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Method for manufacturing immunosensor for detecting chlorpyrifos

An immunosensor, chlorpyrifos technology, applied in the field of agricultural product safety detection, can solve the problems of difficulty in meeting low residue and quantitative detection, high proportion of missed detection, affecting accuracy, etc., achieve high stability and high order, and improve electrical conductivity. and capacitance, the effect of increasing detection accuracy

Inactive Publication Date: 2014-07-23
SHANDONG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the quick test card is to observe the color change of the card with naked eyes, so it can only be used for qualitative measurement of vegetable samples that seriously exceed the standard
The principle of enzyme inhibition spectrophotometry is based on the change of absorbance for detection, but a large number of pigments in vegetables and fruits will affect the accuracy of the detection results
In addition, the above methods have disadvantages such as low recovery rate, high proportion of false detection and missed detection, poor repeatability, and difficulty in meeting the requirements of low residue and quantitative detection.

Method used

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  • Method for manufacturing immunosensor for detecting chlorpyrifos
  • Method for manufacturing immunosensor for detecting chlorpyrifos
  • Method for manufacturing immunosensor for detecting chlorpyrifos

Examples

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Embodiment 1

[0043] Example 1 Preparation of nickel-aluminum hydrotalcite-graphene nanocomposite solution

[0044] The Hummers method is used to prepare graphite oxide. Specific process flow: Assemble a 250mL reaction flask in an ice water bath, add 1g graphite powder, 1g sodium nitrate and 46mL sulfuric acid, stir for 30min, then add 5g potassium permanganate in portions, control the reaction temperature not to exceed 20℃, stir After reacting for a period of time, the temperature is raised to about 35°C, stirring is continued for 30 minutes, and then 80 mL of deionized water is slowly added. After stirring for 20 minutes, 6 mL of 30% hydrogen peroxide is added to reduce the remaining oxidant and the solution becomes bright yellow. Filter while hot and wash with 5% HCl solution and deionized water until no sulfate is detected in the filtrate. Finally, the filter cake is fully dried in a vacuum drying oven at 60°C to obtain graphite oxide. Add 30mg of graphite oxide to 50mL of a mixed alkali...

Embodiment 2

[0045] Example 2 Preparation of empty-shell gold nanoparticles

[0046] Co nanoparticles are used as sacrificial templates to synthesize hollow-shell gold nanoparticles. First pass high purity N into 50mL water 2 Deoxygenate, add 8.4mg citric acid and 15mg sodium borohydride under stirring. Then inject 50μL 0.4M CoCI 2 Solution (The reaction is always carried out under the protection of nitrogen to prevent the generated Co nanoparticles from being oxidized). Stop N after 20min 2， Take 30 mL of cobalt nano colloidal solution and add it to 18 mL of chloroauric acid solution (1 mM), and continue to stir for 30 min. Finally, the product was centrifuged, the supernatant was discarded, and the resulting precipitate was dispersed in 5 mL of pH 7.5 PBS for later use. Take 2mL of the empty-shell nano-gold colloid solution and add it to 5mL of 0.5% chitosan solution, and stir for 2h at room temperature to use as electrode modification material. figure 2 In order to prepare the TEM image...

Embodiment 3

[0047] Example 3 Cleaning, activation and testing of glassy carbon electrode

[0048] Before modification of the glassy carbon electrode, first immerse it in a hot "piranha" solution (H 2 SO 4 : 30% H 2 O 2 = 3:1) Soak in the water for 15 minutes, rinse with water, and then use 0.3μm, 30nm Al 2 O 3 The slurry is polished on the suede to the mirror surface, after polishing, it is cleaned with deionized water to remove the surface dirt, and then 6mol / L HNO 3 , Anhydrous ethanol and deionized water are each ultrasonically cleaned for 5 minutes, and dried in a nitrogen environment.

[0049] Activation of glassy carbon electrode: After thorough cleaning, the electrode is at 0.5mol / L H 2 SO 4 The solution is activated by cyclic voltammetry, and the scanning range is 1.0V~-1.0V, and the scanning is repeated until a stable cyclic voltammogram is reached.

[0050] Test of pre-processed glassy carbon electrode: when containing 1×10 -3 mol / L K 3 Fe(CN) 6 0.20 mol / L KNO 3 Run a cyclic voltammet...

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Abstract

The invention discloses a method for manufacturing an immunosensor for detecting chlorpyrifos, which belongs to the technical field of biologic sensor manufacturing. The method comprises the following steps: dropwise coating the surface of a naked glassy carbon electrode, which is subjected to cleaning, activation and performance testing, with a nickel aluminum hydrotalcite-graphene compound liquid, subsequently modifying by hollow nanogold, further fixing a chlorpyrifos monoclonal antibody on the modified electrode, and finally, sealing by bovine serum albumin. The immunosensor manufactured by the method has the characteristics of high sensitivity, low cost, high detection speed, good stability and high sample recycling rate. The detection limit of the immunosensor manufactured by the method can be 0.052ng / L, the linear range of the immunosensor is 5ng / mL-2 micrograms / mL, the recycling rate of the immunosensor can be 82.0-118.0%, and therefore the immunosensor is applicable to rapid detection on the residual amount of chlorpyrifos in fruits and vegetables.

Description

Technical field [0001] The invention relates to a preparation method of an immunosensor for detecting chlorpyrifos, and belongs to the technical field of agricultural product safety detection. technical background [0002] my country is a large agricultural country, and agricultural development plays a pivotal role in the national economy and people's lives. Pesticides are indispensable substances used in agricultural production to prevent and control diseases, insects and weeds on crops, and play an important role in promoting agricultural production. However, the unreasonable use of pesticides has not only caused pollution of the water environment, soil, and atmosphere, but also caused excessive pesticide residues in agricultural products, causing human poisoning and trade barriers. In addition, my country’s annual economic losses caused by excessive pesticide residues are huge. With the continuous improvement of people's living standards in my country, the quality and safety...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/531
CPCG01N33/54366G01N33/5438G01N33/54393G01N2430/10G01N2430/20
Inventor 郭业民乔璐孙霞王相友刘君峰
Owner SHANDONG UNIV OF TECH
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