CN1238182A - Process for extracting coarse lipo-hepinette from lung of animal - Google Patents
Process for extracting coarse lipo-hepinette from lung of animal Download PDFInfo
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- CN1238182A CN1238182A CN 98112054 CN98112054A CN1238182A CN 1238182 A CN1238182 A CN 1238182A CN 98112054 CN98112054 CN 98112054 CN 98112054 A CN98112054 A CN 98112054A CN 1238182 A CN1238182 A CN 1238182A
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Abstract
A process for extracting coarse heparin sodium from lung of animal includes such technological steps as homogenizing the lung of animal to obtain slurry, holding its temp. constant for enzymolysis, filtering enzymolyzed liquid, and collecting filtrate, ion exchange and adsorption, washing resin and edution, clarifying and filtering, and drying. Its advantages include high potency increased by 15-20%, high output rate increased by more than 10%, low cost of raw material, and low environmental pollution.
Description
The present invention relates to a kind of preparation technology who utilizes animal viscera to extract heparin sodium, particularly a kind of preparation technology who utilizes the pig pulmonis Bovis seu Bubali to extract heparin sodium crude.
Heparin sodium is a kind of anticoagulant efficacy that has, and prevents that tumor focus from shifting and the costly medicine of diffusion.The oozing of blood that is used for the treatment of the nephropathy patient clinically, acute myocardial infarction disease prevents thrombosis, cerebrovascular disease, dermatosis is removed uremic and partner treatment fulminant epidemic encephalitis septicemia that nephropathy forms, nephritis etc.Simultaneously, heparin sodium also has better action at aspects such as blood fat reducing and immunity, and along with it is understood in depth gradually, its medical application enlarges day by day, and supply falls short of demand in market.At present, China utilizes pig small intestine usually, extracts fresh intestinal mucosa liquid, uses traditional salt to separate the explained hereafter heparin sodium, and this preparation method is difficult to carry out scale production because raw material is deficient and technical limitation; Environmental pollution is serious in process of production; And the production cost height, product yield is low, and it is low to tire, and can not satisfy the wilderness demand in market.
The object of the present invention is to provide a kind of productive rate height, the height of tiring, cost is low, and is the preparation technology who extracts heparin sodium crude from the animal lung that raw material is easy to get.
The present invention is achieved in that a kind of preparation technology who utilizes the animal lung to extract heparin sodium crude, it is characterized in that: animal lung homogenize becomes serosity～serosity insulation enzymolysis～enzymolysis solution to filter to collect filtrate～filtrate the is carried out washing and the eluting～clarification filtration～dry heparin sodium crude that gets of ion exchange absorption processings～resin.
Described animal lung homogenize becomes serosity to be meant cleaned animal lung rubbed wears into slurry, adds the entry mixing thereafter under fully stirring and gets serosity, and the addition of water doubly is the best with the 1-1.3 of slurry.
Described serosity insulation enzymolysis is meant and adds gastric enzyme or the agent of pancreatin enzymolysis when the pH value of adjusting serosity is 10-10.5, stirs, and is warming up to 38 ℃-41 ℃, continues to stir, and keeps the pH value 8-8.5 of serosity, 37 ℃ of-40 ℃ of following enzymolysis 3.5-4.5 of temperature hours; Be warming up to 47 ℃-50 ℃ then, keep serosity pH value 8-8.5, add a small amount of enzymolysis agent, continue enzymolysis and got enzymolysis solution in 4.5-5.5 hour.In whole insulation enzymolysis process, adopt sig water such as sodium hydroxide to adjust to the adjustment of serosity pH value; The Pancreas Sus domestica slurry is adopted in the enzymolysis agent, and its addition is advisable according to the 1-1.3% of slurry weight, and additional amount is advisable according to the 0.3-0.5% of slurry weight.
