Aflatoxin biosynthesis pathway targeted metabolome analysis method

A kind of aflatoxin, biosynthesis technology, applied in the field of analysis and detection, to achieve the effect of accurate quantitative and qualitative analysis

Inactive Publication Date: 2019-06-11
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, there is still no analytical method that can effectively and stably quantify aflatoxin biosynthesis pathway compounds simultaneously in the world

Method used

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  • Aflatoxin biosynthesis pathway targeted metabolome analysis method
  • Aflatoxin biosynthesis pathway targeted metabolome analysis method
  • Aflatoxin biosynthesis pathway targeted metabolome analysis method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Aflatoxin Biosynthesis Pathway Targeted Metabolome Analysis Method

[0032] (1) Optimization of D-optimal mixture design extraction solvent:

[0033] The physical and chemical properties of the compounds in the aflatoxin biosynthetic pathway are different. The design of experiments (DOE) method was used to investigate the extraction efficiency of methanol, acetonitrile, ethyl acetate, and dichloromethane four kinds of water-miscible organic solvents to the extraction efficiency of aflatoxin biosynthesis pathway compounds. Ratio 1%) to improve the ionization efficiency of compounds, obtain optimized extraction reagents, and efficiently extract the entire pathway compounds.

[0034] The experimental method is: methanol (CH3OH), acetonitrile ACN, ethyl acetate EA, dichloromethane DCM, mixed solvents with different ratios of four solvents and formic acid are used to model and optimize the extraction efficiency of Aspergillus flavus biosynthetic pathway compounds,...

Embodiment 2

[0065] A targeted metabolome analysis method for aflatoxin biosynthesis pathway was applied to target control and anti-toxin production mechanism.

[0066] Non-decarboxyluckia-WT16 (preserved in the China Center for Type Culture Collection, referred to as CCTCC on June 13, 2017, with the preservation number CCTCC No.M 2017331, recorded in patent 201711228608X), is a biological control bacterium, Inhibits the production of aflatoxins. We used the developed method to reveal the antitoxigenic mechanisms involved.

[0067] Aflatoxin-producing bacteria AF73 (isolated and purified from a peanut sample by the Institute of Oil Plants, Chinese Academy of Agricultural Sciences, and identified as a strain of Aspergillus flavus after species identification) was grown on the culture supernatant of non-decarboxylated Luxobacillus-WT16 and yellow The biosynthesis and metabolism of aflatoxin in Aspergillus liquid culture medium prepared according to the ratio of 0%, 50%, and 100% can be foun...

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Abstract

The invention relates to an aflatoxin biosynthesis pathway targeted metabolome analysis method, comprising the steps of feeding a sample, performing liquid chromatography-mass spectrometry detection to obtain raw data comprising primary and secondary mass spectrometry information, and performing qualitative analysis referring to a mass spectrometry standard library to obtain qualitative results ofcompounds with standard spectrum; generating a multi-stage mass spectrometric fragmentation tree according to mass spectrum information of the compounds with standard spectrum, generating a multi-stage mass spectrometry tree according to other aflatoxin biosynthesis pathway compounds without standard spectrum in a primary mass spectrometry mass-to-charge ratio sample, and determining the qualityof the compounds without standard spectrum based on the mass spectrometric fragmentation rule according to the discipline of similar structures and similar mass spectrometric behaviors of adjacent compounds in a biosynthesis pathway, thereby completing the qualitative analysis of unknown compounds.

Description

technical field [0001] The invention relates to a method for analyzing aflatoxin biosynthetic pathway targeted metabolome, and belongs to the field of analysis and detection. Background technique [0002] Aspergillus flavus produces the notorious strong carcinogenic secondary metabolite aflatoxin. Consuming agricultural products contaminated with aflatoxin can cause adverse reactions such as liver cancer, acute toxicity, and birth deformities in children. All over the world, the occurrence of aflatoxin contamination in food or feed has a huge impact on human health, and at the same time, huge economic losses are caused by aflatoxin contamination every year. Since the turkey incident in the 1960s, researchers have extensively carried out research on aflatoxin analysis methods and risk assessment. Since the 1980s, structural genes and their regulatory genes in aflatoxin biosynthesis have also attracted extensive research. attention. So far, more than 20 genes, a 75kb gene cl...

Claims

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Application Information

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IPC IPC(8): G01N30/86
Inventor 李培武张奇谢华里王秀嫔张良晓姜俊张文
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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