CN103601815A - Extraction of sodium chondroitin sulfate through combination of alkaline hydrolysis-enzymolysis method and flocculation precipitation method - Google Patents
Extraction of sodium chondroitin sulfate through combination of alkaline hydrolysis-enzymolysis method and flocculation precipitation method Download PDFInfo
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- CN103601815A CN103601815A CN201310508887.0A CN201310508887A CN103601815A CN 103601815 A CN103601815 A CN 103601815A CN 201310508887 A CN201310508887 A CN 201310508887A CN 103601815 A CN103601815 A CN 103601815A
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- chondroitin sulfate
- sodium chondroitin
- enzymolysis
- alkaline hydrolysis
- sodium
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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
Abstract
The present invention relates to a sodium chondroitin sulfate extraction method, wherein sodium chondroitin sulfate is a mucopolysaccharide substance extracted from swine nasal bone and other animal cartilages. The traditional process method adopts alkaline hydrolysis, enzymolysis, salt hydrolysis, an ion exchange resin or a combination method thereof to extract, and the extraction solution contains a large amount of proteins, nucleic acids and other impurities so as to cause a certain difficult problem to further purification. According to the invention, the alkaline hydrolysis-enzymolysis method and the flocculation precipitation method are combined to remove impurities remained in the sodium chondroitin sulfate extraction solution, such that the process is reasonable, the removing efficiency is high, the subsequent refinement working hour is shortened, the product yield is improved, and the drug meets standards of the Chinese pharmacopoeia (2000 edition).
Description
Technical field
Under the present invention, field is biochemical pharmacy field, relates to a kind of extracting method of Sodium chondroitin sulfate A, particularly a kind of Sodium chondroitin sulfate A producing and manufacturing technique.
Background technology
Sodium chondroitin sulfate A is from animal cartilaginous tissue, to extract a kind of acid mucopolysaccharides of preparation, belongs to glycosaminoglycan.Clinical lipid regulating agent, sacroiliitis, the eye drop etc. of being mainly used in.Sodium chondroitin sulfate A is a kind of biochemical drug of outbalance in the market, is recorded in < < Chinese Pharmacopoeia (2010 editions) > >.In recent years, Sodium chondroitin sulfate A heats up gradually as the trend of healthcare products consumption, makes its Application Areas more and more extensive.
Traditional process for extracting takes the method for alkaline hydrolysis, enzymolysis, salt solution and ion exchange resin or its combination to extract, extraction efficiency is low, complex operation, still contains the impurity such as a large amount of protein and nucleic acid in extracting solution, give to be further purified and bring certain difficult problem.
Summary of the invention
Object of the present invention provides a kind of method of extracting Sodium chondroitin sulfate A, be that alkaline hydrolysis-enzymolysis process is in conjunction with flocculent precipitation, can effectively remove impurity remaining in extracting solution, effectively simplify refining step, save ethanol consumption in settling step, can in the shorter time, produce the Sodium chondroitin sulfate A product that foreign matter content is low, yield is good.
Object of the present invention reaches by the following technical programs, realizes successively by following step.
(1) cartilage soaks: hog snout diaphyseal bone is joined to the complete submergence of water in retort, soak after 3-5 hour, put the dry water that soaks;
(2) alkaline hydrolysis: add alkali lye, dry bone weight is 1 with the ratio of alkali lye weight: 2-1: 3, stir and extract after 3-5 hour, with 100 order nylon net filters;
(3) enzymolysis: regulate after pH8-9, be warming up to 45 ℃, add pig source pancreatin and sodium-chlor, 45-50 ℃ of insulation 3-5 hour;
(4) temperature that rises suddenly: be warming up to 80-85 ℃, standing 15-20min, filters with filter bag;
(5) ratio of 20%-30% adds 95% ethanol in filtered liquid by volume;
(6) flocculating settling: control temperature 40-50 ℃, regulate pH11-12, in 0.3%-0.5% (mass/volume) ratio, add flocculating settling agent, standing until layering, siphon supernatant, the centrifugal rear merging mother liquor of throw out, to supernatant liquor, discards centrifugal slag;
(7) through decolouring process, after ethanol precipitation, dewatered drying, obtains the Sodium chondroitin sulfate A of separation and purification.
