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Method for extracting unmodified natural collagen from bullfrog skin

A technology of natural collagen and skin collagen, applied in the field of preparation of biomedical materials, can solve the problem of unsatisfactory yield of undenatured natural collagen, and achieve the effects of unique biocompatibility, integrity and excellent quality.

Active Publication Date: 2014-03-19
浙江珂瑞康生物医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the raw materials of the above methods are mostly mammals or fish, and amphibians are rarely involved. At the same time, the yield of undenatured natural collagen extracted by a single method is not ideal

Method used

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  • Method for extracting unmodified natural collagen from bullfrog skin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] (1) Raw material pretreatment

[0033] Remove muscle, fat and blood vessels from the fresh bullfrog skin, wash, squeeze dry, and chop; add NaOH solution with a material-to-liquid ratio of 1:20 and a concentration of 0.05M / L for 48 hours, keep stirring at a constant speed, and change the solution 5 times in the middle. Take out the bullfrog skin, wash it several times, adjust the pH to 8-12, add H 2 o 2 The solution was treated for 12 hours, kept stirring at a constant speed, and the solution was changed twice in the middle, the bullfrog skin was taken out, rinsed several times, the pH was adjusted to 6.0-6.5, the ratio of material to liquid was 1:10, and the volume concentration was 30% isopropanol for 24 hours. Keep stirring at a constant speed, take it out, rinse, soak in pure water overnight, squeeze dry, and set aside;

[0034] (2) Extraction of undenatured natural collagen

[0035] 60g (dry weight 15.6g) of the bullfrog skin obtained by the above pretreatment wa...

Embodiment 2

[0040] (1) Raw material pretreatment

[0041] Remove muscle, fat, and blood vessels from the fresh bullfrog skin, wash, squeeze dry, and chop; add NaOH solution with a material-to-liquid ratio of 1:40 and a concentration of 0.2M / L for 24 hours, keep stirring at a constant speed, and change the liquid twice midway. Take out the bullfrog skin, wash it several times, adjust the pH to 8-12, add H 2 o2 The solution was treated for 48 hours, kept stirring at a constant speed, and the liquid was changed 4 times in the middle, the bullfrog skin was taken out, rinsed several times, and the pH was adjusted to 6.0-6.5. Stir at a constant speed, take it out, rinse, soak in pure water overnight, squeeze dry, and set aside;

[0042] (2) Extraction of undenatured natural collagen

[0043] The bullfrog skin 100g (dry weight 26.0g) that above-mentioned pretreatment gained is that concentration is 2M / L acetic acid solution leaching 48h, keeps stirring at a constant speed, and the concentratio...

Embodiment 3

[0048] (1) Raw material pretreatment

[0049] Remove muscle, fat and blood vessels from the fresh bullfrog skin, wash, squeeze dry, and chop; add NaOH solution with a material-to-liquid ratio of 1:30 and a concentration of 0.1M / L for 36 hours, keep stirring at a constant speed, and change the liquid 4 times in the middle. Take out the bullfrog skin, wash it several times, adjust the pH to 8-12, add H 2 o 2 Treat the solution for 24 hours, keep stirring at a constant speed, change the solution 3 times in the middle, take out the bullfrog skin, rinse several times, adjust the pH to 6.0-6.5, use a material-to-liquid ratio of 1:30, and treat with acetone at a volume concentration of 50% for 18 hours, maintaining a constant speed Stir, take out, rinse, soak in pure water overnight, squeeze dry, set aside;

[0050] (2) Extraction of undenatured natural collagen

[0051] The bullfrog skin 80g (dry weight 20.8g) that above-mentioned pretreatment gained is that concentration is 1M / L...

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Abstract

The invention discloses a method for extracting unmodified natural collagen from bullfrog skin. The method is characterized by comprising the following steps of: treating washed bullfrog skin by using NaOH solution, H2O2 solution and a degreasing agent in the treatment phase to remove non-collagen protein, pigments and fat; leaching acetic acid solution and acetic acid-protease in the extraction phase to obtain acid-soluble collagen and enzyme-soluble collagen; performing salting-out for many times by utilizing a salt in the purification phase; and dialyzing and performing freeze-drying to obtain spongy bullfrog skin collagen. A lauryl sodium sulfate-polyacrylamide gel electrophoretogram indicates that the bullfrog skin collagen contains a gamma chain with the molecular weight of 0.3 million, a beta chain with the molecular weight of 0.2 million and alpha chains with the molecular weight of 0.1 million, wherein the alpha chains are two, and a triple helix structure of the collagen is better reserved. Unless otherwise specified, the operation temperature is below 15 DEG C in the extraction process, the dry weight extraction rate of the obtained product is more than or equal to 80 percent, and the collagen is white in color and high in quality.

Description

technical field [0001] The invention relates to a method for extracting undenatured natural collagen from bullfrog skin, belonging to the field of preparation of biomedical materials. Background technique [0002] Collagen is the most abundant protein in animals, accounting for 25%-30% of the total protein in the body. It is the main component of tendons, cartilage, skin and blood vessels. Type I collagen molecules are composed of three peptide chains and two αI chains , an α∏ chain, three peptide chains are intertwined with a unique left-handed helix to form a right-handed helix, and the molecular structure shows a unique glycine-X-Y (X, Y are other types of amino acids, mostly proline, hydroxyproline, gluten amino acid and alanine) periodic structure. The unique triple helical structure of collagen makes its molecular structure very stable, and has low immunogenicity and good biocompatibility. Therefore, collagen has a wide range of applications in medicine, beauty, cosme...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/78C07K1/36C07K1/34C07K1/30C12P21/06A23L1/305A61K8/65
Inventor 李国英杨波刘文涛
Owner 浙江珂瑞康生物医疗科技有限公司
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