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Method for preparing wood frog skin collagen

A collagen and frog skin technology, applied in the biological field, can solve problems that need to be improved

Active Publication Date: 2015-07-15
ACADEMY OF MILITARY MEDICAL SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, the current technology for preparing Rana skin collagen still needs to be improved

Method used

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  • Method for preparing wood frog skin collagen

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Experimental program
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Effect test

Embodiment 1

[0023] (1) Preprocessing:

[0024] Take the freshly stripped Rana skin, remove the subcutaneous tissue and adhesions, wash with tap water, dry naturally at room temperature, and pulverize. All extraction operations were carried out at 4°C.

[0025] Weigh 10 g of crushed dry Rana skin powder, add it into 100 mL of 0.3 mol / L NaOH aqueous solution, keep stirring at a slow speed for 24 h, and replace the mixed solution every 4 h. Wash the sample with 10 times the volume of distilled water (4°C) until neutral.

[0026] Add the water-washed to neutral wood frog skin sample to 100mL 20% n-butanol solution to degrease, keep stirring at a slow speed for 24h, and replace the mixed solution every 4h. The samples were washed three times with 10 volumes of distilled water (4°C).

[0027] (2) Extraction of acid-soluble collagen:

[0028] Add the pretreated wood frog skin sample to 100mL 1.0mol / L acetic acid solution. Under the power of 200W, microwave irradiation for 1min. Afterwards,...

Embodiment 2

[0035] (1) Preprocessing:

[0036] Take the freshly stripped Rana skin, remove the subcutaneous tissue and adhesions, wash with tap water, dry naturally at room temperature, and pulverize. All extraction operations were carried out at 4°C.

[0037] Weigh 10 g of crushed dry Rana skin powder, add it into 500 mL of 0.3 mol / L NaOH aqueous solution (containing 3% hydrogen peroxide), continue stirring at a slow speed for 120 h, and replace the mixed solution every 8 h. Wash the sample with 10 times the volume of distilled water (4°C) until neutral.

[0038] Add the water-washed to neutral wood frog skin sample into 500mL 20% n-butanol solution to degrease, keep stirring at a slow speed for 72h, and replace the mixed solution every 8h. The samples were washed three times with 10 volumes of distilled water (4°C).

[0039] (2) Extraction of acid-soluble collagen:

[0040] Add the pretreated wood frog skin sample to 500mL 1.0mol / L acetic acid solution. Under the power of 800W, the...

Embodiment 3

[0048] (1) Preprocessing:

[0049] Take the freshly stripped Rana skin, remove the subcutaneous tissue and adhesions, wash with tap water, dry naturally at room temperature, and pulverize. All extraction operations were carried out at 4°C.

[0050] Weigh 10 g of crushed dry Rana skin powder, add it into 300 mL of 0.3 mol / L NaOH aqueous solution (containing 1.5% hydrogen peroxide), continue stirring at a slow speed for 96 h, and replace the mixed solution every 6 h. Wash the sample with 10 times the volume of distilled water (4°C) until neutral.

[0051] Add the water-washed to neutral wood frog skin sample to 300mL 20% n-butanol solution to degrease, keep stirring at a slow speed for 48h, and replace the mixed solution every 6h. The samples were washed three times with 10 volumes of distilled water (4°C).

[0052] (2) Extraction of acid-soluble collagen:

[0053] Add the pretreated Rana skin sample to 300mL 1.0mol / L acetic acid solution. Under the power of 500W, the micro...

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Abstract

The invention discloses a method for preparing wood frog skin collagen, which comprises the following steps: obtaining wood frog skin powder; pretreating; adding into an acetic acid solution, irradiating under microwaves, stirring, and centrifuging to obtain a precipitate and a solution containing acid-soluble collagen; dispersing the precipitate in a pepsin-containing acetic acid solution, centrifuging to obtain a supernate, dialyzing the supernate, centrifuging to obtain a precipitate, and dissolving the precipitate in an acetic acid solution to obtain a solution containing enzyme-soluble collagen; adding NaCl into the solution containing acid-soluble collagen and the solution containing enzyme-soluble collagen, stirring, standing, and adding NaCl and trihydroxymethyl aminomethane to obtain a solution; regulating the pH value of the solution to obtain a precipitate, dissolving the precipitate in an acetic acid solution to obtain a solution, and dialyzing the solution until the pH value is neutral, thereby obtaining the solution; and prefreezing the solution and drying to obtain the wood frog skin collagen. The method can enhance the collagen extraction rate.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for preparing collagen, more specifically, the invention relates to a method for preparing collagen from wood frog skin. Background technique [0002] Collagen is a white, opaque, unbranched triple-helical fiber protein with a very stable molecular structure. It mainly exists in animal skin, bone, cartilage, teeth, muscle tendons, ligaments and blood vessels, accounting for about 100% in mammals. 25% to 30% of the total protein is an extremely important structural protein in connective tissue, which plays a role in supporting organs and protecting the body. It has been reported that collagen can effectively enhance the water storage capacity of human skin tissue cells, improve skin elasticity, and relieve wrinkles; it can promote bone formation and enhance the collagen structure at low calcium levels, thereby improving bone strength and preventing bone loss. Porosis; can ab...

Claims

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Application Information

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IPC IPC(8): C12P21/06C07K14/78C07K1/36C07K1/34C07K1/30
Inventor 南雪裴雪涛单紫筠张迦南姚海雷岳文
Owner ACADEMY OF MILITARY MEDICAL SCI
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