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Extraction method of mixed colored cowpea esterase and application of mixed colored cowpea esterase in detection of organophosphorus pesticide residues

An organophosphorus pesticide and extraction method technology, applied in the field of pesticides, can solve the problems of high detection cost and difficulty in obtaining materials, and achieve the effects of easy acquisition, convenient extraction, and rapid detection and detection.

Inactive Publication Date: 2018-11-16
HUIZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Previously, we mostly used acetylcholinease to detect pesticides, but the enzyme sources were mostly expensive materials such as sardines and fly heads, which also made the detection cost high and difficult to obtain materials.

Method used

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  • Extraction method of mixed colored cowpea esterase and application of mixed colored cowpea esterase in detection of organophosphorus pesticide residues
  • Extraction method of mixed colored cowpea esterase and application of mixed colored cowpea esterase in detection of organophosphorus pesticide residues
  • Extraction method of mixed colored cowpea esterase and application of mixed colored cowpea esterase in detection of organophosphorus pesticide residues

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] 1.1 Experimental materials and reagents

[0062] Cowpea (Taobao), 0.1mol / L, pH 6.5 phosphate buffer, 0.04mol / L, pH 6.5 phosphate buffer, α-naphthyl acetate (chemically pure), absolute ethanol (chemically pure), Fast Blue B salt (chemically pure), ammonium sulfate (chemically pure), distilled water, 1mmol / L α-naphthol absolute ethanol standard solution (chemically pure), α-naphthyl acetate acetone solution: 0.016mol / L: use acetone (99% ) preparation, stored in the dark at 4°C, Coomassie brilliant blue reagent, bovine serum albumin, NaCl solution

[0063] 1.2 Reagent equipment

[0064] (1) Substrate sheet dripping liquid equipment

[0065] Dissolve 14.8mg (0.014g) of acetic acid-α-naphthyl ester in 50mL of acetone, which is the substrate tablet drop solution.

[0066] (2) Chromogenic solution equipment

[0067] Dissolve 15.6mg (0.015g) of Fast Blue B salt in 10mL of distilled water and shake for 30min at a rate of 180 times min to obtain the chromogenic solution.

[...

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Abstract

The invention discloses an extraction method of mixed colored cowpea esterase. The mixed colored cowpea esterase is extracted from mixed colored cowpeas by adopting a crushing centrifugation method; the mixed colored cowpea esterase is salted out through ammonium sulfate solutions at different concentrations, and then mixed colored cowpea esterase raw liquid is purified through an ethanol precipitation purification process; acetic acid and alpha-naphthol are generated from a mixed colored cowpea esterase hydrolysis substrate according to the principle of an enzyme inhibition method; and the alpha-naphthol reacts with fast blue B salt to generate an amaranth azocompound. The influence of pesticide on the activity of the enzyme is detected by manufacturing a fast detection card; after the mixed colored cowpea esterase is inhibited by organophosphorus pesticide, the chromogenic reaction is affected; and therefore, the pesticide residue situation is indicated by the strength of the chromogenic reaction.

Description

technical field [0001] The invention relates to the technical field of pesticides, in particular to a method for extracting cowpea enzyme and its application in the detection of organophosphorus pesticide residues. Background technique [0002] Pesticides are indispensable in agricultural production. my country is a big agricultural country, and it is also a big country in the production and use of pesticides. It is undeniable that pesticides have played an important role in pest control and weed removal. Excessive use of chemical substances that will remain in the soil and vegetables will bring immeasurable harm to human health, and the pesticides left in the soil will destroy the ecological environment and affect the farming environment. Relevant reports show that a large number of pesticides in my country have not been used, mainly diffused into the atmospheric environment through pesticide residues, polluted the system through precipitation, polluted the soil, and finally...

Claims

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Application Information

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IPC IPC(8): C12N9/18C12Q1/44
CPCC12N9/18C12Q1/44
Inventor 谢海伟黄彩景罗沛如易婷
Owner HUIZHOU UNIV
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