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Vomitoxin and derivative aptamer affinity column and preparation method and application thereof

A technology for deoxynivalenol and its derivatives, applied in the field of deoxynivalenol and its derivatives aptamer affinity column and its preparation, can solve the problems of unsatisfactory deoxynivalenol and its derivatives, improve pretreatment efficiency, and be widely used Prospects, the effect of reducing the cost of use

Active Publication Date: 2019-06-04
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] At present, the aptamer affinity column reported at home and abroad can only target a certain toxin, which cannot meet the requirement of one-step purification and enrichment of deoxynivalenol and its derivatives in samples in practical applications. toxin requirements

Method used

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  • Vomitoxin and derivative aptamer affinity column and preparation method and application thereof
  • Vomitoxin and derivative aptamer affinity column and preparation method and application thereof
  • Vomitoxin and derivative aptamer affinity column and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1 Preparation of aptamer affinity column using amino-modified deoxynivalenol and its derivative aptamers

[0058] 1. Preparation of N-hydroxysuccinimide modified agarose. Sepharose 4B was used as the carrier and activated with N-hydroxysuccinimide.

[0059] Mix agarose with an equal volume of water in a fume hood, and add it to a reactor equipped with a pH electrode and a magnetic stirrer, and add 100 mg of N-hydroxysuccinimide per mL of agarose solution to the above agarose solution For N-hydroxysuccinimide, adjust the pH value to 11 with NaOH, control the pH value of the whole reaction at 11, control the temperature at about 20°C, and complete the reaction in 10 minutes; after the reaction, add an equal volume of ice chips to the above reaction quickly solution, and quickly poured into the Buchner funnel, with 10 times the volume of the agarose solution of cold buffer (200mM Na 2 HPO 4 , 5 mM MgCl 2 , pH 8.0) was washed by suction, and the hydroxyl groups ...

Embodiment 2

[0074] Example 2 Purification and detection of deoxynivalenol and its derivatives in wheat samples by using aptamer affinity column for deoxynivalenol and its derivatives

[0075] In this example, quantitative addition of deoxynivalenol and its derivatives standard substance to normal wheat samples, followed by purification with the aptamer affinity column of deoxynivalenol and its derivatives prepared in Example 1, followed by purification by high performance liquid chromatography- Fluorescence detector detection, determination of recovery. details as follows:

[0076] 1. Wheat sample processing

[0077] 1) Pulverize the wheat sample.

[0078] 2) According to the standards of 0.5, 5, and 50 μg per gram of sample, respectively, add deoxynivalenol and its derivative standard to the pulverized wheat sample.

[0079] 3) Weigh 5g of the sample, add 25mL of methanol-water (7:3, v / v), and place it on a homogenizer for 11000rpm high-speed homogenization for 3min.

[0080] 4) Filt...

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Abstract

The invention provides vomitoxin and a derivative aptamer affinity column and a preparation method and application thereof. The affinity column is prepared by the following steps: using N-hydroxysuccinimide modified agarose as a carrier, then carrying out covalent coupling on the carrier and a nucleic acid aptamer capable of high-affinity and high-specificity recognition of deoxynivalenol, deoxynivalenol-3-glucoside, 3-acetyl deoxynivalenol and 15-acetyl deoxynivalenol in a sample, and finally filling an affinity column with the coupled aptamer composite carrier. The affinity column is mainlyused for purification and decontamination of vomitoxin and its derivative in food, feeds, milk, food crops, traditional Chinese medicines and other various samples, which is beneficial to high performance liquid chromatography and rapid detection of toxins in the sample in later period. By one-time loading of the column, four types of toxins can be purified and enriched, the number of reuse can reach 15 times, recovery rate is 80% and above, the cost is reduced, and the sample pre-treatment efficiency of the sample is enhanced.

Description

technical field [0001] The invention belongs to the technical field of food safety detection, and in particular relates to vomitoxin and its derivative aptamer affinity column and its preparation method and application. Background technique [0002] DON and its derivatives belong to a kind of mycotoxin, also known as vomitoxin, which is a kind of trichothecene toxin, which is mainly produced by certain Fusarium fungi. Vesonder et al. isolated such compounds from feed and corn that caused American sows to refuse to eat and vomit in 1973, and named them vomitoxin. Trichothecene toxoids have a double bond between C-9 and C-10 in the chemical structure, an oxygen ring at C-12 and C-13, and other positions also contain hydroxyl and acetyl groups group. DON is composed of a 12,13-epoxy group, an α,β-unsaturated ketone group and 3 -OH functional groups, and its chemical name is 3,7,l5-trihydroxyl-12,l3-epoxy mono Teprosporine-9-en-8-one (3,7,15-trihydroxy-12,13-epoxytrichothec-9...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01D15/38B01J20/281G01N1/34
Inventor 栾云霞刘洪美陆安祥付海龙郭晓军冯晓元
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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