Described enzymolysis solution filters collection filtrate and is meant that the pH value of adjusting enzymolysis solution is 5-5.5, is warming up to 75 ℃-80 ℃, stirs to add the sodium chloride mixing, is warming up to 90 ℃-95 ℃ again, is incubated 1-2 hour, stops to stir the filtration that warms up, collection filtrate; Treat that filtrate is cooled to 40 ℃-45 ℃, the pH value of adjusting filtrate is 10-11, filters once more, collects filtrate and adjust its pH value to be 8-8.5, and is stand-by.In this step diluted acid and diluted alkaline such as dilute hydrochloric acid and diluted sodium hydroxide solution are adopted in the adjustment of the acid-base value of enzymolysis solution and filtrate; The addition of sodium chloride is incorporated as suitable according to the 4.5-5% of enzymolysis solution weight; Warm up and filter the nylon net cloth that adopts 60 order apertures; Filter the nylon net cloth that adopts 80 order apertures once more.
Described ion exchange absorption is handled and is meant the above-mentioned filtrate of cooling, removes the oils and fats lamella, is warmed up to 45 ℃-50 ℃ then, and it is resin dedicated that stirring adds heparin absorption down, in order to the effective ingredient in the absorption filtrate, after stirring and adsorbing, leaves standstill filtration.The consumption of new resin is generally filtrate 3.5-4%, and stirring and adsorbing was handled 7-8 hour.
The washing of described resin and eluting are meant that to be adsorbed with the heparin absorption of heparin compositions with water rinse resin dedicated, and filter is done; Times salinity of reuse resin volume 0.8-0.85 is the washing of 4.5-5 baume sodium chloride solution, and filter is done; Times salinity that continues with resin volume 0.5-0.6 is a 23-24 baume sodium chloride solution, and adds sodium chloride, and addition is 0.8-1 with the amount of resin ratio: 7.5; Then the heparin that washs is adsorbed resin dedicated employing sodium chloride concentrated solution and carry out the eluting operation as eluent; After eluting finished, the filter dried resin was merged eluent, and filtered with 80 order aperture nylon net cloths, collected eluent.
The operation of described eluting is meant that the eluent salinity is the 18-20 baume, when temperature is 30-35 ℃, for the first time with the 0.8-0.85 eluent eluting doubly of resin volume 3-3.5 hour, for the second time with the 0.5-0.6 eluent eluting doubly of resin volume 3-3.5 hour.
Described clarification filtration is meant that the pH value of adjusting eluent is 11-12, after stirring, leaves standstill clarification, take out the supernatant, the bottom precipitation is carried out sucking filtration, merge clear liquid and filtrate, the adjusting pH value is 5-5.5, adds 3-3.5 80-85% ethanol clarifier doubly, leaves standstill till the clarification of upper strata liquid; Aspirate out the supernatant, collecting precipitation thing and sucking filtration are to doing.
Described drying is meant and places buchner funnel to drain sucking filtration to the heparin sodium precipitate of doing, and puts into exsiccator then, regularly replaces desiccant such as phosphorus pentoxide till no longer absorbing water, and smashs into graininess to pieces, promptly gets heparin sodium crude.
The heparin that contains among the known animal organ is by hexanal acid, iduronic acid, the how sticking sugar that glucuronic acid and the group alternate group that contains the hexylamine sugar of sulfonic acid are made into, can think the mixture of polymer analog and polymer homologue, molecular weight is usually at 6000-20000, can become anion by enzymolysis under certain condition, the reuse resin anion (R.A.) extracts, therefore the present invention is on extracting method, adopt enzymolysis, resin is purified, and facts have proved that the heparin sodium color and luster that the present invention produces is whiter, stable yield, average each pig pulmonis Bovis seu Bubali can extract 0.8-1.2 gram heparin sodium, product quality is also better, on average tires all between 90-120, compares with traditional salt solution, product is tired and can be improved 15%-20%, and productive rate improves more than 10%; The present invention adopts pig, pulmonis Bovis seu Bubali cheap and that be easy to get on raw material, cost is reduced widely, is fit to large-scale production; In the preparation process of the present invention, no toxic and harmful produces, though a small amount of waste residue and small amount of acid alkali cleaning water generates are arranged, waste residue can be processed into feedstuff and sell after drying, and discharging was again compared with the salt solution after acidic and alkaline waste water can neutralize, and had reduced environmental pollution.
