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Method for preparing monomers of different polymerization degrees of fully deacetylated chitosan oligosaccharide capable of being detected on line

A technology of acetylated chitosan oligosaccharide and degree of polymerization is applied in the field of preparation of monomers with different degrees of polymerization of fully deacetylated chitosan oligosaccharide, which can solve the problems such as inability to realize continuous production, inability to guarantee accuracy, and influence on the experimental process, etc. The effect of convenient and flexible time, reduction of manpower and material resources, and fast flow rate

Active Publication Date: 2016-04-06
TECHNICAL INST OF PHYSICS & CHEMISTRY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The whole process needs to connect hundreds of components and carry out hundreds of color reactions. The process is cumbersome, the method is rigid, and wastes time, manpower and material resources; on the other hand, when it is extended to the separation and production of different batches, it cannot To ensure accuracy, continuous production cannot be realized, and the production rate is low, which affects the follow-up experiment process

Method used

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  • Method for preparing monomers of different polymerization degrees of fully deacetylated chitosan oligosaccharide capable of being detected on line
  • Method for preparing monomers of different polymerization degrees of fully deacetylated chitosan oligosaccharide capable of being detected on line
  • Method for preparing monomers of different polymerization degrees of fully deacetylated chitosan oligosaccharide capable of being detected on line

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Take 10 g of fully deacetylated chitosan oligosaccharide (molecular weight <1000) and dissolve it in 150 ml of deionized water, add 4 times the volume of ethanol for precipitation, filter after precipitation, and freeze-dry the filtrate to obtain a powder after concentration.

[0033] After dissolving the obtained powder with acetic acid buffer solution with a pH of 4, after filtering through a microporous membrane with a pore size of 0.45 μm, the cation exchange column SP Sepharose HP was used for separation on the AKTAavant150 protein separation system, with 1M concentration of NaCl The solution is eluted at a flow rate of 2mL / min, and the components are collected according to the peak by detecting the ultraviolet absorption at 190nm online: the peak value is >100mAU, and the collection volume of each collection tube is 2ml. The specific elution positions are 9.84cV, 11.83cV, 14.36cV, 17.10cV and 19.74cV according to the combination of the components near the peak tip ...

Embodiment 2

[0037] Take 15g of fully deacetylated chitosan oligosaccharide (molecular weight<1000) and dissolve it in 150ml of deionized water, add 5 times the volume of ethanol for precipitation, filter after precipitation, and freeze-dry the filtrate to obtain a powder after concentration.

[0038] After dissolving the obtained powder with an acetic acid buffer solution with a pH of 5, and filtering through a microporous membrane with a pore size of 0.45 μm, the cation-exchange chromatography column SP Sepharose HP was used for separation on the AKTAavant150 protein separation system. The solution was eluted at a flow rate of 5mL / min, and the components were collected according to the peak by detecting the ultraviolet absorption at 200nm online: the peak value was >120mAU, and the collection volume of each collection tube was 1.5ml. According to the graph, the components near the peak tip were combined, and the specific elution positions were 10.12cV, 12.77cV, 15.41cV, 18.36cV and 21.55c...

Embodiment 3

[0042] Take 20 g of fully deacetylated chitosan oligosaccharide (molecular weight <1000) and dissolve it in 200 ml of deionized water, add 3 times the volume of ethanol for precipitation, filter after precipitation, and freeze-dry the filtrate to obtain a powder after concentration.

[0043] After the obtained powder was dissolved in acetic acid buffer solution with a pH of 6, filtered through a microporous membrane with a pore size of 0.45 μm, and separated on the AKTAavant150 protein separation system with a cation-exchange column SP SepharoseHP, with 3M concentration of NaCl The solution was eluted at a flow rate of 8mL / min, and the components were collected according to the peak by detecting the ultraviolet absorption at 220nm online: the peak value was >140mAU, and the collection volume of each collection tube was 1.0ml. The specific elution positions are 9.57cV, 11.93cV, 14.17cV, 16.32cV and 19.03cV respectively according to the components near the peak tip combined in th...

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Abstract

The invention discloses a method for preparing monometers of different polymerization degrees of fully deacetylated chitosan oligosaccharide capable of being detected on line. The method comprises the steps of performing alcohol precipitation, dissolution and filtration for the fully deacetylated acetylation chitosan oligosaccharide, utilizing an ion exchange chromatographic column SP Sepharose HP to perform separation by NaCI solution with the concentration of 0-3M at the flow rate of 2-8 mL / min, performing ultraviolet on-line detection simultaneously, collecting components according to ultraviolet absorption peaks, utilizing a gel column Sephadex G10 to perform desalination by utilizing deionized water as buffer solution, detecting and collecting salt free components on line through ultraviolet absorption and conductivity and performing freezing and drying to obtain a product. The purity of the prepared fully deacetylated acetylation monometers is above 90% respectively. According to the method, an ultraviolent detector is utilized for the first time to detect on line the separation process of the chitosan oligosaccharide, ion exchange chromatography is adopted in a matched mode to perform separation, the method saves time, reduces manpower and material input and is favorable for further enlarged production, the gel column is utilized to perform desalination treatment on the components, and the content of the oligosaccharide and salt is detected on line. The method has the advantages of being high in flow rate, simple in steps and high in efficiency.

Description

technical field [0001] The invention relates to the field of chitosan oligosaccharide preparation. More specifically, it relates to a method for preparing fully deacetylated chitosan oligosaccharide monomers with different polymerization degrees that can be detected online. Background technique [0002] Chitosan is composed of glucosamine or acetylglucosamine residues linked by β-1,4 glycosidic bonds. It is a naturally occurring linear polysaccharide. Due to its excellent properties such as safety, biocompatibility, and degradability , has been widely used in food, medicine, agriculture, wastewater treatment, biological materials and other fields. Oligochitosan is a degradation product of chitosan, and its degree of polymerization is in the range of 2-20. Due to its low molecular weight, its water solubility is greatly increased, and it is easier to be absorbed by organisms, so it shows superior physiological activity than chitosan. On April 16, 2014, the National Health a...

Claims

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Application Information

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IPC IPC(8): C07H5/06C07H1/06
CPCC07H1/06C07H5/06
Inventor 郭燕川吴玉潇卢伟鹏王毅虎王佳宁
Owner TECHNICAL INST OF PHYSICS & CHEMISTRY - CHINESE ACAD OF SCI
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