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Anion exchange chromatographic filler for purification of low molecular weight heparin, preparation method, chromatographic column filled thereby and purification method

A low-molecular-weight heparin and exchange chromatography technology, which is applied in the field of purifying low-molecular-weight heparin, can solve problems such as poor pressure resistance, low dynamic adsorption capacity of chromatographic columns, and low purification efficiency of low-molecular-weight heparin, and achieve good pressure resistance and dynamic High adsorption capacity and high purification efficiency

Active Publication Date: 2015-01-14
WUXI GALAK CHROMATOGRAPHIC TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, after filling the chromatographic column with the current anion exchange chromatographic filler for purification, the dynamic adsorption capacity of the chromatographic column is low, and the pressure resistance performance is not good, so it can only be used at a low flow rate and cannot meet the requirements of high-efficiency production.
Therefore, the purification efficiency of current low molecular weight heparins is low

Method used

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  • Anion exchange chromatographic filler for purification of low molecular weight heparin, preparation method, chromatographic column filled thereby and purification method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1: the method for preparing anion exchange chromatographic filler.

[0025] Steps.

[0026] 1. Preparation of cross-linked polyglycidyl methacrylate-ethylene glycol diester methacrylate substrate microspheres:

[0027] a. Add 1g of polyvinyl alcohol (average molecular weight: 88-130KDa) and 360g of sodium chloride into a 4L reactor, then add 3000mL of water, and seal the reactor. Stir the materials in the reaction kettle while raising the temperature inside the kettle to 60°C. After stirring continuously for 5 hours, the polyvinyl alcohol and sodium chloride are fully dissolved, stop heating and stirring, and cool the temperature inside the kettle to room temperature to obtain an aqueous phase;

[0028] b. Add 20mL glycidyl methacrylate (GMA), 80mL ethylene glycol diester methacrylate (EDMA), 30mL cumene, 70mL n-dodecyl alcohol and 1g benzoyl peroxide in a beaker and mix them mixing to obtain a mixed material; stirring the mixed material while ultrasonica...

Embodiment 2

[0035] Embodiment 2: the method for preparing anion exchange chromatographic filler.

[0036] Steps.

[0037] 1. Preparation of cross-linked polyglycidyl methacrylate-ethylene glycol diester methacrylate substrate microspheres:

[0038]a. Add 3g of polyvinyl alcohol (average molecular weight: 88-130KDa) and 450g of sodium chloride into a 4L reactor, then add 2700mL of water, and seal the reactor. Stir the materials in the reaction kettle while raising the temperature inside the kettle to 65°C. After 4 hours of continuous stirring, the polyvinyl alcohol and sodium chloride are fully dissolved, stop heating and stirring, and cool the temperature inside the kettle to room temperature to obtain an aqueous phase;

[0039] b. Add 50mL of GMA, 50mL of EDMA, 50mL of toluene, 58mL of isooctyl alcohol and 2g of 2,2'-azobisisovaleronitrile into a beaker and mix them to obtain a mixed material; stir the mixed material to 2, 2'-Azobisisovaleronitrile is completely dissolved to obtain a...

Embodiment 3

[0046] Embodiment 3: the method for preparing anion exchange chromatographic filler.

[0047] Steps.

[0048] 1. Preparation of cross-linked polyglycidyl methacrylate-ethylene glycol diester methacrylate substrate microspheres:

[0049] a. Add 5g of polyvinyl alcohol (average molecular weight: 88-130KDa) and 300g of sodium chloride into a 4L reactor, then add 2000mL of water, and seal the reactor. Stir the materials in the reaction kettle while raising the temperature inside the kettle to 70°C. After continuous stirring for 3 hours, the polyvinyl alcohol and sodium chloride are fully dissolved, stop heating and stirring, and cool the temperature inside the kettle to room temperature to obtain an aqueous phase;

[0050] b. Add 225mL of GMA, 75mL of EDMA, 100mL of toluene and 9g of benzoyl peroxide into a beaker and mix them to obtain a mixed material; stir the mixed material while ultrasonicating until the benzoyl peroxide is completely dissolved to obtain an organic phase; ...

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Abstract

The invention provides an anion exchange chromatographic filler for purification of low molecular weight heparin. The matrix of the anion exchange chromatographic filler is porous spherical particles of crosslinked poly(glycidyl methacrylate)-ethylene glycol dimethacrylate, the particle size range is 30-150 micrometers, the average particle size is 50-80 micrometers, and the aperture range is 30-180nm. The filled chromatographic column has high dynamic adsorption capacity and good pressure resistant performance, is suitable for use under a high flow rate, and can adapt to the requirements of efficient production. The invention also provides a preparation method of the anion exchange chromatographic filler, the anion exchange chromatographic column filled by the filler, and a method for purifying low molecular weight heparin by the anion exchange chromatographic column.

Description

technical field [0001] The invention relates to an anion-exchange chromatographic filler, a preparation method thereof, an anion-exchange chromatographic column filled with the anion-exchange chromatographic column and a purification method using the anion-exchange chromatographic column, and is especially suitable for purifying low-molecular-weight heparin. Background technique [0002] Heparin is a dextran sulfate amino acid, which is an acidic mucopolysaccharide. As an anticoagulant, heparin has anticoagulant effects both in vivo and in vitro. Clinically applied heparin preparations are divided into two categories: common unfractionated heparin and low molecular weight heparin, among which the average molecular weight of low molecular weight heparin (Low Molecular Weight Heparin, LMWH) is less than 5000Da. Compared with unfractionated heparin, low molecular weight heparin also has antithrombotic effect, but the risk of bleeding is lower. Low-molecular-weight heparin als...

Claims

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Application Information

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IPC IPC(8): B01J20/285B01J20/30B01D15/08B01D15/36G01N30/06
CPCB01D15/08B01D15/363B01J20/285B01J2220/52B01J2220/54B01J2220/80C08F220/20C08F220/32G01N30/06
Inventor 穆宁刘辉孙国威
Owner WUXI GALAK CHROMATOGRAPHIC TECH
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