CN104592420A - Method for preparing intermediate HA-VS of cross-linked sodium hyaluronate - Google Patents
Method for preparing intermediate HA-VS of cross-linked sodium hyaluronate Download PDFInfo
- Publication number
- CN104592420A CN104592420A CN201410844380.7A CN201410844380A CN104592420A CN 104592420 A CN104592420 A CN 104592420A CN 201410844380 A CN201410844380 A CN 201410844380A CN 104592420 A CN104592420 A CN 104592420A
- Authority
- CN
- China
- Prior art keywords
- solution
- reaction
- hyaluronic acid
- reaction solution
- acid sodium
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Polysaccharides And Polysaccharide Derivatives (AREA)
Abstract
The invention discloses a method for preparing an intermediate HA-VS of cross-linked sodium hyaluronate. The method comprises the following steps: carrying out a modification reaction between sodium hyaluronate solution and divinyl sulfone solution under alkaline conditions, adding 3-8mol/L of hydrochloric acid solution to stop the reaction, and adding sodium chloride solution into the reaction solution, so that the ion strength of the solution reaches 0.1-0.5mol/L; slowly dripping 40-95 percent of ethanol solution into the reaction solution, wherein the addition amount of ethanol is 1-5 times that of the reaction solution, and the HA-VS is separated out in a particle form; and standing the particles, pouring the supernatant, washing the precipitate by using ethanol, performing suction filtration and washing by using ethanol. The method is easy and convenient to operate, and the production cost is greatly reduced.
Description
Technical field:
The present invention relates to crosslinked hyaluronic acid sodium preparing technical field, particularly the preparation method of a kind of intermediate HA-VS of crosslinked hyaluronic acid sodium.
Background technology:
Hyaluronic Acid (hyaluronic acid, HA) generates HA-VS by NAG and D-glucuronic acid disaccharide bond structure by first hydroxyl and divinyl sulfone generation modification reaction.General preparation method adopts and HA-VS reaction solution is first carried out ultrafiltration suitable multiple by purified water, DVS residual in removing reaction solution, after ultrafiltrated be concentrated to certain concentration carry out lyophilize, production cost is very high.
Summary of the invention
The object of this invention is to provide a kind of simple to operation, the preparation method of the intermediate HA-VS of the crosslinked hyaluronic acid sodium that production cost is low.
For realizing object of the present invention, the technical solution adopted in the present invention is:
A preparation method of the intermediate HA-VS of crosslinked hyaluronic acid sodium, is characterized in that, comprise the following steps:
(1) hyaluronic acid sodium solution and divinyl sulfone generation modification reaction, generates HA-VS solution A;
(2) in the HA-VS solution A of step (1) gained, add hydrochloric acid soln, when pH value reaches 4 ~ 6.5, termination reaction obtains reaction solution B.
(3) in reaction solution B, sodium chloride solution is added, when making the ionic strength of solution reach 0.1 ~ 0.5mol/L, slowly adding mass percent concentration is again 40 ~ 95% ethanolic solns, ethanolic soln add-on is 1 ~ 5 times of reaction solution B, HA-VS is separated out in the form of granules, particle outwells supernatant liquor after leaving standstill, then throw out mass percent concentration is 40 ~ 95% ethanolic soln washings, then is the intermediate HA-V that 40 ~ 95% ethanolic soln filtering and washing obtain crosslinked hyaluronic acid sodium with mass percent concentration.
In a preferred embodiment of the invention, described concentration of hydrochloric acid solution 3 ~ 8mol/L.
In a preferred embodiment of the invention, ethanolic soln add-on is 1 ~ 5 times of reaction solution B.
Because HA-VS is insoluble to ethanol under the condition having certain ionic concn, DVS is soluble in ethanol simultaneously, uses the method for alcohol precipitation that HA-VS both can have been made to separate out in the form of granules, and the DVS that can remove again in reaction solution remains.
Embodiment
The present invention can be understood more specifically by following Examples, the present invention includes but be not limited to following particular content.
