Chromatographic packing for separating water soluble polymer and protein and preparation method of same

A water-soluble polymer and hydrophilic polymer technology, applied in the field of chromatographic packing, can solve the problems of poor stability and insufficient hydrophilicity, and achieve the effects of low hydrophobicity, excellent resolving power, and high theoretical plate number.

Active Publication Date: 2015-04-01
浙江月旭材料科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are problems such as insufficient hydrophilicity and poor stability of the size exclusion chromatography packing materials for preparing isolated proteins, peptides and biological macromolecules such as biological macromolecules by traditional techniques.

Method used

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  • Chromatographic packing for separating water soluble polymer and protein and preparation method of same
  • Chromatographic packing for separating water soluble polymer and protein and preparation method of same
  • Chromatographic packing for separating water soluble polymer and protein and preparation method of same

Examples

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preparation example Construction

[0037] The present invention provides a preparation method of hydrophilic silica gel chromatography packing for separating water-soluble polymers and proteins, which comprises the following steps:

[0038] A. Activation of silica gel to obtain activated silica gel;

[0039] B. Perform epoxy modification on the activated silica gel to obtain epoxy modified silica gel;

[0040] C introduce a hydrophilic polymer into the epoxy modified silica gel to prepare a hydrophilic polymer modified silica gel; and

[0041] D Hydrolyze the hydrophilic polymer-modified silica gel to prepare the hydrophilic silica gel chromatographic packing for separating water-soluble polymers and proteins.

[0042] Step A includes: washing the silica gel with distilled water, then soaking it with an aqueous solution of hydrofluoric acid, then washing with distilled water to neutrality, and finally washing with acetone and drying.

[0043] Wherein, step B includes: using boric acid buffer as a solvent, the activated si...

Embodiment 1

[0048] Example 1: Preparation of hydrophilic silica gel chromatography packing for separating water-soluble polymers and proteins

[0049] (1) Add 50 g of silica gel to a 500 mL reaction vessel, add 200 mL of distilled water, stir for 30 minutes, filter, and repeat this step twice. Then add 400 mL of 0.08% hydrofluoric acid aqueous solution with a mass ratio of 0.08%, stir for 24 hours and wash with distilled water to neutrality, and finally add 200 mL of acetone to wash, filter, and dry at 110°C for 12 hours to obtain activated silica gel;

[0050] (2) Add 400mL pH=9.0 boric acid buffer solution to 500mL reaction vessel (equipped with condenser, mechanical stirrer and thermometer) ( Buffer, purchased from Guangzhou Huayuehang Instrument Co., Ltd.), 50g activated silica gel and 100mL γ-(2,3-glycidoxy)propyltrimethoxysilane, stirred at 80°C for 30 minutes; obtained epoxy Group modified silica gel;

[0051] (3) Add 100g of soluble starch to the reaction vessel, reflux and stir at 97°...

Embodiment 2

[0053] Example 2: Preparation of hydrophilic silica gel chromatography packing for separating water-soluble polymers and proteins

[0054] (1) Add 50 g of silica gel to a 500 mL reaction vessel, add 200 mL of distilled water, stir for 30 minutes, filter, and repeat this step twice. Then add 400 mL of 0.08% hydrofluoric acid aqueous solution with a mass ratio of 0.08%, stir for 24 hours and wash with distilled water to neutrality, and finally add 200 mL of acetone to wash, filter, and dry at 110°C for 12 hours to obtain activated silica gel;

[0055] (2) Add 450mL pH=9.0 boric acid buffer solution, 50g activated silica gel and 150mL γ-(2,3-epoxypropoxy)propyl to the 500mL reaction vessel (equipped with condenser, mechanical stirrer, and thermometer) Trimethoxysilane, stirred and reacted at 80°C for 30 minutes; obtained epoxy group modified silica gel;

[0056] (3) Add 150g of dextran to the reaction vessel, reflux and stir at 97°C for 20-22 hours; after the reaction, cool and reflux ...

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Abstract

The invention discloses a hydrophilic silica gel chromatographic packing for separating a water soluble polymer and a protein and a preparation method of the hydrophilic silica gel chromatographic packing. The preparation method comprises the following steps: (A), activating silica gels to prepare activated silica gels; (B), carrying out epoxy modification on the activated silica gels so as to prepare epoxy-based modified silica gels; (C), introducing a hydrophilic polymer to the epoxy-based modified silica gels so as to prepare hydrophilic polymer modified silica gels; and (D), hydrolyzing the hydrophilic polymer modified silica gels so as to prepare the hydrophilic silica gel chromatographic packing for separating the water soluble polymer and the protein. The hydrophilic silica gel chromatographic packing disclosed by the invention has the advantages of larger pore volume, lower hydrophobicity, higher number of theoretical plates, and larger peak capacity.

Description

Technical field [0001] The invention relates to a chromatographic filler, in particular to a chromatographic filler for separating water-soluble polymers and proteins and a preparation method thereof. Background technique [0002] With the rapid development of chromatographic technology, chromatographic fillers are constantly updated and developed. Various new types of chromatographic fillers have been prepared to solve various problems in separation science. At present, the main problem faced in separation chemistry is the difficulty of separating complex biological samples. Due to the extremely complex composition of biological samples, the content of many biological macromolecules in biological samples is extremely low, so separation interference is large, separation and purification steps are numerous, and the process is long; and many biological macromolecules are separated and purified using organic solvents as mobile phases. It is easy to lose activity; therefore, the sep...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01J20/285B01J20/30B01D15/34
Inventor 赵岳星薛昆鹏姚立新
Owner 浙江月旭材料科技有限公司
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