CN1544645A - Double-enzyme method for preparing chondroitin sulfate - Google Patents
Double-enzyme method for preparing chondroitin sulfate Download PDFInfo
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- CN1544645A CN1544645A CNA2003101153839A CN200310115383A CN1544645A CN 1544645 A CN1544645 A CN 1544645A CN A2003101153839 A CNA2003101153839 A CN A2003101153839A CN 200310115383 A CN200310115383 A CN 200310115383A CN 1544645 A CN1544645 A CN 1544645A
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- chondroitin sulfate
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Abstract
The invention relates to a biochemical medicine chondroitin sulfate extracting method, i.e. it uses a double-enzyme method to produce chondroitin sulfate. It mainly uses animal cartilages as raw material and is made by the working procedures of basic compound enzyme and trypsin enzymolysis, thermal treatment, acid processing, filtering with ultrafiltration membrane, crystallization, drying, etc. It adopts new techniques of double-enzyme enzymolysis and ultrafiltration, therefore the products are pure white in color, and the product powder is loose and the yield of the product is 165-22%, and the products meet oral quality standard. And it has the characters of short cycle, low cost, and simple and convenient and easy operation.
Description
Technical field:
The present invention relates to the biochemical pharmacy field, particularly be chondroitin sulfate and the production method thereof that cardiovascular and cerebrovascular disease etc. is had good effect.Belong to International Classification of Patents C08B37/08 " Preparation of chondroitin sulfate " technical field.
Background technology:
Chondroitin sulfate mainly is the medicine of treatment rheumatism and rheumatism, and it also has effects such as anti-freezing, anti-class, anticancer, antithrombotic, reducing blood-fat.Therefore be subjected to generally paying attention to both at home and abroad.The method of present domestic extraction chondroitin sulfate mainly contains three kinds: salt solution, alkaline hydrolysis method and enzymolysis process.The main separating and purifying technology that adopts has: ethanol precipitation method, Mierocrystalline cellulose partition method, absorption method, quaternary amine complexometry, ion exchange chromatography, zone electrophoresis partition method etc.In the current technology, it is consuming time very long when product volume is big that existence removes acid albumen adjusting pH value 1-2 in the salt solution technology, and the amino in the chondroitin sulfate is easily disintegrated down, thereby has influenced yield and quality; Owing to use atlapulgite or gac to be sorbent material, when removing protein and polypeptide, also the part chrondroitin is adsorbed to fall in the alkaline hydrolysis method technology, thereby yield is reduced; In traditional enzymolysis-resin extraction method, thereby owing to adopt expensive resin that cost is increased, and complex steps; Patent CN1182593A is about in " chondroitin sulfate and extracting method thereof ", adopt hydrochloride to separate, add when filtering calcium chloride, other adds hydrogen peroxide filtrate is handled, improved quality product, but owing to add calcium chloride and two kinds of chemical feedstockss of hydrogen peroxide, thereby cause the increase of recurrent expenditure.
Summary of the invention:
The objective of the invention is to overcome above-mentioned the deficiencies in the prior art, and provide that a kind of cost is low, quality good, chondroitin sulfate production method easy and simple to handle.
Technical scheme of the present invention is as follows.
A kind of chondroitin sulfate is characterized in a kind of that extract, its specific rotation-20 to-35 ℃ from animal cartilage, glucosamine content is the white powder of 24-30%.
A kind of production method of chondroitin sulfate is characterized in:
(1) will place enzymatic vessel through the broken cartilage of pre-treatment cleaning, add a kind of homemade alkaline prozyme, under 55 ℃, enzymolysis stirring and dissolving 12 hours.
(2) above-mentioned enzymolysis solution being regulated pH is 6.5, coarse crystallization in 70% ethanol.
(3) add pancreatin in the enzymolysis solution of process (2), regulating pH was 8.5,53 ℃ of following enzymolysis 4 to 5 hours.
(4) above-mentioned material is heated to 85 ℃, heated 15 to 20 minutes, filter after heat-treating.
(5) process (4) gained filtrate is regulated pH2.5 and is carried out acid treatment, leaves standstill 2 hours after-filtration.
(6) with hollow fiber ultrafiltration membrane process (5) gained filtrate is carried out ultra-filtration and be concentrated into half volume.
