CN103601815B - Alkaline hydrolysis-enzymolysis process extracts Sodium chondroitin sulfate A in conjunction with flocculent precipitation - Google Patents
Alkaline hydrolysis-enzymolysis process extracts Sodium chondroitin sulfate A in conjunction with flocculent precipitation Download PDFInfo
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- CN103601815B CN103601815B CN201310508887.0A CN201310508887A CN103601815B CN 103601815 B CN103601815 B CN 103601815B CN 201310508887 A CN201310508887 A CN 201310508887A CN 103601815 B CN103601815 B CN 103601815B
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- chondroitin sulfate
- sodium chondroitin
- alkaline hydrolysis
- enzymolysis
- 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 kind of extracting method of Sodium chondroitin sulfate A.Sodium chondroitin sulfate A extracts obtained a kind of mucopolysaccharide class material in the animal cartilages such as hog snout bone.Traditional processing method takes the method for alkaline hydrolysis, enzymolysis, salt solution and ion exchange resin or its combination to extract, and still containing the impurity such as a large amount of protein and nucleic acid in extracting solution, brings certain difficult problem to being further purified.The present invention adopts alkaline hydrolysis-enzymolysis process to remove impurity residual in Sodium chondroitin sulfate A extracting solution in conjunction with flocculent precipitation, technological design is reasonable, and removal efficiency is high, shortens follow-up refining man-hour, improve product yield, standard that medicine meets " Chinese Pharmacopoeia 2010 editions ".
Description
Technical field
Field belonging to the present invention 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 a kind of acid mucopolysaccharides extracting preparation from animal cartilaginous tissue, belongs to glycosaminoglycan.Clinically be mainly used in lipid regulating agent, sacroiliitis, eye drop etc.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, the trend that Sodium chondroitin sulfate A is consumed as healthcare products heats up gradually, 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 containing the impurity such as a large amount of protein and nucleic acid in extracting solution, brings certain difficult problem to being further purified.
Summary of the invention
Object of the present invention provides a kind of method extracting Sodium chondroitin sulfate A, namely alkaline hydrolysis-enzymolysis process is in conjunction with flocculent precipitation, can effectively remove impurity remaining in extracting solution, effective simplification refining step, save ethanol consumption in settling step, the Sodium chondroitin sulfate A product that foreign matter content is low, yield is good can be produced in the shorter time.
Object of the present invention reaches by the following technical programs, is realized successively by following step.
(1) cartilage soaks: hog snout diaphyseal bone joined with the complete submergence of water in retort, soak after 3-5 hour, puts dry immersion water;
(2) alkaline hydrolysis: add alkali lye, the ratio of dry bone weight and alkali lye weight is 1: 2-1: 3, stirs and extracts after 3-5 hour, with 100 order nylon net filters;
(3) enzymolysis: after regulating pH8-9, be warming up to 45 DEG C, add pig source pancreatin and sodium-chlor, 45-50 DEG C of insulation 3-5 hour;
(4) rise suddenly temperature: be warming up to 80-85 DEG C, leaves standstill 15-20min, with sock filtration;
(5) ratio of 20%-30% adds 95% ethanol in filtered liquid by volume;
(6) flocculating settling: control temperature 40-50 DEG C, regulates pH11-12, adds flocculating settling agent in 0.3%-0.5% (mass/volume) ratio, leave standstill until layering, siphon supernatant, the centrifugal rear merging mother liquor of throw out, in supernatant liquor, discards centrifugal slag;
(7) after desolventing technology, alcohol settling, dewatered drying, obtains the Sodium chondroitin sulfate A of separation and purification.
The beneficial effect of the technical program is: through alkaline hydrolysis-enzymolysis process in conjunction with the obtained Sodium chondroitin sulfate A of flocculent precipitation, content 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 hydrolysis result be strengthened, Sodium chondroitin sulfate A is 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 added 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, add ethanol in flocculating settling forward direction filtrate, this alcohol concn is 95%, and is 20%-30% with filtering the volume ratio at night.The combination of the impurity such as protein nucleic acid and flocculating settling agent can be increased 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 the effective impurity such as associated proteins nucleic acid form throw out, removes after centrifugal.
Embodiment
Following instance describes the present invention in detail:
(1) cartilage soaks: 100Kg hog snout diaphyseal bone joined with the complete submergence of water in retort, soak after 4 hours, puts dry immersion water;
(2) alkaline hydrolysis: add 1mol/L alkali lye 250L, stirs extraction after 4 hours, with 100 order nylon net filters;
(3) enzymolysis: after regulating pH8.5, be warming up to 45 DEG C, add pancreatin 1Kg, add sodium-chlor 3Kg, is incubated 4 hours under 45-50 DEG C of condition;
(4) rise suddenly temperature: be warming up to 80 DEG C, after leaving standstill 20min, with sock filtration;
(5) in filtered liquid, 95% ethanol 80L is added;
(6) flocculating settling: control temperature 45 DEG C, regulate pH=11, add flocculating settling agent 1.2Kg, siphon supernatant after stratification, throw out is centrifugal, merges mother liquor in supernatant liquor, discards centrifugal slag;
(7) after desolventing technology, alcohol settling, dewatered drying, obtains the Sodium chondroitin sulfate A 32.5Kg of separation and purification, and content is 95%, and yield is 32.5%.
Claims (4)
1. alkaline hydrolysis-enzymolysis process extracts a method for Sodium chondroitin sulfate A in conjunction with flocculent precipitation, it is characterized in that:
(1) cartilage soaks: hog snout diaphyseal bone joined with the complete submergence of water in retort, soak after 3-5 hour, puts dry immersion water;
(2) alkaline hydrolysis: add alkali lye, the ratio of dry bone weight and alkali lye weight is 1: 2-1: 3, stirs and extracts after 3-5 hour, with 100 order nylon net filters;
(3) enzymolysis: after regulating pH8-9, be warming up to 45 DEG C, add pig source pancreatin and sodium-chlor, 45-50 DEG C of insulation 3-5 hour;
(4) rise suddenly temperature: be warming up to 80-85 DEG C, leaves standstill 15-20min, with sock filtration;
(5) ratio of 20%-30% adds 95% ethanol in filtered liquid by volume;
(6) flocculating settling: control temperature 40-50 DEG C, regulate pH11-12, flocculating settling agent is added in mass/volume 0.3%-0.5% ratio, leave standstill until layering, siphon supernatant, the centrifugal rear merging mother liquor of throw out, in supernatant liquor, discards centrifugal slag, and described flocculating settling agent is selected from hydroxyapatite and sodium laurylsulfonate or its mixture;
(7) after desolventing technology, alcohol settling, dewatered drying, obtains the Sodium chondroitin sulfate A of separation and purification.
2. in accordance with the method for claim 1, it is characterized by: in alkaline hydrolysis step, the concentration of alkali lye used is 1-2mol/L.
3. in accordance with the method for claim 1, it is characterized by: the weight sodium chloride added in enzymolysis step (3) and the weight ratio of dry bone are 3%-5%; The pig source pancreatin weight added 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: the Sodium chondroitin sulfate A prepared according to present method meets the requirement of version Chinese Pharmacopoeia in 2010.
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 CN103601815A (en) | 2014-02-26 |
| CN103601815B true 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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Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
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| 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 |
| 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 |
| 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 |
| 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 |
| CN111925461B (en) * | 2020-08-31 | 2021-08-31 | 四川大学 | Fibrocartilage glycosaminoglycan and extraction method thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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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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. |