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Method for extracting hyaluronic acid from eggshell membranes of poultry

A technology of hyaluronic acid and eggshell membrane is applied in the field of extraction of hyaluronic acid in poultry eggshell membrane, which can solve the problems of endotoxin pollution, hyaluronic acid molecular weight (difficult to grasp the length, limited output of animal tissue, etc.) Broad, molecular weight stable, endotoxin-free effect

Active Publication Date: 2016-08-24
ZHEJIANG FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The former method limits the output of hyaluronic acid due to the limited production of animal tissues; the latter method, because the molecular weight (length) of hyaluronic acid produced is difficult to grasp and is easily polluted by endotoxins, affects the further development of hyaluronic acid. application

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] A method for extracting hyaluronic acid in poultry eggshell membranes, the method comprising the following steps:

[0054] 1) Separation of shell membranes and calcified shells: Soak eggshells with 3% acetic acid for 12 hours, then manually peel off the eggshells to quickly separate and obtain a large number of shell membranes;

[0055] 2) freeze-drying the shell membrane for 1 day, and pulverizing the shell membrane with a pulverizer;

[0056] 3) Separating and extracting total glycosaminoglycans from the pulverized shell membrane, specifically comprising the following steps:

[0057] A. to digest protein

[0058] Suspend the shell membrane powder in phosphate buffer (PBS buffer, pH=8.4) containing 0.001wt% sodium dodecylsulfonate (SDS), the mass ratio of the shell membrane to PBS is 1:50;

[0059] Add pronase (Actinase E) and conduct water bath treatment at 38°C for 20 hours, the mass ratio of pronase to shell membrane is 1:10; inactivate Actinase E at 90°C for 15 m...

Embodiment 2

[0070] A method for extracting hyaluronic acid in poultry eggshell membranes, the method comprising the following steps:

[0071] 1) Separation of shell membranes and calcified shells: Ethylenediaminetetraacetic acid disodium salt with a volume concentration of 5% is used to soak eggshells for 36 hours, and then manually peeled off or shaken to separate, and a large amount of shell membranes can be quickly separated;

[0072] 2) Freeze-dry the shell membrane for 2 days, and pulverize the shell membrane with an ultrafine pulverizer;

[0073] 3) Separating and extracting total glycosaminoglycans from the pulverized shell membrane, specifically comprising the following steps:

[0074] A. to digest protein

[0075] Suspend the shell membrane powder in phosphate buffer (PBS buffer, pH=8.5) containing 0.002wt% sodium dodecylsulfonate (SDS), the mass ratio of the shell membrane to PBS is 1:40;

[0076] Add pronase (Actinase E), and conduct water bath treatment at 37°C for 28 hours,...

Embodiment 3

[0087] A method for extracting hyaluronic acid in poultry eggshell membranes, the method comprising the following steps:

[0088] 1) Separation of shell membrane and calcified shell: After soaking the eggshell with 10% acetic acid by volume concentration for 12 hours, peeling off manually can quickly separate and obtain a large amount of shell membrane;

[0089] 2) Freeze-dry the shell membrane for 3 days, and pulverize the shell membrane with an ultrafine pulverizer;

[0090] 3) Separating and extracting total glycosaminoglycans from the pulverized shell membrane, specifically comprising the following steps:

[0091] A. to digest protein

[0092] Suspend the shell membrane powder in sodium dodecylsulfonate (SDS)-free phosphate buffer (PBS buffer, pH=8.2), the mass ratio of the shell membrane to PBS is 1:30;

[0093] Add pronase (Actinase E) and conduct water bath treatment at 39°C for 12 hours, the mass ratio of pronase to shell membrane is 1:10; inactivate Actinase E at 90...

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PUM

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Abstract

The invention provides a method for extracting hyaluronic acid from eggshell membranes of poultry. The method includes steps: separating the eggshell membranes from calcified shells; drying and grinding the eggshell membranes; separating and extracting total glycosaminoglycan from the ground eggshell membranes; removing heparin / heparan sulfate and keratan sulfate from the total glycosaminoglycan, and separating the hyaluronic acid from chondroitin sulfate / dermatan sulfate. By the method, high-purity hyaluronic acid can be separated out, direct extraction of the hyaluronic acid from the eggshell membranes of poultry is realized, extensive sources are available, and the obtained hyaluronic acid which is derived from natural tissues of animals is stable in molecular weight and free of endotoxin.

Description

technical field [0001] The invention belongs to the field of hyaluronic acid extraction, and in particular relates to a method for extracting hyaluronic acid from poultry eggshell membranes. Background technique [0002] Hyaluronic acid is a linear macromolecular polysaccharide rich in anions, which can bind to various cell growth factors, enzymes, cytokines and receptors in animals to induce signal transduction, thereby regulating cell growth, differentiation and apoptosis. Death. Therefore, hyaluronic acid is widely used in the fields of medicine (for example, for the treatment of arthritis, tendon and soft tissue injuries, as an adjuvant for cancer nanomedicine, etc.), food health care and cosmetics (for example, for anti-wrinkle cosmetics, etc.). [0003] At present, there are two main sources of hyaluronic acid. One is animal tissues, such as rooster combs and fetal navels, mostly from slaughterhouses or hospitals; the other is through microbial fermentation. The form...

Claims

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Application Information

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IPC IPC(8): C08B37/08
CPCC08B37/0003C08B37/0072
Inventor 刘长国赵红娟
Owner ZHEJIANG FORESTRY UNIVERSITY
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