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Method for synthesizing gallic acid petunidin agarose hydrophile adsorption chromatography medium

A technology of gallic acid and adsorption chromatography, applied in separation methods, chemical instruments and methods, solid adsorbent liquid separation, etc., can solve the problems of low market share and few types

Inactive Publication Date: 2009-09-16
北京康铭优盛生化技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few types of agarose matrix medium products in my country, and the market share is low. More than 98% of the domestic market is occupied by foreign products. The development of agarose medium with independent intellectual property rights is an urgent strategic problem to be solved.

Method used

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  • Method for synthesizing gallic acid petunidin agarose hydrophile adsorption chromatography medium
  • Method for synthesizing gallic acid petunidin agarose hydrophile adsorption chromatography medium

Examples

Experimental program
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Effect test

Embodiment 1

[0035] Example 1, Synthesis Method 1 of Agarose Gel Hydrophilic Adsorption Chromatography Medium of Gallic Acid Ligand

[0036] 1) 100ml of agarose gel microspheres with a concentration of 12% and an average particle size of 30μm are washed with water on a sand core funnel and replaced with acetone, transferred to a reaction bottle or reactor, and 100ml of acetone is added;

[0037] 2) Add 10g of glycidyl ether and stir for 60min;

[0038] 3) Add 3ml of 20% sodium hydroxide and stir for 8h;

[0039] 4) Add acetone and stir for 1h;

[0040] 5) Wash with 5 times the volume of acetone for 10 minutes, and wash with 5 times the volume of water for 10 minutes;

[0041] 6) Add 200ml of water, add 5g of sodium sulfate, and stir for 2 hours;

[0042] 7) Add 3ml of 20% sodium hydroxide and stir for 1h;

[0043] 8) Add 10ml of epichlorohydrin and stir at 35-60°C for 3h;

[0044] 9) wash with 5 times the volume of water;

[0045] 10) Add 200ml of water, add 5g of sodium sulfate, and...

Embodiment 2

[0056] Embodiment 2, agarose gel hydrophilic adsorption chromatographic medium synthesis method 2 of gallic acid ligand

[0057] 1) 100ml of agarose gel microspheres with a concentration of 12% and an average particle size of 30μm are washed with water on a sand core funnel and replaced with acetone, transferred to a reaction bottle or reactor, and 100ml of acetone is added;

[0058] 2) Add 30g glycidyl ether and stir for 120min;

[0059] 3) Add 3ml of 20% sodium hydroxide and stir for 12h;

[0060] 4) add acetone and stir for 2h;

[0061] 5) Wash with 5 times the volume of acetone for 20 minutes, and wash with 5 times the volume of water for 20 minutes;

[0062] 6) Add 200ml of water, add 15g of sodium sulfate, and stir for 3h;

[0063] 7) Add 3ml of 20% sodium hydroxide and stir for 2h;

[0064] 8) Add 10-30ml of epichlorohydrin and stir at 40°C for 5h;

[0065] 9) wash with 5 times the volume of water;

[0066] 10) Add 200ml of water, add 15g of sodium sulfate, and stir...

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Abstract

The invention relates to a method for synthesizing a hydrophile adsorption chromatography medium which takes gallic acid as the petunidin and the agarose gel with high concentration and high crosslinking degree as the stroma. The high concentration and high crosslinking agarose gel medium surface has high density reactionable hydroxyl group which is activated with allyl bromide followed by reacting with bromine water, and then gallic acid is added, and the prepared medium can be used as a hydrogen bond supplyer in the hydrophile adsorption reaction. The medium prepared by the synthesis method has strong rigidity, high petunidin density, high selectivity, and is fit for separating polyphenol protein, hydrophilic polypeptide and natural small organic molecules.

Description

technical field [0001] The invention relates to a method for synthesizing a chromatographic medium, in particular to a method for synthesizing a gallic acid agarose hydrophilic adsorption chromatographic medium. [0002] The invention belongs to the field of biochemical separation of bioengineering. Background technique [0003] The cost of separation and purification in the production of biopharmaceuticals accounts for the vast majority of the cost, among which the most commonly used is chromatographic technology and its combination. Currently, the media used are mainly gel filtration, ion exchange, hydrophobic interaction, affinity, and reversed phase media. , and its matrix materials include agarose, dextran, polystyrene and silica gel. Agarose medium is the most widely used matrix material in the production of biopharmaceuticals. It is a natural polysaccharide with high porosity; high connectivity effectively ensures intramolecular substance transfer; large specific su...

Claims

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Application Information

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IPC IPC(8): B01J20/286
CPCB01D15/305
Inventor 孟桂凤邢思亮
Owner 北京康铭优盛生化技术有限公司
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