CN105461827A - Alkali-enzymolysis method for extracting sodium chondroitin sulfate - Google Patents
Alkali-enzymolysis method for extracting sodium chondroitin sulfate Download PDFInfo
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- CN105461827A CN105461827A CN201511002245.9A CN201511002245A CN105461827A CN 105461827 A CN105461827 A CN 105461827A CN 201511002245 A CN201511002245 A CN 201511002245A CN 105461827 A CN105461827 A CN 105461827A
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- cartilage
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- chondroitin sulfate
- sodium chondroitin
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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
- C08B37/0069—Chondroitin-4-sulfate, i.e. chondroitin sulfate A; Dermatan sulfate, i.e. chondroitin sulfate B or beta-heparin; Chondroitin-6-sulfate, i.e. chondroitin sulfate C; Derivatives thereof
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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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- Polysaccharides And Polysaccharide Derivatives (AREA)
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Abstract
The invention relates to a novel extraction method of sodium chondroitin sulfate, particularly sodium chondroitin sulfate extracted by alkali-enzymolysis. The method mainly solves the problems of complex process, high alkali/acid or salt consumption, higher production cost, long extraction period, low product quality stability, lower yield and the like in the current like product production technique. The method mainly comprises the following steps: by using animal cartilage as a raw material, carrying out alkaline hydrolysis and one-step enzymolysis, thereby extracting the sodium chondroitin sulfate from the animal cartilage. The product has the advantages of white color, loose powder, high yield (up to 30% or above) and high purity, and conforms to the quality standard for oral administration. The method has the characteristics of simple production procedure, short period, low cost and the like, is simple to operate and easy to implement, is helpful to relieving the environmental pollution, and is beneficial to environment friendliness.
Description
Technical field
The present invention relates to biochemical pharmacy field, particularly direct production technique cardiovascular and cerebrovascular disease etc. being had to the Sodium chondroitin sulfate A of good effect extracted from animal cartilage.
Background technology
Sodium chondroitin sulfate A, English name Chondroitinsulfatesodium, it is the acidic mucopolysaccharide extracted in animal cartilage, be present in the cartilage of animal, larynx bone, nasal bone (41%), ox, Malaysia and China's diaphragm and tracheae, the polysaccharide chain that its molecular structure consists of the repetition disaccharide unit that β 1 → 3 glycosidic link is formed for β-D-Glucose aldehydic acid and N-acetylamino galactosamine more.
Sodium chondroitin sulfate A is mainly used in the medicine for the treatment of rheumatism and rheumatism, also has the effects such as anti-freezing, anticancer, antithrombotic, reducing blood-fat, can be used for the symptoms such as treatment headache, migraine, coronary heart diseases and angina pectoris.Abroad, molecular level is reached to the research of Sodium chondroitin sulfate A pharmacology.
The method of current domestic extraction Sodium chondroitin sulfate A mainly contains three kinds: salt solution, alkaline hydrolysis and enzymolysis process.The separating and purifying technology of main employing has: ethanol precipitation, ion exchange chromatography, Mierocrystalline cellulose partition method, quaternary ammonium salt companion method, absorption method, zone electrophoresis partition method etc.In above concentrated extracting method, although yield can reach very high level, but all there is very large problem in production method, as existed except acidic protein regulates pH value 1-2 consuming time very long when product volume is large in salt solution technique, amino in Sodium chondroitin sulfate A is easily dissociated, thus have impact on yield and quality.
Summary of the invention
The object of the invention is: the extracting method that a kind of Sodium chondroitin sulfate A is provided, adopt this extracting method can not only overcome above-mentioned the deficiencies in the prior art, and can reduce costs, improve the quality of products, shorten the production cycle, enhance productivity.
Technical solution of the present invention is that this preparation method comprises the following steps:
1, soak: be placed in reactor by through the pure white clean animal cartilage block of pre-treatment, and add appropriate water, soak 5-6 hour.
2, alkaline hydrolysis: will soak animal cartilage completely, adds water by the 1:1 of weight in cartilage, and adds alkali sheet by the 4-6% of cartilage weight, stirs 3-5 hour.
3, enzymolysis: above-mentioned alkali solution liquid is adjusted PH8.0-8.5, is heated to 45-50 DEG C, adds Pancreas Sus domestica, and add sodium-chlor by the 12-15% of cartilage weight, in 45-50 DEG C of insulation 3-4 hour.
4, reduce: by above-mentioned enzymolysis solution, adjust PH6.5-7.0, be heated to 65 DEG C, add the V-Brite B of 1% of cartilage weight, stir 15 minutes, leave standstill after 20 minutes and filter.
5, precipitate: the filtrate of upper step gained is lowered the temperature less than 30 DEG C, adds ethanol, at 20 DEG C, make the precipitation number of degrees reach 60 degree, precipitation 6-8 hour.
6, multiple heavy: upper step throw out to be added ethanol, makes the precipitation number of degrees reach 80 degree, leave standstill 10-12 hour.
7, dehydrate: merge throw out, add dehydrated alcohol dehydration, after drying, namely obtain described Sodium chondroitin sulfate A finished product.
