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Method for extracting HA (hyaluronic acid) from tuna eyes

A technology for hyaluronic acid and tuna, which is applied in the field of extracting hyaluronic acid from tuna eyes, can solve the problems of complicated operation and high production process cost, and achieve the effects of high purity, reduced production cost and small loss.

Inactive Publication Date: 2017-12-29
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The hyaluronic acid product produced by this method has the advantages of cheapness, safety, simple technical route, and easy operation, and solves the problems of high cost, original scarcity, and complicated operation of the existing HA production process.

Method used

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  • Method for extracting HA (hyaluronic acid) from tuna eyes
  • Method for extracting HA (hyaluronic acid) from tuna eyes
  • Method for extracting HA (hyaluronic acid) from tuna eyes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1: tuna eye pretreatment prepares fish eye powder

[0023] Take 4314g of frozen fresh fish eye, after thawing at normal temperature, remove the shell, fish tissue, optic nerve, etc., and keep the black fish eye vitreous body, which weighs 1431g. During the operation, the liquid in the vitreous is easy to flow out and should be recovered as much as possible. Cut the black glass into pieces, add 3 times the volume of acetone, and stir at room temperature for 6 h. Then centrifuge at 10,000 rpm at 4°C, take the precipitate and dry it in a vacuum oven at 45°C for 10 hours, and take out and turn over the product from time to time. After the precipitate was dried, it was pulverized and then dried in a vacuum oven at 45°C for 10 hours. The obtained powder was 210g of fish eye powder, which could be used as a raw material for extracting hyaluronic acid, and could also be stored at -20°C for a long time.

Embodiment 2

[0024] Embodiment 2: tuna eye pretreatment prepares fish eye powder

[0025] Take 2932 g of frozen fresh fish eyes, after thawing at normal temperature, remove the shell, fish tissue, optic nerve, etc., and keep the black fish eye vitreous body, which weighs 941 g. During the operation, the liquid in the vitreous is easy to flow out and should be recovered as much as possible. Cut the black glass into pieces, add 3 times the volume of acetone, and stir at room temperature for 4 h. Then centrifuge at 10,000 rpm at 4°C, take the precipitate and dry it in a vacuum oven at 30°C for 6 hours, and take out and turn over the product from time to time. After the precipitate was dried, it was crushed and then dried in a vacuum oven at 30°C for 6 hours. The obtained powder was 154g of fish eye powder, which could be used as a raw material for extracting hyaluronic acid, and could also be stored at -20°C for a long time.

Embodiment 3

[0026] Example 3: Enzymatic Extraction of Fisheye Hyaluronic Acid

[0027] The fish eye powder obtained by the method of Example 1 is used as a raw material, and the process of extracting hyaluronic acid by enzymatic hydrolysis is as follows

[0028] (1) Weigh 25g of fish eye powder, add 500mL of deionized water, and stir at room temperature for 6h.

[0029] (2) Centrifuge the reaction solution of (1), take the supernatant and add 500U trypsin / g fish eye powder, and react at 40°C and pH 8.5 for 120min.

[0030] (3) After inactivating the reaction solution of (2) in a 90°C water bath, cool it down. Then centrifuge at 10000 rpm at 4°C to obtain supernatant.

[0031] (4) Put 500mL of the supernatant in (3) into the separatory funnel, then add 200mL of chloroform, 50mL of n-butanol, stir vigorously for 20min, and put it in the separatory funnel for stratification (divided into upper, middle and lower layers) , release the lower layer of chloroform and the middle layer of protei...

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Abstract

The invention discloses a method for extracting HA (hyaluronic acid) from tuna eyes. The method comprises the following steps: tuna eye powder is added to deionized water, sufficient extraction and centrifugation are performed, and a liquid supernatant is obtained; trypsin is added to the liquid supernatant, the materials react under the conditions of 20-45 DEG C and pH being 7.0-8.5 for 60-180 min, and a crude HA product is obtained after sufficient reaction; an HA solution with mass concentration being 1%-5% is prepared from the crude HA product and deionized water, sodium chloride with final concentration being 0.1-0.3 mol / L is added, and mother liquor is prepared; a cetylpyridinium chloride monohydrate aqueous solution with mass concentration being 1%-10% is added to the mother liquor, the mixed solution is stirred at 30 DEG C and reacts sufficiently, and HA is obtained. The molecular weight of HA extracted from tuna is smaller and about 0.470 MDa, the process route is simple and feasible, the purity of obtained HA is higher, and HA is less lost in the process flow.

Description

(1) Technical field [0001] The invention relates to an extraction process and product purification method for producing medical hyaluronic acid from tuna eyes. (2) Background technology [0002] Hyaluronic acid (HA) is an anionic acidic mucopolysaccharide composed of β-(1→3)-D-N-acetylglucosamine and β-(1→4)-D-glucuronic acid disaccharide units repeatedly linked Become. Hyaluronic acid has unique moisturizing properties and bioaffinity. It exerts its physical functions in the body through its physical functions such as viscoelasticity (macromolecule network structure) and high hydration, as well as its physiological functions by interacting with HA receptors. Due to its important role, these two functions are currently widely used in cosmetics and pharmaceutical industries. At present, the production processes of hyaluronic acid mainly include biological tissue extraction method and streptococcus fermentation method. Research shows that almost all animal tissues contain H...

Claims

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Application Information

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IPC IPC(8): C08B37/08
CPCC08B37/0003C08B37/0072
Inventor 应国清易喻徐骏梅建凤陈建澍张彦璐
Owner ZHEJIANG UNIV OF TECH
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