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Simultaneous determination of tobacco minor alkaloids and tobacco-specific nitrosamines in mainstream smoke by dispersive solid-phase extraction coupled with ultra-performance liquid chromatography/tandem orbitrap mass spectrometry.

RATIONALE: The minor alkaloids in tobacco play an important role in the chemical composition of cigarette smoke, and they are precursors of tobacco-specific nitrosamines (4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), N-nitrosonornicotine (NNN), N-nitrosoanabasine (NAB) and N-nitrosoanatabine (NAT)). NNN and NNK are classified as group 1 carcinogens. A method quantitating both tobacco minor alkaloids and tobacco-specific nitrosamines in mainstream smoke has not been reported.

METHODS: Tobacco minor alkaloids and tobacco-specific nitrosamines in cigarette mainstream smoke were extracted by sonication. The extract was cleaned up by dispersive solid-phase extraction, and separation was achieved via ultra-performance liquid chromatography/tandem orbitrap mass spectrometry.

RESULTS: The method was validated by analysis of six replicate samples spiked with three levels of the analyses. The mean recoveries for the six replicates were from 84.7% to 118% with less than 15% relative standard deviation except myosmine at the low spiked level and the calculated detection limits were 0.066 to 13.2 ng/cig, respectively. The mean concentrations of nicotyrine, anabasine, nornicotine, anatabine, myosmine, 2,3-bipyridine, cotinine, nicotelline, N-formylnornicotine, NNK, NNN, NAB and NAT in 30 different brands of commercial cigarette smoke under the ISO smoking regimen were 2.50 μg/cig, 2.34 μg/cig, 3.21 μg/cig, 5.78 μg/cig, 2.83 μg/cig, 1.05 μg/cig, 1.55 μg/cig, 0.55 μg/cig, 2.48 μg/cig, 6.06 ng/cig, 3.62 ng/cig, 0.40 ng/cig and 6.15 ng/cig, respectively.

CONCLUSIONS: The proposed method was suitable for analysis of tobacco minor alkaloids and tobacco-specific nitrosamines in cigarette mainstream smoke.

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