Detecting method for aflatoxins

A technology of aflatoxin and detection method, which is applied in the field of detection of aflatoxin, can solve the problems of complex operation, dependence on import of ELISA kits, neglect of AFT supervision and detection, etc., and achieve the effect of simple operation

Inactive Publication Date: 2005-03-02
SHANGHAI UNIV
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  • Application Information

AI Technical Summary

Problems solved by technology

Although TLC is simple and economical, it has many steps and low sensitivity
HPLC has high sensitivity, but the sample processing is cumbersome, the operation is complicated, and the instrument is expensive
ELISA has poor repeatability and reagent life is short
In addition, these determination methods need to use highly toxic AFT as a standard substance during operation, and toxic and odorous organ

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] 1. Preparation and preparation of immunoaffinity column:

[0026] 1. Preparation of monoclonal antibodies. The preparation of monoclonal antibodies should be obtained through mouse immunization, cell fusion, hybridoma screening, obtaining hybridoma cell lines, and collecting monoclonal antibodies. The steps are:

[0027] (1). Mouse immunity: use aflatoxin B1 (abbreviated as AFB) 1 ) And the conjugate complex of bovine serum protein BSA as immunogen to immunize mice; 50ug / 50ul antigen AFB 1 -BSA and 50ul Freund's complete adjuvant emulsified and injected intraperitoneally into each 10-week-old BALB / C male mouse. One month later, the booster immunization was carried out. 50ug / 50ul antigen and the emulsified antigen of Freund's incomplete adjuvant were injected intraperitoneally. After another 2 months, the immunization was boosted, and the dose of 50ug / 50ul antigen was injected into the tail vein. After 4 days, the spleen cells of the mice were taken out for the next step of ...

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Abstract

The invention relates to a assaying method for the aflatoxin, specifically saying, it is a method of adopting immunity affinity column and fluorophotometer prepared by monoclonal antibody of aflatoxin to assayt the aflatoxin in the food, which belongs to biocytology and microbacteria toxic metabololic products assaying method technical field.The method is as following: first, preparing monoclonal antibody by the steps involving: immunizing mouse, cell fusion, screening crossing tumour and obtaining the cell line of the crossing tumourc, gathering monoclonal antibody, etc, then using the abovementioned monoclonal antibody to prepare the immunoaffinity column, last, after milling the food and feed, extracting the aflatoxin out of it with methanol/water solution, after filtering, letting the liquid sample flow through the affinity column to be refined, successively, eluting the AFT from the affinity column with methanol, and assaying the content of aflatoxin in the sample with a fluorophotometer.

Description

Technical field [0001] The present invention relates to a method for detecting aflatoxin, more specifically, it is a method for detecting aflatoxin in food and feed by using an immunoaffinity column prepared by monoclonal antibody of aflatoxin and a fluorometer. It belongs to biological cells The field of science and technology for the detection of toxic metabolites of microorganisms. Background technique [0002] Aflatoxin is a group of secondary metabolic toxic products with similar chemical structures of common molds, Aspergillus flarus and A. parasiticus. AFB has been isolated and identified 1 , AFB 2 , AFG 1 , AFG 2 , AFM 1 Wait 12 kinds. Aflatoxins are often found in various nuts such as peanuts and walnuts. They are also often found in soybeans, corn, rice, milk, dairy products, edible oils and grain and oil products, especially in humid tropical and subtropical areas where AFT pollution More serious. [0003] Aflatoxin (AFT) has a strong damaging effect on human and anima...

Claims

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Application Information

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IPC IPC(8): G01N30/88G01N33/52G01N33/53G01N33/532G01N33/558G01N33/577
Inventor 黎双华陈宇光
Owner SHANGHAI UNIV
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