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Detection gel column of aflatoxin M1 and detection method of aflatoxin M1

A technology for the detection of aflatoxin and aflatoxin, which is applied in the field of detection gel column for aflatoxin M1, which can solve the problems of complex pretreatment process, unsuitable for rapid on-site detection, and large manual errors.

Inactive Publication Date: 2018-02-13
SHENZHEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, instrumental methods such as chromatography-mass spectrometry have sufficient sensitivity for the analysis of most chemical pollutant residues, but the detection not only requires expensive analytical instruments, but also requires the operation level of technicians and a large number of pre-treatment reagents
Moreover, before the detection process, it is generally necessary to perform complex derivatization processing on the analytes. The pre-treatment process is complex, time-consuming, and inefficient, and is not suitable for screening and detection of a large number of samples.
High-performance liquid chromatography and liquid chromatography-tandem mass spectrometry are not suitable for rapid detection of a large number of samples on site due to expensive equipment and cumbersome processes
In comparison, the enzyme-linked immunoassay detection method has higher sensitivity, but the traditional enzyme-linked immunosorbent assay (ELISA method) is more complicated to operate, with relatively large manual errors, and is not suitable for on-site rapid detection
[0006] In summary, the traditional aflatoxin M 1 The detection methods are not suitable for the rapid detection of a large number of copies on site

Method used

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  • Detection gel column of aflatoxin M1 and detection method of aflatoxin M1
  • Detection gel column of aflatoxin M1 and detection method of aflatoxin M1
  • Detection gel column of aflatoxin M1 and detection method of aflatoxin M1

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0092] Preparation of aflatoxin M 1 detection gel column

[0093] 1. Preparation of detection gel

[0094] (1) Activation of gel carrier: Take 1.0 g of cyanogen bromide (CN-Br) activated agarose gel (Sepharose4B) after swelling with 1 mmol / L HCl, and then fully wash with 200 mL of HCl solution. The whole process is completed within 15 minutes. Finish. The swollen gel was re-used with 0.1mol / L NaHCO 3 The solution is fully equilibrated. Typically 1.0 g of cyanogen bromide activated Sepharose 4B can swell about 3.5 mL of gel.

[0095] (2) Coupling: Dissolve 0.3 mL of primary polyclonal secondary antibody (goat anti-mouse antibody, secondary antibody, 2.3 g / L) in 5 mL of gel coupling solution, mix with the above activated gel carrier, and stir for 20 h .

[0096] (3) Washing: use 50 mL of gel coupling solution to wash to remove the unbound first polyclonal secondary antibody (goat anti-mouse antibody).

[0097] (4) Blocking: 20 mL of gel blocking solution was added to the ...

Embodiment 2

[0115] Preparation of HRP-labeled aflatoxin M 1 Enzyme-labeled antigen (AFM) 1 -HRP)

[0116] 2 mg of aflatoxin M 1 and 4 mg of aminooxyacetic acid hemi-hydrochloride (CMO) were dissolved in 2 mL of pyridine mixed solution (pyridine: methanol: double distilled water in the mixed solution = 1:4:1), and the temperature was controlled (120° C.) to reflux. Use thin-layer chromatography to monitor the reaction progress. When the reaction is complete, use a rotary evaporator to spin dry the solvent, add 2 mL of ddw (ultra pure water) to dissolve, and use 2 mol / L of H 2 SO 4 Adjust pH=4, and then extract with 5 mL of ethyl acetate for 2 to 3 times, and combine the organic phases. And wash 2 times with ddw (ultrapure water), spin dry the liquid again, the yellow oily substance obtained is the aflatoxin M substituted by aminooxyacetic acid hemihydrochloric acid 1 (AFM 1 -CMO).

[0117] Dissolve AFM with 0.4 mL of N,N-dimethylformamide (DMF) 1 -CMO, take 0.2mL of dissolved AFM ...

Embodiment 3

[0120] Aflatoxin M prepared in Example 1 1 The detection gel column (visualization gel ELISA detection gel column) detects the sample to be tested

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Abstract

The invention relates to a detection gel column of aflatoxin M1 and a detection method of the aflatoxin M1. The detection gel column of the aflatoxin M1 comprises a hollow column as well as a detection layer and a quality control layer that are filled into the hollow column. When in detection, a to-be-detected sample and an aflatoxin M1 enzyme-labelled antigen marked by a chemiluminiscent label are added into the detection gel column of the aflatoxin M1 and flow through the detection layer and the quality control layer. Both the to-be-detected sample and the aflatoxin M1 enzyme-labelled antigen marked by the chemiluminiscent label contend an aflatoxin M1 monoclonal antibody, and whether the to-be-detected sample contains the aflatoxin M1 is judged according to the color of the detection layer and the color of the quality control layer. The detection gel column of the aflatoxin M1 is carried conveniently, can be used for detecting whether the to-be-detected sample contains the aflatoxinM1 rapidly and sensitively and is suitable for on-site rapid detection of a large number of samples.

Description

technical field [0001] The invention relates to the technical field of detection, in particular to a kind of aflatoxin M 1 The detection gel column and aflatoxin M 1 detection method. Background technique [0002] Aflatoxins (AFT) are a class of highly mutagenic, highly teratogenic and highly carcinogenic compounds, which are produced by Aspergillus flavus and A. parasiticus at a certain temperature and relatively produced under high humidity. There are many kinds of aflatoxins, such as aflatoxin B 1 , Aflatoxin B 2 , Aflatoxin G 1 , Aflatoxin G 2 , Aflatoxin M 1 Wait. Among them, aflatoxin B 1 The high carcinogenicity and high mutagenicity of caffeine stem from the direct consumption of food contaminated with it. Aflatoxin M 1 (AFM 1 ) is the body ingested by aflatoxin B 1 After contaminated food, it is metabolized by liver microsomal enzymes and is mainly excreted in milk. Aflatoxin M 1 second only to aflatoxin B in toxicity 1 , and aflatoxin M 1 The natur...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N33/532
CPCG01N33/5302G01N33/532
Inventor 蒋文晓陈献雄柯跃斌翟鹏庄俊钰周芳梅吕子全
Owner SHENZHEN UNIV
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