(7) regulate pH6.5, carry out crystallization in 70% ethanol, the gained crystallization was 55 ℃ of warm air dryings 6 hours, and finished product is pulverized the packing sterilization and preserved.
The add-on of above-mentioned chondroitin sulfate extracting method neutral and alkali prozyme and pancreatin is the 0.5-1.5% of raw material cartilage amount; Adopt the MNWC6000 hollow fiber ultrafiltration membrane during ultrafiltration.
The alkaline prozyme that the present invention uses is self-control, its preparation process can be sketched and be: will contain the Sumizyme MP of Bacillus subtillis or the species of alkaline lipase and carry out fermentation culture,, get its supernatant liquor and concentrate, add adsorbents adsorb, drying obtains alkaline compound enzymic preparation under vacuum state after filtering.
Embodiment:
Embodiment:
1, the hog snout bone 155Kg of cleaning is added in the enzymatic vessel, add alkaline prozyme 1.55Kg, add appropriate amount of deionized water till the complete submergence of bone slag, stirred 12 hours at 55 ℃ of following enzymolysis.
2, above-mentioned alkaline complex enzyme hydrolysis liquid is regulated pH6.5, coarse crystallization in 70% ethanol.
3, the pancreatin that adds 1.55Kg in the enzymolysis solution of process 2 was regulated pH8.5,53 ℃ of following enzymolysis 4 to 5 hours.
4, above material is heated to 85 ℃, heated 15 to 20 minutes, heat-treat after-filtration.
5, process 4 gained filtrates adjusting pH2.5 handles, and leaves standstill 2 hours after-filtration.
6, with hollow fiber ultrafiltration membrane process 5 gained filtrates are carried out ultra-filtration and be concentrated into half volume.
7, regulate pH6.5, crystallization in 70% ethanol, the gained crystallization was 55 ℃ of warm air dryings 6 hours, and finished product is pulverized the packing sterilization and is preserved.
Advantage of the present invention is a lot.Owing to adopt alkaline prozyme in the technology, concentrate with pancreatin enzymolysis, ultrafilter membrane filtering and impurity removing again behind the enzymolysis, product yield height, the outward appearance that obtains is pure white, powder is loose, and production operation is simple and easy to do, and process is easily controlled, and cost is lower, and is with short production cycle.
Adopt above its detected result of method products obtained therefrom to be:
Yield (to dried cartilage) 16-17%
The appearance white powder
Solubleness (2%H
2O) complete
Dry moisture＜10%
Protein＜3.5%
Total count＜1000/g
Sulphur content (dry product meter)＜6.4%
Nitrogen content (dry product meter)＜3.3%
Heavy metal＜1.0 * 10
-5
Glucosamine 24-30%
By above-mentioned detected result analysis, product meets oral level index request.
Claims (5)
1, a kind of production method of chondroitin sulfate is characterized in that: be dissolving fully of extracting from animal cartilage, specific rotation is-20 to-35 ℃, and glucosamine content is the white powder of 24-30%.
2, the production method of chondroitin sulfate according to claim 1 is characterized in that: its extraction step is
(1) will place enzymatic vessel through the broken cartilage of pre-treatment cleaning, add a kind of homemade alkaline prozyme, under 55 ℃, enzymolysis stirring and dissolving 12 hours;
(2) above-mentioned enzymolysis solution being regulated pH is 6.5, coarse crystallization in 70% ethanol;
(3) add pancreatin in the enzymolysis solution of process (2), regulating pH was 8.5,53 ℃ of following enzymolysis 4 to 5 hours;
(4) above-mentioned material is heated to 85 ℃, heated 15 to 20 minutes, filter after heat-treating;
(5) process (4) gained filtrate is regulated pH2.5 and is carried out acid treatment, leaves standstill 2 hours after-filtration;
(6) with hollow fiber ultrafiltration membrane process (5) gained filtrate is carried out ultra-filtration and be concentrated into half volume;
(7) regulate pH6.5, carry out crystallization in 70% ethanol, the gained crystallization was 55 ℃ of warm air dryings 6 hours, and finished product is pulverized the packing sterilization and preserved.