The invention will be further described below in conjunction with embodiment:
Embodiment 1:
At first the fresh Pulmonis Sus domestica of 1000 grams is carefully cleaned with clear water, remove inside and outside dirt and outer cortical fat after, with its rubbing pulp thing of regrinding, you then add 1000 gram water mixings and get serosity under fully stirring.
Serosity is 10 o'clock with its pH value of the meticulous adjusting of diluted sodium hydroxide solution under fully stirring, and adds Pancreas Sus domestica slurry 20 grams, after stirring, slowly is warming up to 40 ℃, continues to stir, and keeps the pH value 8-8.5 of serosity, 37 ℃ of-40 ℃ of following enzymolysis of temperature 4 hours; Be warming up to 47 ℃-50 ℃ then, keep serosity pH value 8-8.5, add 10 gram Pancreas Sus domestica slurries, continue enzymolysis and got enzymolysis solution in 5 hours.In whole insulation enzymolysis process, when check descends to some extent as the pH value of enzymolysis solution, just should in time carefully adjust with dilute sodium hydroxide.
After above-mentioned insulation enzymolysis process finishes, adjust its pH value 5-5.5 with dilute hydrochloric acid, be warming up to 80 ℃ then, under fully stirring, add 100 gram sodium chloride, make it miscible evenly after, be warmed up to 90 ℃ again, be incubated 1 hour, stop to stir, with the filtration that warms up of the nylon net cloth in 60 order apertures, remove impurity, collect filtrate; Treat that filtrate is cooled to 40 ℃, adjust its pH value 11, remove residue with the nylon net cloth fine filtering in 80 order apertures and collect filtrate, filtrate is adjusted back in its PH8-8.5 scope with dilute hydrochloric acid with dilute sodium hydroxide.
Cool off above-mentioned filtrate, carefully drag for and remove the oils and fats lamella that floats on liquid level, control is warmed up to 45 ℃ then, stop heating, it is resin dedicated that adding 80 restrains heparin absorption under stirring, in order to the effective ingredient in the absorption filtrate, after stirring and adsorbing is handled 8 hours, leave standstill filtration.
The resin dedicated water rinse of using of heparin absorption after ion exchange absorption is handled, filter is done; Reuse 70 gram 4.5-5 baume sodium chloride washings are once filtered and are done; Continue with 240 gram 23-24 baume sodium chloride solutions, and add 14 gram sodium chloride; Then the heparin that washs is adsorbed resin dedicated employing sodium chloride concentrated solution and carry out the eluting operation as eluent, the eluent salinity is the 18-20 baume, when temperature is 30-35 ℃, restrain eluent eluting 3.5 hours with 1500 for the first time, restrain eluent eluting 3 hours with 1200 for the second time; After eluting finished, the filter dried resin was merged eluent, and filtered with 80 order aperture nylon cloths, collected eluent.
The pH value of adjusting above-mentioned eluent is 11-12, after stirring, leaves standstill clarification, carefully aspirate out the supernatant, sucking filtration is extremely dried as far as possible to the bottom precipitation, merges clear liquid and filtrate, the adjusting pH value is 5-5.5, adds 3600 gram 80%-85% ethanol, leaves standstill till the clarification of upper strata liquid; Aspirate out the supernatant, collecting precipitation thing and sucking filtration are to doing.
Place buchner funnel to drain sucking filtration to the heparin sodium precipitate of doing, put into phosphorus pentoxide desiccator then, regularly replace till desiccant to phosphorus pentoxide no longer absorbs water, smash into graininess to pieces, promptly get heparin sodium crude.
Embodiment 2:
Take by weighing Pulmonis Sus domestica 1000 grams, water 3000 grams, sodium hydroxide 30 grams, Pancreas Sus domestica slurry 17 grams, sodium chloride 350 grams, hydrochloric acid 15 grams, special-purpose adsorbent resin 93 grams of heparin, ethanol 3000 grams prepare heparin sodium crude 2.0 grams, its every milligram 90 unit of tiring with reference to embodiment 1 described step.
