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Chemiluminescence sensing detection method for organophosphorus pesticide residues and application thereof

An organophosphorus pesticide and chemiluminescence technology, which is applied in the field of analysis and detection, can solve the problems of low sensitivity, time-consuming, rapid blood samples from poisoned patients, and difficulty in high-throughput detection.

Inactive Publication Date: 2015-05-06
FUJIAN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing organophosphorus poison detection methods or instruments have the defects of low sensitivity, low sample processing capacity, and time-consuming. It is difficult to realize rapid and high-throughput detection of pesticide residue samples, environmental samples, and blood samples of poisoned patients.

Method used

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  • Chemiluminescence sensing detection method for organophosphorus pesticide residues and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Standard Curve of Dichlorvos Pesticides

[0028] Draw 50 μL of 1 ng / mL~1 μg / mL series of dichlorvos standard solutions and place them in the wells of a 96-well microtiter plate, then draw another 50 μl of distilled water and add them to other wells of the 96-well microtiter plate as a blank control. Add 50 μl of acetylcholinesterase (pH=7) solution with a concentration of 10 U / mL to all the wells that have been added with the solution, shake and react at 37°C for 10 minutes on a microplate shaker, and then add 1 ×10 -4 50 μl of acetylcholine at a concentration of 10 U / mL and 50 μl of choline oxidase at a concentration of 10 U / mL (pH=7), shake and react at 37 ° C for 10 minutes, then transfer the microplate plate to a chemiluminescent sensor detector, and automatically inject chemical The luminescent probe reagent luminol or oxalate peroxide TCPO, the concentration of luminol solution or TCPO peroxide oxalate was 1×10 -4 mol / L, 1×10 -3 mol / L, while det...

Embodiment 2

[0029] Example 2 Detection of Dichlorvos Pesticides in Apples

[0030] Take 1 fresh apple, squeeze the juice, filter the juice, take 1 mL of each of the three supernatants, add 10 μL, 50 μL, and 100 μL of 1 μg / mL dichlorvos to the above three supernatants with a pipette, and mix well , is the sample solution. Take 50 μl of the sample solution, place them in the wells of the 96-well microplate, and then draw another 50 μl of distilled water and add it to the other wells of the 96-well microplate as a blank control. Add the solution to all the wells of the 96-well microplate Add 50 μl of acetylcholinesterase (pH=7) solution with a concentration of 10 U / mL to each well, shake and react at 37°C for 10 minutes on a microplate shaker, and then add 1×10 -450 μl of acetylcholine at a concentration of 10 U / mL and 50 μl of choline oxidase at a concentration of 10 U / mL (pH=7), shake and react at 37 ° C for 10 minutes, then transfer the microplate plate to a chemiluminescent sensor dete...

Embodiment 3

[0033] Example 3 Detection of Dichlorvos Pesticide Residues in Chinese Cabbage

[0034] 5 pieces of fresh Chinese cabbage, after squeezing the juice, filter with ordinary filter paper, take 50 μL of the filtrate, place it in the well of a 96-well enzyme label plate, and then absorb another 50 μl of distilled water and add it to the other wells of the 96-well enzyme label plate as a blank control. Add 50 μl of acetylcholinesterase (pH=7) solution with a concentration of 10 U / mL to all the wells of the 96-well microplate plate, shake and react at 37°C for 10 minutes on a microplate shaker, and then Each added concentration is 1×10 -4 50 μl of acetylcholine at a concentration of 10 U / mL and 50 μl of choline oxidase at a concentration of 10 U / mL (pH=7), shake and react at 37 ° C for 10 minutes, then transfer the microplate plate to a chemiluminescent sensor detector, and automatically inject chemical The luminescent probe reagent luminol, the concentration of luminol solution...

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Abstract

The invention discloses a chemiluminescence sensing detection method for organophosphorus pesticide residues and application thereof. The method is characterized by comprising the following steps: by taking a chemiluminescent probe as an indicator, indicating the organophosphorus pesticides to inhibit the degree of reducing choline produced by acetylcholine chloride subjected to catalytic hydrolysis of acetylcholin esterase, coupling choline oxidase to catalyze choline to generate hydrogen peroxide, then oxidizing a luminescence probe to generate chemiluminiscence by virtue of hydrogen peroxide, carrying out a multi-step coupling reaction to realize sensing detection of the organophosphorus pesticide residues by utilizing a chemiluminescence apparatus. According to the sensing method, 96 samples can be detected within 20 minutes, and dichlorvos is taken as an example, the detection limit reaches 1nmol / L. The method is sensitive and rapid and can be used for blood sample monitoring of patients suffering from poisoning of foods, water samples and organophosphorus poisons.

Description

technical field [0001] The invention belongs to a new method and a new principle of analysis and detection, and in particular relates to a new principle and application of a high-throughput and rapid detection of organophosphorus pesticide residues using a chemiluminescence microplate reader and a chemiluminescence probe. Background technique [0002] Organophosphorus pesticides are the most widely used and common insecticides in my country. In recent years, due to the abuse, excessive use and misuse of organophosphorus pesticides, organophosphorus poisoning incidents have occurred frequently. Especially in the process of crop pest control, organophosphorus pesticides are often used in excessive amounts, resulting in serious organophosphorus pesticide residues in agricultural products. Long-term consumption of agricultural products with excessive pesticide residues will undoubtedly cause harm to the body. In addition, acute poisoning incidents caused by mistaken intake of o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/76
Inventor 姚宏史培颖林新华
Owner FUJIAN MEDICAL UNIV
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