The beneficial effect of the technical program is: the Sodium chondroitin sulfate A making in conjunction with flocculent precipitation through alkaline hydrolysis-enzymolysis process, content is more than 90%, yield >=30%.
(1) in the technical program, enzymolysis step adds sodium-chlor in (3), and the weight ratio of its weight and dry bone is 3%-5%.Can strengthen hydrolysis result, make Sodium chondroitin sulfate A effectively separated with impurity such as albumen.
(2) in the technical program, in alkaline hydrolysis step (2), the concentration of alkali lye used is 1-2mol/L, can reduce the destruction of alkali lye to Sodium chondroitin sulfate A, improves yield.
(3) in the technical program, the pig source pancreatin weight adding in enzymolysis step (3) and the weight ratio of dry bone are 1%-2%, and soaking time is 3-5 hour, can reduce the destruction of pancreatin to Sodium chondroitin sulfate A, improve yield.
(4) in the technical program, in flocculating settling forward direction filtrate, add ethanol, this alcohol concn is 95%, and with filter night volume ratio be 20%-30%.Can increase the combination of the impurity such as protein nucleic acid and flocculating settling agent like this, increase granularity, contribute to sedimentation.
(5), in the technical program, flocculating settling agent is selected from hydroxyapatite (HAP) and sodium laurylsulfonate (SDBS) or its mixture, can be effectively in conjunction with impurity such as protein nucleic acids and form throw out, after centrifugal, remove.
Embodiment
Following instance describes the present invention in detail:
(1) cartilage soaks: 100Kg hog snout diaphyseal bone is joined to the complete submergence of water in retort, soak after 4 hours, put the dry water that soaks;
(2) alkaline hydrolysis: add 1mol/L alkali lye 250L, stir and extract after 4 hours, with 100 order nylon net filters;
(3) enzymolysis: regulate after pH8.5, be warming up to 45 ℃, add pancreatin 1Kg, add sodium-chlor 3Kg, be incubated 4 hours under 45-50 ℃ of condition;
(4) temperature that rises suddenly: be warming up to 80 ℃, after standing 20min, filter with filter bag;
(5) in filtered liquid, add 95% ethanol 80L;
(6) flocculating settling: control temperature 45 C, regulate pH=11, add flocculating settling agent 1.2Kg, siphon supernatant after stratification, throw out is centrifugal, merges mother liquor to supernatant liquor, discards centrifugal slag;
(7) through decolouring process, after ethanol precipitation, dewatered drying, obtains the Sodium chondroitin sulfate A 32.5Kg of separation and purification, content is 95%, yield is 32.5%.
Claims (6)
1. alkaline hydrolysis-enzymolysis process extracts Sodium chondroitin sulfate A in conjunction with flocculent precipitation, it is characterized in that:
(1) cartilage soaks: hog snout diaphyseal bone is joined to the complete submergence of water in retort, soak after 3-5 hour, put the dry water that soaks;
(2) alkaline hydrolysis: add alkali lye, dry bone weight is 1 with the ratio of alkali lye weight: 2-1: 3, stir and extract after 3-5 hour, with 100 order nylon net filters;
(3) enzymolysis: regulate after pH8-9, be warming up to 45 ℃, add pig source pancreatin and sodium-chlor, 45-50 ℃ of insulation 3-5 hour;
(4) temperature that rises suddenly: be warming up to 80-85 ℃, standing 15-20min, filters with filter bag;
(5) ratio of 20%-30% adds 95% ethanol in filtered liquid by volume;
(6) flocculating settling: control temperature 40-50 ℃, regulate pH11-12, in 0.3%-0.5% (mass/volume) ratio, add flocculating settling agent, standing until layering, siphon supernatant, the centrifugal rear merging mother liquor of throw out, to supernatant liquor, discards centrifugal slag;
(7) through decolouring process, after ethanol precipitation, dewatered drying, obtains the Sodium chondroitin sulfate A of separation and purification.
2. it is characterized by accordance with the method for claim 1: in alkaline hydrolysis step, the concentration of alkali lye used is 1-2mol/L.
