Gas Chromatography-Tandem Mass Spectrometry Method for Selective Detection of 2-Nitropropane in Mainstream Cigarette Smoke.

Sensitive and selective gas chromatography-tandem mass spectrometry method for the detection of nitrobenzene in tobacco smoke

The particle size distribution of cigarette smoke is an important factor in predicting the deposition fraction of the inhaled particles in various regions of the respiratory tract. Mainstream cigarette smoke is a direct concern for smokers, while the exhaled and side stream smoke contribute to passive (second hand) smoking. The particle size distribution of tobacco smoke for mainstream and exhaled smoke has been studied in this work. This study investigates how smoking behavior, including puff volume and number of puffs, affect the particle size distribution of mainstream cigarette smoke. Scanning Mobility Particle Sizer (SMPS) was used to measure the particle size distribution of mainstream and exhaled cigarette smoke. The values of count median diameter (CMD) mobility size for the first four consecutive puffs were found to be 193 nm, 198 nm, 194 nm and 186 nm. As the volume of a puff increases from 35 mL to 85 mL, the CMD shifted slightly towards the higher particle size ranges, because of increased probability of coagulation and other combination processes. For the exhaled cigarette smoke, the growth factor was found to be 1.5 ± 0.3 with respect to mainstream cigarette smoke. The experimental results of particle size distributions were used in a Multiple Path Particle Dosimetry (MPPD) model to predict deposition patterns in the human respiratory tract. The MPPD model results show the deposition fractions for mainstream cigarette smoke were 0.163, 0.152 and 0.298 for the head, trachea and bronchi (TB) and pulmonary region, and those for exhaled cigarette smoke were 0.273, 0.064 and 0.134 respectively. The total calculated deposition fraction for mainstream and exhaled cigarette smoke was found to be 0.613 and 0.471, respectively.

Nitromethane is a volatile organic compound categorized as a Group 2B carcinogen by the International Agency for Research on Cancer. It has been detected in mainstream cigarette smoke, but few reliable methods have been reported for accurate quantification. We developed, a sensitive, selective, fully validated method for the targeted determination of nitromethane in mainstream tobacco smoke in ten U.S. domestic brands and two quality control materials (3R4F and CM6). The vapor phase portion of machine-generated cigarette mainstream smoke, under modified ISO 3308:2000 regime (ISO) and modified intense regime (HCI), from single cigarettes was collected using airtight polyvinylfluoride sampling bags. The bags' contents were extracted using methanol containing an isotopically labeled internal standard followed by gas chromatography-tandem mass spectrometry. This approach is sufficiently sensitive to measure nitromethane levels in the nanogram range, with a method limit of detection of 72.3 ng/cig. Within-product variability estimated from the replicate analysis of 10 products ranged from 4.6%-16.3% (n = 6) over the two different smoking regimes, and method reproducibility estimated from two products used as quality control materials (3R4F and CM6) yielded intermediate precision values ranging from 16.6 to 20.8% (n = 20). Under HCI, nitromethane yields in machine-generated cigarette smoke from ten different domestic cigarette products ranged from 3.2 to 12 μg/cig; under ISO yields ranged from 1.6 to 4.9 μg/cig under standardized smoking machine conditions. Nitromethane yields are related to both the smoke regime (blocking of vent holes, puff duration and puff volume) and the heterogeneity of tobacco mixtures. This method provides a selective and fully validated technique to accurately quantify nitromethane in mainstream cigarette smoke, with minimal waste generation. It is an improvement over previous methods with regards to specificity, throughput, and simplicity of the sample collection process.

The long-term health effects of using e-cigarette, or vaping, products (EVPs; also known as e-cigarettes, electronic nicotine delivery systems, and vape pens) remain largely unknown. The inhalation of excipients, such as propylene glycol (PG) and glycerin (GLY), may have long-term health effects. In addition to the direct health effects of PG and GLY, glycerin-containing products can be contaminated with toxic ethylene glycol (EG) and diethylene glycol (DEG). To assess this issue, we developed a simple, versatile, high-throughput isotope dilution gas chromatography-tandem mass spectrometry method for quantifying these common excipients and contaminants. The method is applicable to both the liquid contents and machine-generated aerosols of EVPs. Our rigorous method validation demonstrates that the new method is specific, precise, accurate, and rugged/robust. The calibration range is linear from 0.1–7 mg for the excipients and 2.5–1,000 µg for the contaminants. These ranges encompass expected excipients levels in EVP e-liquids and their machine-generated aerosols and the relevant maximum residue safety limit of 1 mg/g, or 0.1% (w/w), for the contaminants. The calculated limits of detection for PG, GLY, EG, and DEG were determined as 0.0109 mg, 0.0132 mg, 0.250 µg, and 0.100 µg, respectively. The method was applied to the aerosol emissions analysis of 141 EVPs associated with the 2019 lung injury outbreak, and found typical levels of PG (120.28–689.35 mg/g of aerosol) and GLY (116.83–845.96 mg/g of aerosol) in all nicotine-containing products; PG (81.58–491.92 mg/g of aerosol) and GLY (303.86–823.47 mg/g of aerosol) in 13% of cannabidiol (CBD) products; PG (74.02–220.18 mg/g of aerosol) and GLY (596.43–859.81 mg/g of aerosol) in products with neither nicotine nor CBD; and none detected in tetrahydrocannabinol (THC) products. No products contained glycol contaminants above the recommended maximum residue safety limit.

