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Method for extracting collagen from bovine achilles tendons

A technology of bovine Achilles tendon and collagen, applied in the direction of animal/human protein, connective tissue peptide, specific peptide, etc., can solve the problems of not being able to meet medical purposes, reduce mechanical strength, easy to break, etc., achieve good integrity, simple method, The effect of low extraction cost

Inactive Publication Date: 2017-04-26
BEIJING HOTWIRE MEDICAL TECH DEV CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In the process of realizing the present invention, the inventors have found that the prior art has at least the following problems: in the process of extracting collagen from raw materials, the triple helix of collagen maintained by hydrogen bonds is easily broken, and is easily hydrolyzed into collagen, while The mechanical strength is greatly reduced, which cannot meet many medical purposes

Method used

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  • Method for extracting collagen from bovine achilles tendons
  • Method for extracting collagen from bovine achilles tendons
  • Method for extracting collagen from bovine achilles tendons

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1-4

[0048] Example 1-4 extracts collagen

Embodiment 1

[0050] Select 1 kg of fresh bovine Achilles tendon tissue, wash it with water for 6 times, and rinse it with 75% alcohol for 20 minutes; remove fascia and impurities with scissors, wash with water, and freeze overnight at -20°C; slice with a microtome to about 1 mm, and set aside;

[0051] Weigh 100g tendon slices, rinse them with purified water for 5 times; soak in 5L of purified water at room temperature for 5 hours; rinse them with purified water for 5 times, and drain the water for later use;

[0052] The cleaned tendon piece (i.e. sectioned bovine Achilles tendon) is soaked in 0.05mol / L phosphate buffered saline solution containing 0.05% ficin with a pH value of 6, after soaking for 24 hours at 4°C; Mix hydrogen peroxide equal to 3‰ of the volume of phosphate buffer, let stand for 4 hours to inactivate ficin, wash with purified water; then soak in 1M sodium hydroxide and sodium chloride solution for 4 days at 4°C, Shake gently in the middle of standing; add 0.05mol / L hydr...

Embodiment 2

[0055] Select 1 kg of fresh bovine Achilles tendon tissue, wash it with water for 9 times, and rinse it with 45% alcohol for 30 minutes; remove fascia and impurities with scissors, wash with water, and freeze overnight at -20°C; slice with a microtome to about 2mm, and set aside;

[0056] Weigh 100g tendon slices, rinse them with purified water for 5 times; soak in 5L of purified water at room temperature for 3 hours; rinse them with purified water for 5 times, and drain the water for later use;

[0057] The cleaned tendon slices (i.e. sectioned bovine Achilles tendon) were soaked in 0.01mol / L phosphate buffered saline solution containing 0.005% ficin with a pH value of 5, after soaking for 24 hours at 4°C; Sodium chlorite, which is 1‰ of the volume of the phosphate buffer solution, was mixed evenly and left to stand for 4 hours to inactivate ficin, rinsed with purified water; then soaked in 1.5M sodium hydroxide and sodium chloride solution for 4 After 1 day, shake gently in ...

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Abstract

The invention discloses a method for extracting collagen from bovine achilles tendons. The method includes the following steps that the bovine achilles tendons are sliced; the sliced bovine achilles tendons are soaked in a phosphate buffer solution with the mass fraction of ficin proteinase being 0.005%-0.2%, and the mixture is kept for 12 h to 24 h at the temperature of 4 DEG C to 15 DEG C; then an oxidizing agent is added into the phosphate buffer solution, and the sliced bovine achilles tendons are washed with water, wherein the oxidizing agent is selected from at least one of chlorite and hydrogen peroxide; finally, the sliced bovine achilles tendons are soaked in a saline solution with the molar concentration of sodium hydroxide being 1 mol / L to 2 mol / L, the mixture is kept for 1 day to 4 days at the temperature of 4 DEG C to 15 DEG C, then the pH value of the saline solution is adjusted to be 4 to 7, a product is collected, repeatedly washed and dehydrated, and the collagen is obtained. The collagen obtained with the method for extracting the collagen from the bovine achilles tendons is good in triple-helical structure integrity and high in purity; the method is simple, a high-end device is not required, and the extracting cost is low.

Description

technical field [0001] The invention relates to the technical field of biomedical materials, in particular to a method for extracting collagen from bovine Achilles tendon. Background technique [0002] There are many types of collagen, the common types are type Ⅰ, type Ⅱ, type Ⅲ, type Ⅴ and type Ⅺ. Collagen has good biocompatibility, biodegradability and biological activity, such as low antigenicity and easy to be absorbed in the body. It can be absorbed by the human body, can promote cell survival and growth, and promote platelet aggregation, etc., and has been widely used in food, medicine, tissue engineering, cosmetics and other fields. In the medical field, collagen can be made into collagen sponge, collagen membrane, artificial skin and capsules due to its low immunogenicity, fiber re-formation, high mechanical properties and biodegradability. Type I collagen in collagen is ubiquitous in animals, accounting for about 90% of the total amount of collagen in animals and n...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/78C07K1/14
Inventor 富勇王丽君邓贺颖周月芬
Owner BEIJING HOTWIRE MEDICAL TECH DEV CO LTD
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