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Preparation of pesticide-carbaryl molecular-imprinting bionic rapid-detection test strip and application thereof

A technology of molecular imprinting and detection test paper, applied in biological testing, analytical materials, measuring devices, etc., can solve the problems of difficult to guarantee antibody specificity, long preparation period of biological antibody, limited application, etc. The results are obvious and intuitive, and the preparation process is simple.

Inactive Publication Date: 2016-05-11
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main problem facing the current immunoassay method is the preparation of biological antibodies (especially biological antibodies of small molecular compounds). Molecular compounds, it is difficult to prepare biological antibodies with stable quality
In addition, biological antibodies are more sensitive to the external environment, easily lose their activity, and easily cause false positive results, which limits their application. Bionic Enzyme-linked Immunoassay (BELISA)

Method used

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  • Preparation of pesticide-carbaryl molecular-imprinting bionic rapid-detection test strip and application thereof
  • Preparation of pesticide-carbaryl molecular-imprinting bionic rapid-detection test strip and application thereof
  • Preparation of pesticide-carbaryl molecular-imprinting bionic rapid-detection test strip and application thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0033] figure 1 It is a flow chart for the preparation of carbaryl molecularly imprinted bionic test strips.

[0034] figure 2 It is a schematic diagram of detection of carbaryl molecularly imprinted bionic test strip.

[0035] image 3 Schematic diagram for optimizing the detection of enzyme-labeled antigens, from image 3 It can be seen that the color gradually becomes lighter with the increase of enzyme-labeled antigen dilution.

[0036] Figure 4 Schematic diagram of reaction time optimization, from Figure 4 It can be seen that the color gradually becomes darker with the increase of reaction time.

[0037] Figure 5 Schematic diagram of the results of molecularly imprinted bionic test strips for different concentrations of carbaryl (from A to C, the color of the test paper gradually becomes lighter as the concentration of carbaryl increases).

Embodiment 1

[0039] Preparation of biomimetic rapid test strips for molecularly imprinted pesticide carbaryl:

[0040] A. Pretreatment of solid phase carrier: WhantmanNO.5 filter paper is pre-dried in a vacuum drying oven to remove moisture, immersed in 80% ethanol containing 0.5-0.6% γ-methacrylic acid oxypropyltrimethoxysilane (γ-MAPS) solution, fixed in a drying oven and cut into circular paper pieces with a certain diameter.

[0041] B. Preparation of carbaryl molecularly imprinted bionic test strip: template molecule (carbaryl), functional monomer (MAA), cross-linking agent (EGDMA) with a molar ratio of 1:4:4, initiator (content of 2-2.5% of the total mass of functional monomers and cross-linking agents) dissolved in chloroform, added to pre-treated paper sheets, degassed, thermally polymerized in a water bath at 55-60°C for 10-20h, and obtained carbaryl molecularly imprinted bionic test strips.

[0042] C. Removal of template molecules: Soxhlet extraction of template molecules with...

Embodiment 2

[0050] Green vegetables were selected as test samples, and carbaryl molecularly imprinted bionic test strips were prepared to detect the effect in actual samples by adding different concentrations of carbaryl.

[0051] Sample addition and recovery experiment 2: Weigh three 0.5g vegetable blank samples, add 1mL methanol, add 0.25, 0.5, 1mgkg -1 After shaking, react with carbaryl molecularly imprinted bionic test strips respectively, and at the same time add enzyme-labeled antigen diluent with a volume ratio of 1:1 (enzyme-labeled antigen dilution is 1:100-1:32000), react 1-50min. After washing, add substrate for color development and read analysis.

[0052] Using the test strip of Example 2 is easy to operate, and the test results are intuitive and easy to distinguish. The color of the carbaryl molecularly imprinted bionic test strips with different concentrations of carbaryl changed significantly compared with the green vegetable blank sample without adding, and the color be...

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PUM

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Abstract

The invention belongs to the molecular-imprinting technology, and discloses preparation of a pesticide-carbaryl molecular-imprinting bionic rapid-detection test strip and an application thereof. Whatman NO.5 filter paper serves as a solid phase carrier, and silanization is carried out on the surface of the Whatman NO.5 filter paper to introduce double bonds; carbaryl, methacrylic acid, ethylene glycol dimethacrylate and azodiisobutyronitrile are dissolved into chloroform, the pretreated carrier is added, triggering is carried out in a water bath, and a molecular imprinting layer of the carbaryl is formed on the filter paper fiber carrier in a polymerization mode; the biological antibody and antigen and enzyme-labeled-antigen competitiveness immunoassay conjugation reaction principle is simulated, a carbaryl enzyme-labeled antigen serves as a reaction marker to compete with the carbaryl for binding with an imprinting hole locus of a carbaryl molecularly imprinted polymer, and after a substrate is added for displaying, the content of the carbaryl in a sample is qualitatively detected by judging changes in color in the paper. Compared with a biological antibody rapid-detection test strip, production cost is low, a detection method is simple, convenient and rapid, the detection result is obvious and visible, and the test strip is suitable for on-site rapid identification.

Description

technical field [0001] The invention belongs to the molecular imprinting technology, and in particular relates to the preparation of a bionic rapid detection test strip for the molecular imprinting of the pesticide carbaryl based on filter paper as a solid phase carrier and its application. Background technique [0002] Carbaryl, also known as carbaryl and carbaryl, is the world's first commercialized and widely used carbamate insecticide with high efficiency and low toxicity. It is widely used in the control of wheat, rice, cotton, vegetables and fruit trees. Pests of crops such as crops, ectoparasites of livestock and poultry, etc., mixed with organophosphorus pesticides have obvious synergistic effects on various agricultural pests. Carbaryl contains an N-methyl group, is a white crystal, hardly soluble in water, easily soluble in organic solvents such as acetone, dichloromethane, etc., and is easily hydrolyzed under alkaline and high temperature conditions. Carbaryl has...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/53G01N21/78
CPCG01N21/78G01N33/53G01N33/543G01N2430/10
Inventor 张灿崔涵雨韩玉凤蔡健荣
Owner JIANGSU UNIV
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