Embodiment 1
The preparation method of a kind of intermediate HA-VS of crosslinked hyaluronic acid sodium, first by hyaluronic acid sodium solution in the basic conditions with divinyl sulfone solution generation modification reaction, after add 3mol/L hydrochloric acid soln, termination reaction when reacting liquid pH value is 4, sodium chloride solution is added in backward reaction solution, after making the ionic strength of solution reach 0.1mol/L, the ethanolic soln that mass percent concentration is 40% is slowly instilled in reaction solution, ethanolic soln add-on is 5 times of reaction solution, and HA-VS is separated out in the form of granules.Particle outwells supernatant liquor after leaving standstill, then throw out mass percent concentration be 40% ethanolic soln wash twice, be then the ethanolic soln filtering and washing 3 times of 40% with mass percent concentration.
Embodiment 2
The preparation method of a kind of intermediate HA-VS of crosslinked hyaluronic acid sodium, first by hyaluronic acid sodium solution in the basic conditions with divinyl sulfone solution generation modification reaction, after add 5mol/L hydrochloric acid soln, reacting liquid pH value is 5 termination reactions, sodium chloride solution is added in backward reaction solution, the ionic strength of solution is made to reach 0.3mol/L, the ethanolic soln that mass percent concentration is 50% is slowly instilled in backward reaction solution, ethanolic soln add-on is 4 times of reaction solution, HA-VS is separated out in the form of granules, particle outwells supernatant liquor after leaving standstill, then throw out mass percent concentration be 50% ethanolic soln wash twice, then be the ethanolic soln filtering and washing 3 times of 50% with mass percent concentration.
Embodiment 3
The preparation method of a kind of intermediate HA-VS of crosslinked hyaluronic acid sodium, first by hyaluronic acid sodium solution in the basic conditions with divinyl sulfone solution generation modification reaction, after add 8mol/L hydrochloric acid soln, reacting liquid pH value is 6.5 termination reactions, sodium chloride solution is added in backward reaction solution, the ionic strength of solution is made to reach 0.5mol/L, the ethanolic soln that mass percent concentration is 70% is slowly instilled in backward reaction solution, ethanolic soln add-on is 3 times of reaction solution, and HA-VS is separated out in the form of granules.Particle outwells supernatant liquor after leaving standstill, then throw out mass percent concentration be 70% ethanolic soln wash twice, be then the ethanolic soln filtering and washing 3 times of 70% with mass percent concentration.
Embodiment 4
The preparation method of a kind of intermediate HA-VS of crosslinked hyaluronic acid sodium, first by hyaluronic acid sodium solution in the basic conditions with divinyl sulfone solution generation modification reaction, after add 8mol/L hydrochloric acid soln, reacting liquid pH value is 6.5 termination reactions, sodium chloride solution is added in backward reaction solution, the ionic strength of solution is made to reach 0.5mol/L, the ethanolic soln that mass percent concentration is 80% is slowly instilled in backward reaction solution, ethanolic soln add-on is 2 times of reaction solution, and HA-VS is separated out in the form of granules.Particle outwells supernatant liquor after leaving standstill, then throw out mass percent concentration be 80% ethanolic soln wash twice, be then the ethanolic soln filtering and washing 3 times of 80% with mass percent concentration.
Embodiment 5
The preparation method of a kind of intermediate HA-VS of crosslinked hyaluronic acid sodium, first by hyaluronic acid sodium solution in the basic conditions with divinyl sulfone solution generation modification reaction, after add 3mol/L hydrochloric acid soln, reacting liquid pH value is 4 termination reactions, sodium chloride solution is added in backward reaction solution, the ionic strength of solution is made to reach 0.1mol/L, the ethanolic soln that mass percent concentration is 95% is slowly instilled in backward reaction solution, ethanolic soln add-on be reaction solution 1 times, HA-VS is separated out in the form of granules.Particle outwells supernatant liquor after leaving standstill, then throw out mass percent concentration be 95% ethanolic soln wash twice, be then the ethanolic soln filtering and washing 3 times of 95% with mass percent concentration.
