High-strength polysaccharide gel microsphere used for chromatographic separation filler

A technology of gel microspheres and separation fillers, applied in separation methods, solid adsorbent liquid separation, alkali metal oxides/hydroxides, etc., can solve the problems of inability to meet the requirements of the medium skeleton, slow reaction speed, and low solubility , to achieve fast and efficient chromatographic separation and improved mechanical strength

Inactive Publication Date: 2018-11-30
江苏珐玛赛谱生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the cross-linking agent is generally oil-soluble, the solubility in water is very low, and the cross-linking agent can only react with the hydroxyl groups on the polysaccharide chain by diffusing into the gel network structure that has been formed, so the reaction speed is very slow. It is difficult to further improve the degree of rigidity, and cannot meet the requirements of mass production on the rigidity of the medium skeleton
Cross-linking in the oil phase is beneficial to improve the solubility of the cross-linking agent in the reaction system, but the polysaccharide gel is difficult to swell in the oil phase, that is, the microspheres are cross-linked in the shrinking state. Although the degree of cross-linking is high, the resulting gel Microspheres no longer have a macroporous network structure, and their applications in the separation and purification of biomacromolecules are limited

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Example 1 Blending method to prepare high-strength agarose microspheres with a concentration of 4wt%

[0062] (1) Modification of agarose raw material

[0063]Weigh 4g of agarose powder, add 40mL of water, heat and dissolve to obtain a 10% agarose solution, cool down to about 65°C, slowly add 0.5mL of 40% NaOH solution and bifunctional cross-linking agent allyl glycidyl ether to it 6mL. Wherein, the concentration of allyl glycidyl ether in the water phase is 15%, and the concentration of OH- is 0.21mol / L. After stirring and reacting at 65° C. for 8 h, the reaction was terminated by adjusting the pH value to 7 with 60% glacial acetic acid solution to obtain an allylated agarose solution. Add 4 times the volume of protective agent and polyethylene glycol to the solution, centrifuge to collect the precipitate, pre-freeze at -70°C for 2 hours, and freeze-dry for 72 hours to obtain the modified agarose raw material.

[0064] (2) Preparation of agarose microspheres modifie...

Embodiment 2

[0070] Example 2 Blending method to prepare high-strength agarose microspheres with a concentration of 4wt%

[0071] In order to further increase the strength of the agarose gel microspheres, on the basis of the cross-linking process described in Example 1, the traditional method was used to continue cross-linking once. That is, take 20 g of cross-linked agarose microspheres obtained in step (3) of Example 1, disperse them in 40 mL of deionized water, and gradually raise the temperature to 47.5° C. for 2 hours. After that, 1.6mL epichlorohydrin and 2.4mL 40% NaOH solution (containing 3% NaBH4) were slowly added dropwise to the system, and the reaction was continued for 12h in a constant temperature water bath shaker. After the cross-linking was completed, it was washed with water to neutrality, and the obtained 4wt% highly cross-linked agarose microspheres had an average particle size of 86.41 μm and a maximum flow rate of 2180 cm / h in the linear range.

Embodiment 3

[0081] Example 3 Blending method to prepare high-strength agarose microspheres with a concentration of 6wt%

[0082] (1) Modification of agarose raw material

[0083] Prepare 40 mL of agarose solution with a concentration of 18%, cool down to about 80 °C, and slowly add 3 mL of 40% NaOH solution and 8 mL of cross-linking agent allyl glycidyl ether into it. Wherein, the concentration of allyl glycidyl ether in the water phase is 20%, and the concentration of OH- is 1.25mol / L. After stirring and reacting at 80° C. for 3 h, the reaction was terminated by adjusting the pH value to 7 with 60% glacial acetic acid solution to obtain an allylated agarose solution. Add 4 times the volume of protective agent and polyethylene glycol to the solution, centrifuge to collect the precipitate, pre-freeze at -70°C for 2 hours, and freeze-dry for 72 hours to obtain the modified agarose raw material.

[0084] (2) Preparation of agarose microspheres modified by cross-linking agent by blending

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Abstract

The invention discloses a high-strength polysaccharide gel microsphere used for a chromatographic separation filler; firstly a polysaccharide raw material is modified with a bifunctional crosslinkingagent, and then the modified polysaccharide raw material is mixed with an unmodified raw material for emulsifying into microspheres, and post-activation is carried out to realize cross-linking insidethe microspheres; modified polysaccharide chains greatly improve the mechanical strength of the gel microspheres by forming of covalent cross-linking bonds in gel fiber bundles and among the fiber bundles; unmodified polysaccharide chains contain a large amount of hydroxyl groups to facilitate formation of hydrogen bonds during the gelation process, the hydrogen bonds play a role of skeleton support, maintain a macroporous network structure formed by gel solidification, and effectively avoid shrinkage and deformation of the microspheres. The obtained gel microsphere not only has the excellentproperties imparted by natural polysaccharides, but also has significant advantages in skeleton rigidity and operating flow rate, and is an ideal industrialized chromatographic separation filler.

Description

technical field [0001] The invention relates to the technical field of chromatographic separation materials, in particular to a high-strength polysaccharide gel microsphere that can be used as a chromatographic separation filler. Background technique [0002] Natural polysaccharides are rich in hydroxyl groups, highly hydrophilic, and have good compatibility with biomacromolecules, occupying a core position in the field of biomacromolecules separation. Especially the gel-type polysaccharide separation medium, which has a macroporous network structure in the swollen state, has special advantages for the separation and purification of biological macromolecules. However, the skeleton structure of polysaccharide gel is mainly maintained by hydrogen bonds. Although it has certain mechanical strength, compared with inorganic microspheres and other organic polymer microspheres, the particles are relatively soft, so it is called "soft matrix". When used as a separation medium, unde...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/24B01J20/28B01J20/30B01D15/20
CPCB01D15/20B01J20/24B01J20/28004B01J20/28021B01J20/28047
Inventor 樊丰林
Owner 江苏珐玛赛谱生物科技有限公司
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