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An electrochemical sensor for detecting acetamiprid, its preparation method and application

A technology for detecting acetamiprid and acetamiprid, which is applied in the field of electrochemical sensors for detecting acetamiprid and the field of preparation thereof, can solve the problems of small molecular weight, inability to detect, insufficient to combine two kinds of antibodies at the same time, etc., and achieves easy portability, Simple electrode, accurate and reliable detection results

Inactive Publication Date: 2020-06-02
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These substances to be detected usually contain multiple recognition sites and can be detected by conventional sandwich immunoassays, but pesticides such as acetamiprid cannot be detected by sandwich immunoassays due to their small molecular weights and insufficient binding to two antibodies at the same time

Method used

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  • An electrochemical sensor for detecting acetamiprid, its preparation method and application
  • An electrochemical sensor for detecting acetamiprid, its preparation method and application
  • An electrochemical sensor for detecting acetamiprid, its preparation method and application

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preparation example Construction

[0044] The present invention also provides a method for preparing the detection electrode, which includes the following steps: 1) The cleaned gold electrode is immersed in an ethanol solution of zinc dihydrogen sulfate, and the Au-S bond self-assembly is performed to form a surface of the gold electrode. The zinc dihydrogen sulfate self-assembled layer; 2) the zinc dihydrogen sulfate self-assembled layer obtained in step 1) is coated with an MES buffer containing NHS and EDC, and the zinc dihydrogen sulfate self-assembled layer is modified Activation, forming a dihydrolipoic acid activation layer on the surface of the gold electrode; 3) coating the acetamiprid capture antibody solution on the surface of the zinc dihydrogen sulfate activation layer for incubation, forming acetamiprid on the outer surface of the DHLA activation layer An amidine antibody layer; 4) The outer surface of the acetamiprid antibody layer is coated with MCH solution for sealing, and an MCH sealing layer i...

Embodiment 1

[0086] A preparation and verification of the sandwich electrochemical sensor of the present invention includes the following steps:

[0087] a. The gold electrode is first polished in 0.3 and 0.05μm alumina slurry until it has a mirror surface, and then rinsed with secondary water;

[0088] b. Soak the gold electrode in H 2 SO 4 (98%) and H 2 O 2 (30%) In the mixed solution (volume ratio) with a ratio of 3:1, after 20 minutes, it is washed several times with secondary water ultrasonically, and the mixture is washed with 0.25mM KCl and 5.0mM K 3 [Fe(CN) 6 ] 10mM PBS as the base solution, using EIS and CV technology to read K 3 [Fe(CN) 6 ] Signal changes;

[0089] c. Configure 1mM DHLA ethanol solution, soak the pretreated gold electrode in the DHLA ethanol solution for two hours, and DHLA self-assembles on the gold electrode surface through Au-S bond;

[0090] d. Configure 0.1M MES buffer (pH=6.5) containing 5mM NHS and 2.5mM EDC, and take 8uL to modify the carboxyl group on the surface...

Embodiment 2

[0097] A control experiment verification of the sandwich electrochemical sensor of the present invention includes the following steps:

[0098] a. Six gold electrodes (named a, b, c, d, e, f) are first polished in 0.3 and 0.05 μm alumina slurry until they are mirrored, and rinsed with water twice;

[0099] b. Soak the gold electrode in H 2 SO 4 (98%) and H 2 O 2 (30%) In a mixture with a ratio of 3:1 (volume ratio), after 20 minutes, it will be washed with secondary water for several times and then dried with nitrogen for use;

[0100] c. Configure 1mM DHLA ethanol solution, soak the pretreated gold electrode in the DHLA ethanol solution for two hours, and DHLA self-assembles on the gold electrode surface through Au-S bond;

[0101] d. Configure 0.1M MES buffer (pH=6.5) containing 5mM NHS and 2.5mM EDC, and take 8uL to modify the carboxyl group on the surface of the electrode to activate the DHLA surface;

[0102] d. Add 8μL 10μg mL -1 The capture antibody of Ⅱ was dropped on the surfac...

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Abstract

The invention provides an electrochemical sensor for detecting acetamiprid, and a preparation method thereof. The electrochemical sensor comprises a detection electrode and a matching reagent, whereinthe detection electrode comprises a gold electrode, a DHLA activation layer, an acetamiprid antibody layer and an MCH blocking layer, the exterior of the gold electrode is sequentially wrapped with the DHLA activation layer, the acetamiprid antibody layer and the MCH blocking layer, and the matching reagent comprises an acetamiprid hapten probe solution and a methylene blue probe solution. According to the present invention, the electrochemical sensor achieves the immunodetection of small molecules, the detection is rapid, the detection limit is 3.2 ng / L, the detection is sensitive, and the use method is simple and rapid, and it is suitable for the online detection of acetamiprid in food safety.

Description

Technical field [0001] The invention relates to the technical field of electrochemical sensors, in particular to an electrochemical sensor for detecting acetamiprid and a preparation method thereof. Background technique [0002] The new insecticide Acetamiprid belongs to the nitromethylene heterocyclic compound, which can act on the nicotinic acetylcholine receptors in the synapses of the insect nervous system, interfere with the stimulation conduction of the insect nervous system, cause the nervous system pathways to be blocked, and cause nerves The accumulation of the transmitter acetylcholine at the synaptic site causes the insect to paralyze and eventually die. Acetamiprid has replaced conventional pesticides such as organophosphorus, carbamate, etc., and is widely used worldwide. Acetamiprid is similar to many other pesticides, and residues inevitably exist in the application process. Food safety issues caused by pesticide residues have become the focus of public attention ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327G01N27/30
CPCG01N27/307G01N27/3276G01N27/3277
Inventor 徐霞红王新全王祥云齐沛沛汪志威徐浩王强
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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