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Weak anion chromatography medium, preparation method and application thereof

A chromatographic medium, weak anion technology, applied in separation methods, chemical instruments and methods, solid adsorbent liquid separation, etc.

Active Publication Date: 2020-06-26
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Rigid resin has good mechanical strength and chemical stability, but it involves hydrophilic modification and surface graft modification, the preparation process is complicated, the production cycle is long, and the reagents used are more harmful to the environment
[0004] In order to avoid the complicated operation of grafting modification, there are also patent reports on the preparation of spherical ion exchange media by one-step copolymerization, such as: U.S. Patent No. 6,423,666 discloses a method using quaternary ammonium salts, such as dimethyl diallyl ammonium chloride, in one step The method of preparing strong anion exchange medium by copolymerization is used for the separation and purification of proteins. However, in the application, because this kind of packing has strong anion exchange groups and shows strong basicity, it is very useful for the separation and purification of sensitive proteins. big limitations
The weak anionic separation medium has better applicability, but due to the inhibitory effect of weak basic functional groups on free radical polymerization, the polymerization speed is slow and the polymerization process is difficult to control, so it is difficult to prepare weak anionic chromatography medium
Although U.S. Patent No. 5,645,717 discloses a method of utilizing N-allylmethylamine and N,N-methylenebisacrylamide (BIS) mixed reaction to form a monolithic column chromatographic medium in a 6×350mm column, but This method can only obtain a continuous phase medium, but not a dispersed filler, and the prepared chromatographic medium is not suitable for large-scale use in factories

Method used

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  • Weak anion chromatography medium, preparation method and application thereof
  • Weak anion chromatography medium, preparation method and application thereof
  • Weak anion chromatography medium, preparation method and application thereof

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preparation example Construction

[0024] The invention provides a preparation method of a weak anion chromatography medium, comprising the steps of:

[0025] Dissolving tetrachlorethylene and Span 80 in liquid paraffin to obtain an oil phase;

[0026] mixing allylamine salt, hydroxy methacrylate, N,N-methylenebisacrylamide, porogen, buffer solution with a pH value of 7, and an initiator to obtain an aqueous phase;

[0027] The oil phase and the water phase are mixed, and a copolymerization reaction is carried out under a protective atmosphere to obtain a weak anion chromatographic medium.

[0028] In the present invention, tetrachlorethylene and Span 80 are dissolved in liquid paraffin to obtain an oil phase.

[0029] In the present invention, the mass concentration of tetrachlorethylene in the oil phase is preferably 30-40%, more preferably 36-38%, and the mass concentration of Span 80 is preferably 0.35-0.41%, more preferably 0.41% . The present invention uses tetrachlorethylene to adjust the specific gra...

Embodiment 1

[0055] Mix 95g of tetrachlorethylene, 800.9g of Span, and 160g of liquid paraffin, heat up to 70°C, dissolve the solid, and obtain 230mL of oil phase;

[0056] N-methyldiallylamine hydrochloride 6.25g, N,N-methylene bisacrylamide 5.50g, hydroxypropyl methacrylate 2g, ammonium sulfate 1.5g and ethylene glycol 4g and the pH value 7 buffer solution (in which the concentration of sodium phosphate is 1mol / L, and the concentration of hydrogen chloride is 2mol / L) 18g is mixed and heated to 70°C. After the solid is dissolved, add 45mg of 4,4-azobiscyanovaleric acid, After 4,4-azobiscyanovaleric acid is completely dissolved, 39.3mL of water phase is obtained;

[0057] Mix the water phase and the oil phase, under stirring conditions, maintain the particle size of the oil phase droplets at 30-150 μm, raise the temperature to 85°C, pass in nitrogen, react at 83-85°C for 2.1h, and then cool to room temperature; The reaction solution cooled to room temperature was filtered, and the obtaine...

Embodiment 2

[0064] Mix 95g of tetrachlorethylene, 800.9g of Span, and 160g of liquid paraffin, heat up to 70°C, dissolve the solid, and obtain 230mL of oil phase;

[0065] 6.13g of diallylamine hydrochloride with a purity of 98%, 4.40g of N,N-methylenebisacrylamide, 2g of hydroxyethyl methacrylate, 1.8g of ammonium sulfate, 4g of ethylene glycol and the pH Mix 23g of 7 buffer solution (in which the concentration of sodium phosphate is 1mol / L and the concentration of hydrogen chloride is 2mol / L) and heat up to 70°C. After the solid dissolves, add 20mg of 4,4-azobiscyanovaleric acid , until the 4,4-azobiscyanovaleric acid is completely dissolved to obtain 40mL of the aqueous phase;

[0066]Mix the water phase and the oil phase, under stirring conditions, maintain the particle size of the oil phase droplets at 30-150 μm, raise the temperature to 85°C, pass in nitrogen, react at 83-85°C for 2.1h, and then cool to room temperature; The reaction solution cooled to room temperature was filtered...

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Abstract

The invention provides a weak anion chromatography medium, a preparation method and application thereof, and belongs to the technical field of ion exchange chromatography. The preparation method of the weak anion chromatography medium comprises the following steps: dissolving tetrachloroethylene and span 80 in liquid paraffin to obtain an oil phase; mixing an allyl amine salt, hydroxyl methacrylate, N, N-methylene bisacrylamide, a pore-foaming agent, a buffer solution with a pH value of 7 and an initiator to obtain a water phase; and mixing the oil phase and the water phase, and carrying out copolymerization reaction in a protective atmosphere to obtain the weak anion chromatography medium. The weak anion chromatography medium obtained by the preparation method provided by the invention has pressure resistance and chromatographic broadening behavior equivalent to those of medium modified filler, and is large in ion exchange capacity, so that the weak anion chromatography medium has higher adsorption capacity.

Description

technical field [0001] The invention relates to the technical field of ion exchange chromatography, in particular to a weak anion chromatography medium and its preparation method and application. Background technique [0002] Chromatography is one of the key technologies for protein separation and purification. As the core of chromatography technology, chromatography medium has always been a research hotspot. In the production process of biological products, due to the advantages of fast exchange speed, large adsorption capacity, stable structure and long service life, chromatography media are widely used in the separation and purification of nucleic acids, peptides, proteins, antibiotics and natural products. [0003] Most of the traditional separation and purification media use cellulose, dextran and cross-linked agarose gel. This kind of soft gel matrix is ​​characterized by good hydrophilicity and high biocompatibility, but its bed volume changes greatly and is not stabl...

Claims

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Application Information

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IPC IPC(8): C08F214/04C08F226/02C08F222/38C08F220/20C08F2/14B01D15/20
CPCB01D15/20C08F2/14C08F214/04C08F220/20C08F222/385C08F226/02
Inventor 马利腾訾文娟王春红雷晓菊
Owner NANKAI UNIV
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