212149B (en) | 2012-11-28 |
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Families Citing this family (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102344502B (en) * | 2011-11-11 | 2012-12-12 | 宁发子 | Method for extracting heparin sodium by utilizing pork lungs |
| CN103183745B (en) * | 2012-09-19 | 2015-08-12 | 杭州龙扬生物科技有限公司 | Using alkali proteinase method extracts the technique of heparin sodium from intestinal mucosa |
| CN103755836A (en) * | 2013-11-25 | 2014-04-30 | 青岛九龙生物医药有限公司 | Preparation technology for extracting heparin sodium crude product by utilizing animal lung |
| CN103951768B (en) * | 2014-04-29 | 2016-05-11 | 贵州慧静生物科技有限公司 | Utilize pig, ox lung to extract the method for sodium heparin and co-producing polypeptide albumen powder, amino acid protein powder |
| CN103980387B (en) * | 2014-05-26 | 2016-03-23 | 安徽科宝生物工程有限公司 | A kind of biological enzyme prepares the method for heparin sodium |
| CN104098717A (en) * | 2014-07-21 | 2014-10-15 | 南通恒阳生物科技有限公司 | Method for extracting heparin sodium |
| CN104530262B (en) * | 2015-01-26 | 2016-08-24 | 安徽科宝生物工程有限公司 | A kind of production method extracting heparin from Pulmonis Sus domestica |
| CN106317259A (en) * | 2016-09-19 | 2017-01-11 | 陈石良 | Joint production technology for extracting heparin sodium and protein peptide powder from pork lung |
| CN107446067A (en) * | 2017-08-10 | 2017-12-08 | 如皋市永兴肠衣有限公司 | A kind of intestinal mucosa comprehensive processing and utilization technology |
| CN107216412A (en) * | 2017-08-10 | 2017-09-29 | 如皋市永兴肠衣有限公司 | The Hydrolysis kinetics technology of liquaemin in pig intestinal mucosa |
| CN112574327A (en) * | 2019-09-27 | 2021-03-30 | 成都祁连山生物科技股份有限公司 | Method for preparing crude heparin sodium by utilizing pig lungs through biological enzyme method |
| CN112142873A (en) * | 2020-10-20 | 2020-12-29 | 德清亚泰肠衣有限公司 | Preparation process for efficiently extracting high-quality heparin sodium |
| CN113999330B (en) * | 2021-12-07 | 2022-08-19 | 潢川县鹏升畜产品有限公司 | Separation and co-production process of low-salt-concentration heparin sodium and active intestinal protein peptide |
| CN113980155A (en) * | 2021-12-17 | 2022-01-28 | 揭阳市润达肠衣有限公司 | Method for extracting heparin in animal lungs |
| 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 |
Citations (3)
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| WO1989004328A1 (en) * | 1987-11-06 | 1989-05-18 | Opocrin S.P.A. Laboratorio Farmacobiologico | Non-anticoagulant heparan sulfate, process for extraction from organs, and pharmaceutical compositions thereof |
| CN1238182A (en) * | 1998-06-05 | 1999-12-15 | 何德惠 | Process for extracting coarse lipo-hepinette from lung of animal |
| CN1566162A (en) * | 2003-07-07 | 2005-01-19 | 张国良 | Heparin sodium and its preparing process |
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2011
- 2011-06-15 CN CN2011101611386A patent/CN102212149B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1989004328A1 (en) * | 1987-11-06 | 1989-05-18 | Opocrin S.P.A. Laboratorio Farmacobiologico | Non-anticoagulant heparan sulfate, process for extraction from organs, and pharmaceutical compositions thereof |
| CN1238182A (en) * | 1998-06-05 | 1999-12-15 | 何德惠 | Process for extracting coarse lipo-hepinette from lung of animal |
| CN1566162A (en) * | 2003-07-07 | 2005-01-19 | 张国良 | Heparin sodium and its preparing process |
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