Present method a little and effect: present method is by reformed AHP to current technique, alkaline hydrolysis is adopted to produce Sodium chondroitin sulfate A in conjunction with the method for enzymolysis, optimized production process condition: the alkaline hydrolysis time is 4 hours, alkaline hydrolysis temperature is 48 DEG C, Pancreas Sus domestica consumption is 14% of animal cartilage weight, and enzymolysis time is 4 hours, and hydrolysis temperature is 48 DEG C, be mainly alkaline hydrolysis temperature to alkaline hydrolysis influence degree in five kinds of factors, topmost on enzymolysis impact is Pancreas Sus domestica consumption.Whole reaction process pH value is between 8.0-8.5.The combination of this optimum level can make Sodium chondroitin sulfate A yield reach more than 35 or higher.After adopting new technology, greatly shorten the production cycle, improve production efficiency, save production cost, improve the quality of production.
Embodiment
Embodiment 1
(1) soak: be placed in reactor by through the pure white clean animal cartilage block 100kg of pre-treatment, and add appropriate water, soak 5 hours;
(2) alkaline hydrolysis: will soak animal cartilage completely, adds water by the 1:1 of weight in cartilage, and adds alkali sheet by 4% of cartilage weight, stirs 3 hours;
(3) enzymolysis: above-mentioned alkali solution liquid is adjusted PH8.0, is heated to 45 DEG C, adds Pancreas Sus domestica, and add sodium-chlor by 12% of cartilage weight, in 45 DEG C of insulations 3 hours;
(4) reduce: by above-mentioned enzymolysis solution, adjust PH6.5, be heated to 65 DEG C, add the V-Brite B of 1% of cartilage weight, stir 15 minutes, leave standstill after 20 minutes and filter;
(5) precipitate: the filtrate of upper step gained is lowered the temperature less than 30 DEG C, adds ethanol, at 20 DEG C, make the precipitation number of degrees reach 60 degree, precipitate 6 hours;
(6) multiple heavy: upper step throw out to be added ethanol, makes the precipitation number of degrees reach 80 degree, leave standstill 10 hours;
(7) dehydrate: merge throw out, add dehydrated alcohol dehydration, after drying, namely obtain described Sodium chondroitin sulfate A finished product.
Embodiment 2
(1) soak: be placed in reactor by through the pure white clean animal cartilage 100kg block of pre-treatment, and add appropriate water, soak 6 hours;
(2) alkaline hydrolysis: will soak animal cartilage completely, adds water by the 1:1 of weight in cartilage, and adds alkali sheet by 5% of cartilage weight, stirs 4 hours;
(3) enzymolysis: above-mentioned alkali solution liquid is adjusted PH8.5, is heated to 50 DEG C, adds Pancreas Sus domestica, and add sodium-chlor by 15% of cartilage weight, in 50 DEG C of insulations 4 hours;
(4) reduce: by above-mentioned enzymolysis solution, adjust PH7.0, be heated to 65 DEG C, add the V-Brite B of 1% of cartilage weight, stir 15 minutes, leave standstill after 20 minutes and filter;
(5) precipitate: the filtrate of upper step gained is lowered the temperature less than 30 DEG C, adds ethanol, at 20 DEG C, make the precipitation number of degrees reach 60 degree, precipitate 7 hours;
(6) multiple heavy: upper step throw out to be added ethanol, makes the precipitation number of degrees reach 80 degree, leave standstill 11 hours;
(7) dehydrate: merge throw out, add dehydrated alcohol dehydration, after drying, namely obtain described Sodium chondroitin sulfate A finished product.
Embodiment 3
(1) soak: be placed in reactor by through the pure white clean animal cartilage 100kg block of pre-treatment, and add appropriate water, soak 6 hours;
(2) alkaline hydrolysis: will soak animal cartilage completely, adds water by the 1:1 of weight in cartilage, and adds alkali sheet by 6% of cartilage weight, stirs 4 hours;
(3) enzymolysis: above-mentioned alkali solution liquid is adjusted PH8.5, is heated to 48 DEG C, adds Pancreas Sus domestica, and add sodium-chlor by 14% of cartilage weight, in 50 DEG C of insulations 4 hours;
(4) reduce: by above-mentioned enzymolysis solution, adjust PH7.0, be heated to 65 DEG C, add the V-Brite B of 1% of cartilage weight, stir 15 minutes, leave standstill after 20 minutes and filter;
(5) precipitate: the filtrate of upper step gained is lowered the temperature less than 30 DEG C, adds ethanol, at 20 DEG C, make the precipitation number of degrees reach 60 degree, precipitate 8 hours;
(6) multiple heavy: upper step throw out to be added ethanol, makes the precipitation number of degrees reach 80 degree, leave standstill 12 hours
(7) dehydrate: merge throw out, add dehydrated alcohol dehydration, after drying, namely obtain described Sodium chondroitin sulfate A finished product.
