Monolithic Column Combining deep eutectic solvents Monomer and Hybrid Monomer

A deep eutectic solvent and monolithic column technology, applied in the field of separation and analysis, can solve the problems affecting the life and durability of the monolithic column, and achieve the effects of easy control of reaction conditions, good mechanical strength, and high enrichment effect

Active Publication Date: 2019-08-16
TIANJIN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no literature report on the use of DES monomer for the preparation of monolithic columns. During the experiment, we found that the monolithic columns prepared with DES monomers are relatively soft, which affects the life and durability of monolithic columns.

Method used

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  • Monolithic Column Combining deep eutectic solvents Monomer and Hybrid Monomer
  • Monolithic Column Combining deep eutectic solvents Monomer and Hybrid Monomer
  • Monolithic Column Combining deep eutectic solvents Monomer and Hybrid Monomer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Preparation of functional monomers

[0025] 1. Preparation of DES monomer: Choline chloride (60 mmol, 8.4 g) and methacrylic acid (120 mmol, 10.2 mL) were added into a threaded vial at a molar ratio of 1:2, followed by an oil bath at 90 °C for 1 h , a homogeneous transparent solution can be obtained. placed in a desiccator for later use;

[0026] 2. Preparation of hybrid monomer: Take 1498 μL (6.4 mmol) of 3-aminopropyltriethoxysilane (APTES) and 688 μL (8.1 mmol) of methacrylic acid (MAA) into a reaction flask and mix , sonicated for 10 min to dissolve. Then put it into a 60 °C water bath, take it out after 24 hours of reaction, and obtain a yellow transparent liquid, which is the hybrid monomer APTES-MAA;

Embodiment ２

[0028] Preparation of monolithic columns

[0029] Add functional monomer APTES-MAA (64 μL) and DES monomer ChCl-MAA (144 μL) to 670 μL methanol and 70 mg PEG 2000 binary porogen, crosslinker ethylene glycol dimethacrylate ( 62 μL) and the initiator azobisisobutyronitrile (2.6 mg), mixed with ultrasonic to form a pre-polymerization solution, injected into the treated capillary, sealed both ends of the capillary with rubber stoppers, and placed in a constant temperature water bath at 60°C. After 2 h of reaction, the capillary was taken out, and unreacted substances and other soluble substances in the monolithic column were washed with acetonitrile.

Embodiment 3

[0031] In order to determine the effect of functional monomer type on the protein extraction effect, monolithic columns of single monomer (APTES-MAA monolithic column, DES monolithic column) and traditional monomer (MAA monolithic column) were synthesized as controls. The specific operation steps are as follows:

[0032] The preparation of the DES / APTES-MAA monolithic column is the monolithic column prepared in Example 2;

[0033] Preparation of control monolithic columns

[0034] Preparation of APTES-MAA monolithic column: except not adding DES monomer, synthesized with the same method and experimental conditions as in Example 2; Preparation of DES monolithic column: except not adding APTES-MAA monomer, using the same method as in Example 2 Synthesis by method and experimental conditions; preparation of MAA monolithic column: except that APTES-MAA monomer and DES monomer were not added, 210 μL of MAA monomer was added, and synthesized by the same method and experimental cond...

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Abstract

The invention relates to an integral column combining a deep eutectic solvent monomer and a hybrid monomer. DES monomer (choline chloride-methacrylic acid) and hybrid monomer (3-aminopropyltriethoxysilane-methacrylic acid) as binary functional monomers, ethylene glycol dimethacrylate as a cross-linking agent, methanol and PEG 20000 as a binary porogen, and an in-situ thermal polymerization processare used to prepare the a novel monolithic column. The monolithic column has the advantages of low cost and simple synthesis process, and the prepared monolithic column has good permeability, strongmechanical strength and biocompatibility, and can successfully separate bovine serum albumin and cytochrome C, and simultaneously can achieve better enrichment of two proteins of the bovine serum albumin and the cytochrome C. The method provides a new idea for the separation and purification of proteins.

Description

technical field [0001] The invention belongs to the technical field of separation and analysis, and in particular relates to a new monolithic column combining deep eutectic solvent monomer and hybrid monomer. Background technique [0002] Protein is a biologically active macromolecule that is closely related to various life activities. With the development of proteomics and the Human Genome Project, people hope to understand the biology of cells at the molecular level. Protein biochemistry attempts to reveal the role and biological function of gene products. Separating desired proteins from complex matrices while maintaining their biological activity and chemical integrity is of great significance for the development of life sciences, molecular biology and other fields. Intact protein separation techniques mainly include polyacrylamide gel electrophoresis, capillary electrophoresis, and liquid chromatography. However, protein separation using particle-packed column chroma...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/26B01J20/28B01J20/30B01D15/22G01N23/2251G01N27/447G01N33/68C07K14/765C07K1/14
CPCB01D15/22B01J20/261B01J20/28014B01J20/28042B01J20/3092B01J2220/4812C07K1/14C07K14/765G01N23/2251G01N27/44717G01N33/6803G01N2550/00
Inventor 刘照胜柴美红黄艳萍
Owner TIANJIN MEDICAL UNIV
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