CN1563109A - Method for preparing hyaluronic acid - Google Patents
Method for preparing hyaluronic acid Download PDFInfo
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- CN1563109A CN1563109A CN 200410024014 CN200410024014A CN1563109A CN 1563109 A CN1563109 A CN 1563109A CN 200410024014 CN200410024014 CN 200410024014 CN 200410024014 A CN200410024014 A CN 200410024014A CN 1563109 A CN1563109 A CN 1563109A
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- hyaluronic acid
- water
- aqueous solution
- organic solvent
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Abstract
The invention discloses a method for preparing hyaluronic acid. said method includes the following steps: converting sodium hyaluronate solution into hyaluronic acid and free sodium ions, remving free low-molecular inorganic salt from the above-mentioned material, then utilizing emulsification principle to make the hyaluronic acid be precipitated and separated out from the solution in the form of amorphous powder or floccule; then using organic solvent mixed with water to dehydrate the hyaluronic acid solid for several times, making centrifugal separation and vacuum drying so as to obtain the invented hyaluronic acid product. Said invented preparation process is simple, its production is easy to control, and suitable for industrial large-scale production.
Description
1, technical field
The present invention relates to a kind of complete processing of macromole acidic mucopolysaccharide, the hyaluronic method of specifically a kind of preparation.
2, technical background
Hyaluronic acid is a kind of macromole acidic mucopolysaccharide, and its aqueous solution presents typical non-Newtonian fluid.Hyaluronic acid variation with concentration in the aqueous solution presents different structures, presents primary structure during lower concentration, i.e. linear structure; Present secondary structure during intermediate concentration, i.e. spirane structure; Present tertiary structure during high density, i.e. stepped construction.Hyaluronic acid exists with the polyanion state in the aqueous solution, because its molecular weight is huge, viscosity-average molecular weight is generally greater than 5 * 10
5Dalton when no settling agent, is difficult to precipitate and separate from the aqueous solution.The hyaluronic technology of existing preparation is by the powder process of freeze-drying mode, do not only complete processing complexity like this, and tooling cost is very high, is a kind of unfavorable working method.
3, summary of the invention
The purpose of this invention is to provide the hyaluronic method of a kind of preparation.
The objective of the invention is to realize in the following manner: the hyaluronate sodium dissolving crude product that will make from fermentation method or extraction method is in water, obtain the aqueous solution of 0.01%-10%, regulate its pH value 3.0-6.0, the gac (m/V) that adds 0.01%-2% is in 25-45 ℃ of decolouring, add and bury as flocculating aids, filter, filtrate is regulated pH value 8.0-9.0, clarification filtration, filtrate is as the hyaluronic precursor filtrate of preparation; Hyaluronate sodium precursor filtrate is regulated pH value 3.0-6.0, and stirring reaction is abundant, changes hyaluronate sodium into hyaluronic acid and free sodium ion, with molecular weight cut-off 5 * 10
4-1 * 10
5The ultrafiltration post filter, remove free low molecule inorganic salt wherein, what retain in the remaining liq is hyaluronic acid, its pH value is 2.0-5.0; Perhaps aqueous solution of sodium hyaluronate is regulated pH value 3.0-6.0, stirring reaction is abundant, change hyaluronate sodium into hyaluronic acid and free sodium ion, Zeo-karb is passed through in the aqueous solution of sodium hyaluronate following current, fully absorption exchange positively charged ion wherein changes aqueous solution of sodium hyaluronate into hyaluronic acid aqueous solution; In hyaluronic acid aqueous solution, add 1%-100% (V/V) with the not miscible organic solvent of water, dispersed with stirring, make solution become water-in-oil type emulsion, under agitation condition, stream adds and the miscible organic solvent of water in solution, behind the above-mentioned organic solvent that adds 100-300% (V/V), hyaluronic acid comes out with unformed powder or cotton-shaped form precipitate and separate from solution; The hyaluronic acid solid that makes use with the miscible dehydration of organic solvent of water for several times after, centrifugation, vacuum-drying promptly obtain hyaluronic acid product.
This preparation method's complete processing is simple relatively, produces to be easy to control, can obtain Powdered amorphous precipitatedly, and the cycle is short, and the cycle is short, is fit to large-scale industrialization production, and has reduced production cost greatly.
