Preparation method for organic/inorganic hybrid quartz capillary monolithic column modified by single-chain DNA nucleic acid aptamer

A nucleic acid aptamer and quartz capillary technology, applied in chemical instruments and methods, separation methods, solid adsorbent liquid separation, etc., can solve the problems of low selectivity, easy breakage of SPME, complex and highly selective separation and enrichment, etc. problem, to achieve good permeability and increase the effect of immobilized capacity

Active Publication Date: 2015-12-09
SOUTH CHINA NORMAL UNIVERSITY
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Problems solved by technology

[0006] The present invention aims at the problems of easy breakage of SPME, insufficient extraction amount, low selectivity and poor compatibility of biological samples as mentioned above, and combines the characteristics of high affinity, strong specificity and good compatibility of biological samples of nucleic acid aptamers Combined with the advantages of In-tubeSPME technology, such as high efficiency, easy to use with instruments, and simple operation, the organic/i

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  • Preparation method for organic/inorganic hybrid quartz capillary monolithic column modified by single-chain DNA nucleic acid aptamer
  • Preparation method for organic/inorganic hybrid quartz capillary monolithic column modified by single-chain DNA nucleic acid aptamer
  • Preparation method for organic/inorganic hybrid quartz capillary monolithic column modified by single-chain DNA nucleic acid aptamer

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Embodiment Construction

[0024] This example takes the preparation of biotin-labeled ochratoxin A nucleic acid aptamer-modified organic / inorganic hybrid quartz capillary monolithic column In-tubeSPME extraction column as an example to describe the present invention in detail, but it does not limit the present invention scope of protection.

[0025] Such as figure 1 As shown, the preparation steps of ochratoxin A nucleic acid aptamer-modified organic / inorganic hybrid quartz capillary monolithic column In-tubeSPME extraction column are as follows:

[0026] (1) Take a hollow fused silica capillary with an inner diameter of 0.53 mm and a length of 15 cm (hereinafter referred to as "the capillary") to prepare a piranha alkali lotion—30% H 2 o 2 -H 2 SO 4(浓) (1:4, v / v), adopt the pressure difference circulation flow method, wash at 60°C for 1.5h; cool to room temperature, wash with absolute ethanol for 3-5 times, and then wash with pure water for 3-5 times, N 2 Blow dry and activate in an oven at 120°...

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Abstract

The invention discloses a preparation method for an organic/inorganic hybrid quartz capillary monolithic column modified by single-chain DNA nucleic acid aptamer. The preparation method is performed successively through the following steps: (1) performing activation processing on the internal surface of a hollow quartz capillary; (2) weighing CTAB (hexadecyltrimethylammonium bromide) and putting into a centrifugation tube, respectively adding TEOS (tetraethyl orthosilicate), APTES (3-aminopropyltriethoxysilane), anhydrous ethanol and the like, and performing multi-step processing, so as to obtain an organic/inorganic hybrid monolithic column; (3) subsequently using methanol and pure water for flushing; (4) activating, reacting at room temperature for a period, and using a PBS (phosphate buffer saline) for repeated flushing; and (5) injecting an ochratoxin A aptamer solution into the organic/inorganic hybrid monolithic column, and performing circular reaction at room temperature and storing at a low temperature, so as to obtain the organic/inorganic hybrid quartz capillary monolithic column modified by single-chain DNA nucleic acid aptamer. The product is applied to high-selectivity separation and enrichment of trace quantity or ultra trace quantity of ochratoxin, aflatoxin and other substances in complex biological samples and food.

Description

technical field [0001] The invention belongs to the field of chemical analysis and testing instruments, and relates to an organic / inorganic hybrid capillary monolithic column modified based on nucleic acid aptamers, and a preparation method for a novel in-tube solid-phase microextraction monolithic column. The monolithic column is suitable for complex food and biological Rapid, efficient and highly selective separation and enrichment of trace and ultra-trace ochratoxin components in samples. Background technique [0002] Due to the interference of the component to be measured by its coexisting components or due to the limitation of the sensitivity of the determination method itself and the requirements of the state of the component to be measured, most chemical detection and analysis methods require effective and reasonable treatment of the sample in advance. That is, before the analysis and determination, the sample should be physically or chemically treated to extract the ...

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Application Information

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IPC IPC(8): B01J20/286B01J20/30B01D15/22
Inventor 胡小刚郭璇陈时远刘炉英刘忠勇
Owner SOUTH CHINA NORMAL UNIVERSITY
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