Method for simultaneously detecting multiple fungal toxins in wheat

A mycotoxin and wheat technology, which is applied in the field of simultaneous detection of multiple mycotoxins in wheat, can solve the problems of great harm to human body, complicated treatment, high content, etc., and achieve the effect of accurate and reliable content, simplified operation process and good repeatability

Active Publication Date: 2018-01-26
NANJING UNIV OF FINANCE & ECONOMICS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

AFB1, ZEN, and DON are high in content in wheat and are very harmful to the human body. The separate determination of each toxin is not only complicated to deal with, but also takes a long time. There is no simultaneous detection of these three mycotoxins in the current national standards. method

Method used

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  • Method for simultaneously detecting multiple fungal toxins in wheat
  • Method for simultaneously detecting multiple fungal toxins in wheat
  • Method for simultaneously detecting multiple fungal toxins in wheat

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Take 2.5±0.01g of wheat samples, pulverize them with a pulverizer and pass through a 2mm round hole sieve, mix them evenly, put them into a clean container, add 12.5ml of 80% acetonitrile aqueous solution, vortex and mix for 3 times during ultrasonic extraction for 15min, and centrifuge at 5000rpm for 5min ; Take 5ml supernatant and add 35ml phosphate buffer solution, pH = 7.1, filter with glass fiber filter paper, collect the filtrate in a clean container, and use a composite immunoaffinity column to purify. The eluate was blown with nitrogen at 50°C, redissolved in 1ml of 50% methanol solution, and passed through a 0.22µm filter membrane for determination by high performance liquid chromatography, such as figure 1 shown.

[0026] Mobile phase: phase A ultrapure water; phase B methanol (chromatographic grade), flow rate: 1ml / min, column temperature: 35°C, injection volume: 20µl. Mobile phase program: 0-6min, 80%A+20%B; 7min, 50%A+50%B; 17min, 20%A+80%B; 18min 80%A+20%...

Embodiment 2

[0029] Take 2.5±0.01g of wheat sample, pulverize it with a pulverizer and pass it through a 2mm round hole sieve, mix well, put it into a clean container, add 7.5ml of 60% acetonitrile-water solution, vortex and mix three times during ultrasonic extraction for 30min, and centrifuge at 5000rpm 5min; take 5ml of supernatant and add 35ml of phosphate buffer solution, pH=6.9, filter with glass fiber filter paper, collect the filtrate in a clean container, and use a composite immunoaffinity column to purify. The eluate was blown with nitrogen at 50°C, redissolved in 3ml of 40% methanol solution, and passed through a 0.22µm filter membrane for determination by high performance liquid chromatography.

[0030] Mobile phase: phase A ultrapure water; phase B methanol, flow rate: 0.5ml / min, column temperature: 25°C, injection volume: 10µl. Mobile phase program: 0-6min, 60%A+40%B; 7min, 30%A+70%B; 17min, 10%A+90%B; 18min 60%A+40%B; 23min, 50%A +50%B.

Embodiment 3

[0032] Take 2.5±0.01g of wheat samples, pulverize them with a pulverizer and pass through a 2mm round hole sieve, mix them evenly, put them into a clean container, add 17.5ml of 90% acetonitrile-water solution, vortex and mix for 3 times during ultrasonic extraction for 10min, and centrifuge at 5000rpm 5min; take 5ml supernatant and add 35ml phosphate buffer solution, pH=7.3, filter with glass fiber filter paper, collect the filtrate in a clean container, and use composite immunoaffinity column to purify. The eluate was blown with nitrogen at 50°C, redissolved in 2ml of 60% methanol solution, and passed through a 0.22µm filter membrane for determination by high performance liquid chromatography.

[0033] Mobile phase: phase A ultrapure water; phase B methanol (chromatographic grade), flow rate: 1ml / min, column temperature: 40°C, injection volume: 50µl. Mobile phase program: 0-6min, 90%A+10%B; 7min, 60%A+40%B; 17min, 30%A+70%B; 18min 90%A+10%B; 23min, 20%A +80%B.

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Abstract

The invention discloses a method for simultaneously detecting multiple fungal toxins in wheat, and concretely relates to a method for detecting vomitoxin (DON), aflatoxin B1 (AFB1) and zearalenone (ZEN) in wheat through immunoaffinity purification-high performance liquid chromatography. The method comprises the following steps: extracting a sample by using an acetonitrile-water solution having a certain ratio, purifying the obtained sample solution by a composite immunoaffinity column, simultaneously detecting the three fungal toxins through adopting the high performance liquid chromatography,preparing a standard working solution to qualitatively and quantitatively detect the three fungal toxins. Compared with the prior art, the method adopting the immunoaffinity chromatography to greatlyimprove the purifying degree of the sample and adopting the high performance liquid chromatography to realize simultaneous detection of multiple toxins has the advantages of operating time saving, high sensitivity, accuracy in quantification and good repeatability.

Description

technical field [0001] The invention relates to a method for simultaneously detecting multiple mycotoxins in wheat, belonging to the technical field of instrument detection. Background technique [0002] Mycotoxins are toxic metabolites produced by microorganisms during grain production, harvesting, and storage. The most polluted ones are aflatoxins and fusarium toxins, while aflatoxins (AFB1), zearalenone (zearalenone) , ZEN) and deoxynivalenol (DON) are serious and frequent toxins in wheat mycotoxin contamination. Aflatoxins are mainly divided into AFB1, AFB2, AFG1, AFG2, AFM1, AFM2, etc., which mainly pollute corn, peanuts, wheat, etc., among which AFB1 is extremely toxic, causing the greatest damage to human or animal liver and strong carcinogenicity . my country's General Administration of Quality Supervision, Inspection and Quarantine has stipulated that AFB1 is one of the mandatory inspection items for most foods. Zearalenone is mainly produced by Fusarium graminea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/34G01N30/74
Inventor 方勇张盈月裴斐李彭沈飞胡秋辉
Owner NANJING UNIV OF FINANCE & ECONOMICS
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