Triazophos detection kit based on quantum dot probe

A technology of quantum dots and triazophos, which is applied in the field of triazophos detection kits, can solve the problems of long detection time, difficult preparation of specific antibodies, and expensive instruments, and achieves high sensitivity, high sensitivity and high selectivity. Effect

Inactive Publication Date: 2019-01-11
INST OF QUALITY STANDARD & TESTING TECH FOR AGRO PROD OF CAAS
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  • Application Information

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Problems solved by technology

[0006] The purpose of the present invention is to overcome the shortcomings of the traditional triazophos detection method, such as long detection time, expensive equipment, and difficult preparation of specific antibodies, to provide a biomimetic immunosorbent detection method based on quantum dot-labeled triazophos

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  • Triazophos detection kit based on quantum dot probe

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preparation example Construction

[0023] In some embodiments, the preparation method of the triazophos hapten labeled with the quantum dot probe comprises:

[0024] The triazophos hapten is sequentially activated and reacted with NHS and EDC to obtain an activated ester, and the activated ester is coupled with the quantum dot probe.

[0025] In some embodiments, the molar ratio of the triazophos hapten to NHS and EDC is 1: (1.7-2.3): (2.7-3.3);

[0026] In some embodiments, the molar ratio of the triazophos hapten to NHS and EDC is 1:2:3.

[0027] In some embodiments, the molar ratio of the activated ester to the quantum dot probe is (460-500):1; (470-490):1, or 480:1 can also be selected.

[0028] In some embodiments, the quantum dot probes are CdSe / ZnS QDs.

[0029] In some embodiments, the preparation of the molecularly imprinted polymer membrane is carried out on the solid phase carrier, and the method includes:

[0030] The template molecule is pre-reacted with methacrylic acid (MAA) in an organic solv...

Embodiment 1

[0062] This embodiment provides a biomimetic immunoassay method for triazophos quantum dot labeling, comprising the following steps:

[0063] 1. Synthesis of hapten:

[0064] (1) Synthesis of O-ethylthiophosphoryl dichloride (TZM-1)

[0065] Weigh the phosphorus trichloride (PSCl 3 ) 68g (about 0.4mol) was placed in a three-necked flask with a low-temperature thermometer, cooled to -10~-5°C with an ice-salt water bath, and 55g (about 1.2mol) of absolute ethanol was added dropwise under vigorous stirring, strictly Control the rate of addition so that the temperature of the reaction solution is always not greater than 0°C. After the dropwise addition, the reaction was continued at 10°C for 2h. After the reaction, the reaction solution (100ml×2) was washed with (0±5)°C distilled water, the oil layer was separated and washed with anhydrous Na 2 SO 4 After drying, it was distilled under reduced pressure by a water pump, and the fraction at 65-75°C was collected to obtain 51.8 ...

Embodiment 2

[0087] This embodiment provides a biomimetic immunoassay method for triazophos quantum dot labeling, comprising the following steps:

[0088] 1. Same as embodiment 1

[0089] 2. Preparation of quantum dot-labeled hapten probes:

[0090] (1) Preparation of activated ester triazophos hapten (15mmol L -1 , dissolved in DMF) and NHS (12.3 mg / mL, dissolved in DMF) were added to the centrifuge tube and vortexed for 1 min. Then, EDC (5 mg / mL, dissolved in DMF) was added, and the volume was adjusted to 1 ml with DMF. The mixture was shaken and reacted at room temperature for more than 12 hours. Hapten, NHS, EDC molar ratio is 1:2.3:2.7.

[0091] (2) Before the labeling reaction, 1.5 μL Tween-20 was added to reduce the aggregation of quantum dots. 50 μL CdSe / ZnS QDs (dissolved in 340 μL BBS solution) and 80 μL activated ester were added to a centrifuge tube and shaken at room temperature for more than 12 hours. The molar ratio of activated ester to quantum dots is 500:1.

[0092...

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Abstract

The invention relates to the field of pesticide detection, in particular to a triazophos detection kit based on a quantum dot probe. The kit comprises a solid-phase carrier coated with a molecularly imprinted polymer membrane and a triazophos hapten labeled by the quantum dot probe, wherein the molecularly imprinted polymer membrane is prepared by taking triazolone as a template molecule. A triazophos adsorption functional material provided by the invention has relatively high selectivity, the problem of template leakage can be solved by adopting a virtual template, and the material can replace a biological antibody to be applied to a fluorescence immunoassay technology; and the adsorption functional material is prepared with a chemical method, has relatively high stability, relatively long service life and relatively strong severe environment resistance, and overcomes the shortcomings of long preparation period, easy inactivation, high cost and the like of a traditional biological antibody.

Description

technical field [0001] The invention relates to the field of pesticide detection, in particular to a quantum dot probe-based triazophos detection kit. Background technique [0002] Organophosphate insecticides are the most widely used insecticides and include some of the most toxic chemicals used in agriculture. Triazophos has been widely used as an excellent substitute for highly toxic organophosphates to control pests on grains, fruits, and vegetables. Due to chemical and optical stability and long half-life, triazophos may persist in the natural environment for a long time. According to literature reports, triazophos induces oxidative stress and histopathological changes in rats, which may also be potentially harmful to human health. Since the end of 2016, the Chinese government has banned the use of triazophos on vegetables. Therefore, it is necessary to develop an effective analytical method for the regular monitoring of triazophos residues in agricultural and enviro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/64
Inventor 佘永新王静洪思慧王淼王珊珊郑鹭飞曹振
Owner INST OF QUALITY STANDARD & TESTING TECH FOR AGRO PROD OF CAAS
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