Table 1
Can be reached a conclusion by upper table, alcohol precipitation condition prepared by the intermediate HA-VS being cross-linked hyaluronic acid sodium: add 0.3 ~ 0.8mol hydrochloric acid in HA-VS reaction solution, the pH value modulation 4.0 ~ 6.5 of reaction solution, after add sodium chloride solution, regulator solution ionic strength 0.1 ~ 0.5mol, after slowly drip 40% ~ 95% ethanol, amount of alcohol added is 1 ~ 5 times of reaction solution, and HV-VS separates out in the form of granules.
Claims (3)
1. a preparation method of the intermediate HA-VS of crosslinked hyaluronic acid sodium, is characterized in that, comprise the following steps:
(1) hyaluronic acid sodium solution and divinyl sulfone generation modification reaction, generates HA-VS solution A;
(2) in the HA-VS solution A of step (1) gained, add hydrochloric acid soln, when pH value reaches 4 ~ 6.5, termination reaction obtains reaction solution B.
(3) in reaction solution B, sodium chloride solution is added, when making the ionic strength of solution reach 0.1 ~ 0.5mol/L, slowly adding mass percent concentration is again 40 ~ 95% ethanolic solns, ethanolic soln add-on is 1 ~ 5 times of reaction solution B, HA-VS is separated out in the form of granules, particle outwells supernatant liquor after leaving standstill, then throw out mass percent concentration is 40 ~ 95% ethanolic soln washings, then is the intermediate HA-V that 40 ~ 95% ethanolic soln filtering and washing obtain crosslinked hyaluronic acid sodium with mass percent concentration.
2. the preparation method of the intermediate HA-VS of a kind of crosslinked hyaluronic acid sodium as claimed in claim 1, is characterized in that, described concentration of hydrochloric acid solution 3 ~ 8mol/L.
3. the preparation method of the intermediate HA-VS of a kind of crosslinked hyaluronic acid sodium as claimed in claim 1, is characterized in that, described ethanolic soln add-on is 1 ~ 5 times of reaction solution B.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410844380.7A CN104592420A (en) | 2014-12-25 | 2014-12-25 | Method for preparing intermediate HA-VS of cross-linked sodium hyaluronate |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410844380.7A CN104592420A (en) | 2014-12-25 | 2014-12-25 | Method for preparing intermediate HA-VS of cross-linked sodium hyaluronate |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104592420A true CN104592420A (en) | 2015-05-06 |
Family
ID=53118509
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410844380.7A Pending CN104592420A (en) | 2014-12-25 | 2014-12-25 | Method for preparing intermediate HA-VS of cross-linked sodium hyaluronate |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104592420A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105440159A (en) * | 2015-12-22 | 2016-03-30 | 上海景峰制药有限公司 | Alcohol precipitation method for preparation of cross-linked sodium hyaluronate intermediate HA-VS |
| CN109223708A (en) * | 2018-10-24 | 2019-01-18 | 上海景峰制药有限公司 | A kind of antineoplastic pharmaceutical compositions and its preparation method and application being crosslinked sodium hyaluronate |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6521223B1 (en) * | 2000-02-14 | 2003-02-18 | Genzyme Corporation | Single phase gels for the prevention of adhesions |
| CN101611063A (en) * | 2006-11-10 | 2009-12-23 | 施泰福实验室股份有限公司 | Cross-linked-hyaluronic acid and preparation method thereof |
| CN101759881A (en) * | 2008-10-08 | 2010-06-30 | 上海建华精细生物制品有限公司 | Medical cross-linking sodium hyaluronate gel derivative product and preparation method thereof |
| CN101952325A (en) * | 2007-12-19 | 2011-01-19 | 诺维信生物制药丹麦公司 | Crosslinked hyaluronic acid in emulsion |
| CN102942699A (en) * | 2012-10-26 | 2013-02-27 | 暨南大学 | Self-reinforced bi-crosslinking hyaluronic acid hydrogel and preparation method thereof |
| CN102952275A (en) * | 2011-08-19 | 2013-03-06 | 上海建华精细生物制品有限公司 | Hyaluronic acid gel employing biphasic technology, and preparation method thereof |
