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Comprehensive extracting process for collagen type II and chondroitin sulfate

A chondroitin sulfate, comprehensive extraction technology, applied in the direction of fermentation, etc., can solve the problems of waste of resources, reduce production costs, discarding, etc., and achieve the effect of saving resources and avoiding denaturation

Inactive Publication Date: 2012-12-19
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the quality of the product produced by the alkaline hydrolysis-enzyme method is high, and the production cycle is greatly shortened, but the optimal process conditions have not been determined, and it is necessary to further optimize the extraction conditions, improve product quality, and reduce production costs.
[0005] At present, the extraction, separation, and purification processes of type II collagen and chondroitin sulfate are relatively mature, but the extraction of these two substances is mostly carried out separately at home and abroad. When one of the substances is extracted, the other will be denatured. processing or discarding, resulting in a great waste of resources

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Take fresh bovine articular cartilage, cut it into small pieces, put it into 500mL of 5% NaCl solution, stir at 20°C for 48h, filter, wash twice with distilled water, then put it into 500mL of 0.5mol / L acetic acid solution, at 20 Stir at ℃ for 48h, wash with distilled water twice, and pulverize with a pulverizer. Weigh 20 g of the pretreated pulverized material, disperse it in 400 mL of 0.5 mol / L acetic acid solution, add 0.4 g of pepsin, stir at room temperature for 98 h, and centrifuge at 3000 r / min for 10 min to obtain Supernatant A and pellet B. Take the supernatant A, add NaCl solution to a final concentration of 0.9mol / L, let it stand overnight for 12 hours for salting out, and then centrifuge to obtain white flocs, put the above white flocs into a dialysis bag, and dialyze with distilled water for 3 days , desalted to obtain a purified type II collagen solution, and the purified type II collagen solution was vacuum freeze-dried for 24 hours to obtain a dry type ...

Embodiment 2

[0032] Take fresh bovine articular cartilage, cut into small pieces, put into 400mL of 7.5% NaCl solution, stir at 25°C for 36h, filter, wash twice with distilled water, then put into 400mL of 0.75 mol / L acetic acid solution, Stir at ℃ for 36h, wash with distilled water twice, and pulverize with a pulverizer. Weigh 30 g of the pretreated pulverized material, disperse it in 1500 mL of 0.5 mol / L acetic acid solution, add 1.0 g of pepsin, stir at room temperature for 86 h, and centrifuge at 3000 r / min for 10 min to obtain Supernatant A and pellet B. Take the supernatant A, add NaCl solution to a final concentration of 1.2 mol / L, let stand overnight for 12 hours for salting out, and then centrifuge to obtain white flocs, put the above white flocs into a dialysis bag, and dialyze with distilled water for 4 days , desalted to obtain a purified type II collagen solution, and the purified type II collagen solution was vacuum freeze-dried for 24 hours to obtain a dry type II collagen ...

Embodiment 3

[0034]Take fresh bovine articular cartilage, cut into small pieces, put into 300mL of 10% NaCl solution, stir at 25°C for 24h, filter, wash twice with distilled water, then put into 300mL of 0.5mol / L acetic acid solution, Stir at ℃ for 24h, wash with distilled water twice, and pulverize with a pulverizer. Weigh 40 g of the pretreated pulverized material, disperse it in 4000 mL of 0.5 mol / L acetic acid solution, add 4.0 g of pepsin, stir at room temperature for 72 h, and centrifuge at 3000 r / min for 10 min to obtain Supernatant A and pellet B. Take the supernatant A, add NaCl solution to a final concentration of 1.5mol / L, let stand overnight for 12 hours for salting out, and then centrifuge to obtain white flocs, put the above white flocs into a dialysis bag, and dialyze with distilled water for 5 days , desalted to obtain a purified type II collagen solution, and the purified type II collagen solution was vacuum freeze-dried for 24 hours to obtain a dry type II collagen produ...

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Abstract

The invention discloses a comprehensive extracting process for collagen type II and chondroitin sulfate. The comprehensive extracting process comprises the following steps of: using fresh cattle articular cartilages as raw materials; degreasing and de-calcifying the fresh cattle articular cartilages; dispersing the degreased and de-calcified fresh cattle articular cartilages into an acetic acid solution; performing enzymolysis and centrifuging the mixture to acquire liquid supernatant; salting out, dialyzing, freezing and drying the liquid supernatant to acquire collagen type II; dissolving precipitate acquired from centrifuging to a NaOH solution, and adding trypsin to the mixture; and de-coloring, ethanol- precipitating, dialyzing, freezing and drying the mixture after the enzymolysis at 40 DEG C to acquire the chondroitin sulfate. Collagen type II is the main organic component of a cartilage matrix, and is widely applied in the fields of tissue engineering, food and the like. The chondroitin sulfate widely exists in the cartilage tissues of the human and animals, has such functions of reducing blood fat, preventing tumors and the like, and has an extensive application prospect. According to the invention, bovine cartilage collagen type II and chondroitin sulfate are comprehensively used, so as to provide a novel idea for recycling waste.

Description

technical field [0001] The invention relates to a process for comprehensively extracting type II collagen and chondroitin sulfate from bovine articular cartilage. Background technique [0002] Collagen is a class of proteins with a unique structure that is widely found in animal tissues. Type II collagen belongs to fibrillar collagen, mainly composed of three identical α 1 (II) The chain constitutes the basic structure of cartilage in a helical manner, is the main organic component of the cartilage matrix, and has the function of maintaining the morphology and tensile strength of articular cartilage. The unique structure and chemical composition of type II collagen make it widely used in the fields of biomedical materials, drug delivery vehicles, tissue engineering, cosmetics and food. Recent studies have shown that type II collagen with a complete triple helix structure can also be used as medicine and health care products to prevent and treat rheumatoid arthritis, so the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C08B37/08
Inventor 李德富毕彩霞穆畅道李新莹王瑞瑞葛黎明
Owner SICHUAN UNIV
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