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.
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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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