CN104650263A - Method for improving stability of refined heparin sodium 10% water solution by virtue of stabilizer adding process - Google Patents
Method for improving stability of refined heparin sodium 10% water solution by virtue of stabilizer adding process Download PDFInfo
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- CN104650263A CN104650263A CN201410819358.7A CN201410819358A CN104650263A CN 104650263 A CN104650263 A CN 104650263A CN 201410819358 A CN201410819358 A CN 201410819358A CN 104650263 A CN104650263 A CN 104650263A
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Abstract
The invention discloses a method for improving stability of a refined heparin sodium 10% water solution by virtue of a stabilizer adding process. The method can meet requirements proposed by customers from America on our company that the refined heparin sodium 10% water solution, after boiled in boiling water for 3h, is free from significant change in color, absorbance and active ingredients. The technical scheme is as follows: an appropriate stabilizer is added according to a certain proportion since the intermediate control of a heparin sodium refining process is qualified, so as to protect heparin from being damaged; and the produced 10% water solution, after being boiled in boiling water for 3h, is free from significant change in color, absorbance and active ingredients. Therefore, the method disclosed by the invention, which can meet customer requirements, has promoted export and won a good reputation for our company.
Description
Technical field
The present invention relates to biological technical field, relate in particular to and add suitable stablizer, the method that heparin is not destroyed can be protected.
Background technology
Heparin sodium is a kind of biochemical drug extracted from pig intestinal mucosa, be irreplaceable in operation, save the choice drug that life, market can not be out of stock.From nineteen forties be used for clinical since, its range of application constantly expands, and especially since nineteen nineties, clinical being mainly used in of this product prevents thrombosis, treatment cardiovascular diseases, hemopathy, uremia etc.Western countries have begun one's study the preventive and therapeutic effect of heparin sodium to cancer, and its novelty teabag constantly increases.
Since entering nineteen seventies, China's heparin sodium production technique is updated, and becomes the country that world's heparin sodium output is maximum.The heparin sodium product of world market has more than 70% from China.Refined heparin sodium is produced and is developed into hydrogen peroxide oxidation method by potassium permanganate oxidation method, in conjunction with alcohol wash partition method, is taken away by a large amount of impurity ethanol.On February 11st, 2008, the U.S. is because of the death of injecting heparin sodium appearance 4 example, and 350 many cases untoward reactions, become " the heparin sodium event " that cause a sensation the world.Through being expounded through peer review, above-mentioned death and untoward reaction are owing to containing caused by chondroitin polysulfate in heparin sodium.Chondroitin polysulfate is the one of heparitin, its character and heparin sodium basic simlarity, is: 1, heparin sodium is right-handed spiral configuration through detecting with the difference of heparin sodium; And chondroitin polysulfate is left-handed spiral configuration; 2, absolute molecular weight: chondroitin polysulfate is larger than the molecular weight of heparin sodium after testing; 3, its peakedness ratio heparin sodium of capillary electrophoresis first occurs.
The Chinese government attaches great importance to and pays close attention to impact development, April 7, Bureau of Drugs Supervision of country holds in Beijing " about reinforcement heparin sodium quality of production management symposium " national heparin sodium bulk drug and preparation manufacturing enterprise responsible official meeting, meeting discloses international expert and U.S. FDA about oversulfated chondroitin sulfate in sodium heparin to be people be add and indissociable identification, and require that every heparin sodium containing chondroitin polysulfate come into the market is recalled without exception.April 17 to 18 and April 24 to 25 have held again heparin sodium international symposium in the U.S., May 4, U.S. FDA official website announces, and the major cause of causing death is exactly chondroitin polysulfate, and also do not report so far relevant from heparin sodium by the technology of chondroitin polysulfate separation.
Summary of the invention
The invention provides a kind of method adding stablizer method protection heparin and be not destroyed; to make refined heparin sodium 10% aqueous solution boil 3 hours in boiling water; its color, absorbancy, effective constituent are all without larger change; utilize the character of heparin sodium in stablizer; when making it boil in boiling water, effective constituent is all without larger change, thus realizes adding stablizer method and making refined heparin sodium 10% aqueous solution in boiling water, boil still stable existence.
