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Method for preparing micro-fiber collagen by utilizing ion liquid in different steps

An ionic liquid and microfiber technology, which is applied in the preparation methods of peptides, chemical instruments and methods, animal/human proteins, etc., can solve the problems of loss of biological activity, difficult removal, toxicity, etc., so as to reduce the amount of use and ensure biological activity. , the effect of reducing the cost of industrialization

Active Publication Date: 2019-01-25
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] 1. The temperature in the process of preparing microfibrous collagen is 90-130°C, far exceeding the denaturation temperature of collagen, which loses its biological activity and cannot be used in the field of biomedicine;
[0007] 2. It adopts ionic liquid / dimethyl sulfoxide system to prepare microfibril collagen. Dimethyl sulfoxide has certain toxicity and is difficult to completely remove, so the prepared microfibril collagen cannot be applied to biomedical materials;
[0008] 3. The prepared microfibrous collagen is not sterilized and has certain biological toxicity;
[0009] 4. The content of short fibers (fiber length ≤ 1 mm) in the prepared microfibrous collagen is insufficient, does not meet the standards of biomedical materials, and the yield is not high

Method used

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  • Method for preparing micro-fiber collagen by utilizing ion liquid in different steps

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] (1) Raw material pretreatment

[0058] In parts by weight, add 1000 parts of cowhide to 5000 parts of isopropanol, shake for 10 hours, remove the isopropanol when the time is up, then mix with 1500 parts of water and 0.2 parts of DTM198 enzyme, and use Na 2 CO 3 Adjust the pH to 9.0, stir for 30 minutes, and let it stand. Stir for 1-2 min for every 1 hour of standing. After standing for a total of 16 hours, filter the solid product, and then add the solid product to 3000 parts of peroxide with a mass concentration of 60g / L In hydrogen, it acts for 4.5h as a raw material for microfibrillar collagen.

[0059] (2) Step-by-step ionic liquid treatment

[0060] Cut the raw material of microfibrillar collagen into blocks with a diameter of 10mm, immerse them in a 30% mass concentration of 1-ethyl-3-methylimidazole acetate solution, and stir for 13h at a temperature of 4-15℃ After the action time is reached, the precipitate is centrifuged and washed, and then mechanically cut to a bl...

Embodiment 2

[0066] (1) Raw material pretreatment

[0067] In parts by weight, add 1000 parts of pig skin to 5200 parts of isopropanol, shake for 11h, remove the isopropanol when the time is up, then mix with 1550 parts of water and 0.25 parts of DTM198 enzyme, and use Na 2 CO 3 Adjust the pH to 9.1, stir for 35 minutes, and let stand for 1 min for every 1 hour of standing. After standing for a total of 16 hours, filter the solid product, and then add the solid product to 3100 parts of hydrogen peroxide with a mass concentration of 60g / L , Acting for 4.5h, as the raw material of microfibrous collagen.

[0068] (2) Preparation of microfibrous collagen

[0069] Cut the raw material of microfibrous collagen into blocks with a diameter of 10mm, immerse them in a 70% mass concentration of 1-butyl-3-methylimidazolium dinitrile salt solution, and stir at a temperature of 4-15℃ 11h, after the action time is reached, centrifuge to collect the precipitate and wash it, and then mechanically cut it to a blo...

Embodiment 3

[0075] (1) Raw material pretreatment

[0076] In parts by weight, add 1000 parts of sheepskin to 5400 parts of isopropanol, shake for 12h, remove the isopropanol when the time is up, then mix with 1540 parts of water and 0.22 parts of DTM198 enzyme, and use Na 2 CO 3 Adjust the pH to 9.0, stir for 35 minutes, and let it stand for 2 minutes for every 1 hour of standing. After standing for a total of 16 hours, filter the solid product, and then add the solid product to 3000 parts of hydrogen peroxide with a mass concentration of 60g / L , Acting for 4.5h, as the raw material of microfibrous collagen.

[0077] (2) Preparation of microfibrous collagen

[0078] Cut the raw material of microfibrous collagen into blocks with a diameter of 10mm, immerse them in a 50% mass concentration of 1-ethyl-3-methylimidazole tetrafluoroborate solution, and stir at 4-15℃ Act for 8h, after the time of effect is reached, centrifuge to take the precipitate and wash it, then mechanically cut it to a block wi...

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Abstract

The invention provides a method for preparing micro-fiber collagen by utilizing ion liquid in different steps. The method comprises the following steps: separately preparing at least three ion liquidwith the mass concentration of 5 to 95 percent, sequencing the ion liquid according to the polarity of the ion liquid from small to big or from big to small, successively adding a raw material of micro-fiber collagen into the ion liquid in different steps, performing the ion liquid treatment, and performing the after-treatment, thus obtaining the micro-fiber collagen. The preparation method is mild in conditions, and by regulating and controlling the polarity of the ion liquid at the temperature less than or equal to a denaturation temperature of the collagen, the micro-fiber collagen with thebiological activity can be obtained under a non-denaturation of the collagen; and the yield of the preparation method can reach 80 percent, the ion liquid can be recycled and reused, the prepared micro-fiber collagen has the characteristics of high short fiber content, high purity, good biological compatibility and nontoxicity, and can be directly used as a biological medical material in the field of biological medicine.

Description

Technical field [0001] The invention belongs to the technical field of biological material preparation, and relates to a method for preparing microfibrous collagen by stepwise use of the gradient polarity of an ionic liquid. Background technique [0002] Collagen is a water-insoluble fibrous protein that constitutes the skeleton of the extracellular matrix. It is widely present in the connective tissues of vertebrates (including soft tissues, skin, tendons, bones and cartilage). It plays an important role in regulating the normal physiological functions of organisms. The role of. At the same time, as a natural polymer material, collagen has biocompatibility, biodegradability and biological activity that other synthetic materials cannot match. It can promote cell growth, promote platelet coagulation, and has relatively low immunogenicity. The denaturation temperature of solution collagen is 38-40°C, and the denaturation temperature of mammalian skin collagen varies slightly with ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/78C07K1/02
CPCC07K14/78
Inventor 李国英刘思聪黄雨琳
Owner SICHUAN UNIV
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