A method using liquid chromatography bilayer stationary phase

A bilayer, liquid chromatography technology, applied in material separation, analytical materials, measurement devices, etc., can solve the problems of stationary phase contamination, conflicting research purposes, proteins affecting separation, etc., to avoid non-specific binding, reduce time and desired effect

Active Publication Date: 2017-01-04
SHANDONG MEASUREMENT SCI RES INST
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  • Abstract
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AI Technical Summary

Problems solved by technology

Since proteins are macromolecules, they are likely to bind non-specifically to the stationary phase, resulting in "contamination" of the stationary phase, and even residual protein affecting separation
Generally, the chromatographic column will first pretreat the sample and remove some pollutants through the guard column. However, some proteins are components of interest to researchers and should not be removed. This method is undoubtedly contrary to the purpose of the research.
Failure to carry out these protection measures may result in reduced service life of the guard column or chromatographic column

Method used

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  • A method using liquid chromatography bilayer stationary phase
  • A method using liquid chromatography bilayer stationary phase
  • A method using liquid chromatography bilayer stationary phase

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Embodiment Construction

[0039] Select silica particles as the stationary phase, wash with anhydrous solvent, and put it into an organic solvent after drying. The organic solvent is preferably free of water (toluene can be used), and it can react with the modified amphoteric substance (Y) by adding Silicon coupling agent (ATPS):

[0040]

[0041] Amino groups are formed on the surface of the silica.

[0042] After the reaction is completed, centrifuge to separate the activated silica on the surface, and react with Y and fatty acid after drying (the fatty acid used is a straight-chain carboxylic acid containing 42 carbon atoms), and react in an organic solvent under heating conditions to form an amide bond Modify the amphoteric substances and non-polar groups on the surface of silica, centrifuge, wash and dry, mix with the unmodified amphoteric substance (X) in an organic solvent, the solvent is preferably chloroform, after uniform mixing, use an inert gas Dry as vibes.

[0043] The Y is 16:0 Glut...

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Abstract

The invention relates to a method using liquid chromatography to prolong the service life of a separated biological substance and to reduce and lower the preparation time and requirements for a separated biological substance sample. According to the method, a series of chemical modifications are performed on the surface of a stationary phase, a bimolecular layer stabilized by a polymer is added to the surface of the stationary phase, and the bimolecular layer cannot generate nonspecific binding with biomass such as a protein, so that the risk that the stationary phase is polluted and absorbed by a complicated macromolecule protein is reduced greatly.

Description

technical field [0001] The invention belongs to the field of liquid chromatography, and mainly relates to the improvement of the liquid chromatography method, including improving the tolerance of the chromatographic column packing (stationary phase), reducing the requirement for sample pretreatment so as to save time and increase the detection speed , while also reducing the replacement frequency of the guard column in front of the chromatographic column. Background technique [0002] There are many examples of protein separations using liquid chromatography, especially reversed-phase liquid chromatography. However, due to the complex molecular structure and different compositions of proteins, generally successful separation requires a lot of early attempts: such as the selection of stationary phase, mobile phase selection, and optimization of mobile phase concentration ratio. [0003] In addition, preparations for purification of the protein to be isolated are essential. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/60
Inventor 郭波许爱华隋峰林振强孙倩芸何云馨许思思李锋丽
Owner SHANDONG MEASUREMENT SCI RES INST
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