For solving the problems of the technologies described above, the present invention is achieved by the following technical solutions:
Add the method that stablizer method protection heparin is not destroyed, it is characterized in that: employing adds stablizer method makes heparin not be destroyed in boiling water.
In the technical scheme of invention, also there is following technical characteristic: described in add stablizer method and comprise the steps:
One, extract
1, dissolving crude product:
Open water valve, add a certain amount of tap water, then heparin sodium crude is joined in retort, stir while add, in raw material and tap water 1:(6-7) ratio dissolves, stirs after 5-10 hour, whether all dissolves bottom trying by stirring rod;
2, enzymolysis:
By previous step lysate, first adjust PH7.0-8.0 with hydrochloric acid soln, when being warming up between 50-55 DEG C, add stomach en-5-10g/ hundred million unit, then add 10-20g/ hundred million unit pancreatin, 50-55 DEG C of insulation 2-4 hour; According to 1:(500-700) ratio add heparin sodium stablizer.
3, rise suddenly temperature:
Previous step enzymolysis solution was warming up to 85-90 DEG C in 30 ~ 40 minutes, and static 10-30 minute, opens stirring, passes into circulating water cooling, when temperature is down to 50-55 DEG C, adjusts PH10.0-12.0, static layering 20-30 hour with 2-6mol/L sodium hydroxide solution;
4, the impurity of bottom settlings is centrifugal:
Siphon supernatant, with 40-100 order sock filtration, the solution after filtration is divided into supernatant liquor and lower sediment, and stay supernatant liquor to be precipitated, lower sediment puts centrifuge, stays upper strata centrifugate after centrifugal;
Two, refining
1, first time is precipitated:
Treat that above-mentioned supernatant liquor and centrifugate temperature are down to 20-30 DEG C, add 20-30g/L sodium-chlor, after stirring and dissolving, with hydrochloric acid, filtrate is adjusted PH10.0-12.0, stir while add the ethanolic soln that concentration is 95 ~ 97%, make the concentration of ethanol reach 45 ~ 50% 20 DEG C time, precipitation 10-12 hour;
2, first time oxidation: discard upper strata waste ethanol, lower sediment thing purified water is dissolved, and with sodium hydroxide solution, solution is adjusted PH10.0-12.0, then adds volume 3-5% hydrogen peroxide, oxidation 10-12 hour;
3, second time precipitation: solution adds 20-30g/L sodium-chlor, after stirring and dissolving, adjust solution PH 6.0-7.0 with hydrochloric acid soln, stir while add the ethanolic soln that concentration is 95 ~ 97%, the concentration of ethanol is made to reach 45 ~ 50% 20 DEG C time, precipitation 10-12 hour;
4, second time oxidation: discard upper strata waste ethanol, lower sediment thing purified water is dissolved, and with sodium hydroxide solution, solution is adjusted PH10.0-12.0, then adds volume 3-5% hydrogen peroxide, oxidation 10-12 hour;
5, third time is precipitated: solution adds 20-30g/L sodium-chlor, with hydrochloric acid soln, solution is adjusted PH10.0-12.0, stirs while add the ethanolic soln that concentration is 95 ~ 97%, makes the concentration of ethanol reach 45 ~ 50% 20 DEG C time, precipitation 10-12 hour;
6, third time oxidation: discard upper strata waste ethanol, lower sediment thing purified water is dissolved, and with sodium hydroxide solution, solution is adjusted PH10.0-12.0, then adds volume 3-5% hydrogen peroxide, oxidation 10-12 hour;
7, 4th precipitation: if previous step solution has throw out to separate out, then use centrifuge, do not separate out and then directly add 20-30g/L sodium-chlor, with hydrochloric acid soln, solution is adjusted PH6.0-7.0, solution is let cool in storehouse and be refrigerated to-10 DEG C--5 DEG C, while stir the acetone of Bian Jia-10 DEG C--5 DEG C, the mass percentage making acetone account for solution is 40-45%,-5 DEG C-0 DEG C insulation 24-30 hour, then waste acetone is discarded, chondroitin polysulfate is taken away by acetone, portion's throw out purified water of keeping on file is dissolved, solution adds 20-30g/L sodium-chlor again, adjust solution PH 6.0-7.0 with hydrochloric acid soln, stir while add ethanol, alcohol concn is made to be 45-50% 20 DEG C time, precipitation 10-12 hour,
8, after throw out being pressed the dissolving of 2-5L/ hundred million unit by purified water, with aperture 0.22-0.3 micron membrane filter board and frame machine micro-filtration, with alcohol settling, alcohol concn is made to reach 75-80%, add the sodium chloride solution of 23-26% again, every 1 liter of solution adds 6-10ml sodium chloride solution, quiescent setting 10-12 hour;
9, upper strata ethanol is discarded, add dehydrated alcohol dehydration, stir while add, make alcohol concn be 97-99%, static 24-30 hour, discards upper strata ethanol, stainless steel core rod is put in the product, by pipeline and filter flask, residue ethanol is drained with vacuum pump, to be dried;
10, the product drained evenly is placed in Stainless Steel Disc, is placed in vacuum drying oven and vacuumizes, heat with recirculated water, temperature 35-60 DEG C, dry 70-80 hour;
11, pack: product is taken out, weighs, make every packing bag 5kg, with sealing machine sealing, be placed in Aluminum Drum to be tested.
