Method for preparing undenatured natural collagen from sucking pig skin

A technology of natural collagen and skin collagen, which is applied in the field of preparation of biomedical materials, can solve the problems of lack of collagen fibers, difficulty in removing fat, and inapplicability to the preparation of undenatured natural suckling pig skin collagen, so as to ensure biological activity and low immunity original effect

Active Publication Date: 2014-02-26
SICHUAN UNIV
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  • Claims
  • Application Information

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Problems solved by technology

The collagen fibers in the dermis do not have a fixed texture, the collagen fiber bundles are smaller than those of adult pig skin, and the gaps between fiber bundles are larger than those of adult pig skin collagen. fiber destruction
And the fat content in the dermis is equivalent to that of adult pigskin, close to 15%. The presence of a large amount of fat seriously affects the penetration of chemicals. Therefore, it is difficult to remove fat w

Method used

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  • Method for preparing undenatured natural collagen from sucking pig skin
  • Method for preparing undenatured natural collagen from sucking pig skin

Examples

Experimental program
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Effect test

Embodiment 2

[0043] (1) Raw material pretreatment

[0044] Take 100g of fresh suckling pig skin, remove the subcutaneous fat, and treat it with isopropanol with a material-to-liquid ratio of 1:10 (w / v) and a volume concentration of 45% for 36 hours, accompanied by an intermittent power of 150W ultrasonic wave every 20 minutes and stop for 10 minutes. Repeat 8 times, change the medium 4 times in the middle, rinse with tap water, add the protease Nowolase NG solution with a material-to-liquid ratio of 1:2.5 (w / v), and a volume concentration of 1.5%, keep the temperature at 35°C, rotate for 45 minutes to remove hair, and then Chop the skin, add a solution of sodium deoxycholate with a ratio of material to liquid of 1:30 (w / v), and a concentration of 0.3M / L sodium deoxycholate, stir for 36 hours, change the solution 4 times in the middle, rinse with tap water, adjust the pH to 8.0, and measure Ash content 0.2%. Then use a material-liquid ratio of 1:10 (w / v), volume concentration of 35% aceton...

Embodiment 3

[0051] (1) Raw material pretreatment

[0052] Take 100g of fresh suckling pigskin, remove the subcutaneous fat, and treat it with AXAUT AP with a solid-liquid ratio of 1:5 (w / v) and a volume concentration of 60% for 48 hours, accompanied by intermittent power of 200W ultrasonic waves every 10 minutes and stop for 5 minutes, repeat 10 times, change the liquid 5 times in the middle, rinse with tap water, add the protease Basozyme L10 solution with a solid-liquid ratio of 1:1.5 (w / v) and a volume concentration of 3%, keep the temperature at 45°C, and rotate for 60 minutes for hair removal, then put Cut the skin into pieces, add a solution of sodium deoxycholate with a ratio of material to liquid of 1:40 (w / v) and a concentration of 0.05M / L, stir for 48 hours, change the solution 5 times in the middle, rinse with tap water, adjust the pH to 9.0, and measure the ash content Content 0.2%. Then use a material-liquid ratio of 1:6 (w / v), volume concentration of 50% acetone and AXAUT-5...

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Abstract

The invention discloses a method for extracting undenatured natural collagen from sucking pig skin. The method is characterized in that treatment is carried out by sub-step repeated different combinations in a manner that a degreasing agent is assisted by intermittent ultrasonic waves, an unhairing agent and a non-collagen removing agent are respectively used for unhairing and removing non-collagen, and no hair root exists in the skin after unhairing, wherein shown by determination, the fat content is lower than 0.5% and the ash content is lower than 0.3%; enzyme-soluble collagen is obtained through leaching by acetic acid-protease and is purified by high-speed centrifugation, sodium chloride salting-out and dialysis, and spongy sucking pig skin collagen is obtained after freeze-drying, wherein the collagen extraction ratio exceeds 50% (dry weight), and shown by electrophoretogram, the sucking pig skin collagen contains a beta chain and an alpha chain, of which the molecular weights are 200000 and 100000 respectively, and is free from other hybrid protein bands, so that the collagen reserves three helical structures and has relatively high purity.

Description

technical field [0001] The invention relates to a method for preparing undenatured natural collagen from suckling pig skin, belonging to the field of preparation of biomedical materials. Background technique [0002] Undenatured natural collagen still maintains its unique triple helical structure, which is a right-handed superhelical structure, and its molecular structure is a unique glycine-X-Y (X, Y are other types of amino acids, mostly proline and hydroxyproline) periodic structure . It is mainly distributed in the skin, bone, cornea, tendon and other parts. It has the functions of low antigenicity, good biocompatibility and biodegradability that cannot be compared with synthetic polymers. Its application has been widely penetrated into burn wounds, orthopedics and Hard tissue repair and other fields. [0003] The wide application of undenatured natural collagen will inevitably lead to an increasing demand for it, and higher and higher requirements for its quality. Co...

Claims

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Application Information

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IPC IPC(8): C07K14/78C07K1/30
CPCC07K14/78
Inventor 李国英吴论文刘文涛
Owner SICHUAN UNIV
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