3. it is characterized by accordance with the method for claim 1: the weight sodium chloride adding in enzymolysis step (3) and the weight ratio of dry bone are 3%-5%; The pig source pancreatin weight adding and the weight ratio of dry bone are 1%-2%; Soaking time is 3-5 hour.
4. in accordance with the method for claim 1, it is characterized by: before flocculating settling step, must in filtrate, add ethanol, this alcohol concn is 95%, and with filter night volume ratio be 20%-30%.
5. it is characterized by accordance with the method for claim 1: flocculating settling agent is selected from hydroxyapatite (HAP) and sodium laurylsulfonate (SDBS) or its mixture.
6. it is characterized by accordance with the method for claim 1: the Sodium chondroitin sulfate A of preparing according to the inventive method meets Chinese Pharmacopoeia (version in 2010) requirement.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201511000950.5A CN105504090A (en) | 2013-10-21 | 2013-10-21 | Chondroitin sulfate sodium extracted by combining alkaline hydrolysis-enzyme hydrolysis method and flocculation precipitation method |
| CN201310508887.0A CN103601815B (en) | 2013-10-21 | 2013-10-21 | Alkaline hydrolysis-enzymolysis process extracts Sodium chondroitin sulfate A in conjunction with flocculent precipitation |
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| CN201310508887.0A CN103601815B (en) | 2013-10-21 | 2013-10-21 | Alkaline hydrolysis-enzymolysis process extracts Sodium chondroitin sulfate A in conjunction with flocculent precipitation |
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| CN201511000950.5A Division CN105504090A (en) | 2013-10-21 | 2013-10-21 | Chondroitin sulfate sodium extracted by combining alkaline hydrolysis-enzyme hydrolysis method and flocculation precipitation method |
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| CN103601815A true CN103601815A (en) | 2014-02-26 |
| CN103601815B CN103601815B (en) | 2016-04-13 |
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| CN201511000950.5A Pending CN105504090A (en) | 2013-10-21 | 2013-10-21 | Chondroitin sulfate sodium extracted by combining alkaline hydrolysis-enzyme hydrolysis method and flocculation precipitation method |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104558245A (en) * | 2014-12-24 | 2015-04-29 | 青岛九龙生物医药有限公司 | Diluted alkali-enzymolysis extraction method for extracting chondroitin sulfate in pig nasal bones |
| CN105504094A (en) * | 2016-01-20 | 2016-04-20 | 定陶县地元生化制品有限公司 | Preparation method of sodium-ion-free chondroitin enzymolysis solution |
| CN105866109A (en) * | 2016-03-28 | 2016-08-17 | 北京倍肯恒业科技发展股份有限公司 | Rapid detection method of phenolphthalein in weight reducing health foods, and detection card developing method |
| CN105866111A (en) * | 2016-03-29 | 2016-08-17 | 北京倍肯恒业科技发展股份有限公司 | Rapid detection method of nifedipine in blood pressure reducing health foods, and detection card developing method |
| CN106397629A (en) * | 2016-08-30 | 2017-02-15 | 集美大学 | Method for extracting chondroitin sulfate from sturgeon bones, chondroitin sulfate extracted through method, and application of chondroitin sulfate |
| CN109280094A (en) * | 2018-05-18 | 2019-01-29 | 山阳县恒桓生物科技有限公司 | A kind of method that chondroitin sulfate purification rate is high |
| CN109384861A (en) * | 2018-09-30 | 2019-02-26 | 临沂新程金锣肉制品集团有限公司 | A kind of method of heparin sodium pulp thickening dermatan sulfate |
| CN109651529A (en) * | 2018-11-14 | 2019-04-19 | 宁波绿之健药业有限公司 | A kind of chondroitin sulfate preparation method of high bioavilability |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111925461B (en) * | 2020-08-31 | 2021-08-31 | 四川大学 | Fibrocartilage glycosaminoglycan and extraction method thereof |