In the technical scheme of invention, also there is following technical characteristic: the solution after the 4th precipitation carries out QA sample examination, surveys absorbancy: 260nm < 0.1; 400nm < 0.02; If defective, continue to repeat process above, if qualified transfer subsequent processing.
Compared with prior art, advantage of the present invention and positively effect are:
Refined heparin sodium of the present invention is produced and is adopted hydrogen peroxide oxidation method, in conjunction with alcohol wash partition method, a large amount of impurity ethanol is taken away, with extraction by propanone, the chondroitin polysulfate in heparin sodium is removed again, its fine work yield reaches more than 90%, activity is tired at more than 180usp/mg, amounts to WHO international standard at 200 iu/mg, and its quality standard meets the indices of Chinese Pharmacopoeia, American Pharmacopeia, British Pharmacopoeia and West Europe pharmacopeia defined.
Embodiment
Below in conjunction with specific embodiment, the invention will be further described.
Embodiment 1
One, extract
1, dissolving crude product:
Open water valve, add 900L tap water, then 15,000,000,000 units of heparin sodium crude products are joined in retort, stir while add, stir after 6 hours, whether all dissolve with stirring rod examination bottom;
2, enzymolysis:
By previous step lysate, first adjust PH7.5 with hydrochloric acid soln, add stomach en-1500g when being warming up to 53 DEG C, then add 300g pancreatin, 53 DEG C are incubated 3 hours;
3, rise suddenly temperature:
Previous step enzymolysis solution is warming up to 88 DEG C in 35 minutes, static 15 minutes, opens stirring, pass into circulating water cooling, when temperature is down to 50 DEG C, adjust PH11.0 with 6mol/L sodium hydroxide solution, static layering 24 hours;
4, the impurity of bottom settlings is centrifugal:
Siphon supernatant, with 80 order sock filtration, the solution after filtration is divided into supernatant liquor and lower sediment, and stay supernatant liquor to be precipitated, lower sediment puts centrifuge, stays upper strata centrifugate after centrifugal;
Two, refining
1, first time is precipitated:
Treat that above-mentioned supernatant liquor and centrifugate temperature are down to 25 DEG C, add 22.5kg sodium-chlor, after stirring and dissolving, with hydrochloric acid, filtrate is adjusted PH10.5, stir while add the ethanolic soln 920L that concentration is 95 ~ 97%, precipitate 10 hours;
2, first time oxidation: discard upper strata waste ethanol, lower sediment thing and product 640L purified water are dissolved, and with sodium hydroxide solution, solution are adjusted PH11.0, then add 20L hydrogen peroxide, be oxidized 10 hours;
3, second time precipitation: solution adds 12.8kg sodium-chlor, after stirring and dissolving, adjusts solution PH 6.0 with hydrochloric acid soln, stirs while add the ethanolic soln 640L that concentration is 95 ~ 97%, precipitate 10 hours;
4, second time oxidation: discard upper strata waste ethanol, lower sediment thing 640L purified water is dissolved, and with sodium hydroxide solution, solution is adjusted PH11.0, then adds 20L hydrogen peroxide, be oxidized 10 hours;
5, third time is precipitated: solution adds 12.8kg sodium-chlor, with hydrochloric acid soln, solution is adjusted PH6.0, stirs while add the ethanolic soln 640L that concentration is 95 ~ 97%, precipitates 10 hours;
6, third time oxidation: discard upper strata waste ethanol, lower sediment thing 640L purified water is dissolved, and with sodium hydroxide solution, solution is adjusted PH11.0, then adds 21L hydrogen peroxide, be oxidized 10 hours;