4, embodiment
Embodiment 1:
Get hyaluronate sodium crude product 30g, be dissolved in the 4L deionized water, abundant stirring and dissolving, regulator solution pH value 4.0-5.0 adds the 9-12g gac, fully stirring decolouring in 25-40 ℃; Add the kaolin flocculating aids in the solution, after-filtration stirs; Filtrate is regulated pH value 8.0-9.0, stirs, and clarification filtration, filtrate is regulated pH value 4.5-6.0, fully stirs; Filtrate is smooth by Zeo-karb (as 001 * 7), collects exchange liquid; In exchange liquid, add 100-400ml ether (or CCl
4, CHCl
3, CH
2Cl
2, CH
3COOC
2H
5, benzene, toluene etc. and the not miscible organic solvent of water), fully stirring and emulsifying makes solution change oil-in-water emulsion into.Under agitation condition, stream adds the acetone (or organic solvent miscible with water such as ethanol, acetonitrile) of 1-3 times of volume in the phase emulsion, and hyaluronic acid can be adorned amorphous precipitated separating out with powder.Fully after the sedimentation, the hyaluronic acid solid precipitation is dewatered 2-3 time with 100-200ml acetone (ethanol or acetonitrile), centrifugation, 40 ± 5 ℃ of thorough dryings of vacuum obtain 17.5g hyaluronic acid solid phase prod.Yield 58.3%.
Preparation example 2
Get hyaluronate sodium crude product 25g, separate in the 5L deionized water, abundant stirring and dissolving, regulator solution pH value 4.0-5.0 adds the 8-10g gac, fully stirring decolouring in 25-40 ℃; Add super-cell in the solution, after-filtration stirs; Filtrate is regulated pH value 8.0-9.0, stirs clarification filtration.Filtrate is regulated pH value 4.5-6.0, fully stirs, and positively charged ion is discharged.Filtrate is smooth to be the ultrafiltration post of 5 * 104-1 * 105 by molecular weight cut-off, and fully desalination makes the small molecules inorganic salt break away from solution, obtains 3.8L filtering and concentrating liquid.In filtering and concentrating liquid, add 100-400ml ether (or CCl
4, CHCl
3, CH
2Cl
2, CH
3COOC
2H
5, benzene, toluene etc. and the not miscible organic solvent of water), fully stirring and emulsifying makes solution change oil-in-water emulsion into.Under agitation condition, stream adds the acetone (or organic solvent miscible with water such as ethanol, acetonitrile) of 1-3 times of volume in the phase emulsion, and hyaluronic acid can be adorned amorphous precipitated separating out with powder.Fully after the sedimentation, the hyaluronic acid solid precipitation is dewatered 2-3 time with 100-200ml acetone (ethanol or acetonitrile), centrifugation, 40 ± 5 ℃ of thorough dryings of vacuum obtain 15.3g hyaluronic acid solid phase prod.Yield 61.2%.
Claims (4)
1, the hyaluronic method of a kind of preparation, it is characterized in that the preparation method is: the hyaluronate sodium dissolving crude product that will make from fermentation method or extraction method is in water, obtain the aqueous solution of 0.01%-10%, regulate its pH value 3.0-6.0, the gac (m/V) that adds 0.01%-2% decolours under 25-45 ℃ of temperature, adds and buries as flocculating aids, filter, filtrate is regulated pH value 8.0-9.0, clarification filtration, and filtrate is as the hyaluronic precursor filtrate of preparation;
Hyaluronate sodium precursor filtrate is regulated pH value 3.0-6.0, and stirring reaction is abundant, changes hyaluronate sodium into hyaluronic acid and free sodium ion, with molecular weight cut-off 5 * 10
4-1 * 10
5The ultrafiltration post filter, filtrate is hyaluronic acid aqueous solution, its pH value is 2.0-5.0; Perhaps aqueous solution of sodium hyaluronate is regulated pH value 3.0-6.0, stirring reaction is abundant, by Zeo-karb, aqueous solution of sodium hyaluronate is changed the aqueous solution of sodium hyaluronate following current into hyaluronic acid aqueous solution;
In hyaluronic acid aqueous solution, add 1%-100% (V/V) with the not miscible organic solvent of water, dispersed with stirring, make solution become water-in-oil type emulsion, under agitation condition, stream adds and the miscible organic solvent of water in solution, behind the above-mentioned organic solvent that adds 100-300% (V/V), hyaluronic acid comes out with unformed powder or cotton-shaped form precipitate and separate from solution; The hyaluronic acid solid that makes use with the miscible dehydration of organic solvent of water for several times after, centrifugation, vacuum-drying promptly obtain hyaluronic acid product.
2, method according to claim 1, it is characterized in that being used to help the soil of filter is kaolin, white bole or diatomite.
