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Kit for detecting vomitoxin in food

A technology for detection kits and vomitoxin, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of easy inactivation of marker enzymes, cumbersome operations, complex environmental interference factors, etc., and achieve high sensitivity and specificity

Inactive Publication Date: 2017-05-31
BEOSON JIANGSU FOOD SAFETY TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods have their own advantages and disadvantages, and cannot meet the needs of on-site rapid detection
The pretreatment process of physical and chemical methods is complicated, the degree of instrumentation is high, the operation is cumbersome, and the efficiency is low
The ELISA method has gone through multiple elution processes, the marker enzyme is easy to inactivate, the substrate is easy to decompose when exposed to light, and the environmental interference factors are complicated, etc., and can no longer meet the lower and lower limit standards.
Colloidal gold chromatography can meet the requirements of on-site rapidity, but only qualitative detection can not obtain quantitative results

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1: Preparation of specific components of the kit

[0024] 1. Deoxynivalenol hapten synthesis

[0025] A mixture of 1.0 g of vomitoxin and 0.08 g of ethylenediamine in 100 ml of methanol was reacted at room temperature for 5 to 6 hours, and the solvent was evaporated to obtain the vomitoxin hapten quantitatively.

[0026] 2. Preparation of enzyme-labeled antigen

[0027] Take 10~15mg vomitoxin hapten, dissolve in 1~1.5ml N,N-dimethylformamide (DMF); take 27~32mg dichloroethane (EDC) and N-hydroxysuccinimide (NHS ) was fully dissolved in 0.1-0.3ml of water, then added to the hapten dissolving solution, and stirred at room temperature for 24 hours to obtain the reaction solution A; weigh 30-50 mg of horseradish peroxidase (HRP) to fully dissolve it in In the pH value of 7.2, 3.8ml of phosphate buffer solution, the reaction solution A was slowly added dropwise to the HRP solution, and stirred at room temperature for 24h; dialyzed with 0.01mol / L phosphate buffer...

Embodiment 2

[0035] Embodiment 2: the formation of kit

[0036] Construct a test kit for the detection of vomitoxin in food so that it contains the following components:

[0037] Horseradish peroxidase-labeled deoxynivalenol hapten marker

[0038] Enzyme-labeled Antigen Diluent

[0039] Deoxynivalenol monoclonal antibody

[0040]Vomitoxin standard solution, the concentrations are: 0mg / L, 0.25mg / L, 0.50mg / L, 0.75mg / L, 1.0mg / L, 2.0mg / L, 4.0mg / L, 8.0mg / L, standard The diluent is a phosphate buffer solution containing 0.03wt.% Tween-20, pH 7.2, and 0.05mol / L.

[0041] Concentrated complex solution is 10 times concentrated complex solution, specifically containing 100-120g NaH per liter 2 PO 4 2H 2 Aqueous solution of O.

[0042] The concentrated washing solution is a 10-fold concentrated washing solution, specifically containing 0.5-0.6 wt.% Tween-20 by volume, pH 7.4-7.5, and 1.2-1.6 mol / L phosphate buffer.

Embodiment 3

[0043] Embodiment 3: Detection of the residual amount of vomitoxin in the sample

[0044] 1. Sample pretreatment method

[0045] (1) milk

[0046] Take 35 μL of fresh milk sample and add 950 μL of sample diluent (dilute the concentrated complex solution to 10 times the volume with deionized water), vortex and mix well, and take the solution for sample analysis.

[0047] (2) milk powder

[0048] Weigh 0.5g±0.05g milk powder sample, add 5ml sample diluent, vortex mix, take 200μL from it and add to 600μL sample diluent, vortex mix, take the solution for sample analysis.

[0049] 2. Detection with kit and analysis of results

[0050] Dilute the enzyme-labeled antigen and the enzyme-labeled antigen diluent at a volume ratio of 1:20 to obtain the enzyme-labeled antigen working solution; take 50-55 μL enzyme-labeled antigen, 50-55 μL sample extract and 50-55 μL vomitoxin monoclonal Add antibodies to the container one by one, react at room temperature for 22 minutes, discard the s...

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PUM

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Abstract

The invention discloses a kit for detecting vomitoxin in food. The kit is prepared from an enzyme-labelled antigen, a thinning liquid of the enzyme-labelled antigen, a vomitoxin monoclonal antibody, a vomitoxin series standard product solution, a chemical fluorescent primer solution A, a chemical fluorescent primer solution B, a concentration compound solution, and a concentration detergent. The kit has the advantages that the sensitivity and specificity are higher, and the detection sensitivity on the vomitoxin is 0.25mg / L.

Description

technical field [0001] The invention relates to the technical field of food detection, in particular to a detection kit for vomitoxin in food. Background technique [0002] Deoxynivalenol was first discovered in Japan in 1970 in a scab barley poisoning virus, which can cause food refusal and vomiting in animals. As a common mycotoxin, deoxynivalenol exists widely in nature and mainly contaminates wheat, corn and other food crops and their products. Due to the strong toxicity of vomitoxin, countries all over the world attach great importance to its monitoring and set strict limit standards: the US Food and Drug Administration (FDA) stipulates that the limit in wheat products is 1000μg / kg, and the European Union stipulates that grain flour The limit of vomitoxin in corn flour is 750μg / kg, and my country's national standard stipulates that the limit of vomitoxin in grain is 1000μg / kg. [0003] The existing detection methods of DON mainly include physical and chemical methods, ...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/535G01N21/76
CPCG01N33/577G01N21/76G01N33/535
Inventor 周朱晨张根义胡彬张进吴念绮周合
Owner BEOSON JIANGSU FOOD SAFETY TECH CO LTD
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