An organic polymeric monolithic column that specifically recognizes ochratoxin and its preparation method

A technology for ochratoxin and specific recognition, which is applied in separation methods, chemical instruments and methods, and other chemical processes, and can solve problems such as large material consumption, polar solvent interference, and rigid recognition "holes are easily destroyed or deformed" , to achieve more sites, improve stability, and improve the efficiency of modification and bonding

Active Publication Date: 2018-12-25
FUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Due to the characteristics of molecular imprinting technology, there are disadvantages such as large material consumption during synthesis, polar solvent interference, rigid recognition "holes" are easily destroyed or deformed, and non-specific binding sites are inevitably generated during the synthesis of MIP, resulting in Compared with specific biological recognition systems such as antibody antigens and enzyme substrates, there is a big gap in its selectivity

Method used

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  • An organic polymeric monolithic column that specifically recognizes ochratoxin and its preparation method
  • An organic polymeric monolithic column that specifically recognizes ochratoxin and its preparation method
  • An organic polymeric monolithic column that specifically recognizes ochratoxin and its preparation method

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Experimental program
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Embodiment 1

[0031] An organic polymeric monolithic column that specifically recognizes ochratoxin. The monolithic column first thermally initiates polymerization in an alkenylated capillary with epoxy cyclosiloxane acrylate monomer and acrylate crosslinking agent to form a Highly cross-linked, epoxy-based cyclosiloxane polymerized monolithic column matrix stationary phase with a branched structure of epoxy groups. The ligand is prepared; the surface of the monolithic column stationary phase is bonded with a nucleic acid aptamer that specifically recognizes ochratoxin A.

[0032] Table 1 Contents of different components of epoxy cyclosiloxane acrylate polymerized monolithic stationary phase

[0033]

[0034] * : Epoxy cyclosiloxane acrylate monomer is 2-hydroxy-3-{3-[2,4,6,8-tetramethyl-4,6,8-tri(propyl glycidyl ether)- 2-Cyclotetrasiloxane]propoxy}propylmethacrylate.

[0035] Prepare the capillary monolithic column matrix stationary phase according to the component content table sho...

Embodiment 2

[0043] Apply the base stationary phase of the unmodified nucleic acid aptamer and the epoxy-based hydrolysis stationary phase, and implement according to the following steps: (1) Equilibration: For the monolithic column prepared by formula B, first equilibrate with binding buffer for 1 hour, and the chromatographic conditions are: Flow rate 0.10 mL / min, pressure 250psi, the pH of the binding buffer is 8.00, containing 10mmol / L Tris-HCl, 120 mmol / L NaCl, 5 mmol / L KCl and 20mmol / L CaCl 2 ; (2) Enrichment: Inject 20 microliters of 0.50 μg / mL ochratoxin A (OTA) solution, enrich on the monolithic column for 1 hour, the chromatographic conditions are 0.05 mL / min, pressure 250 psi; (3) Cleaning : Install the enrichment column on the liquid chromatography pump, wash the matrix stationary phase and the epoxy hydrolysis stationary phase of the unmodified nucleic acid aptamer with binding buffer for 14 hours and 10 hours respectively; the elution condition is a flow rate of 0.03 mL / min,...

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Abstract

The invention discloses an organic polymerized monolithic column capable of specially recognizing ochratoxin, and a preparation method of the organic polymerized monolithic column. According to the monolithic column, a polymerized monolithic column fixed phase having a branched structure of an epoxy group is firstly formed by polymerizing an epoxy cyclosiloxane acrylate monomer and an acrylate cross-linking agent, then post-column ring-opening addition is carried out, and an aptamer capable of specially recognizing the ochratoxin is prepared by covalent bonding. The aptamer and an epoxy group, distributed on the surface of the epoxy cyclosiloxane polymerized monolithic column matrix fixed phase in a branching way, are subjected to further ring-opening addition, so that condensation coupling is realized; the prepared polymerized monolithic column matrix fixed phase has a cyclosiloxane octatomic ring-shaped structure and the epoxy group distributed in a branching way, thus being stable in structure; the organic polymerized monolithic column is more in sites acting with 5'-amino aptamer and small in steric hindrance, and can realize efficient bonding with ochratoxin A aptamer, thus being suitable for the specific recognition of the ochratoxin A.

Description

technical field [0001] The invention belongs to the field of preparation of monolithic column functional materials, and in particular relates to an organic polymeric monolithic column for specifically recognizing ochratoxin and a preparation method thereof. Background technique [0002] Capillary monolithic columns are a new type of chromatographic micro-separation columns. Currently, capillary monolithic columns are mainly based on the principle of reversed-phase interaction or hydrophilic interaction for separation. In terms of specific analysis, affinity chromatographic columns use ligands with specific recognition functions immobilized on chromatographic stationary phases to make specific interactions with analytes and achieve separation from other substances in the sample. [0003] Molecularly imprinted polymers (MIPs) have become the main research object of selective monolithic column materials due to their recognition principle similar to the "key-lock" interaction. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01D15/22B01J20/285B01J20/30
CPCB01D15/22B01J20/285
Inventor 林旭聪王敏宋红亮朱春玲谢增鸿
Owner FUZHOU UNIV
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