Embodiment 3:
Take by weighing pulmonis Bovis seu Bubali 1000 grams, water 3000 grams, sodium hydroxide 40 grams, Pancreas Sus domestica slurry 30 grams, sodium chloride 410 grams, hydrochloric acid 10 grams, special-purpose adsorbent resin 80 grams of heparin, ethanol 3600 grams prepare heparin sodium crude 2.4 grams, its every milligram 107 unit of tiring with reference to embodiment 1 described step equally.
In the above-described embodiments, it is resin dedicated that the special-purpose adsorbent resin of heparin all adopts the win honour for board heparin absorption that wins honour for of biological product company limited production of Hang Zhoutang Qi Zhen.
Claims (11)
1. preparation technology who utilizes the animal lung to extract heparin sodium crude is characterized in that: animal lung homogenize becomes serosity～serosity insulation enzymolysis～enzymolysis solution to filter to collect filtrate～filtrate the is carried out washing and the eluting～clarification filtration～dry heparin sodium crude that gets of ion exchange absorption processings～resin.
2. the preparation technology who utilizes the animal lung to extract heparin sodium crude as claimed in claim 1, it is characterized in that: described animal lung homogenize becomes serosity to be meant cleaned animal lung rubbed wears into slurry, thereafter add the entry mixing under fully stirring and get serosity, the addition of water doubly is the best with the 1-1.3 of slurry.
3. the preparation technology who utilizes the animal lung to extract heparin sodium crude as claimed in claim 1, it is characterized in that: described serosity insulation enzymolysis is meant when the pH value of adjusting serosity is 10-10.5, add gastric enzyme or the agent of pancreatin enzymolysis, stir, be warming up to 38 ℃-41 ℃, continue to stir, and keep the pH value 8-8.5 of serosity, 37 ℃ of-40 ℃ of following enzymolysis 3.5-4.5 of temperature hours; Be warming up to 47 ℃-50 ℃ then, keep serosity pH value 8-8.5, add a small amount of enzymolysis agent, continue enzymolysis and got enzymolysis solution in 4.5-5.5 hour.
4. the preparation technology who utilizes the animal lung to extract heparin sodium crude as claimed in claim 3 is characterized in that: in whole insulation enzymolysis process, adopt sig water such as sodium hydroxide to adjust to the adjustment of serosity pH value; The Pancreas Sus domestica slurry is adopted in the enzymolysis agent, and its addition is advisable according to the 1-1.3% of slurry weight, and additional amount is advisable according to the 0.3-0.5% of slurry weight.
5. the preparation technology who utilizes the animal lung to extract heparin sodium crude as claimed in claim 1, it is characterized in that: described enzymolysis solution filters collection filtrate and is meant that the pH value of adjusting enzymolysis solution is 5-5.5, be warming up to 75 ℃-80 ℃, stir and add the sodium chloride mixing, be warming up to 90 ℃-95 ℃ again, be incubated 1-2 hour, stop to stir, filtrate is collected in the filtration that warms up; Treat that filtrate is cooled to 40 ℃-45 ℃, the pH value of adjusting filtrate is 10-11, filters once more, collects filtrate and adjust its pH value to be 8-8.5, and is stand-by.
6. the preparation technology who utilizes the animal lung to extract heparin sodium crude as claimed in claim 5 is characterized in that: filter adjustment employing diluted acid and diluted alkaline such as dilute hydrochloric acid and the diluted sodium hydroxide solution of collecting in the filtrate step the acid-base value of enzymolysis solution and filtrate at enzymolysis solution; The addition of sodium chloride is incorporated as suitable according to the 4.5-5% of enzymolysis solution weight; Warm up and filter the nylon net cloth that adopts 60 order apertures; Filter the nylon net cloth that adopts 80 order apertures once more.
7. the preparation technology who utilizes the animal lung to extract heparin sodium crude as claimed in claim 1, it is characterized in that: described ion exchange absorption is handled and is meant the above-mentioned filtrate of cooling, remove the oils and fats lamella, be warmed up to 45 ℃-50 ℃ then, it is resin dedicated that stirring adds heparin absorption down, in order to the effective ingredient in the absorption filtrate, after stirring and adsorbing, leave standstill filtration; The consumption of new resin is generally filtrate 3.5-4%, and stirring and adsorbing was handled 7-8 hour.