Analyzed by the Sodium chondroitin sulfate A that above-mentioned 3 embodiments are produced, result is as follows:
| Project | Example 1 result | Example 2 result | Example 3 result |
| Specific optical rotation | -25.4o | -26.2o | -27.1o |
| 420nm absorbancy | 0.21 | 0.185 | 0.155 |
| Acidity | 6.76 | 6.55 | 6.25 |
| Total nitrogen | 3.2% | 2.8% | 2.6% |
| Weight loss on drying | 9.6% | 8.5% | 7.9% |
| Residue on ignition | 28.5% | 27.6% | 23.4% |
| Organic residue | 4210ppm | 3514ppm | 2010ppm |
| CPC content | 93.1% | 94.2% | 95.8% |
| HPLC content | 92.8% | 94.0% | 96.3% |
Claims (6)
1. extract a method for Sodium chondroitin sulfate A, it is characterized in that: be dissolving completely of extracting from animal cartilage, specific rotation is-25 ° to-35 °, and itself HPLC and CPC content is all greater than the white powder of 95%.
2. method according to claim 1, is characterized in that the method comprises successively:
(1) be placed in reactor by through the pure white clean animal cartilage block of pre-treatment, and add appropriate water, soak 5-6 hour;
(2) animal cartilage completely will be soaked, add water by the 1:1 of weight in cartilage, and add the alkali sheet of the 4-6% of cartilage weight, stir 3-5 hour;
(3) above-mentioned alkali solution liquid is adjusted PH8.0-8.5, be heated to 45-50 DEG C, after adding the Pancreas Sus domestica of the 12-15% of cartilage weight, add sodium-chlor, in 45-50 DEG C of insulation 3-4 hour;
(4) by above-mentioned enzymolysis solution, adjust PH6.5-7.0, be heated to 65 DEG C, add the V-Brite B of 1% of cartilage weight, stir 15 minutes, leave standstill after 20 minutes and filter;
(5) filtrate of upper step gained is lowered the temperature less than 30 DEG C, add ethanol, at 20 DEG C, make the precipitation number of degrees reach 60 degree, precipitation 6-8 hour;
(6) upper step throw out is added ethanol, make the precipitation number of degrees reach 80 degree, leave standstill 10-12 hour;
(7) merge throw out, add dehydrated alcohol dehydration, after drying, namely obtain described Sodium chondroitin sulfate A finished product.
3. method according to claim 1 and 2, is characterized in that in (2) step, add the alkali sheet that weight is 4-6%, to strengthen the alkaline hydrolysis effect of Sodium chondroitin sulfate A.
4. method according to claim 1 and 2, is characterized in that in (4) step, add weight 1% V-Brite B saltouts, and has strengthened the sedimentation effect of Sodium chondroitin sulfate A.
5. method according to claim 1 and 2, is characterized in that best alkaline hydrolysis temperature is 48 DEG C.
6. method according to claim 1 and 2, is characterized in that best Pancreas Sus domestica consumption is the dry cartilage weight of 12-15%.
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| CN201310615924.8A CN103665188B (en) | 2013-11-23 | 2013-11-23 | Alkali-enzymolysis process extracts Sodium chondroitin sulfate A |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1310186A (en) * | 2001-01-19 | 2001-08-29 | 华北工学院 | pectin producing method |
| CN101348815A (en) * | 2008-09-03 | 2009-01-21 | 南京工业大学 | Process for preparing chondroitin sulfate by natural enzyme and protease synergistic method |
| CN103130913A (en) * | 2013-02-17 | 2013-06-05 | 暨南大学 | Calcium pectinate and production method and application thereof |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1182593A (en) * | 1996-11-18 | 1998-05-27 | 王立杰 | Chondroitiln sulfate and extrating method |
| JP4580688B2 (en) * | 2004-05-27 | 2010-11-17 | 株式会社日本バリアフリー | Method for producing chondroitin sulfate |
| CN1273497C (en) * | 2005-06-29 | 2006-09-06 | 上海大学 | Process for preparing chondroitin sulfate |
| CN101348814B (en) * | 2008-09-03 | 2012-03-14 | 南京工业大学 | Method for improving quality of chondroitin sulfate by using isoelectric points of proteins |
| CN101696246B (en) * | 2009-10-23 | 2011-09-28 | 于燕莉 | Method for producing chondroitin sulfate |
| CN103214595B (en) * | 2013-04-11 | 2015-09-23 | 重庆奥力生物制药有限公司 | The preparation method of Sodium chondroitin sulfate A |
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2013
- 2013-11-23 CN CN201511002245.9A patent/CN105461827A/en active Pending
- 2013-11-23 CN CN201310615924.8A patent/CN103665188B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1310186A (en) * | 2001-01-19 | 2001-08-29 | 华北工学院 | pectin producing method |
| CN101348815A (en) * | 2008-09-03 | 2009-01-21 | 南京工业大学 | Process for preparing chondroitin sulfate by natural enzyme and protease synergistic method |
| CN103130913A (en) * | 2013-02-17 | 2013-06-05 | 暨南大学 | Calcium pectinate and production method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 崔英德等: "《漂白剂及其应用》", 28 February 1999, 中山大学出版社 * |
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| CN103665188A (en) | 2014-03-26 |
| CN103665188B (en) | 2016-04-13 |
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Application publication date: 20160406 |