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Application of fenpropathrin artificial antigen in enzyme linked immunosorbent assay kit

An enzyme-linked immunosorbent reagent and a technology of fenothrin, which can be used in testing organic pollutants in water, testing water, biological testing, etc., and can solve problems such as harming ecological environment safety and agricultural product quality.

Active Publication Date: 2020-10-23
BEIJING KWINBON BIOTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The long-term large-scale production and use of fenpropathrin has seriously endangered the safety of the ecological environment and the quality of agricultural products

Method used

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  • Application of fenpropathrin artificial antigen in enzyme linked immunosorbent assay kit
  • Application of fenpropathrin artificial antigen in enzyme linked immunosorbent assay kit
  • Application of fenpropathrin artificial antigen in enzyme linked immunosorbent assay kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] The preparation of embodiment 1 kit components

[0023] 1. Preparation of fenpropathrin hapten

[0024] Take 3.61g of vinylfenpropathrin, add 30ml of acetonitrile to dissolve, add 1ml of 1mol / L HCl, stir well, add 3ml of aqueous solution containing 1.58g of potassium permanganate, react at 50°C for 3h, stop the reaction, add 200ml of water, add ethyl acetate The ester was extracted three times with 100ml×3, the organic phases were combined, dried over anhydrous sodium sulfate, and evaporated to dryness to obtain a red oil, which was applied to a silica gel column, eluted with petroleum ether / ethyl acetate (v / v, 5 / 1), and separated to obtain The hapten product of carboxyfenpropathrin was 3.4 g, and the yield was 89.7%.

[0025] 2. Antigen preparation

[0026] Immunogen preparation—Fenpropathrin hapten was coupled with bovine serum albumin (BSA) to obtain immunogen.

[0027] Take 14mg of carboxyfenpropathrin hapten product, add 1ml of DMF to dissolve, add 12mg of HOBT,...

Embodiment 2

[0040] Embodiment 2 detects the formation of the ELISA kit of fenpropathrin

[0041] The ELISA kit for detecting fenpropathrin was set up to include the following components:

[0042] (1) A microtiter plate coated with fenpropathrin-conjugated antigen;

[0043] (2) 6 bottles of fenpropathrin standard solution, the concentrations were 0 μg / L, 1 μg / L, 3 μg / L, 9 μg / L, 27 μg / L, 81 μg / L;

[0044] (3) fenpropathrin antibody labeled with horseradish peroxidase;

[0045](4) Substrate chromogenic solution is made up of A liquid and B liquid, and A liquid is carbamide peroxide, and B liquid is tetramethylbenzidine;

[0046] (5) The stop solution is 2mol / L sulfuric acid;

[0047] (6) 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 weight volume percentage;

Embodiment 3

[0048] The detection of fenpropathrin in embodiment 3 vegetables and fruits

[0049] 1. Detection with kit

[0050] Number the corresponding microwells of the samples and standards in sequence, make 2 parallel wells for each sample and standard, and record the positions of the standard wells and sample wells. Add 50 μl of standard / sample to corresponding microwells, then add 50 μl / well of enzyme conjugate working solution, and react for 30 minutes at 25°C in a dark environment. Shake the liquid in the hole dry, wash 4-5 times, and pat dry. Add 50 μl / well of substrate solution A, then add 50 μl / well of substrate solution B, mix well, and react for 15 minutes at 25°C in a dark environment. Add 50 μl / well of stop solution, mix well, set the microplate reader at 450 nm, and measure the OD value of each well.

[0051] 2. Analysis of test results

[0052] The percent absorbance of the standard or sample is equal to the average of the absorbance values ​​of the standard or sample...

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Abstract

The invention provides an enzyme linked immunosorbent assay kit for detecting fenpropathrin. The enzyme linked immunosorbent assay kit comprises an ELISA plate coated with a coating antigen, a fenpropathrin standard substance solution, a fenpropathrin antibody, an enzyme conjugate concentrated solution, an enzyme conjugate diluent, a substrate developing solution, a terminating solution and a washing solution, wherein the coating antigen is a fenpropathrin coupled antigen, the enzyme conjugate is an enzyme-labeled fenpropathrin antibody, and the fenpropathrin antibody is obtained by immunizinganimals with immunogen. The invention further discloses a method for detecting fenpropathrin by applying the enzyme linked immunosorbent assay kit. The method comprises the following steps of: pretreating a sample, detecting the sample by using the kit, and finally analyzing a detection result. The enzyme linked immunosorbent assay kit provided by the invention can be used for detecting the content of fenpropathrin in vegetable and fruit samples, is simple and convenient to operate, low in cost and high in sensitivity, can be monitored on site, and is suitable for screening a large number ofsamples.

Description

technical field [0001] The present invention relates to the application technology of an artificial antigen of fenpropathrin in an ELISA kit, in particular to a molecular structure of an artificial antigen of fenpropathrin and an ELISA kit for detecting fenpropathrin. Qualitative and quantitative detection of fenpropathrin drug residues in vegetables and fruits. Background technique [0002] Pesticides play an extremely important role in modern agricultural production. Pyrethroid pesticides are widely used all over the world due to their stable properties, wide insecticidal spectrum, high insecticidal activity, and short residual period. They are also widely used in China. The fastest growing pesticide, of which fenpropathrin is the most widely used one. The long-term large-scale production and use of fenpropathrin has seriously endangered the safety of the ecological environment and the quality of agricultural products. [0003] The standard analytical methods commonly us...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/58
CPCG01N33/5308G01N33/543G01N33/581G01N33/184
Inventor 万宇平王兆芹贾芳芳崔海峰屈秀玲张洪亮
Owner BEIJING KWINBON BIOTECH
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