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

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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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 / inorganic hybrid capillary monolithic column is prepared, and the nucleic acid aptamer is synthesized by the high affinity between streptavidin and biotin In the fixed capillary monolithic column, a new in-tube SPME extraction column based on nucleic acid aptamer modified organic / inorganic hybrid quartz capillary monolithic column was developed, which is used for trace and ultra-trace ochratoxin, Highly selective separation and enrichment of aflatoxin and other substances

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