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| CN101280027A (en) * | 2007-10-18 | 2008-10-08 | 日照众山生物科技有限公司 | Extracting method of chondroitin sulfate |
| CN101851300A (en) * | 2010-06-19 | 2010-10-06 | 江西海鑫贸易有限公司 | Process for extracting chondroitin sulfate |
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| 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 |
| CN102816809B (en) * | 2012-07-20 | 2014-09-17 | 王世强 | Preparing process for calcium chondroitin sulfate |
| CN103030715B (en) * | 2012-12-07 | 2015-05-06 | 青岛九龙生物医药有限公司 | Method for separating purified heparin sodium |
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2013
- 2013-10-21 CN CN201310508887.0A patent/CN103601815B/en active Active
- 2013-10-21 CN CN201511000950.5A patent/CN105504090A/en active Pending
Patent Citations (2)
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| CN101280027A (en) * | 2007-10-18 | 2008-10-08 | 日照众山生物科技有限公司 | Extracting method of chondroitin sulfate |
| CN101851300A (en) * | 2010-06-19 | 2010-10-06 | 江西海鑫贸易有限公司 | Process for extracting chondroitin sulfate |
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104558245A (en) * | 2014-12-24 | 2015-04-29 | 青岛九龙生物医药有限公司 | Diluted alkali-enzymolysis extraction method for extracting chondroitin sulfate in pig nasal bones |
| CN105504094B (en) * | 2016-01-20 | 2017-09-29 | 定陶县地元生化制品有限公司 | Liquid and preparation method thereof is digested without sodium ion chondroitin |
| CN105504094A (en) * | 2016-01-20 | 2016-04-20 | 定陶县地元生化制品有限公司 | Preparation method of sodium-ion-free chondroitin enzymolysis solution |
| CN105866109A (en) * | 2016-03-28 | 2016-08-17 | 北京倍肯恒业科技发展股份有限公司 | Rapid detection method of phenolphthalein in weight reducing health foods, and detection card developing method |
| CN105866109B (en) * | 2016-03-28 | 2018-06-12 | 北京倍肯恒业科技发展股份有限公司 | Phenolphthalein detection method and detection blocking Preparation Method in weight-reducing class health food |
| CN105866111B (en) * | 2016-03-29 | 2018-05-08 | 北京倍肯恒业科技发展股份有限公司 | Nifedipine detection method and detection blocking Preparation Method in antihypertensive health care food |
| CN105866111A (en) * | 2016-03-29 | 2016-08-17 | 北京倍肯恒业科技发展股份有限公司 | Rapid detection method of nifedipine in blood pressure reducing health foods, and detection card developing method |
| CN106397629A (en) * | 2016-08-30 | 2017-02-15 | 集美大学 | Method for extracting chondroitin sulfate from sturgeon bones, chondroitin sulfate extracted through method, and application of chondroitin sulfate |
| CN106397629B (en) * | 2016-08-30 | 2019-08-20 | 集美大学 | Method, the chondroitin sulfate by this method extraction and the application of chondroitin sulfate are extracted from sturgeon fish-bone |
| CN109280094A (en) * | 2018-05-18 | 2019-01-29 | 山阳县恒桓生物科技有限公司 | A kind of method that chondroitin sulfate purification rate is high |
| CN109384861A (en) * | 2018-09-30 | 2019-02-26 | 临沂新程金锣肉制品集团有限公司 | A kind of method of heparin sodium pulp thickening dermatan sulfate |
| CN109651529A (en) * | 2018-11-14 | 2019-04-19 | 宁波绿之健药业有限公司 | A kind of chondroitin sulfate preparation method of high bioavilability |
| CN109651529B (en) * | 2018-11-14 | 2020-10-30 | 宁波绿之健药业有限公司 | Preparation method of chondroitin sulfate with high bioavailability |
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| CN105504090A (en) | 2016-04-20 |
| CN103601815B (en) | 2016-04-13 |
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Address after: 266100 Zhuzhou Road, Laoshan District, Shandong, No. 97, No. Patentee after: Qingdao Jiulong biological medicine group Co., Ltd. Address before: 266100 Zhuzhou Road, Laoshan District, Shandong, No. 97, No. Patentee before: Qingdao Jiulong Bio-Pharmaceutical Co., Ltd. |
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