-
2014
- 2014-12-25 CN CN201410844380.7A patent/CN104592420A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6521223B1 (en) * | 2000-02-14 | 2003-02-18 | Genzyme Corporation | Single phase gels for the prevention of adhesions |
| CN101611063A (en) * | 2006-11-10 | 2009-12-23 | 施泰福实验室股份有限公司 | Cross-linked-hyaluronic acid and preparation method thereof |
| CN101952325A (en) * | 2007-12-19 | 2011-01-19 | 诺维信生物制药丹麦公司 | Crosslinked hyaluronic acid in emulsion |
| CN101759881A (en) * | 2008-10-08 | 2010-06-30 | 上海建华精细生物制品有限公司 | Medical cross-linking sodium hyaluronate gel derivative product and preparation method thereof |
| CN102952275A (en) * | 2011-08-19 | 2013-03-06 | 上海建华精细生物制品有限公司 | Hyaluronic acid gel employing biphasic technology, and preparation method thereof |
| CN102942699A (en) * | 2012-10-26 | 2013-02-27 | 暨南大学 | Self-reinforced bi-crosslinking hyaluronic acid hydrogel and preparation method thereof |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105440159A (en) * | 2015-12-22 | 2016-03-30 | 上海景峰制药有限公司 | Alcohol precipitation method for preparation of cross-linked sodium hyaluronate intermediate HA-VS |
| CN109223708A (en) * | 2018-10-24 | 2019-01-18 | 上海景峰制药有限公司 | A kind of antineoplastic pharmaceutical compositions and its preparation method and application being crosslinked sodium hyaluronate |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104448075B (en) | For the method preparing heat stability Kynoar | |
| CN112724266A (en) | Preparation method of lithium carboxymethyl cellulose for lithium battery | |
| CN105062455A (en) | Water-based fracturing fluid thickening agent and synthetic method thereof | |
| CN103570844B (en) | Preparation method of carboxyethyl welan gum | |
| CN103059163B (en) | Method for preparing alginate oligosaccharide monomers by using microwave radiation | |
| CN104592420A (en) | Method for preparing intermediate HA-VS of cross-linked sodium hyaluronate | |
| CN110237824A (en) | The preparation method of the boron chelating adsorbent of meglumine functionalization cross-linked chitosan base | |
| CN107759735B (en) | Water-insoluble hemicellulose grafted polyacrylamide and preparation and application thereof | |
| CN110642357B (en) | Flocculating agent for microalgae capture and preparation method and application thereof | |
| CN103319418A (en) | Method for preparing sulfamonomethoxine sodium | |
| CN104475755A (en) | Preparation method of spheroidal ultrafine silver powder for front surface of solar cell | |
| CN105440159A (en) | Alcohol precipitation method for preparation of cross-linked sodium hyaluronate intermediate HA-VS | |
| CN102489261B (en) | Method for preparing heavy metal ion capture agent with environmental protection and high efficiency by using semicellulose | |
| CN103275338B (en) | A kind of method adopting glutaraldehyde to improve gracilaria agar gel strength | |
| CN103059144B (en) | Preparation method of high-viscosity pelletizing sodium carboxymethylcellulose | |
| CN104909987A (en) | Method for extracting mannitol in kelp | |
| CN102702390A (en) | Method for preparing low molecular weight chitosan through acidic ionic liquid aqueous solution degradation method | |
| CN104211088A (en) | Preparation method of cuprous cyanide | |
| CN102992293A (en) | Rod-shaped zirconium phosphate and preparation method thereof | |
| CN103467625B (en) | A kind of preparation method of agar-agar Polygalactan sulfuric ester | |
| CN104445303A (en) | Preparation method of magnesium hydrate | |
| CN104437416A (en) | Method for adsorbing trace lead ions in water | |
| CN104448048A (en) | Method for improving dissolving efficiency of crude heparin sodium | |
| CN109431972B (en) | Gelling method of sucralfate gel with particle size of 100 nm-500 nm | |
| CN118725148A (en) | Preparation method of lithium carboxymethyl cellulose |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20150506 |