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Method for rapid preparation of sodium hyaluronate from sodium hyaluronate fermentation broth

A technology of sodium hyaluronate and fermentation liquid, which is applied in the field of preparation of sodium hyaluronate, can solve the problems of low protein content, low cost, and high purity of powdered HA, reduce the yield of HA, reduce production time, and save complex The effect of melting time

Inactive Publication Date: 2014-07-23
SHANGHAI HAOHAI BIOLOGICAL TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention aims to overcome the time-consuming and laborious step of ethanol precipitate or CPC precipitate redissolution in the prior art, and provides a method for directly obtaining powdered sodium hyaluronate from fermentation broth, which is time-consuming Less, low cost, the obtained powdered HA has high purity and low protein content

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] 1) Take 1000ml of fermented broth, containing 4g of HA, with a molecular weight of 2.26MDa.

[0015] 2) The temperature of the fermentation broth is raised to 100° C., maintained for 20 minutes, and filtered to obtain a filtrate.

[0016] 3) The filtrate was diluted to 3000 ml with purified water, 20 g / L of activated carbon and 2 g / L of diatomaceous earth were added, stirred and adsorbed for 4 hr, and filtered to obtain the filtrate.

[0017] 4) Add 500 g of D113 resin to the filtrate, stir and absorb for 1 hour, then filter off the resin to obtain the filtrate.

[0018] 5) Add sodium chloride to the filtrate to make the final concentration 0.4mol / L, use 2-3 times of ethanol to precipitate HA, then wash, solidify and dry to obtain powdered HA.

[0019] The whole process took 8 hours, and the obtained HA dry powder was 3.8g, the purity of glucuronic acid was about 43.6%, the molecular weight was 1.67MDa, protein / HA (g / g)=0.08%.

Embodiment 2

[0021] 1) Take 1000ml of fermented broth, containing 4g of HA, with a molecular weight of 2.26MDa.

[0022] 2) The temperature of the fermentation broth was raised to 121° C., maintained for 10 minutes, and filtered to obtain a filtrate.

[0023] 3) The filtrate was diluted to 4000 ml with purified water, 50 g / L of activated carbon and 2 g / L of diatomaceous earth were added, stirred and adsorbed for 2 hr, and filtered to obtain the filtrate.

[0024] 4) Add 500 g of D113 resin to the filtrate, stir and absorb for 2 hours, then filter off the resin to obtain the filtrate.

[0025] 5) Add sodium chloride to the filtrate to make the final concentration 0.4mol / L, use 2-3 times of ethanol to precipitate HA, then wash, solidify and dry to obtain powdered HA.

[0026] The whole process takes 7 hours, and the obtained HA dry powder is 3.6g, the glucuronic acid purity is about 44.5%, the molecular weight is 1.76MDa, protein / HA (g / g)=0.06%.

Embodiment 3

[0028] 1) Take 1000ml of fermented broth, containing 4g of HA, with a molecular weight of 2.26MDa.

[0029] 2) The temperature of the fermentation broth was raised to 110° C., maintained for 15 minutes, and filtered to obtain a filtrate.

[0030] 3) The filtrate was diluted to 3000 ml with purified water, 40 g / L of activated carbon and 2 g / L of diatomaceous earth were added, stirred and adsorbed for 5 hours, and filtered to obtain the filtrate.

[0031] 4) Add 500 g of D113 resin to the filtrate, stir and absorb for 1 hour, then filter off the resin to obtain the filtrate.

[0032] 5) Add sodium chloride to the filtrate to make the final concentration 0.4mol / L, use 2-3 times of ethanol to precipitate HA, then wash, solidify and dry to obtain powdered HA.

[0033] The whole process took 9 hours, and the obtained HA dry powder was 3.7g, the purity of glucuronic acid was about 43.8%, the molecular weight was 1.70MDa, protein / HA (g / g)=0.09%.

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Abstract

The invention provides a method for rapid preparation of sodium hyaluronate from a sodium hyaluronate fermentation broth, and the method comprises the following steps: 1) heating the sodium hyaluronate fermentation broth to 100-121 DEG C, maintaining for 10 to 30 minutes, and then quickly performing plate-frame pressure filtration; 2) diluting the filtrate with purified water of 2-4 times, adding 1%-5% of activated carbon for adsorption for 1-5 hours, performing the plate-frame pressure filtration; 3) adsorbing the filtrate with resin for 1 to 2 hours, proportionally adding 0.4mol / L sodium chloride into the adsorption solution; 4) precipitating the sodium hyaluronate with 2-3 times of ethanol, washing, curing and drying to obtain powdery sodium hyaluronate. The method for extraction and preparation of the sodium hyaluronate has the advantages that: redissolving of a crude product is omitted in the whole extraction process, the extraction time is greatly reduced, and the obtained sodium hyaluronate is high in purity, low in protein content, simple in process, less in preparation time, and conducive to large-scale industrial production.

Description

technical field [0001] The invention relates to a preparation method of sodium hyaluronate, in particular to a method for separating and extracting sodium hyaluronate from a sodium hyaluronate fermentation broth. Background technique [0002] Hyaluronic acid (Hyaluronic Acid, HA) is a linear polymer formed by repeating units of β-D-glucuronic acid and N-acetamidoglucose through β, 1-3 and β, 1-4 glycosidic bonds. Mucopolysaccharide, the number of disaccharide units is 300-1100, and the molecular weight range is 10 5 -10 7 Da. [0003] Since Mayer first extracted HA from the vitreous of animal eyes in the 1930s, the production and application research of HA has a history of more than 70 years. HA has unique viscoelasticity and physiological functions. It is the main component of the extracellular matrix. It has the functions of water retention, lubrication, and regulation of osmotic pressure. differentiation etc. Therefore, HA is widely used in orthopedics, ophthalmology...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/08
Inventor 肖刚薛勇侯永泰任彩霞李映严
Owner SHANGHAI HAOHAI BIOLOGICAL TECH
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