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Reagent and card for detecting organophosphorus and carbamates pesticide residues

A technology of methylumbelliferyl butyrate and reagents, which can be used in measuring devices, fluorescence/phosphorescence, and material analysis by observing the influence of chemical indicators, and can solve problems such as limited applications

Active Publication Date: 2019-12-20
GUANGZHOU JUBIOS BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, although 4-methylumbelliferyl acetate or 4-methylumbelliferyl butyrate can be hydrolyzed to 4-methylumbelliferone under the action of cholinesterase, it can theoretically be applied to choline The detection of esterase activity, but in fact, the application is limited because it is easy to self-degrade and degrade rapidly in the detection environment. There is no report showing that 4-methylumbelliferone acetate or 4-methylumbelliferone butyrate Esters as Enzyme Activity Indicators for Pesticide Residue Detection

Method used

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  • Reagent and card for detecting organophosphorus and carbamates pesticide residues
  • Reagent and card for detecting organophosphorus and carbamates pesticide residues

Examples

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preparation example Construction

[0039] A kind of preparation method of stable 4-methylumbelliferyl ketone ester reagent in aqueous solution, comprises the steps:

[0040] 1) Dissolving 4-methylumbelliferyl ester and molecules with internal hydrophobic spaces in an anhydrous solvent, fully mixing and reacting;

[0041] 2) The solvent is removed to obtain a stable 4-methylumbelliferone ester reagent.

[0042] The anhydrous solvent can be various solvents that can effectively dissolve the 4-methylumbelliferone ester, preferably an anhydrous solvent with a higher solubility.

[0043] In some examples of the preparation method of the present invention, the 4-methylumbelliferone ester includes 4-methylumbelliferone acetate or 4-methylumbelliferone butyrate.

[0044] In some examples of the preparation method of the present invention, the molecule having an internal hydrophobic space is selected from calixarene, cyclodextrin or cucurbit ring and derivatives thereof.

[0045] In some examples of the preparation me...

Embodiment 1

[0059]1) Dissolve 10 mg of 4-methylumbelliferyl acetate in 10 mL of methanol, and add 0.05% calixarene to form a fluorescent substrate solution;

[0060] 2) Take a bottle of prepared cholinesterase, the enzyme activity of which is 0.5U;

[0061] 3) When using, first add 100 μL cholinesterase to 2.5 mL sample extract, after inhibiting for 10-15 minutes, add 100 μL fluorescent substrate solution, and measure the increase in fluorescence value within 2 minutes at 450 nm.

[0062] At the same time, a blank control was made to calculate the enzyme inhibition rate.

[0063] Enzyme inhibition rate calculation formula:

[0064]

[0065] In the formula:

[0066] ΔR 0 ───The change value of the fluorescence value of the control solution after 2 minutes of reaction;

[0067] ΔR t ───The change value of the fluorescence value of the sample solution after 2 minutes of reaction.

Embodiment 2

[0069] 1) Fix the chromatography filter paper with circular holes on the bottom plate;

[0070] 2) Soak the 11mm circular filter paper in the fluorescent substrate solution containing 0.08% cucurbitan and 0.1% 4-methylumbelliferyl acetate, dry it and fix it in the circular hole on the 22×50mm chromatographic filter paper;

[0071] 3) Embedding 0.1U cholinesterase at a distance of 30mm from the edge of the circular hole on the chromatography filter paper, drying and sealing the upper cover film, and assembling to obtain a rapid detection card for pesticides by fluorescence method.

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Abstract

The invention discloses a reagent and card for detecting organophosphorus and carbamates pesticide residues. Through including 4-methylumbelliferone ester in a molecule with a hydrophobic space, the stability is improved. The 4-methylumbelliferone ester reagent of some examples of the invention can be stably present in an aqueous solution, and can also be hydrolyzed by a reaction with cholinesterase, so that the 4-methylumbelliferone ester reagent can be used as a color developer for the rapid detection of organophosphorus and carbamates pesticide residues.

Description

technical field [0001] The invention relates to the field of rapid detection of pesticide residues, in particular to a reagent and a card for detecting residues of organic phosphorus and carbamate pesticides. Background technique [0002] Organophosphorus and carbamate pesticides can bind to the enzymatic center of cholinesterase and compete with the substrate for the binding site, thereby inhibiting the activity of cholinesterase. The color change process of the free chromogenic group of 2,6-dichloroindophenol acetate hydrolyzed by cholinesterase from red to blue is used to judge the degree of enzyme inhibition according to the degree of color change. The pesticide quick test cards currently on the market are basically based on this color rendering principle. The enzyme inhibition rate method in the national standard GB / T 5009.199-2003 uses cholinesterase to decompose the substrate acetylcholine, and the decomposition product reacts with the chromogenic agent 5,5-dithiobis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78G01N21/64
CPCG01N21/6486G01N21/78
Inventor 雷达张联裕
Owner GUANGZHOU JUBIOS BIOTECH CO LTD
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