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POSS (polyhedral oligomeric silsesquioxane)-based organic-inorganic hybrid silica gel monolithic column applied to ochratoxin specific recognition and preparation method thereof

A technology for specific recognition of ochratoxin, applied in chemical instruments and methods, separation methods, and other chemical processes, to achieve the effect of increasing the site of action, reducing steric hindrance, and increasing the amount of bonding

Inactive Publication Date: 2018-06-29
FUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the coverage densities of nucleic acid aptamers in the hybrid silica-based affinity monolithic columns prepared by the above methods are all at the level of 277-342 pmol / μL

Method used

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  • POSS (polyhedral oligomeric silsesquioxane)-based organic-inorganic hybrid silica gel monolithic column applied to ochratoxin specific recognition and preparation method thereof
  • POSS (polyhedral oligomeric silsesquioxane)-based organic-inorganic hybrid silica gel monolithic column applied to ochratoxin specific recognition and preparation method thereof
  • POSS (polyhedral oligomeric silsesquioxane)-based organic-inorganic hybrid silica gel monolithic column applied to ochratoxin specific recognition and preparation method thereof

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Effect test

Embodiment 1

[0029] A method for preparing a POSS-based organic-inorganic hybrid silica monolithic column for specific recognition of ochratoxin. The specific steps are as follows:

[0030] (1) Amination of the inner wall of the capillary:

[0031] The capillary empty column was sequentially treated with 1.0 mol / L HCl solution, 1.0 mol / L NaOH solution and H 2 Rinse with O for 30 min, 3 h and 15 min respectively, then rinse with methanol for 30 min, blow dry with nitrogen at 25 °C and 0.4 MPa; inject methanol and 3-aminopropyltriethoxysilane at a volume ratio of 1:1 The mixture was reacted in a constant temperature water bath at 60 °C for 24 h, so that the silanized substance with amino groups was bonded to the capillary wall; then washed with methanol for 30 min, and blown with nitrogen at 70 °C and 0.4 MPa for 3 h to obtain Capillary with amination of column wall surface;

[0032] (2) Preparation of POSS-based hybrid silica monolithic column matrix:

[0033] Weigh PEI monomer, Poss-epo...

Embodiment 2

[0042] Select formula D to prepare blank column of unmodified nucleic acid aptamer, control column modified with nucleic acid aptamer without anti-ochratoxin A and affinity monolithic column modified with anti-ochratoxin A nucleic acid aptamer, The length of the column is 10cm, and the equilibration, enrichment, cleaning and elution are carried out respectively. The specific steps are: (1) Equilibration: first equilibrate with a buffer solution of pH 8.50 for 30min, with a flow rate of 0.10 mL / min and a pressure of 250psi. From 10mmol / L Tris-HCl, 120 mmol / L NaCl, 5 mmol / L KCl and 20mmol / L CaCl 2 Composition; (2) Enrichment: Inject 20 μL of 10 ng / mL ochratoxin A (OTA) solution separately to enrich it on the monolithic column for 1 hour, the injection conditions are flow rate 0.05 mL / min, pressure 250psi; (3) Cleaning: Install the enrichment column on the liquid chromatographic pump, wash it separately with pH 8.50 buffer solution, the cleaning flow rate is 0.1 mL / min, the press...

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Abstract

The invention discloses a POSS (polyhedral oligomeric silsesquioxane)-based organic-inorganic hybrid silica gel monolithic column applied to ochratoxin specific recognition and a preparation method thereof. The POSS-based organic-inorganic hybrid silica gel monolithic column is characterized in that the hybrid silica gel monolithic column prepared by nucleophilic reaction between a polyhedral oligomeric silsesquioxane (POSS) crosslinking agent and polyamine monomer is utilized as a matrix, the matrix has a high-crosslinking 3D skeleton, the surface of the matrix contains rich amino, the matrixcan load anti-ochratoxin aptamer surface modified gold nano particles in high density, and the monolithic column is prepared by nano gold amino bonding and self assembling. The prepared POSS-based monolithic column matrix disclosed by the invention has the characteristics of high-crosslinking structure and multiple acting sites and can efficiently load the nano gold particles; thus, the aptamer with specific recognition on ochratoxin A can be loaded and fixed to the POSS-based monolithic column in high density, and high-efficiency specific recognition separation on the ochratoxin A is achieved.

Description

technical field [0001] The invention belongs to the field of monolithic polymer material preparation, and in particular relates to a POSS-based organic-inorganic hybrid silica monolithic column for specific recognition of ochratoxin and a preparation method thereof. Background technique [0002] Monolithic column, also known as continuous bed (continuous bed), is a continuous bed stationary phase that uses organic or inorganic polymerization methods to perform in-situ polymerization and similar casting in the chromatographic column. Aptamer-modified affinity monolithic columns have high selectivity, high resolution and high target recognition ability by combining nucleic acid aptamers and monolithic column materials, and have attracted attention as emerging functional materials. [0003] The more mature way to prepare nucleic acid aptamer-modified monolithic columns is to immobilize the aptamer on the stationary phase based on the bridging effect between biotin and streptavi...

Claims

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Application Information

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IPC IPC(8): B01D15/20B01D15/22B01D15/38B01J20/24B01J20/30G01N30/02G01N30/08
CPCB01D15/22B01D15/206B01D15/3804B01J20/0233B01J20/103B01J20/22B01J20/24G01N30/02G01N30/08
Inventor 林旭聪於霞陈怡琼谢增鸿
Owner FUZHOU UNIV
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