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A method for the simultaneous determination of four aflatoxins in tobacco and tobacco products based on liquid chromatography-tandem mass spectrometry

A technique of aflatoxin and tandem mass spectrometry, which is applied in the field of determination of tobacco and tobacco products, can solve the problems of simultaneous detection of multiple aflatoxins, high purity requirements of the components to be tested, complicated method operations, etc., and achieve high extraction Purification effect, increase the recovery rate of standard addition, and realize the effect of extraction and separation

Active Publication Date: 2016-08-24
CHINA TOBACCO HUNAN INDAL CORP +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

RIA is also prone to false positives
[0006] Gas chromatography and high performance liquid chromatography have the advantages of high accuracy, good reproducibility, low limit of quantification, and high efficiency, but they usually need to be derivatized before detection with a fluorescence detector, and derivatization reagents (such as three Fluoroacetic acid, etc.) are often highly toxic, and the method is complicated to operate, and the purity requirements of the components to be tested in the sample are high, so it is not suitable for the simultaneous detection of multiple aflatoxins

Method used

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  • A method for the simultaneous determination of four aflatoxins in tobacco and tobacco products based on liquid chromatography-tandem mass spectrometry
  • A method for the simultaneous determination of four aflatoxins in tobacco and tobacco products based on liquid chromatography-tandem mass spectrometry
  • A method for the simultaneous determination of four aflatoxins in tobacco and tobacco products based on liquid chromatography-tandem mass spectrometry

Examples

Experimental program
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Effect test

Embodiment 1

[0063] 1. Reagents and instruments

[0064] Aflatoxin B1, B2, G1 and G2 standard products were all purchased from Toronto Research Chemicals Inc. (Canada), with a purity of ≥99%; methanol and acetonitrile were chromatographically pure reagents, purchased from Dikma Company in the United States; the rest of the reagents used were analytically pure ; Experimental water is ultrapure water.

[0065] Myco6in1 TM Mycotoxin immunoaffinity purification column, Vicam, USA; Thermo TSQ Quantum Discovery Max liquid chromatography-tandem mass spectrometry, Thermo Electric Corporation, USA; Nova-Pak reversed-phase C18 HPLC column, 3.9mm×150mm, 4μm , Waters Company of the United States; Beckman Coulter Allegra64R desktop centrifuge, Beckman Coulter, Inc. of the United States; ultrapure water preparation instrument, Millipore Company of the United States.

[0066] 2. Sample pretreatment, extraction and purification of aflatoxin: collect finished cigarette samples that have been visually con...

Embodiment 2

[0069] 1. Sample pretreatment, extraction and purification of aflatoxin: as in Example 1, collect the blank tobacco sample of cigarette cut tobacco, and carry out sample pretreatment according to the aforementioned method steps; in the aflatoxin extraction step, first weigh 4g through pretreatment Place the blank tobacco powder sample in 85mL centrifuge tube with stopper, add 0.4g sodium chloride, accurately add 400μL mixed standard solution (wherein the concentration of aflatoxin B1, B2, G1 and G2 is 20ng / mL), and then Accurately add 40mL of 80% methanol aqueous solution, and then carry out the extraction and purification of aflatoxin according to the aforementioned method steps. After the purification solution was blown dry with nitrogen gas, 0.5mL of acetonitrile was accurately added to dissolve to constant volume, and passed through a 0.22μm organic filter membrane before testing.

[0070] 2. As mentioned above, establish a standard curve for quantitative analysis, and use...

Embodiment 3

[0075] 1. Sample pretreatment, extraction and purification of aflatoxin: 3 commercially available flue-cured tobacco cigarettes of a certain brand were purchased, the filter tip and cigarette paper were stripped off, and the shredded tobacco sample of this cigarette was subjected to sample pretreatment according to the method described in Example 1. And the extraction of aflatoxin, carry out the purification of aflatoxin extract by the method described in embodiment 2.

[0076] 2. As mentioned above, establish a standard curve for quantitative analysis, and use liquid chromatography-tandem mass spectrometry to detect the cigarette sample. The results showed that the contents of aflatoxin B1, B2, G1 and G2 in this cigarette sample were all 0.0 μg / kg.

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Abstract

The invention discloses a method for simultaneously measuring the contents of four aflatoxins in tobaccos and tobacco products by liquid chromatography-tandem mass spectrometry combination. The method comprises the steps of firstly, carrying out pretreatment such as drying and crushing on tobaccos or a cut tobacco sample, and then extracting the aflatoxins B1, B2, G1 and G2 in the tobaccos or the cut tobacco sample; purifying extract by adopting an immunoaffinity column; establishing a quantitative analysis standard curve, and detecting by using a liquid chromatography-tandem mass spectrometry combination method. After the method is used, the four aflatoxins B1, B2, G1 and G2 can be measured at the same time; furthermore, the method has the advantages of being high in sensitivity, low in limit of quantitation, high in recovery rate, good in repeatability, simple and convenient to operate, etc.

Description

technical field [0001] The invention relates to a method for simultaneously determining the contents of four aflatoxins in tobacco and tobacco products based on liquid chromatography-tandem mass spectrometry, and belongs to the technical field of tobacco and tobacco product determination. Background technique [0002] In the process of tobacco primary processing, storage and transportation, tobacco factory production and tobacco product sales, due to improper storage and storage, mildew may occur in tobacco or tobacco products, which not only causes major economic losses, but also endangers the safety of cigarette products sex. Therefore, the tobacco industry attaches great importance to the control of mildew in tobacco and tobacco products. [0003] The metabolites produced by tobacco mildew mainly include mycotoxins such as aflatoxins. There are more than 20 kinds of aflatoxins, among which aflatoxins B1, B2, G1 and G2 are the four most important. [0004] At present, m...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/88
Inventor 刘巍毛友安杜文牛承岗
Owner CHINA TOBACCO HUNAN INDAL CORP
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