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Method for simultaneously detecting parathion-methyl, chlorpyrifos and trichlorfon based on quantum dot labeled biomimetic fluorescence immunoassay

A methyl parathion and quantum dot technology, applied in the field of biomimetic fluorescence immunoassay, can solve the problems of single pesticide detection and increase the cost of pesticide residue detection, and achieve the effects of high sensitivity, low detection limit and short analysis process.

Active Publication Date: 2018-11-30
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are often multiple pesticide residues in agricultural products, and most biomimetic immunoassay methods are usually used for single pesticide detection
To achieve comprehensive detection, it is necessary to use a variety of biomimetic antibodies, which will greatly increase the cost of pesticide residue detection

Method used

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  • Method for simultaneously detecting parathion-methyl, chlorpyrifos and trichlorfon based on quantum dot labeled biomimetic fluorescence immunoassay
  • Method for simultaneously detecting parathion-methyl, chlorpyrifos and trichlorfon based on quantum dot labeled biomimetic fluorescence immunoassay

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1: Fe 3 o 4 @SiO 2 Preparation of @MIP:

[0054] 1 mmol DPA was dissolved in 5 mL tetrahydrofuran, followed by the addition of 2 mmol APTES. After stirring for 20min, add 60.00mg Fe 3 o 4 @SiO 2 And 6mmol TEOS, stirred for 30min. When 0.3mL (0.1mol L -1 ) after ammonium hydroxide was added, the mixture was placed at 60° C. for 10 h, filtered and washed with methanol to remove unreacted reagents and template molecules, and then dried in a vacuum oven at 60° C. Treat with 150mL methanol / acetic acid (4 / 1, V / V) for 24h and 150mL methanol for 12h to remove DPA. The imprinted polymer was then vacuum-dried at 60 °C for 12 h to obtain Fe 3 o 4 @SiO 2 @MIP.

Embodiment 2

[0055] Example 2: Synthesis of Quantum Dot Labeled Haptens:

[0056] CdSe / ZnS quantum dots with different emission wavelengths (525nm, 585nm and 625nm) were used to label methyl parathion, chlorpyrifos and trichlorfon hapten respectively. The marking steps are as follows:

[0057] Methyl parathion, chlorpyrifos, trichlorfon hapten, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and three CdSe / ZnS quantum dots with different emission wavelengths ( 525nm, 585nm, 625nm) were added into the reaction vessel at a molar ratio of 200:4000:1, and mixed thoroughly; 80 μL of borate buffer (pH 7.4) was added and reacted at room temperature in the dark for 2 hours. After the reaction, centrifugation was performed to remove aggregates, and the reactant was concentrated by ultrafiltration for separation and purification; the final product was stored at 4°C.

Embodiment 3

[0058] Embodiment 3: the drawing of standard curve

[0059] with Fe 3 o 4 @SiO 2 @MIP as a public biomimetic antibody, methyl parathion, chlorpyrifos and trichlorfon mixed standard solution and quantum dot-labeled haptens of three pesticides (diluted 500 times with methanol / water solution (3 / 17, V / V)) competition Combining a certain amount of polymer, the reaction process is as follows:

[0060] 1mL quantum dot-labeled haptens with different emission wavelengths and 3mL mixed standard solution of methyl parathion, chlorpyrifos and trichlorfon (0.000033-333.33mg L -1 )mix. Then add 5.00mg of Fe 3 o 4 @SiO 2 @MIP. Shake at room temperature for 120 min; centrifuge at 8000 r / min for 15 min, and measure the fluorescence value of the supernatant with a fluorescence spectrometer. Calculate the inhibition rate at different emission wavelengths based on the fluorescence values.

[0061]

[0062] In the formula:

[0063] IC%: the inhibition rate of antigen-antibody binding...

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Abstract

The invention discloses a method for simultaneously detecting parathion-methyl, chlorpyrifos and trichlorfon based on quantum dot labeled biomimetic fluorescence immunoassay. According to the invention, the multi-recognition-site Fe3O4@SiO2@MIP is used as the common biomimetic antibody, and three organophosphorus pesticide haptens such as parathion-methyl, chlorpyrifos and trichlorfon are respectively labeled with three kinds of quantum dots with different emission wavelengths at the same excitation wavelength, such that the simple and rapid biomimetic fluorescence immunoassay method for simultaneously detecting the three organophosphorus pesticides such as parathion-methyl, chlorpyrifos and trichlorfon is established; and the method has advantages of low cost, simple experiment operation,accurate measurement result and low detection limit, and is suitable for the simultaneous detection of various organophosphorus pesticides in various foods.

Description

technical field [0001] The invention relates to a bionic fluorescent immunoassay method, in particular to a method for simultaneously detecting methyl parathion, chlorpyrifos and trichlorfon based on quantum dot-labeled bionic fluorescent immunoassay, and belongs to the technical field of food safety detection. Background technique [0002] Organophosphorus pesticides have the advantages of high insecticidal efficiency, wide range of pest control, low cost, and high selectivity, and have broad application prospects in agricultural production. However, unreasonable use of pesticides will lead to pesticide residues and threaten people's food safety. [0003] As people pay more and more attention to food safety, more and more people begin to pay attention to pesticide residues. At present, most of the methods used for the simultaneous detection of multiple organophosphorus pesticides in food at home and abroad are gas chromatography coupled with mass spectrometry; high performa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/533
CPCG01N33/533
Inventor 徐志祥高绘菊姜名荻徐龙华孙玉奉
Owner SHANDONG AGRICULTURAL UNIVERSITY
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