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Extraction method of hyaluronic acid in poultry eggshell membrane

A hyaluronic acid and eggshell membrane technology, which is applied in the field of extraction of hyaluronic acid from poultry eggshell membranes, can solve the problems of endotoxin pollution, hyaluronic acid molecular weight (difficult to grasp the length, limited production of animal tissues, etc.), and reach the source Broad, stable molecular weight, endotoxin-free effects

Active Publication Date: 2018-07-24
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 from poultry eggshell membranes, the method comprising the following steps:

[0054] 1) Separation of shell membrane and calcified shell: After soaking the egg shell with 3% acetic acid in volume concentration for 12 hours, manually peel it off, and then quickly separate and obtain a large amount of shell membrane;

[0055] 2) Freeze-dry the shell membrane for 1 day, and crush the shell membrane with a pulverizer;

[0056] 3) Separating and extracting total glycosaminoglycans from crushed shell membranes, specifically including the following steps:

[0057] A. Digest protein

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

[0059] Add Pronase E, and conduct water bath treatment at 38°C for 20 hours. The mass ratio of pronase to shell membrane is 1:10; Actinase E is inactivated at 90°C fo...

Embodiment 2

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

[0071] 1) Separation of shell membranes and calcified shells: Use 5% ethylenediaminetetraacetic acid disodium salt to soak the egg shells for 36 hours, then manually peel or shake them to separate, and then quickly separate and obtain a large amount of shell membranes;

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

[0073] 3) Separating and extracting total glycosaminoglycans from crushed shell membranes, specifically including the following steps:

[0074] A. Digest protein

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

[0076] Add Pronase E, and perform water bath treatment at 37°C for 28 hours. The mass ratio of pronase to shell membrane is 1:40...

Embodiment 3

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

[0088] 1) Separation of shell membranes and calcified shells: After soaking the egg shells with 10% acetic acid in volume concentration for 12 hours, peel them off manually to quickly separate and obtain a large amount of shell membranes;

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

[0090] 3) Separating and extracting total glycosaminoglycans from crushed shell membranes, specifically including the following steps:

[0091] A. Digest protein

[0092] The shell membrane powder is suspended in a phosphate buffer (PBS buffer, pH=8.2) without sodium dodecyl sulfonate (SDS), and the mass ratio of the shell membrane to PBS is 1:30;

[0093] Add Pronase E (Actinase E), carry out 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°C ...

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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 extraction of hyaluronic acid, and specifically relates to a method for extracting hyaluronic acid from poultry eggshell membranes. Background technique [0002] Hyaluronic acid is a linear linear macromolecular polysaccharide rich in anions. It can combine with a variety of cell growth factors, enzymes, cytokines and receptors in animals to induce signal transduction, thereby regulating cell growth, differentiation and apoptosis. Die. Therefore, hyaluronic acid is widely used in the fields of medicine (for example, for the treatment of arthritis, tendon and soft tissue injuries, as a cancer nanomedicine adjuvant, etc.), food health 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 belly buttons, mostly from slaughter workshops or hospitals; the other is through microbial fermentation. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08B37/08
CPCC08B37/0003C08B37/0072
Inventor 刘长国赵红娟
Owner ZHEJIANG FORESTRY UNIVERSITY
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