Hapten for isoprothiolane content detection and preparation method and application thereof

A technology of rice blastin and hapten, which is applied in chemical instruments and methods, biological testing, measuring devices, etc., can solve the problem of lack of hapten, etc., and achieve the effect of strong specificity, efficient detection method, and good affinity

Active Publication Date: 2021-12-14
YUNNAN TOBACCO QUALITY SUPERVISION MONITORING STATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The invention provides a hapten for detecting the content of rice blastling, a preparation method and its application, which are used to solve the lack of antibodies with specific reactions to the ingredients of rice blastling and the technology of hapten for synthesizing antibodies existing in the prior art question

Method used

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  • Hapten for isoprothiolane content detection and preparation method and application thereof
  • Hapten for isoprothiolane content detection and preparation method and application thereof
  • Hapten for isoprothiolane content detection and preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0061] Preparation of the above-mentioned hapten comprises the following steps:

[0062] Take 1.88g of diisopropyl malonate, add 20mL of carbon disulfide, stir at room temperature, mix thoroughly, add 13mL of 2mol / L KOH solution, stir at room temperature for 1h, then add 1.56g of 3,4-dichlorobutyric acid, add 2mol / L KOH solution 5mL, tetrabutylammonium bromide 3.8g, acetonitrile 80mL, heat to reflux for 4h, stop the reaction, rotary evaporate, remove acetonitrile and carbon disulfide, add water 80mL, add 6mol / L HCl to adjust the pH value to 6, add acetic acid Ethyl ester 100mL×3 extracted three times, combined the organic phases, washed with water, dried over anhydrous sodium sulfate and evaporated to dryness to obtain a yellow oil, put it on a silica gel column, and eluted and separated it with a mixed solution of petroleum ether-ethyl acetate with a volume ratio of 3:1. After purification, 0.76 g of acetic acid-blastidin was obtained, which was the above-mentioned rice blas...

Embodiment 2

[0064] The preparation of the antigen for the detection of rice blastin content comprises the following steps:

[0065] Take 12.9 mg of the hapten prepared in Example 1, add 1 mL of tetrahydrofuran to dissolve and clarify, add N-hydroxysuccinimide (NHS) 8.5 mg, 1-(3-dimethylaminopropyl)-3-ethylcarbodi Imine hydrochloride (EDC) 14.8 mg, fully dissolved and mixed, reacted at room temperature for 2 hours to obtain hapten activation solution A liquid; take 50 mg of bovine serum albumin (BSA), add 0.1mol / L CB9.1 8mL to dissolve, and obtain Liquid B: Add liquid A dropwise to liquid B, react at room temperature for 4 hours, stop the reaction, dialyze and purify with 0.02mol / L PBS buffer for 3 days, change the liquid 3 times a day, and centrifuge to obtain the bovine serum albumin conjugate. Linked rice blastin artificial antigen, aliquoted and stored at -20°C.

Embodiment 3

[0067] The preparation of the antigen for the detection of rice blastin content comprises the following steps:

[0068] Take 7.7 mg of the rice blastin hapten prepared in Example 1, add 1 mL of dimethyl sulfoxide (DMSO) to dissolve and clarify, add NHS 5.1 mg, EDC 9.7 mg, fully dissolve and mix, and react at room temperature for 2 hours to obtain a hapten activation solution Liquid A; take 50 mg of ovalbumin (OVA), add 0.1mol / L CB9.1 7mL to dissolve, and obtain liquid B; add liquid A dropwise to liquid B, react at room temperature for 4 hours, stop the reaction, and use 0.02mol / L L PBS buffer was purified by dialysis for 3 days, the medium was changed 3 times a day, and centrifuged to obtain the artificial antigen of rice blastilin coupled with ovalbumin, which was aliquoted and stored at -20°C.

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Abstract

The invention discloses a hapten for isoprothiolane content detection and a preparation method and application thereof, particularly relates to a hapten for isoprothiolane enzyme-linked immunosorbent assay, and further discloses a preparation method of the hapten and application of the hapten in antibodies and kits. The enzyme linked immunosorbent assay kit established on the basis of the isoprothiolane hapten can rapidly detect products, and is convenient to use and low in detection cost, and the detection method is efficient, accurate and rapid, can simultaneously detect a large batch of samples, and is suitable for on-site monitoring of isoprothiolane in agricultural products, especially tobacco and screening of a large number of samples.

Description

technical field [0001] The invention relates to the technical field of pesticide residue detection, in particular to a hapten for detecting the content of blastilin, a preparation method and application thereof. Background technique [0002] Isoprothiolane (Isoprothiolane), also known as isopropylthiolane, Fuji No. 1, chemical name 1,3-dithiolane-2-ylidene diisopropyl malonate, is a highly efficient systemic fungicide, Its bactericidal spectrum is narrow, and it is a special drug for preventing and treating rice blast, especially for panicle blast. It can be systemically absorbed and conducted through the roots, stems, and leaves of plants, and can reach the heart leaves. It is also effective against rice seedling blast and Sclerotinia sclerotiorum, and it can also treat rice planthoppers and leafhoppers when used in a large area, and has a regulating effect on plant growth. However, with the large-scale application of rice blastin, it may pollute the environment, increase ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D339/06C07K14/765C07K14/77C07K14/795C07K16/44G01N33/53
CPCC07D339/06C07K14/765C07K14/77C07K14/795C07K16/44G01N33/5308
Inventor 王春琼陈丹张燕秦世春李苓蔡洁云张轲张晓伟彭丽娟孙浩巍杨艺敏张庆刚
Owner YUNNAN TOBACCO QUALITY SUPERVISION MONITORING STATION
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