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Ultrasonic auxiliary acid extracting method for optimizing collagen through response surface method

A collagen, ultrasound-assisted technology, applied in the preparation methods of peptides, chemical instruments and methods, animal/human proteins, etc., can solve the problems of a large amount of hazardous waste, large amount of reagents, long extraction time, etc., and achieve short processing time. , Solubility improvement, the effect of improving solubility

Inactive Publication Date: 2016-12-14
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It takes a long time to extract with acid and alkali, the amount of reagents is large, a large amount of harmful waste is produced, and the extraction rate is low
The enzymatic extraction method can maintain the integrity of the collagen structure, but the extraction cost is high, the extraction time is long, and the extraction rate is low
Although the cost of hot water extraction is very low, the extraction rate is also low

Method used

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  • Ultrasonic auxiliary acid extracting method for optimizing collagen through response surface method
  • Ultrasonic auxiliary acid extracting method for optimizing collagen through response surface method
  • Ultrasonic auxiliary acid extracting method for optimizing collagen through response surface method

Examples

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Comparison scheme
Effect test

Embodiment 1

[0037] An ultrasonic-assisted acid extraction method utilizing response surface methodology to optimize collagen, comprising the following steps:

[0038] Step 1: Take fresh or frozen soft-shelled soft-shelled turtle skirt and chop it, then add NaCl solution with a mass fraction of 2.0% at a material-to-liquid ratio of 1:20, and stir at 20°C for 9 hours. After homogenizing at 8000rpm at high speed for 2min, the precipitate was collected once, and 0.5mol / L of Na was added with a solid-liquid ratio of 1:20 2 CO 3 The solution was stirred at 20°C for 9h. Then, after high-speed homogenization at 8000 rpm for 2 minutes, the secondary precipitate was collected, and 0.3 mol / L EDTA aqueous solution was added to soak and remove impurities at a solid-liquid ratio of 1:25. Stir at 20°C for 10 h, then homogenize at 8000 rpm for 2 min at high speed, collect three precipitates, wash repeatedly with distilled water, drain fully and set aside. Then add isopropanol solution with a mass frac...

Embodiment 2

[0051] An ultrasonic-assisted acid extraction method utilizing response surface methodology to optimize collagen, comprising the following steps:

[0052] Step 1: Take fresh or frozen soft-shelled soft-shelled turtle skirt and chop it, then add NaCl solution with a mass fraction of 1.0% at a material-liquid ratio of 1:10, and stir at 20°C for 6 hours. Then, after high-speed homogenization at 5000rpm for 1min, the precipitate was collected once, and 0.3mol / L Na was added with a solid-liquid ratio of 1:10. 2 CO 3 The solution was stirred at 20°C for 6h. Then, after high-speed homogenization at 5000 rpm for 1 min, the secondary precipitate was collected, and 0.2 mol / L EDTA aqueous solution was added to soak and remove impurities at a solid-liquid ratio of 1:20. Stir at 20°C for 6 hours, then homogenize at 5,000 rpm for 1 minute at high speed, collect three precipitates, wash repeatedly with distilled water, drain well and set aside. Then add isopropanol solution with a mass fr...

Embodiment 3

[0055] An ultrasonic-assisted acid extraction method utilizing response surface methodology to optimize collagen, comprising the following steps:

[0056] Step 1: Take fresh or frozen soft-shelled soft-shelled turtle skirt and chop it, then add NaCl solution with a mass fraction of 3.0% at a material-to-liquid ratio of 1:30, and stir at 20°C for 2 hours. Then, after high-speed homogenization at 10000rpm for 3min, the precipitate was collected once, and 0.6mol / L Na was added with a solid-liquid ratio of 1:30. 2 CO 3 The solution was stirred at 20°C for 12h. Then, after high-speed homogenization at 10,000 rpm for 3 minutes, the secondary precipitate was collected, and 0.4 mol / L EDTA aqueous solution was added to soak and remove impurities at a solid-liquid ratio of 1:30. Stir at 20°C for 12 hours, then homogenize at 10,000 rpm for 3 minutes at high speed, collect three precipitates, wash repeatedly with distilled water, drain fully and set aside. Then add isopropanol solution...

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Abstract

The invention relates to a collagen extracting method, in particular to an ultrasonic auxiliary acid extracting method for optimizing collagen through a response surface method. The method includes the steps of conducting chopping and impurity removal processing on fresh or frozen Chinese softshell turtle outskirts, adding an acetic acid solution with concentration of 0.3-0.6 mol / L to an impurity-removed sample according to the material-liquid ratio of 1:(10-30), putting the materials into an ultrasonic reactor for intermittent ultrasonic extraction in the mode that ultrasonic processing is conducted for the first preset time and the intermitting processing is conducted for the second preset time repeatedly, taking supernate after centrifuging, adding NaCl to the supernate, conducting filtering after standing and salting out, redissolving protein precipitates with acetic acid with the same concentration, sequentially dialyzing acetic acid and distilled water, and finally conducting freeze-drying to obtain a sample, wherein the first preset time is shorter than or equal to the second preset time. The method is high in extracting efficiency, high in collagen solubility and high in raw material utilization value and collagen utilization rate.

Description

technical field [0001] The invention relates to a method for extracting collagen. More specifically, the present invention relates to an ultrasound-assisted acid extraction method for optimization of collagen using response surface methodology. Background technique [0002] Collagen is a kind of structural protein in animal tissues, which has the characteristics of high viscosity, low solubility and low denaturation temperature. The above characteristics limit its application to a certain extent. Its extraction methods mainly include acid extraction, alkali extraction, enzyme extraction and hot water extraction. It takes a long time to extract with acid and alkali, the amount of reagent is large, a large amount of harmful waste is produced, and the extraction rate is low. Enzyme extraction can maintain the integrity of the collagen structure, but the extraction cost is high, the extraction time is long, and the extraction rate is low. Although the cost of hot water extra...

Claims

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Application Information

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IPC IPC(8): C07K14/78C07K1/14
CPCC07K14/78
Inventor 邹烨王道营卞欢李鹏鹏王立蔡盼盼孙冲诸永志徐为民
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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