3, the production method of chondroitin sulfate according to claim 1, it is characterized in that: the alkaline prozyme of described self-control, its preparation process can be sketched and be: will contain the Sumizyme MP of Bacillus subtillis or the species of alkaline lipase and carry out fermentation culture, getting its supernatant liquor concentrates, add adsorbents adsorb, drying obtains alkaline compound enzymic preparation under vacuum state after filtering.
4, the production method of chondroitin sulfate according to claim 2 is characterized in that: the add-on of alkaline prozyme and pancreatin is the 0.5-1.5% of raw material cartilage amount.
5, the production method of chondroitin sulfate according to claim 2 is characterized in that: adopt the MNWC6000 hollow fiber ultrafiltration membrane during ultra-filtration.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2003101153839A CN1544645A (en) | 2003-11-21 | 2003-11-21 | Double-enzyme method for preparing chondroitin sulfate |
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| Application Number | Priority Date | Filing Date | Title |
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| CNA2003101153839A CN1544645A (en) | 2003-11-21 | 2003-11-21 | Double-enzyme method for preparing chondroitin sulfate |
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| CN1544645A true CN1544645A (en) | 2004-11-10 |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100417728C (en) * | 2006-06-01 | 2008-09-10 | 扬州日兴生物化工制品有限公司 | Deacetylate chondroitin sulfate preparation technology |
| CN101348815B (en) * | 2008-09-03 | 2011-09-07 | 南京工业大学 | Process for preparing chondroitin sulfate by natural enzyme and protease synergistic method |
| CN102838691A (en) * | 2012-08-29 | 2012-12-26 | 宁波绿之健药业有限公司 | Extracting and purifying method of chondroitin sulfate |
| WO2016090704A1 (en) * | 2014-12-08 | 2016-06-16 | 张木良 | Chondroitin sulfate preparation process and equipment thereof |
| CN109402205A (en) * | 2018-09-04 | 2019-03-01 | 如皋市桦儒肠衣有限公司 | A kind of technique for extracting heparan or albumen from heparin byproduct |
| CN112301079A (en) * | 2020-11-09 | 2021-02-02 | 湖南夏盛酶技术有限公司 | Method for producing halal chondroitin sulfate by microbial enzyme method |
| CN113429494A (en) * | 2021-08-15 | 2021-09-24 | 四川农业大学 | Method for extracting chondroitin sulfate from pig trachea |
| WO2023082523A1 (en) * | 2021-11-15 | 2023-05-19 | 广东海洋大学 | Method for improving extraction rate of chondroitin sulfate prepared from tilapia skull |
-
2003
- 2003-11-21 CN CNA2003101153839A patent/CN1544645A/en active Pending
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100417728C (en) * | 2006-06-01 | 2008-09-10 | 扬州日兴生物化工制品有限公司 | Deacetylate chondroitin sulfate preparation technology |
| CN101348815B (en) * | 2008-09-03 | 2011-09-07 | 南京工业大学 | Process for preparing chondroitin sulfate by natural enzyme and protease synergistic method |
| CN102838691A (en) * | 2012-08-29 | 2012-12-26 | 宁波绿之健药业有限公司 | Extracting and purifying method of chondroitin sulfate |
| WO2016090704A1 (en) * | 2014-12-08 | 2016-06-16 | 张木良 | Chondroitin sulfate preparation process and equipment thereof |
| CN109402205A (en) * | 2018-09-04 | 2019-03-01 | 如皋市桦儒肠衣有限公司 | A kind of technique for extracting heparan or albumen from heparin byproduct |
| CN109402205B (en) * | 2018-09-04 | 2022-01-28 | 如皋市桦儒肠衣有限公司 | Process for extracting heparinoid or protein from heparin by-product |
| CN112301079A (en) * | 2020-11-09 | 2021-02-02 | 湖南夏盛酶技术有限公司 | Method for producing halal chondroitin sulfate by microbial enzyme method |
| CN113429494A (en) * | 2021-08-15 | 2021-09-24 | 四川农业大学 | Method for extracting chondroitin sulfate from pig trachea |
| WO2023082523A1 (en) * | 2021-11-15 | 2023-05-19 | 广东海洋大学 | Method for improving extraction rate of chondroitin sulfate prepared from tilapia skull |
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