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In-vitro gradient degrading method for multi-crosslinked chitosan or derivative gel of multi-crosslinked chitosan

A chitosan gel, cross-linked chitosan technology, applied in biochemical equipment and methods, microbial determination/inspection, chemical method analysis, etc., can solve problems such as few reports on the degradation of cross-linked chitosan , to achieve the effect of simple operation and accurate results

Active Publication Date: 2017-09-19
IMEIK TECH DEV CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although cross-linked chitosan is based on chitosan, there are few reports on the degradation of cross-linked chitosan

Method used

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  • In-vitro gradient degrading method for multi-crosslinked chitosan or derivative gel of multi-crosslinked chitosan
  • In-vitro gradient degrading method for multi-crosslinked chitosan or derivative gel of multi-crosslinked chitosan
  • In-vitro gradient degrading method for multi-crosslinked chitosan or derivative gel of multi-crosslinked chitosan

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1: Free Radical Degradation of Triple Crosslinked Chitosan Gels

[0058] Prepare an ascorbic acid solution with a concentration of 0.01mol / L, and adjust it to pH=3 with 3 mol / L hydrochloric acid; take cross-linked chitosan (deacetylation degree 90%, molecular weight 20KDa, three times cross-linked, mass concentration 30mg / mL, cross-linked chitosan Place 2 mL of the gel in a 50 mL centrifuge tube, add 2 mL of the above ascorbic acid solution, and cool to 0°C, where the concentration of ascorbic acid is 0.01 mol / L; keep the environment at 0°C, add hydrogen peroxide solution 2 mL, the concentration of hydrogen peroxide is 0.01 mol / L; the system is placed in a constant temperature water bath shaker, the shaking temperature is set at 37°C, samples are taken every 10 minutes for the first 100 minutes, and samples are taken every 20 minutes thereafter, taking out 100 μL of degradation solution each time At the same time, the above-mentioned ascorbic acid solution was 1...

Embodiment 2

[0059] Example 2: Lysozyme degradation of triple cross-linked chitosan gels

[0060] Take 2 mL of cross-linked chitosan (deacetylation degree 90%, molecular weight 20KDa, three times of cross-linking, mass concentration 30mg / mL, cross-linking degree 2%) in a 50 mL centrifuge tube, add 2 mL concentration of 0.01 mol / L phosphate buffer (pH 7.2), add 800 u lysozyme, and cool to 0°C; place the system in a constant temperature water bath shaker, set the shaking temperature to 37°C, and take samples every 1 h for the first 12 hours After that, samples were taken every 2 hours, and 100 μL of the degradation solution was taken out each time, and 100 μL of the above-mentioned phosphate buffer solution was added at the same time. The total degradation time was 14 hours; the degradation solution was sealed in a dialysis bag with a molecular weight cut-off of 14 KDa, Quantitative dialysis was carried out with 5 mL of pure water for 2 h; 4.0 mL of the dialysate was taken, added with 1 mL o...

Embodiment 3

[0061] Example 3: Free radical degradation of six-fold cross-linked carboxymethyl chitosan gel

[0062] Prepare an ascorbic acid solution with a concentration of 0.01mol / L, and adjust it to pH=3 with 3 mol / L hydrochloric acid; take cross-linked carboxymethyl chitosan (deacetylation degree 90%, molecular weight 50KDa, substitution degree 1.2, mass concentration 30mg / mL, equal volumes of five gels with cross-linking degrees of 2%, 4%, 8%, 12%, and 16% and non-cross-linked carboxymethyl chitosan) gel 2 mL were placed in a 50 mL centrifuge tube, Add 2 mL of the above ascorbic acid solution, and cool to 0°C, where the concentration of ascorbic acid is 0.01 mol / L; keep the environment at 0°C, add 2 mL of hydrogen peroxide solution, the concentration of hydrogen peroxide is 0.01 mol / L; place the system in a constant temperature water bath In the oscillator, the shaking temperature was set at 37°C, samples were taken every 10 minutes for the first 100 minutes, and samples were taken ...

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Abstract

The invention relates to an in-vitro gradient degrading method for multi-crosslinked chitosan or derivative gel of the multi-crosslinked chitosan. Free radicals are utilized to degrade the multi-crosslinked chitosan or the derivative gel of the multi-crosslinked chitosan, and a kjeldahl apparatus is utilized to quantify a degradation product; or lysozyme is utilized to degrade the multi-crosslinked chitosan or the derivative gel of the multi-crosslinked chitosan, and a third-method potassium ferricyanide reduced sugar measurement method in GB / T 5009.7-2016 National standard for food safety-Determination of reduced sugars in foods is utilized to quantify the degradation product. The method can simulate an in-vivo enzyme environment or a free radical environment and can quantitatively degrade the multi-crosslinked chitosan gel through the enzyme or the free radicals and can determine the degradation degree of the chitosan gel by determining the reduced sugar content in the degradation product or measuring the nitrogen content in the degradation product; thus, a degradation curve of the multi-crosslinked chitosan gel is obtained, and a degradation period of the chitosan gel is determined.

Description

technical field [0001] The invention relates to an in vitro gradient degradation method of multiple cross-linked chitosan or its derivative gel. The method simulates the enzymatic environment or free radical environment in vivo, and quantitatively degrades multiple cross-linked chitosan or its derivative gel by enzyme or free radical, by determining the content of reducing sugar in the degradation product or the nitrogen content in the degradation product. Determination, determine the degradation degree of the gel, thereby obtain the degradation curve of the multiple cross-linked chitosan or its derivative gel, and determine the degradation period of the gel. Background technique [0002] Chitosan (also known as glucosaminoglycan, English name and abbreviation: chitosan, CS) is the product obtained after deacetylation of chitin, so it is also called "deacetylated chitin". Chitosan is a straight-chain polymer compound connected by N-acetyl-D-glucosamine monomers through β-1,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N31/00G01N1/28C12Q1/34
Inventor 简军李睿智
Owner IMEIK TECH DEV CO LTD
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