4-(methylintrosamino)-1-(3-pyridyl)-1-butanone (NNK) and N\'-nitrosonornicotine (NNN) are the most prevalent and toxic tobacco specific nitrosamines (TSNAs). Due to their carcinogenicity, knowledge of the composition of NNK and NNN in tobacco is necessary. Herein, a sensitive and rapid method, which employs autoclave extraction-supercritical fluid chromatography/tandem mass spectrometry (SFC-MS/MS), has been developed for the analysis of NNK and NNN in tobacco. Both water-soluble and matrix-bound NNK and NNN were extracted with 100 mM ammonium acetate in an autoclave (130 °C, 4 h), and the aqueous extract was subjected to solvent replacement prior to SFC-MS/MS analysis. NNK and NNN were effectively separated within 5 min by using supercritical CO2 as the main mobile phase coupled with a co-solvent of methanol. Excellent linearity was obtained with coefficients of determination (R2) greater than 0.9997 in the range of 1-160 ng/mL and 5-800 ng/mL for NNK and NNN, respectively. The recoveries were in the range of 92.5-110.0% at different spiked levels of real samples. 12 tobacco samples which include 3 typical tobacco varieties of burley, flue-cured, and oriental tobaccos had been analyzed, and the fraction of matrix-bound NNK was determined as well. In addition, a comparison between the proposed SFC-MS/MS method and a validated liquid chromatography tandem mass spectrometry (LC-MS/MS) internal standard method was conducted. Both techniques exhibit comparable analysis results, but peak splitting of NNN was observed by LC-MSMS due to the existence of E/Z isomers, while SFC-MS/MS offers great improvement through elution condition optimization, demonstrating the applicability of SFC-MS/MS as an alternative tool for NNK and NNN analysis.

New shape-selectivity of graphene-based materials was discovered on this article. To explore the new selectivity, the structure and surface state of graphene and carbon nanotube were examined firstly, and their specific selectivity was verified and was compared with that of ZSM-5 zeolite in aqueous solutions of tobacco specific nitrosamines (TSNA) along with dyes. These two adsorbents trapped about 55% and 70% of 4-methylnitrosamino-1-3-pyridyl-1-butanone (NNK) but only 3% of N\'-nitrosonornicotine (NNN) in solution, having an obvious selectivity for the former, due to its stronger interaction with graphene. NNK on graphene sheet obtained more electrons (0.015 e) and owned larger adsorption energy (15.63 kcal mol-1) than that of NNN (0.003 e, 9.19 kcal mol-1), according to theoretical calculation and FTIR results. More 95 or 136 mg g -1 acid red 88 than methyl orange was captured by graphene or carbon nanotube, demonstrating this special and abnormal selectivity again. With new selectivity, graphene showed a higher capacity (6.9%) and shorter adorption equilibrium time (5 min) for TSNA than the typical selecive sorbent ZSM-5 zeolite (1.7% and 20 min) in tobacco solution but kept the similar selctivity to NNK, paving a new way to control the carcinogens like TSNA in environment.