Method for synthesizing integral bed for cracking solution analysis

A synthesis method and monolithic bed technology, applied in the field of biomolecular analysis technology and chromatographic analysis, can solve the problems of chromatographic columns such as easy pollution, low back pressure, and poor mass transfer performance, and achieve the effects of not easy to be polluted, low back pressure, and low cost

Inactive Publication Date: 2010-01-13
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Aiming at the problems of pretreatment, poor mass transfer performance and easy contamination of the chromatographic column in the current analysis of the lysate, the purpose of the present invention is to provide a channel with a suitable modification density, which is not easy to be polluted, and has a large number of plasmids that can enter, Fast mass transfer speed, low back pressure, can be directly used in the synthesis method of monolithic bed for lysate analysis

Method used

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preparation example Construction

[0020] The present invention provides a kind of synthetic method for the integral bed of lysate analysis, it is characterized in that the method comprises the steps:

[0021] 1) Mixing methacrylates containing epoxy end groups or acrylate monomers containing epoxy end groups with a crosslinking agent, a porogen and an initiator to obtain a reaction mixture, wherein:

[0022] a. The ratio of the amount of the crosslinking agent to the amount of the monomer is 0.05 to 0.2;

[0023] b. The amount of the porogen accounts for 40% to 80% of the volume content of the reaction mixture;

[0024] c. The ratio of the amount of the initiator to the monomer is 0.005 to 0.03;

[0025] 2) Ultrasonically mix the above reaction mixture evenly, add it into the chromatographic column tube, and polymerize in situ to form a monolithic bed, the polymerization temperature is controlled between 50-60°C, and the reaction time is 12-72 hours;

[0026] 3) after the end of the polymerization reaction, ...

Embodiment 1

[0034]Weigh monomer glycidyl methacrylate, crosslinking agent divinylbenzene and trimerized isocyanurate triallyl urate (1: 0.04: 0.01mol / mol / mol); porogen toluene and n-heptane ( Pore ​​agent / reaction mixture 80vol%, toluene / n-heptane 3: 2vol / vol); Initiator azobisisobutyronitrile (initiator / monomer 0.005: 1mol / mol), mix well, pack into a 50 ×4.6mm stainless steel tube, the other end was sealed, and reacted at 50°C for 72 hours. Connect the monolithic bed to the pump and flush with ethanol. Then diethylamine was used as a modifier, ethanol was used as a solvent, the volume content of diethylamine was 25%, the modification reaction was controlled at 50° C., and the reaction time was 9 hours. After the reaction, the monolithic bed was rinsed with ethanol. More than 75% of pores in the monolithic bed synthesized by the present invention have a diameter of more than ten microns, and the modification density of the monolithic bed is about 1.1 mmol / g monolithic bed.

Embodiment 2

[0036] Weigh monomer epoxy ethylene methacrylate, crosslinking agent ethylene glycol dimethacrylate and trivinylbenzene (1: 0.03: 0.06mol / mol / mol); porogen ethylbenzene and n-octane ( Porogen / reaction mixture 75vol%, ethylbenzene / n-octane 1:1vol / vol); Initiator azobisisobutyronitrile (initiator / monomer 0.01:1mol / mol), mix well, put into one end and seal In a 50×4.6mm stainless steel tube, the other end was sealed and reacted at 55° C. for 36 hours. Connect the monolithic bed to the pump and flush with THF. Then, dimethylamine was used as a modifier, tetrahydrofuran was used as a solvent, the volume content of dimethylamine was 75%, the modification reaction was controlled at 60° C., and the reaction time was 3 hours. After the reaction, the monolithic bed was flushed with tetrahydrofuran. More than 75% of pores in the monolithic bed synthesized by the present invention have a diameter of more than ten microns, and the modification density of the monolithic bed is about 0.5 m...

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Abstract

The invention relates to a method for synthesizing an integral bed for cracking solution analysis, which belongs to the technical fields of biomelecule analysis and chromatographic analysis. The method comprises the following steps: mixing metacrylic acid ester containing an epoxy terminal group or an acrylic ester monomer containing the epoxy terminal group with cross-linking agent, pore-forming agent and initiating agent to obtain a reaction mixture; uniformly mixing the mixture in an ultraphonic way, adding the mixture into a chromatographic column pipe and carrying out in-situ polymerization to form an integral bed; after polymerization reaction is finished, washing the integral bed by organic solvent; modifying the integral bed by amine modifying agent dissolved in the organic solvent; after modification reaction is finished, washing the integral bed by the organic solvent. The invention has the key point of controlling the hole structure and the modifying density of the integral bed. The integral bed synthesized by the method can analyze cracking solution which is not preprocessed and contains a plurality of impurities, the analyzing process is convenient and rapid, and the cost is low.

Description

technical field [0001] The invention relates to a method for synthesizing an integral bed for lysate analysis, belonging to the fields of biomolecular analysis technology and chromatographic analysis. Background technique [0002] The application research of gene therapy and DNA vaccine in the diagnosis and treatment of major diseases such as cancer, single-gene genetic disease, AIDS, cardiovascular disease and arthritis is attracting more and more attention. At present, plasmids have gradually become the most important gene delivery system for gene therapy, so the large-scale preparation of pharmaceutical plasmids used in gene therapy has become its basis. One of the keys to the large-scale preparation process of pharmaceutical plasmids is how to monitor the purity of the plasmid, monitor the performance of the plasmid preparation process, and evaluate the final product quality and product specifications, which are very important for process development, certification, and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/50G01N30/56C08F220/32C08J9/12C08F8/32
Inventor 张敏莲徐青花秀夫
Owner TSINGHUA UNIV
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