8. the preparation technology who utilizes the animal lung to extract heparin sodium crude as claimed in claim 1 is characterized in that: the washing of described resin and eluting are meant that to be adsorbed with the heparin absorption of heparin compositions with water rinse resin dedicated, and filter is dried; 0.8-0.85 times of salinity of reuse resin volume is the washing of 4.5-5 baume sodium chloride, and filter is done; Continuing with 0.5-0.6 times of salinity of resin volume is 23-24 baume sodium chloride solution, and adds sodium chloride, and addition is 0.8-1 with the amount of resin ratio: 7.5; Then the heparin that washs is adsorbed resin dedicated employing sodium oxide concentrated solution and carry out the eluting operation as eluent; After eluting finished, the filter dried resin was merged eluent, and filtered with 80 order aperture nylon net cloths, collected eluent.
9. the preparation technology who utilizes the animal lung to extract heparin sodium crude as claimed in claim 8, it is characterized in that: described eluting operation is meant that the eluent salinity is the 18-20 baume, when temperature is 30-35 ℃, for the first time with the 0.8-0.85 eluent eluting doubly of resin volume 3-3.5 hour, for the second time with the 0.5-0.6 eluent eluting doubly of resin volume 3-3.5 hour.
10. the preparation technology who utilizes the animal lung to extract heparin sodium crude as claimed in claim 1, it is characterized in that: described clarification filtration is meant that the pH value of adjusting eluent is 11-12, after stirring, leave standstill clarification, take out the supernatant, the bottom precipitation is carried out sucking filtration, merge clear liquid and filtrate, the adjusting pH value is 5-5.5, adds 3-3.5 80-85% ethanol clarifier doubly, leaves standstill till the clarification of upper strata liquid; Aspirate out the supernatant, collecting precipitation thing and sucking filtration are to doing.
11. the preparation technology who utilizes the animal lung to extract heparin sodium crude as claimed in claim 1, it is characterized in that: described drying is meant and places buchner funnel to drain sucking filtration to the heparin sodium precipitate of doing, put into exsiccator then, regularly replace till desiccant such as phosphorus pentoxide no longer absorb water, smash into graininess to pieces, promptly get heparin sodium crude.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 98112054 CN1238182A (en) | 1998-06-05 | 1998-06-05 | Process for extracting coarse lipo-hepinette from lung of animal |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 98112054 CN1238182A (en) | 1998-06-05 | 1998-06-05 | Process for extracting coarse lipo-hepinette from lung of animal |
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|---|---|
| CN1238182A true CN1238182A (en) | 1999-12-15 |
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| CN 98112054 Pending CN1238182A (en) | 1998-06-05 | 1998-06-05 | Process for extracting coarse lipo-hepinette from lung of animal |
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Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1111171C (en) * | 2000-09-07 | 2003-06-11 | 上海惠海生化制品厂 | Heparin and its preparing process |
| CN102212149A (en) * | 2011-06-15 | 2011-10-12 | 天津宝迪农业科技股份有限公司 | Preparation process for extracting crude sodium heparin from pig lungs |
| CN102295711A (en) * | 2011-08-23 | 2011-12-28 | 湖北宝迪农业科技有限公司 | Technology for extracting heparin sodium from pig lungs with polypeptide protein powder as combined product |
| CN102344502A (en) * | 2011-11-11 | 2012-02-08 | 宁发子 | Method for extracting heparin sodium by utilizing pork lungs |
