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Method for detecting alkaloids and nitrosamines in tobaccos simultaneously

A detection method and alkaloid technology, applied in measuring devices, instruments, scientific instruments, etc., can solve problems such as unsatisfactory detection results of Mesmin, unsystematic research on the composition and content of alkaloids and nitrosamines, etc. Sample preparation time, simplified pre-treatment process, quantitative and accurate effect

Active Publication Date: 2013-08-21
YUNNAN ACAD OF TOBACCO AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But there are still many places to be improved: as can not ensure that each component of tobacco alkaloids can be extracted completely and the detection effect of mesmin with extremely low content is not ideal, and in the quantification of tobacco alkaloids, the solution preparation of standard curve needs Do it separately to reduce interference etc.
As far as the current research status is concerned, the research on the composition and content of alkaloids and nitrosamines in tobacco in my country is still not systematic, and further optimization of the relevant technical parameters of extraction and separation technology and determination methods is needed. More importantly, at present, my country There is no report on a method capable of simultaneously detecting alkaloids and nitrosamines in tobacco

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  • Method for detecting alkaloids and nitrosamines in tobaccos simultaneously
  • Method for detecting alkaloids and nitrosamines in tobaccos simultaneously
  • Method for detecting alkaloids and nitrosamines in tobaccos simultaneously

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] ——Determination of alkaloid and nitrosamine content in flue-cured tobacco leaf samples and determination of conversion ratio of alkaloid to nitrosamine

[0051] Experimental materials: 10 groups of prepared flue-cured tobacco leaf samples.

[0052] experimental method:

[0053] Accurately weigh 100 mg of prepared flue-cured tobacco leaf powder passed through a 40-mesh sieve, transfer it into a 50 ml glass conical flask, add 1 ml of 1 μg / ml internal standard solution, and the internal standard solution contains D4-NNN, D4-NAT, D4-NAB and D4-NNK, then add 9ml to ultrapure water. Then, ultrasonically extract for 20min under the condition of 20Hz, pass the extract through a 0.22μm filter membrane, pipette 500μl into a 1.5ml centrifuge tube, add 1ml of methanol, vortex shake for 30s, and centrifuge at 9000r / min for 4min Finally, take 1ml of the supernatant and transfer it into a 1.5ml injection vial for liquid chromatography-tandem mass spectrometry analysis.

[0054] Th...

Embodiment 2

[0063] ——Determination of alkaloid and nitrosamine content in Burley tobacco leaf samples and determination of conversion ratio of alkaloid to nitrosamine

[0064] Experimental materials: 10 groups of Burley tobacco leaf samples.

[0065] experimental method:

[0066] Accurately weigh 90 mg of prepared flue-cured tobacco leaf powder passed through a 50-mesh sieve, transfer it into a 50 ml glass conical flask, add 0.9 ml of 1 μg / ml internal standard solution, and the internal standard solution contains D4-NNN, D4-NAT, and D4-NAB respectively and D4-NNK, then add 9.1ml to ultrapure water. Then, ultrasonically extract at 20Hz for 18min, pass the extract through a 0.22μm filter membrane, pipette 450μl into a 1.5ml centrifuge tube, add 0.9ml methanol, vortex shake for 25s, and centrifuge at a speed of 8000r / min After 5 minutes, take 1ml of the supernatant and transfer it into a 1.5ml injection vial for liquid chromatography-tandem mass spectrometry analysis.

[0067] The analys...

Embodiment 3

[0072] ——Determination of alkaloid and nitrosamine content in oriental tobacco leaf samples and determination of conversion ratio of alkaloid to nitrosamine

[0073] Experimental materials: 10 groups of oriental tobacco leaf samples.

[0074] experimental method:

[0075] Accurately weigh 110 mg of prepared flue-cured tobacco leaf powder that has passed through a 60-mesh sieve, transfer it into a 50 ml glass conical flask, add 1.1 ml of 1 μg / ml internal standard solution, and the internal standard solution contains D4-NNN, D4-NAT, and D4-NAB respectively and D4-NNK, then add 8.9ml to ultrapure water. Then, ultrasonically extract at 20Hz for 22min, pass the extract through a 0.22μm filter membrane, pipette 550μl into a 1.5ml centrifuge tube, add 1.1ml of methanol, vortex shake for 35s, and centrifuge at a speed of 10000r / min After 3 minutes, take 1ml of the supernatant and transfer it into a 1.5ml injection vial for liquid chromatography-tandem mass spectrometry analysis.

...

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Abstract

The invention discloses a method for detecting alkaloids and nitrosamines in tobaccos simultaneously, and belongs to the technical field of tobacco chemical composition detection. The method comprises the working procedures of: sample extraction, purification, analysis and quantification. Firstly an internal standard solution and ultrapure water are added into tobacco leaf powder, extract liquor passes through a filter film after ultrasonic extraction, then methanol is added, protein is removed through centrifugation after shaking, and supernatant is subjected to liquid chromatography-mass spectrometry. After standard solutions of compounds to be detected with different concentration gradients are prepared by adopting a mixing preparation method, and the liquid chromatography-mass spectrometry is carried out, standard curves of instrument responses of the compounds to the actual concentrations of the solutions are drawn, curve equations are fitted, and the detection concentrations are calculated according to measured values of the instrument responses of the compounds to be detected and converted to the actual concentrations of the compounds to be detected in tobacco samples. The detection method provided by the invention can be used for carrying out the qualitative and quantitative analysis on alkaloids and nitrosamines in the tobacco samples simultaneously, and is easy and convenient to operate, good in reproducibility, high in analytic sensitivity and accurate in quantification.

Description

technical field [0001] The invention belongs to the technical field of detection of tobacco chemical components, and in particular relates to a method for simultaneous detection of alkaloids and nitrosamines in tobacco with simple operation, good reproducibility, high analysis sensitivity and accurate quantification. Background technique [0002] my country is a big country in the production and consumption of cigarettes. The number of smokers and the consumption of cigarettes account for one-third of the number of smokers and consumption of cigarettes in the world respectively. As a special commodity, cigarettes occupy a very important position in my country's national economy. As the direct relationship between smoking and health problems has attracted more and more attention, it is an undeniable fact that smoking is harmful to health. Alkaloid compounds and tobacco-specific nitrosamine compounds in tobacco have always been the focus of tobacco chemistry research. Because...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/06
Inventor 李勇逄涛师君丽孔光辉吴玉萍
Owner YUNNAN ACAD OF TOBACCO AGRI SCI
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