3, method according to claim 1, it is characterized in that with the not miscible organic solvent of water be ethyl acetate, chloroform, dichloride charcoal, tetracol phenixin, ether, benzene, toluene or dimethylbenzene.
4, method according to claim 1, it is characterized in that with the miscible organic solvent of water be ethanol, acetone or acetonitrile.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410024014 CN1563109A (en) | 2004-04-13 | 2004-04-13 | Method for preparing hyaluronic acid |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410024014 CN1563109A (en) | 2004-04-13 | 2004-04-13 | Method for preparing hyaluronic acid |
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| Publication Number | Publication Date |
|---|---|
| CN1563109A true CN1563109A (en) | 2005-01-12 |
Family
ID=34480431
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|---|---|---|---|
| CN 200410024014 Pending CN1563109A (en) | 2004-04-13 | 2004-04-13 | Method for preparing hyaluronic acid |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100478360C (en) * | 2006-10-31 | 2009-04-15 | 山东福瑞达生物化工有限公司 | Process for producing hyaluronic acid or its salt by concentration |
| CN102154402A (en) * | 2010-12-22 | 2011-08-17 | 东辰控股集团有限公司 | Pre-fermentation seed culture method and feeding process for sodium hyaluronate |
| CN102174610A (en) * | 2010-12-22 | 2011-09-07 | 东辰控股集团有限公司 | Method and process for drying sodium hyaluronate finished product |
| CN101146830B (en) * | 2005-03-22 | 2011-11-16 | 丘比株式会社 | Low molecular weight hyaluronic acid and/or salt thereof, method for producing same, and cosmetic preparation and food composition containing same |
| CN102630231A (en) * | 2009-09-15 | 2012-08-08 | 一栋制药株式会社 | Method for manufacturing low molecular weight hyaluronic acid |
| CN107522796A (en) * | 2017-08-31 | 2017-12-29 | 浙江工业大学 | A kind of method that hyaluronic acid is extracted in the eye from tuna |
| CN108348660A (en) * | 2015-09-03 | 2018-07-31 | 株式会社真友生物 | It is coated with the manufacturing method and its high performance sewing yarn of the high performance sewing yarn of hyaluronate |
| CN110590972A (en) * | 2019-09-19 | 2019-12-20 | 华熙生物科技股份有限公司 | Drying method of hyaluronic acid |
-
2004
- 2004-04-13 CN CN 200410024014 patent/CN1563109A/en active Pending
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101146830B (en) * | 2005-03-22 | 2011-11-16 | 丘比株式会社 | Low molecular weight hyaluronic acid and/or salt thereof, method for producing same, and cosmetic preparation and food composition containing same |
| US8933054B2 (en) | 2005-03-22 | 2015-01-13 | Q.P. Corporation | Low molecular weight hyaluronic acid and/or salt thereof, method for producing same, and cosmetic preparation and food composition containing same |
| CN100478360C (en) * | 2006-10-31 | 2009-04-15 | 山东福瑞达生物化工有限公司 | Process for producing hyaluronic acid or its salt by concentration |
| CN102630231A (en) * | 2009-09-15 | 2012-08-08 | 一栋制药株式会社 | Method for manufacturing low molecular weight hyaluronic acid |
| CN102630231B (en) * | 2009-09-15 | 2015-05-27 | 一栋制药株式会社 | Method for manufacturing low molecular weight hyaluronic acid |
| CN102154402A (en) * | 2010-12-22 | 2011-08-17 | 东辰控股集团有限公司 | Pre-fermentation seed culture method and feeding process for sodium hyaluronate |
| CN102174610A (en) * | 2010-12-22 | 2011-09-07 | 东辰控股集团有限公司 | Method and process for drying sodium hyaluronate finished product |
| CN108348660A (en) * | 2015-09-03 | 2018-07-31 | 株式会社真友生物 | It is coated with the manufacturing method and its high performance sewing yarn of the high performance sewing yarn of hyaluronate |
| CN108348660B (en) * | 2015-09-03 | 2021-04-06 | 株式会社真友生物 | Method for manufacturing high-performance suture coated with hyaluronate and high-performance suture thereof |
| CN107522796A (en) * | 2017-08-31 | 2017-12-29 | 浙江工业大学 | A kind of method that hyaluronic acid is extracted in the eye from tuna |
| CN110590972A (en) * | 2019-09-19 | 2019-12-20 | 华熙生物科技股份有限公司 | Drying method of hyaluronic acid |
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