Analysis Of Tobacco-Specific Nitrosamines Research Articles

Direct analysis of tobacco specific nitrosamines in tobacco products using a molecularly imprinted polymer-packed column.

Tobacco specific nitrosamines (TSNAs) are highly carcinogenic by-products in tobacco samples, and their presence is regulated by the Food and Drug Administration. Molecularly imprinted polymers (MIPs) are synthetic polymers that have been "imprinted" with a template analyte in a co-polymer system, and can selectively extract analytes from complex matrices. MIPs can be incorporated into online systems, replacing traditional high performance liquid chromatography (HPLC) columns. MIP material specific for TSNAs was packed into an empty HPLC column using a slurry packing technique. The developed method with the MIP-packed HPLC column was validated on a LC-MS/MS system for the quantitation of N-nitrosonornicotine (NNN) and 4- (methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) in commercial tobacco products. The method was linear over .1-10 ng/ml (.4-10 μg/g) for NNN and NNK. The limit of detection (LOD) was .03 ng/ml (12 μg/g) and the limit of quantitation (LOQ), .1 ng/ml (.4 μg/g). All column uniformity parameters with the exception of theoretical plate number were within the accepted criteria (% RSD values <15%). Theoretical plate number was <250, owing to the large (50 μm) sized MIP particles. Twenty-six tobacco products contained TSNA concentrations that were consistent with reported literature values. The TSNA-MIP based HPLC column effectively replaced a traditional reverse phase HPLC column, and was used for the direct analysis of nicotine and tobacco products without extensive sample preparation prior to instrumental analysis.

Analytical Method for Measurement of Tobacco-Specific Nitrosamines in E-Cigarette Liquid and Aerosol

An experimental method was developed and validated for the collection and analysis of tobacco-specific nitrosamines (TSNAs) that are present in electronic cigarette (EC) liquid or are released from aerosol samples using a liquid chromatography-tandem mass spectrometry (LC-MS/MS) system. As part of this study, the relative recovery of four target TSNAs was assessed by spiking standards in a mixture of propylene glycol and vegetable glycerin. Recovery was assessed against two major variables: (1) the chemical media (solution) selected for sample dilution (acetonitrile [ACN] vs. ammonium acetate [AA]) and (2) the type of sampling filter used (Cambridge filter pad [CFP] vs. quartz wool [QW] tube). The average recovery of TSNAs in terms of variable 1 was 134 ± 22.1% for ACN and 92.6 ± 8.27% for AA. The average recovery in terms of variable 2 was 83.4 ± 7.33% for QW and 58.5 ± 12.9% for CFP. Based on these conditions, the detection limits of N′-nitrosonornicotine (NNN), 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), N′-nitrosoanatabine (NAT), and N′-nitrosoanabasine (NAB) were calculated as 4.40, 4.47, 3.71, and 3.28 ng mL−1, respectively. The concentration of TSNAs in liquid and aerosol samples of six commercial EC solutions was measured as below the detection limit.

Analysis of Traces of Tobacco-Specific Nitrosamines (TSNAs) in USP Grade Nicotine, E-Liquids, and Particulate Phase Generated by the Electronic Smoking Devices

Summary The present study describes the development of a liquid chromatography tandem mass spectrometry (LC-MS/MS) technique for the analysis of trace levels of four tobaccospecific nitrosamines (TSNAs): nitrosoanabasine (NAB), nitrosoanatabine (NAT), 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), and nitrosonornicotine (NNN). The technique can be applied for the analysis of TSNAs in USP grade nicotine. Nicotine used in e-liquids for the electronic smoking devices is typically obtained from tobacco plant materials (Nicotiana tabacum, Nicotiana rustica) and, although it is purified, it contains besides nicotine low levels of several contaminants such as minor alkaloids. It also contains traces of TSNAs. Analysis of TSNAs in USP grade nicotine is a challenging task since the analyzed samples contain about 10+7–10+8 times more nicotine than individual TSNAs. Because the analyzed solutions cannot be diluted too much in order to keep the TSNAs level above the limit of quantitation (LOQ), even for apparently good chromatographic separations, the peak tailing of nicotine may generate interferences. The new method of analysis uses a Luna Omega 1.6 μm particles chromatographic column for separation and detection on a LC-MS/MS instrument with scheduled multiple reaction monitoring (Scheduled MRM). The levels of TSNAs in nicotine of USP purity from four commercial sources varied between 3 to 8 ng/g NAB, 4 to 20 ng/g NAT, 30 to 50 ng/g NNK, and 0.5 to 2 ng/g for NNN. Besides the analysis of TSNAs in nicotine, the technique has been applied successfully in the analysis of TSNAs in e-liquids and in particulate phase generated by the electronic smoking devices.

Analysis of Tobacco-Specific Nitrosamines in the Common Smokeless Tobacco Afzal in Oman.

There is a lack of awareness regarding the carcinogenicity of Afzal, an illegal smokeless tobacco product (STP) widely used among the Omani youth. Previous research has shown that certain types of tobacco-specific nitrosamines (TSNAs) are associated with oral and lung cancers. This study therefore aimed to assess levels of four common TSNAs in a randomly selected sample of Afzal. This study was carried out at Sultan Qaboos University in Muscat, Oman, between April and September 2013. A random sample of Afzal was analysed for four types of TSNAs using high-performance liquid chromatography-tandem mass spectrometry. The four types of TSNAs analysed were 4-(N-nitrosomethylamino)-1-(3-pyridyl)-1-butanone (NNK), N-nitrosonornicotine (NNN), N-nitrosoanatabine (NAT) and N-nitrosoanabasine (NAB). As a reference product, a sample of laboratory-manufactured American moist snuff (Centers for Disease Control and Prevention, Atlanta, Georgia, USA) was also used to confirm the accuracy and precision of the analysis. The analysis revealed total TSNA levels of 3.573 μg/g in the Afzal sample. Mean levels of NNN, NNK, NAT and NAB were 1.205, 1.015, 0.809 and 0.545 μg/g, respectively. Levels of the two most abundant TSNAs (NNN and NNK) found in the Afzal sample exceeded international regulatory limits. Afzal users therefore need to be educated regarding the potential health risks associated with their STP use. Stricter implementation of current legislation is recommended to reduce the availability and usage of Afzal in Oman.

The Application of Solid Phase Extraction to the Analysis of Tobacco-Specific Nitrosamines

A new isolation and separation method has been developed for the alkaloid-related nitrosamines in tobacco, called tobacco-specific nitrosamines (TSNA). The improved isolation procedure involves the extraction by sonication of TSNA from cured tobacco with toluene and an aqueous citrate buffer solution, while purification is achieved by solid phase extraction on small silica cartridges. Interfering extract materials are eluted from the silica cartridges with the toluene and a chloroform-methylene chloride solvent, while subsequent elution with chloroform yields the TSNA in a purified fraction. Gas chromatographic analyses on an SE-54-coated glass capillary column and detection by a nitrogen-phosphorous detector produce very good data on the four TSNAs, which include N-nitrosanatabine (NATB), N-nitrosoanabasine (NAB), 4-(N-methyl-N-nitrosamino)-1-(3-pyridyl)-1-butanone (called NNK), and N-nitrosonornicotine (NNN). The method is shown to be quantitative, reproducible, and applicable to the determination of TSNA in various tobacco samples, such as stems and lamina of cured tobacco leaves, as well as to smokeless tobacco.