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Method for detecting aflatoxin M1 in cheese

A kind of aflatoxin and cheese technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of high cost of affinity column, influence of immunoaffinity column efficiency, etc.

Pending Publication Date: 2019-12-17
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are problems such as the detection results are easily affected by the column efficiency of the immunoaffinity column, and the cost of the affinity column is too high

Method used

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  • Method for detecting aflatoxin M1 in cheese
  • Method for detecting aflatoxin M1 in cheese
  • Method for detecting aflatoxin M1 in cheese

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0025] Weigh 5g of cheese sample into a homogenizer cup, add 30mL of acetonitrile with a concentration of 1ng / mL, place in a homogenizer, homogenize for 5min, collect the liquid phase, centrifuge (8000r / min, 10min), collect the supernatant, Filter through filter paper. The filtrate was collected and transferred to a 250 mL separatory funnel. Add 30 mL of petroleum ether into the separatory funnel, shake for 2 minutes, and discard the petroleum ether layer after the layers are separated. Repeat the extraction with petroleum ether twice. Transfer the lower layer solution to a 100mL round bottom flask, concentrate under reduced pressure at 45°C to about 2mL, pour the concentrated solution into a 50mL volumetric flask and add water to the mark.

[0026] Rinse the HLB solid-phase extraction column with 5mL acetonitrile first, and then wash the column with 5mL water. Then put the extract liquid of AFM1 on the column for extraction. After the sample liquid is completely extracted,...

example 2

[0028] Weigh 5g of cheese sample into a homogenizer cup, add 30mL of acetonitrile with a concentration of 4ng / mL, place in a homogenizer, homogenize for 5min, collect the liquid phase, centrifuge (8000r / min, 10min), collect the supernatant, Filter through filter paper. The filtrate was collected and transferred to a 250 mL separatory funnel. Add 30 mL of petroleum ether into the separatory funnel, shake for 2 minutes, and discard the petroleum ether layer after the layers are separated. Repeat the extraction with petroleum ether twice. Transfer the lower layer solution to a 100mL round bottom flask, concentrate under reduced pressure at 45°C to about 2mL, pour the concentrated solution into a 50mL volumetric flask and add water to the mark.

[0029] Rinse the HLB solid-phase extraction column with 5mL acetonitrile first, and then wash the column with 5mL water. Then put the extract liquid of AFM1 on the column for extraction. After the sample liquid is completely extracted,...

example 3

[0031] Weigh 5g of cheese sample into a homogenizer cup, add 30mL of acetonitrile with a concentration of 1ng / mL, place in a homogenizer, homogenize for 5min, collect the liquid phase, centrifuge (8000r / min, 10min), collect the supernatant, Filter through filter paper. The filtrate was collected and transferred to a 250 mL separatory funnel. Add 30 mL of petroleum ether into the separatory funnel, shake for 2 minutes, and discard the petroleum ether layer after the layers are separated. Repeat the extraction with petroleum ether twice. Transfer the lower layer solution to a 100mL round bottom flask, concentrate under reduced pressure at 45°C to about 2mL, pour the concentrated solution into a 50mL volumetric flask and add water to the mark.

[0032] Rinse the HLB solid-phase extraction column with 5mL acetonitrile first, and then wash the column with 5mL water. Then put the extract liquid of AFM1 on the column for extraction. After all the sample liquid has been extracted, ...

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Abstract

The invention discloses a method for detecting aflatoxin M1 in cheese. Extraction, purification and enrichment methods for aflatoxin in cheese are studied, and the aflatoxin M1 in the cheese is detected by combining extraction via hydrophilic-lipophilic balance column with high performance liquid chromatography. The hydrophilic-lipophilic balance column is used to extract the aflatoxin M1. Compared with a aflatoxin M1 test method in the present standard, an OASIS HLB solid-phase extraction purification column has a better separation and purification effect for the aflatoxin M1; the aflatoxin M1 in the cheese can be rapidly detected within 8 min, the average recovery rate is high, the relative standard deviation (RSD) is 1.4%-5.7%, and the lowest detection limit is 0.037 microgram / kg. Compared with the analysis time (100 min) of a second method in the current national standard GB5413.37-2010, detection time is shortened by 40min. The detection method is characterized by short analysis time, simple and rapid operation, low detection cost and accurate detection.

Description

technical field [0001] The invention relates to the fields of biotechnology and food safety detection, in particular to a rapid detection method for aflatoxin M1 in cheese. Background technique [0002] Cheese is a food with high nutritional value made from fermented milk. In recent years, due to the improvement of people's living standards in our country, the per capita consumption of cheese is also increasing year by year. However, during the production and maturation of cheese, due to the control of raw materials and process parameters, biotoxins that are harmful to human health are likely to be produced. Among them, aflatoxin M1 (AFM1) is a relatively common biotoxin in cheese, and it is the second-order of Aspergillus flavus and Aspergillus parasiticus. Grade metabolites are more harmful to the human body. [0003] The production of aflatoxins is mainly due to the secretion of food and feed substrates containing these fungi during processing or preservation under suit...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06G01N2030/062
Inventor 李晓东张宇白杰刘璐郝欣悦
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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