Solid enzyme agent for quick detection of residual pesticide toxicity and preparation method and using method thereof

A technology for pesticide residues and reagents, applied in biochemical equipment and methods, microbial determination/inspection, and analysis by chemical reaction of materials, etc., can solve problems such as reduction of enzyme activity, and achieve low cost and good water solubility. , to ensure the effect of detection sensitivity

Inactive Publication Date: 2006-02-08
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The solid enzyme reagent for rapid detection of organophosphorus and carbamate pesticide residues of the present invention can be stored stably for more than 20 days at room temperature (20-25° C.), thereby overcoming the shortcomings of existing solid enzyme reagents and overcoming the enzyme solution Due to the significant reduction in enzyme activity caused by repeated thawing and freezing at -20°C, it effectively solves the current problems in the rapid detection of organophosphorus and carbamate pesticide residues

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] 1) Preparation of enzyme solution A: put 5 μL of acetylcholinesterase with an activity of 1000 units / mL into a beaker, add 5 mL of 0.1M phosphate buffer solution with pH=7.8-8.2, and dilute the acetylcholinesterase to obtain enzyme solution A;

[0032] 2) Preparation of enzyme solution B: Add 20 mL of 0.1M Tris-HCl buffer solution with pH=7.8-8.2 to the prepared enzyme solution A above to dilute enzyme solution A to obtain enzyme solution B;

[0033]3) Dissolution of the carrier: Take 2.0g of the carrier polyvinylpyrrolidone K30 (PVP) and put it into the above-mentioned prepared enzyme solution B, stir to dissolve the carrier polyvinylpyrrolidone K30 (PVP); put it in the refrigerator at 0-4 ℃ for more than 1 hour to combine the enzyme with the carrier;

[0034] 4) Freeze-drying: transfer the above enzyme-carrier conjugate to a freeze-drying bottle, place the freeze-drying bottle in a low-temperature ethanol bath below -20°C and gently rotate the freeze-drying bottle to ...

Embodiment 2

[0037] 1) Preparation of enzyme solution A: 50 μL of butyrylcholinesterase with an activity of 1000 units / mL; add 30 mL of 0.1M phosphate buffer solution with pH=7..8-8.2 to dilute the enzyme solution to obtain enzyme solution A ;

[0038] 2) Preparation of enzyme solution B: Add 50 mL of 0.1M Tris-HCl buffer solution with pH7=7.8-8.2 to the prepared enzyme solution A above to dilute enzyme solution A to obtain enzyme solution B;

[0039] 3) Dissolution of the carrier: Take 10g of the carrier polyvinylpyrrolidone K30 (PVP) and put it into the prepared enzyme solution B above, stir to dissolve the carrier polyvinylpyrrolidone K30 (PVP); place it in a refrigerator at 0-4°C for reaction More than 1 hour, the enzyme is combined with the carrier;

[0040] 4) Freeze-drying: Transfer the above-mentioned enzyme-carrier conjugate to a freeze-drying bottle, place the freeze-drying bottle in a low-temperature ethanol bath below -20°C and gently turn it to freeze-dry, so that the solute ...

Embodiment 3

[0043] 1) Preparation of solution A: put 10 μL of acetylcholinesterase with an activity of 1000 units / mL into a beaker, add 8 mL of 0.1M pH=7.8=8.2 phosphate buffer solution, dilute the enzyme solution, and obtain enzyme solution A;

[0044] 2) Preparation of enzyme solution B: add 25 mL of 0.1M Tris-HCl buffer solution with pH=7.8-8.2 to the prepared enzyme solution A above, dilute enzyme solution A to obtain enzyme solution B;

[0045] 3) Dissolution of the carrier: Take 5.0g of the carrier polyvinylpyrrolidone K30 (PVP) and put it into the prepared enzyme solution B above, stir to dissolve the carrier polyvinylpyrrolidone K30 (PVP); put it in the refrigerator for more than 1.0 hours Binding the enzyme to the carrier;

[0046] 4) Freeze-drying: Transfer the above-mentioned enzyme-carrier conjugate to a freeze-drying bottle, place the freeze-drying bottle in a low-temperature ethanol bath below -20°C and gently turn it to freeze-dry, so that the solute can be coated on the bo...

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PUM

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Abstract

The invention provides a method for preparing a solid enzyme reagent which is used to fast measure the organophosphorus and carbamate toxicity of pesticide residue and its using method. The agent is formed by choline esterase, buffer solution and carrier. The solid enzyme reagent can be stored at indoor temperature above 20 days.

Description

technical field [0001] The invention relates to a composition, in particular to an enzyme-containing solid composition reagent, in particular to an enzyme-containing solid composition reagent for rapid detection of organophosphorus and carbamate pesticide residues. Background technique [0002] The harm of residual pesticides to human health is one of the hot spots in food safety issues. Worldwide, poisoning incidents caused by excessive residues of organophosphorus and carbamate pesticides have occurred frequently, of which 69.5% are caused by organophosphorus caused by pesticides. Therefore, it is very important to monitor pesticide residues in food and environment. At present, the enzyme inhibition method is usually used for the rapid detection of organophosphorus and carbamate pesticide residues, and the degree of enzyme inhibition is used to measure by enzyme-catalyzed kinetic photometry with the aid of a pesticide residue rapid tester, or by color change, or by acidit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78C12Q1/00
Inventor 邹明强王大宁陈彦长张晓华李锦丰齐小花
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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