7, the 4th precipitation: if previous step solution has throw out to separate out, then use centrifuge, do not separate out and then directly add 12.8kg sodium-chlor, with hydrochloric acid soln, solution is adjusted PH6.0, solution is let cool in storehouse and is refrigerated to-6 DEG C, while stir the acetone 600L of Bian Jia-10 DEG C,-3 DEG C are incubated 24 hours, then discarded by waste acetone, chondroitin polysulfate is taken away by acetone, and portion's throw out purified water of keeping on file is dissolved, solution adds 12.8kg sodium-chlor again, adjust solution PH 6.0 with hydrochloric acid soln, stir while add ethanol 640L, precipitate 10 hours;
8, after throw out being dissolved by 300L purified water, with aperture 0.3 micron membrane filter board and frame machine micro-filtration, use 840L alcohol settling, make alcohol concn reach 75-80%, then add the sodium chloride solution 11L of 26%, quiescent setting 10 hours;
9, upper strata ethanol is discarded, add the dehydration of 500L dehydrated alcohol, stir while add, make alcohol concn be 97-99%, static 24 hours, discard upper strata ethanol, stainless steel core rod is put in the product, by pipeline and filter flask, residue ethanol is drained with vacuum pump, to be dried;
10, the product drained evenly is placed in Stainless Steel Disc, is placed in vacuum drying oven and vacuumizes, heat with recirculated water, temperature 50 C, dry 72 hours;
11, pack: product is taken out, weighs, make every packing bag 5Kg, with sealing machine sealing, be placed in Aluminum Drum to be tested.
The above is only preferred embodiment of the present invention, and be not restriction the present invention being made to other form, any those skilled in the art may utilize the technology contents of above-mentioned announcement to be changed or be modified as the Equivalent embodiments of equivalent variations.But everyly do not depart from technical solution of the present invention content, any simple modification, equivalent variations and the remodeling done above embodiment according to technical spirit of the present invention, still belong to the protection domain of technical solution of the present invention.
Claims (4)
1. add the method that suitable stablizer makes refined heparin sodium 10% aqueous solution boiled products still stable existence, it is characterized in that: in heparin sodium production process, add stablizer.
2. according to claim 1ly add suitable stablizer method and make refined heparin sodium 10% aqueous solution in boiling water, boil 3 hours, its color, absorbancy, effective constituent, all without the method for larger change, is characterized in that adding suitable stabilizers in heparin sodium abstraction process.
3. add stablizer in the production process of heparin sodium more according to claim 1, it is characterized in that the stablizer adding a kind of mixing element.
4. add stablizer in the production process of heparin sodium more according to claim 1, it is characterized in that the ratio added is 1:(500-700).
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103145879A (en) * | 2012-12-08 | 2013-06-12 | 青岛九龙生物医药有限公司 | Method of improving thermal stability for heparin sodium with reduction method |
| CN103694373A (en) * | 2013-11-23 | 2014-04-02 | 青岛九龙生物医药有限公司 | Method for removing DS impurity in refined heparin sodium |
| CN103804520A (en) * | 2013-11-22 | 2014-05-21 | 青岛九龙生物医药有限公司 | Method for improving test index of heparin sodium competitive product 260nm absorbancy |
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- 2014-12-25 CN CN201410819358.7A patent/CN104650263A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103145879A (en) * | 2012-12-08 | 2013-06-12 | 青岛九龙生物医药有限公司 | Method of improving thermal stability for heparin sodium with reduction method |
| CN103804520A (en) * | 2013-11-22 | 2014-05-21 | 青岛九龙生物医药有限公司 | Method for improving test index of heparin sodium competitive product 260nm absorbancy |
| CN103694373A (en) * | 2013-11-23 | 2014-04-02 | 青岛九龙生物医药有限公司 | Method for removing DS impurity in refined heparin sodium |
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