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Conventional pesticide residue quick detecting method

A detection method and pesticide technology, applied in biochemical equipment and methods, microbe determination/inspection, color/spectral characteristic measurement, etc., can solve problems affecting reaction balance, no correction, color interference, etc.

Inactive Publication Date: 2003-04-30
江苏省农业科学院植物保护研究所
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Problems solved by technology

This detection method mainly has the following problems: First, after the plants are dipped in ethanol, the plant pigments are extracted at the same time, and color interference will occur if they are not treated; the substrate and chromogenic agent are added at the same time, although it is more convenient, it will affect the reaction balance. , due to the slightly yellow color of the developer, this method does not correct for

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0009] Embodiment 1: Methamidophos inhibitory rate-the preparation method of concentration standard curve

[0010] 1. Take a 96-well microplate

[0011] 2. Add 50ul of enzyme diluent (30 times) and 50ul of methamidophos serial diluent (control group plus PB solution) and 50ul of substrate to the wells, and incubate at 37°C-38°C for about 15 minutes. Only add 50ul PB solution and 50ul substrate.

[0012] 3. After warming, add 50ul DTNB chromogen to each well.

[0013] 4. Measure the absorbance value (OD value) at 412 nm on a microplate reader.

[0014] 5. Calculate the standard curve of each pesticide inhibition rate under the series concentrations (25.02%, 49.45%, 74.21%, 91.83%) with the inhibition rate formula.

Embodiment 2

[0015] Embodiment 2: the preparation method of triazophos inhibition rate-concentration standard curve.

[0016] 1. Take a 96-well microplate

[0017] 2. Add 50ul of enzyme diluent (30 times) and 50ul of triazophos serial diluent (control group plus PB solution) and 50ul of substrate into the well, and incubate at 37°C-38°C for about 15 minutes. Only add 50ul PB solution and 50ul substrate.

[0018] 3. After warming, add 50ul DTNB chromogen to each well.

[0019] 4. Measure the absorbance value (OD value) at 412 nm on a microplate reader.

[0020] 5. Calculate the standard curve of each pesticide inhibition rate under the series concentrations (11.15%, 31.43%, 55.22%, 79.72%) with the inhibition rate formula.

Embodiment 3

[0021] Example 3: Rapid detection method for leaf samples.

[0022] 1. Sampling: Take 2 grams from 10-15 pieces of leafy vegetables as the test sample.

[0023] 2. Extract with 3ml of acetone and 50ml of bromine water (1%) for 3 minutes, pass through a 2cm high activated carbon column in a 20ml syringe, and let stand for 2 minutes.

[0024] 3. Add 50ul of acetylcholinesterase and 50ul of substrate ATCI (as a blank control) into a 96-well microwell plate, and incubate at 37°C-38°C for 15 minutes.

[0025] 4. After warming, add 50ul extract solution and 50ul chromogen DTNB to each well, and measure the absorbance value (OD value) at 412nm on a microplate reader.

[0026] 5. Calculate the inhibition rate, and compare it with the standard curve to determine the pesticide residue concentration and inhibition rate in the leaf samples.

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Abstract

A quick residual detecting method for conventional pesticide features that the acetylcholine esterase from the liver of white rats is used as the source of the enzyme. The acetylcholine is used as the substrate. Under the action of acetylcholine esterase, the substrate is hydrolyzed so as to generate the sulfocholine and DTNB. The peak of the maximum absorption in the color reaction is at 412 nm.With concentration of enzyme being fixed, comparing OD412 value of the pesticide in different concentrations establishes the standard curvature. With being dipped and extracted, the sample to be measured is tested so as to obtain the value of OD412. Base on the value of OD412 of the sample to be measured, the residual concentration and the depression rate of the pesticide in the sample to be measured in determined.

Description

Technical field: [0001] The invention relates to a rapid detection technology for conventional pesticide residues. Background technique: [0002] With the improvement of people's living standards, people's demand for agricultural and sideline products such as fruits and vegetables is getting higher and higher. In order to meet market needs, prevent pesticide products such as vegetables and fruits from being attacked by diseases and insect pests, and promote their growth, various pesticides and chemical fertilizers are widely used. . The harmful substances stranded in agricultural products directly threaten people's normal life. The technical trade measures (TBT) of countries all over the world to participate in the production of pesticide products emphasize the continuous improvement of the safety of agricultural products, which has become a hot issue of global concern. Among them, pesticide residues in agricultural products have become an important restrictive index for ag...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04G01N21/25
Inventor 刘贤金廖海丰张存政王冬兰
Owner 江苏省农业科学院植物保护研究所
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