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Enzyme-linked immunosorbent assay kit for dibutyl phthalate detection, and application thereof

A technology of dibutyl phthalate and enzyme-linked immunosorbent reagents, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of inability to operate on-site, high cost of detection, complicated pretreatment, etc., and achieve high accuracy , high accuracy and simple pre-processing

Active Publication Date: 2014-06-18
BEIJING KWINBON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method is sensitive, accurate, strong specificity, good separation, and can simultaneously determine a variety of drugs, but requires expensive instruments, complex sample pretreatment, tedious and time-consuming, high cost of detection, can not be operated on site, and requires professionals operation, so limits its application

Method used

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  • Enzyme-linked immunosorbent assay kit for dibutyl phthalate detection, and application thereof
  • Enzyme-linked immunosorbent assay kit for dibutyl phthalate detection, and application thereof
  • Enzyme-linked immunosorbent assay kit for dibutyl phthalate detection, and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Preparation of kit components

[0029] 1. Preparation of dibutyl phthalate hapten

[0030] Dissolve 1.48g of phthalic anhydride in 50ml of chloroform, add 3ml of 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) and 1.0ml of n-butyl while stirring. A mixture of alcohol in 10ml of chloroform, stirring at room temperature for 6h, after evaporating the solvent, purification by column chromatography to obtain monobutyl phthalate; dissolve 1.11g of monobutyl phthalate in 40ml of chloroform , Add 2.06g of dicyclohexylcarbodiimide (DCC), a catalytic amount of 4-dimethylaminopyridine (DMAP), slowly add 0.8g of 4-aminobutyric acid in a suspension of 20ml of chloroform under stirring, After adding, continue to react at room temperature for 8 hours, filter, wash with water, evaporate the solvent, and purify by column chromatography to obtain o-butoxyyl carboxybutyl benzamide, which is the dibutyl phthalate hapten.

[0031] Take the above-mentioned product and measure by hydrogen nucle...

Embodiment 2

[0050] Example 2 Establishment of an enzyme-linked immunoassay kit for detecting dibutyl phthalate

[0051] Establish an enzyme-linked immunoassay kit for detecting dibutyl phthalate, which contains the following components:

[0052] (1) Enzyme-labeled plate coated with dibutyl phthalate coupled antigen;

[0053] (2) 6 bottles of dibutyl phthalate standard solution with concentrations of 0mg / L, 0.02mg / L, 0.06mg / L, 0.18mg / L, 0.54mg / L, 1.62mg / L;

[0054] (3) Dibutyl phthalate monoclonal antibody working solution;

[0055] (4) Goat anti-mouse anti-antibody labeled with horseradish peroxidase;

[0056] (5) The substrate color-developing solution is composed of A solution and B solution, A solution is carbamide peroxide, and B solution is tetramethylbenzidine;

[0057] (6) The stop solution is 2mol / L sulfuric acid;

[0058] (7) The washing solution is pH 7.4, containing 0.5%~1.0% Tween-20, 0.01‰~0.03‰ sodium azide preservative, 0.1~0.3mol / L phosphate buffer, the percentage is Weight volume per...

Embodiment 3

[0060] Example 3 Detection of plasticizer in liquor samples

[0061] 1. Sample pretreatment

[0062] Pipette 100μl liquor sample into a 2ml polystyrene centrifuge tube, then add 400μl sample diluent, shake and mix thoroughly; take 50μl for analysis.

[0063] 2. Detect with the kit

[0064] Add 50μl / well of dibutyl phthalate standard solution or pre-treated sample solution to the microwells of the ELISA plate coated with dibutyl phthalate coupled antigen, and then add dibutyl phthalate 50μl / well of butyl monoclonal antibody working solution, gently shake and mix, cover with a cover membrane and place at 25℃ to avoid light for 30min; pour out the liquid in the well, add 250μl of washing solution to each well and wash it 4~5 times , Each time 10s interval, pat dry with absorbent paper; then add horseradish peroxidase labeled goat anti-mouse anti-antibody 100μl / well, gently shake and mix, cover the plate with a cover membrane and place at 25℃ to avoid light for reaction 30min, take out ...

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Abstract

The present invention provides an enzyme-linked immunosorbent assay kit for dibutyl phthalate detection. The enzyme-linked immunosorbent assay kit comprises: a coating antigen-coated enzyme label plate, specific dibutyl phthalate antibody, an enzyme marker, a dibutyl phthalate standard substance solution, a substrate coloration solution, a stop buffer, a cleaning solution and a sample diluent, wherein the coating antigen is a conjugate formed by conjugating dibutyl phthalate to antigen, and the enzyme marker is an enzyme-labeled anti-antibody. The invention further discloses a method for applying the enzyme-linked immunosorbent assay kit to detect dibutyl phthalate, wherein the method comprises: carrying out a sample pretreatment, adopting the kit to detect, and analyzing the detection result. The enzyme-linked immunosorbent assay kit can be provided for detect the dibutyl phthalate content in the white spirit, has characteristics of simple operation, low cost and high sensitivity, can be provided for on-site monitoring, and is suitable for screening large number of samples.

Description

Technical field [0001] The invention relates to an enzyme-linked immunoassay technology, in particular to an enzyme-linked immunoassay kit for detecting dibutyl phthalate, which is particularly suitable for detecting the content of dibutyl phthalate in liquor. Background technique [0002] Dibutyl phthalate (DBP) belongs to the category of phthalates and is currently the most used plasticizer (also known as plasticizer) in industrial production at home and abroad. It is widely used in food packaging, cosmetics, medical equipment, and Environmental water bodies. Such plasticizers are not food or food additives, but they enter food through environmental migration, food packaging migration and illegal addition, etc., and have toxic effects on human health. For example, they cause endocrine disorders, affect normal fertility, and accumulate in the body for a long time. Lead to deformities, cancers and mutations, and also have great damage to the cardiovascular, digestive and urinary...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/544
CPCG01N33/544G01N33/577G01N33/581
Inventor 何方洋万宇平冯才伟孙震杨昌松李勇崔廷婷杨学林
Owner BEIJING KWINBON BIOTECH
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