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Pesticide residue fast detection card based on horse serum cholinesterase and production method and use method thereof

A cholinesterase and production process technology, which is applied in the direction of chemical reaction of materials and material analysis by observing the impact on chemical indicators, can solve the problem that the results of enzymatic reactions are not easy to be judged by the naked eye, pesticide residues The quick test card is cumbersome to operate, and the visual discrimination is not high. It achieves the effect of low testing cost, convenient on-site testing, and low cost of reagent consumables.

Active Publication Date: 2018-06-15
WUHAN ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although acetylcholine is the natural substrate of cholinesterase, the substrate has no color and is not easy to induce discoloration reaction, so the result of the enzymatic reaction is not easy to judge with the naked eye; although the enzymatic reaction product of thioacetylcholine can induce disulfide The substituted dinitrobenzoic acid changed from colorless to yellow, but the naked eye discrimination of this color change reaction was not high, especially with the milky white to light yellow enzyme tablet as the color background, it was not easy to identify; The introduction of nitrobenzoic acid as a chromogenic agent will make the operation of the rapid test card for pesticide residues more cumbersome
The yield of the traditional method for preparing indophenol acetate is not high, and the final product has more impurities. After the improved process, the yield can reach more than 90%, the impurities are less, and it is easy to purify, and the complex solution of the final product is improved to make it Easier to spread on filter paper discs

Method used

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  • Pesticide residue fast detection card based on horse serum cholinesterase and production method and use method thereof
  • Pesticide residue fast detection card based on horse serum cholinesterase and production method and use method thereof
  • Pesticide residue fast detection card based on horse serum cholinesterase and production method and use method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] A kind of pesticide residue rapid test card production process based on horse serum cholinesterase, comprises the steps:

[0037] S1, the preparation of cholinesterase substrate:

[0038] Add 45mL of acetic anhydride and 4.5mL of acetyl chloride into a three-necked round-bottomed flask equipped with mechanical stirring and a thermometer, and slowly add 10 g of indophenol sodium solid in batches at room temperature. . After the reaction, the reaction solution was concentrated to dryness under reduced pressure at 60°C, and the concentrated solid was redissolved with a mixed solution of 3L methanol and acetone (the volume ratio of methanol to acetone was 1:5) to obtain indophenol acetic acid Methanol solution of ester, standby;

[0039] S2, preparation of horse serum:

[0040] S21. Before blood collection, wrap the required blood collection needles, rubber tubes, centrifuge bottles, and infusion bottles with kraft paper, and strictly sterilize them for 1 hour under the ...

Embodiment 1

[0058] The detection method and judgment standard of the pesticide residue quick test card prepared by the method of Example 1

[0059] 1. Select the sample to be tested (for fruits and vegetables), and carry out the following pretreatment:

[0060] (1) Sampling method: take fresh melon and fruit epidermis and 1 mm thick tissue under the epidermis, cut into about 1 square centimeter in size;

[0061] (2), take 5g of the tissue obtained in step (1), add 10mL of 0.1mol / L, PH8.5 phosphate buffer, shake well, filter, and finally take the filtrate as a test sample and set aside;

[0062] 2. Detection method and judgment standard

[0063] Add 60 μL of the sample to be tested prepared in step 1 dropwise to the horse blood cholinesterase tablet of the Pesticide Residue Rapid Test Card, and react at 37°C for 10 minutes; then press the substrate sheet onto the horse blood cholinesterase sheet, and continue at 37°C React for 3 minutes, observe the color change of the enzyme tablet.

...

Embodiment 3

[0066] The detection test of the pesticide residue quick test card prepared by embodiment one method:

[0067] (1) Sensitivity test of pesticide residue quick test card:

[0068] The standard pesticides such as methamidophos, parathion, dichlorvos, trichlorfon, carbaryl, and haoniandong were respectively diluted by multiple ratios, and tested with the pesticide residue rapid test card respectively. The detection limits of these pesticides are shown in Table 1:

[0069] Table 1. Detection limits of some pesticides by the Pesticide Residue Rapid Test Card

[0070]

[0071] (2), repeatability test of pesticide residue quick test card:

[0072] Under the same conditions, 20 known positive samples and known negative samples were tested repeatedly within the batch every day, and 3 different batches were taken for testing between batches. The detection continued for 7 days, and the color change of the quick test card was used to determine the difference. Repeatability, the resu...

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Abstract

The invention relates to a production method of a pesticide residue fast detection card based on horse serum cholinesterase. The method comprises precipitating and setting cholinesterase in horse serum through a salting-out method, carrying out purification through centrifugation, redissolution and dialysis to obtain cholinesterase satisfying the requirements, carrying out dilution according to the activity of the cholinesterase, adjusting pH and a salt ion concentration, adding a stabilizer into the solution to obtain a pesticide residue fast detection card enzyme work solution, pressing twocircular filter papers on a support substrate at a high temperature, adding a cholinesterase working solution and a substrate working solution on the two filter papers drop by drop, carrying out airing and carrying out plastic packaging to obtain the pesticide residue fast detection card. The pesticide residue fast detection card has good sensitivity, stability, repeatability, reproducibility, simplicity, rapidness, intuitiveness and low cost and is suitable for rapid screening of pesticide residues on fruits and vegetables.

Description

technical field [0001] The invention relates to the technical field of detection of pesticide residues in agricultural products, in particular to a rapid detection card for pesticide residues based on horse serum cholinesterase and its production process and use method. Background technique [0002] As an important means of agricultural production, pesticides play an important role in preventing and controlling pests and diseases, removing weeds and regulating the growth of crops. Due to the failure to apply pesticides according to the regulations or the lack of awareness of the potential hazards of pesticides, pesticides are directly or indirectly enriched in agricultural by-products or excessive residues through the food chain. There are hundreds of acute poisoning incidents caused by eating agricultural by-products contaminated by pesticides every year across the country. Long-term consumption of agricultural and sideline products with excessive pesticide residues will n...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78
CPCG01N21/78
Inventor 刘家飞夏定刘国松韩艳云朱运锋战艺芳肖康飞周琦
Owner WUHAN ACADEMY OF AGRI SCI
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