| CN102462699A (en) * | 2010-11-01 | 2012-05-23 | 台湾糖业股份有限公司 | Use of porcine lung extract as matrix metalloproteinase inhibitor |
| CN102993336A (en) * | 2011-09-14 | 2013-03-27 | 浦江亚太肠衣有限公司 | Crude heparin sodium purification technology |
| CN103173512A (en) * | 2013-03-07 | 2013-06-26 | 安徽丰润生物技术有限公司 | Process for extracting Penis et Testis Canis polypeptide through enzymatic method, and obtained Penis et Testis Canis polypeptide extract |
| CN103450370A (en) * | 2013-08-31 | 2013-12-18 | 青岛九龙生物医药有限公司 | Method for separating high molecular weight heparin sodium by enzyme hydrolysis method |
| 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 |
| CN104448046A (en) * | 2014-12-24 | 2015-03-25 | 青岛九龙生物医药有限公司 | Production process for extracting crude heparin sodium products from animal lungs |
| CN104448049A (en) * | 2014-12-24 | 2015-03-25 | 青岛九龙生物医药有限公司 | Method for extracting heparin sodium from casing |
| CN106215983A (en) * | 2016-08-19 | 2016-12-14 | 山东万邦赛诺康生化制药股份有限公司 | The heparin sodium ion exchanger of air mixing and exchange method thereof |
| 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 |
-
1998
- 1998-06-05 CN CN 98112054 patent/CN1238182A/en active Pending
Cited By (19)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1111171C (en) * | 2000-09-07 | 2003-06-11 | 上海惠海生化制品厂 | Heparin and its preparing process |
| CN102462699B (en) * | 2010-11-01 | 2013-05-08 | 台湾糖业股份有限公司 | Use of porcine lung extract as matrix metalloproteinase inhibitor |
| CN102462699A (en) * | 2010-11-01 | 2012-05-23 | 台湾糖业股份有限公司 | Use of porcine lung extract as matrix metalloproteinase inhibitor |
| CN102212149A (en) * | 2011-06-15 | 2011-10-12 | 天津宝迪农业科技股份有限公司 | Preparation process for extracting crude sodium heparin from pig lungs |
| CN102212149B (en) * | 2011-06-15 | 2012-11-28 | 天津宝迪农业科技股份有限公司 | Preparation process for extracting crude sodium heparin from pig lungs |
| CN102295711A (en) * | 2011-08-23 | 2011-12-28 | 湖北宝迪农业科技有限公司 | Technology for extracting heparin sodium from pig lungs with polypeptide protein powder as combined product |
| CN102993336B (en) * | 2011-09-14 | 2014-06-25 | 浦江亚太肠衣有限公司 | Crude heparin sodium purification technology |
| CN102993336A (en) * | 2011-09-14 | 2013-03-27 | 浦江亚太肠衣有限公司 | Crude heparin sodium purification technology |
| CN102344502A (en) * | 2011-11-11 | 2012-02-08 | 宁发子 | Method for extracting heparin sodium by utilizing pork lungs |
| CN102344502B (en) * | 2011-11-11 | 2012-12-12 | 宁发子 | Method for extracting heparin sodium by utilizing pork lungs |
| CN103173512A (en) * | 2013-03-07 | 2013-06-26 | 安徽丰润生物技术有限公司 | Process for extracting Penis et Testis Canis polypeptide through enzymatic method, and obtained Penis et Testis Canis polypeptide extract |
| CN103450370A (en) * | 2013-08-31 | 2013-12-18 | 青岛九龙生物医药有限公司 | Method for separating high molecular weight heparin sodium by enzyme hydrolysis method |
| CN105461829A (en) * | 2013-08-31 | 2016-04-06 | 青岛九龙生物医药有限公司 | Method for separating high-molecular-weight heparin sodium by enzymolysis process |
| CN103450370B (en) * | 2013-08-31 | 2016-09-28 | 青岛九龙生物医药有限公司 | The method of enzymatic isolation method separation high molecular weight heparin sodium |
| 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 |
| CN104448046A (en) * | 2014-12-24 | 2015-03-25 | 青岛九龙生物医药有限公司 | Production process for extracting crude heparin sodium products from animal lungs |
| CN104448049A (en) * | 2014-12-24 | 2015-03-25 | 青岛九龙生物医药有限公司 | Method for extracting heparin sodium from casing |
| CN106215983A (en) * | 2016-08-19 | 2016-12-14 | 山东万邦赛诺康生化制药股份有限公司 | The heparin sodium ion exchanger of air mixing and exchange method thereof |
| 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 |
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