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ELISA (enzyme linked immunosorbent assay) kit for detecting carbofuran and application of ELISA kit

An enzyme-linked immunosorbent reagent and kit technology, which is applied in the direction of testing food, measuring devices, material inspection products, etc., can solve the problems of difficulty in meeting the needs of rapid detection of large numbers of samples and on-site samples, complex processing process, and time-consuming, etc. Simple structure, low pretreatment requirements, and convenient use

Active Publication Date: 2015-12-09
BEIJING KWINBON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The use of these analytical methods requires expensive instruments and specialized technicians. The sample pretreatment process is complex, costly, and time-consuming, and it is difficult to meet the needs of a large number of samples and rapid detection of on-site samples.

Method used

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  • ELISA (enzyme linked immunosorbent assay) kit for detecting carbofuran and application of ELISA kit
  • ELISA (enzyme linked immunosorbent assay) kit for detecting carbofuran and application of ELISA kit
  • ELISA (enzyme linked immunosorbent assay) kit for detecting carbofuran and application of ELISA kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The preparation of embodiment 1 kit components

[0029] 1. Synthesis of carbofuran hapten (see appendix for the synthetic route figure 1 ) and identification

[0030] A. Take 1.0g of furanol, add 30mL of dichloromethane to dissolve, add carbonyl chloride, react at 0-5°C for 4h, add 90mL of petroleum ether, let it stand still, there is oily substance at the bottom, transfer it to a separatory funnel, and separate the upper organic phase , The oil was dissolved in tetrahydrofuran, then 2 mL of tetrahydrofuran solution containing methylamine was added, and the reaction was carried out at 0-5°C for 2 h. The reaction was stopped, and the tetrahydrofuran was removed by rotary evaporation to obtain a red oil, which was applied to a silica gel column and eluted with petroleum ether / ethyl acetate (20 / 1, v / v) to obtain the product Ⅰ;

[0031] B. Take 1.2g of zinc powder, add distilled water and 0.5mL of glacial acetic acid, heat and activate at 80°C for 30min, add 30mL of ethan...

Embodiment 2

[0063] Embodiment 2 detects the formation of the ELISA kit of Carbofuran

[0064] An enzyme-linked immunosorbent assay kit for detecting carbofuran was constructed to include the following components:

[0065] (1) A microtiter plate coated with carbofuran-conjugated antigen;

[0066] (2) 6 bottles of carbofuran standard solution, the concentrations are 0μg / L, 0.5μg / L, 1.5μg / L, 4.5μg / L, 13.5μg / L, 40.5μg / L;

[0067] (3) Carbofuran monoclonal antibody working solution;

[0068] (4) goat anti-mouse anti-antibody labeled with horseradish peroxidase;

[0069] (5) Substrate chromogenic solution is made up of A liquid and B liquid, and A liquid is carbamide peroxide, and B liquid is tetramethylbenzidine;

[0070] (6) The stop solution is 2mol / L sulfuric acid;

[0071] (7) The washing solution has a pH value of 7.4, contains 0.5% to 1.0% Tween-20, 0.01‰ to 0.03‰ sodium azide preservative, and 0.1 to 0.3mol / L phosphate buffer, and the percentage is mass volume percentage;

[0072]...

Embodiment 3

[0073] Example 3 Detection of Carbofuran in Vegetables and Tea Samples

[0074] 1. Sample pretreatment

[0075] Vegetable samples:

[0076] Weigh 1.0g±0.05g of the homogenized vegetable sample into a 10mL polystyrene centrifuge tube, add 5mL of sample extractant (weigh 4.4g of disodium hydrogen phosphate dodecahydrate and 13.68g of sodium dihydrogen phosphate dihydrate into 500mL Dissolve and mix in deionized water), oscillate with an oscillator for 5min, mix well, centrifuge at 3000g room temperature (20-25℃) for 5min; pipette 200μL supernatant into a 2mL polystyrene centrifuge tube, add 600μL complex solution, and vortex Vortex for 2 min and mix thoroughly; take 50 μL for analysis.

[0077] Tea samples:

[0078] Weigh 1g of tea leaves into a 50mL polystyrene centrifuge tube, add 10mL of acetonitrile, vortex for 1min with a vortexer, mix well, centrifuge at 3000g at room temperature (20-25°C) for 5min; pipette 1mL of the upper organic phase to 10mL of clean and dry In the...

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Abstract

The invention provides an ELISA (enzyme linked immunosorbent assay) kit for detecting carbofuran. The ELISA kit comprises an ELISA plate coated with a carbofuran conjugated antigen, a carbofuran monoclonal antibody, an enzyme labeled antibody, a carbofuran standard solution, a substrate color development solution, a stop buffer, a cleaning solution and a re-dissolution solution. The invention further discloses a method for applying the ELISA kit to detection of the carbofuran. The method comprises the following steps: a sample is pretreated firstly, then the kit is used for detection, and a detection result is analyzed finally. The ELISA kit can be used for detecting the content of the carbofuran in vegetables and tea leaves, the operation is simple and convenient, the cost is low, the sensitivity is high, on-site supervision can be realized, and the kit is suitable for screening of a large quantity of samples.

Description

technical field [0001] The invention relates to an enzyme-linked immunosorbent detection technology, in particular to an enzyme-linked immunosorbent assay kit for detecting carbofuran, which is particularly suitable for detecting the content of carbofuran in vegetables and tea. Background technique [0002] Carbofuran, chemical name 2,3-dihydro-2,2-dimethyl-7-benzofuran-N-methylcarbamate, is a highly efficient broad-spectrum carbamate Systemic insecticide and nematicide are widely used in the control of cotton, rice, sugarcane, peanut, soybean, tobacco and other crop pests. Carbofuran is not easy to degrade in soil, and it is easy to cause surface and groundwater pollution through runoff and infiltration. Although my country's pesticide residue standards stipulate that carbofuran should not be detected in vegetables and fruits, the phenomenon of illegal use of carbofuran is still relatively common. Poisoning incidents caused by eating vegetables and other foods contaminated...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/02G01N33/577
Inventor 冯才伟崔海峰万宇平罗晓琴何方洋朱亮亮崔廷婷杨烁
Owner BEIJING KWINBON BIOTECH
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