In order to investigate the effects of cigarette paper properties on the migration ratios of heavy metal elements in mainstream and sidestream cigarette smoke, 13 cigarette samples were prepared according to single factor design, the effects of permeability, grammage and combustion improver content of cigarette paper on the migration ratio of Cr, Ni, As, Se, Cd and Pb were studied. The results showed that: 1) The air permeability of cigarette paper significantly positively correlated to the migration ratio of Cd in sidestream cigarette smoke in the range of20-80 CU, while significantly negatively correlated to the migration ratios of As, Cd and Pb in mainstream cigarette smoke in the range of 20-50 CU. 2) The grammage of cigarette paper significantly positively correlated to the migration ratio of Cd in sidestream cigarette smoke, and had no obvious correlations with the migration ratios of the other elements. 3) The content of combustion improver in cigarette paper significantly negatively correlated to the migration ratios of Cr and As in mainstream cigarette smoke and that of As in sidestream cigarette smoke. In conclusion, cigarette paper properties have some, but not the same, effects on the migration ratios of Cr, Ni, As, Se, Cd and Pb in mainstream and sidestream cigarette smoke.

An international literature survey of “IARC group I carcinogens” reported in mainstream cigarette smoke

These are curious times. The Canadian government has passed legislation that requires cigarette manufacturers to routinely test and publish the amounts of 44 toxic substances in cigarette mainstream smoke (MSS). Followingin the footsteps of their northern neighbor, various US legislators and regulators are considering modifications to their cigarette testing and reporting programs that will also list toxicants in MSS. Across the Atlantic Ocean, the European Commission has passed a directive that may also follow the North American lead for public disclosure of MSS toxic chemicals for each brand of cigarette sold in the marketplace. United Kingdom authorities have also expressed their intention to follow this mandate. It is difficult to understand the motivation and value of these existing or potentially forthcoming legislative actions. Although there is nearly total agreement among the world's scientists that cigarette smoking is a health hazard, few are bold enough to say with credibility which smoke chemicals or classes of chemicals are responsible for the adverse effects. Therefore, if the specialists are unable to interpret the smoke toxicant data, how is the general public to use their newfound knowledge? The posting of smoke chemical toxicant data is also problematic for the Tobacco Industry for several reasons. First, no standard analytical methods exist for most suspected toxicants. Second, the listing of smoke toxicant yields may ignite a 21 s t Century version of the "tar" wars in the USA during the 1960s; we have already seen evidence of such competition beginning in the US. Third, and most important of all, no one knows whether or not reducing the yield of one or more publicized MSS toxicant will result in a "less hazardous" cigarette. Assuming that the current situation is approximately as described above, the authors of this paper critically examined the existing lists of MSS toxicants. They discarded chemicals that are no longer relevant, e. g., DDT, N-nitrosodiethanolamine, added known smoke constituents that are glaringly absent, e.g., dioxins, and replaced the existing 1950-60s era nonfiltered cigarette MSS yields with those more representative of the present-day marketplace. Data for the Kentucky reference 1R4F cigarette smoked under standardized smoking conditions, i.e., those established by the International Organization for Standardization (ISO) and the Federal Trade Commission (FTC), are used as a surrogate for the modern-day cigarette whenever possible. A list of smoke toxicants and their approximate concentrations in today's cigarettes is nearly useless without an appropriate ranking of their relative toxicity. Unfortunately, the toxicological data for ranking importance are available for fewer than 5% of the approximately 4800 reported smoke constituents. Although neither of this paper's authors presumes to be a toxicologist, we cite in our discussion several published attempts at ranking smoke toxicants. Specifically, ranking by US Occupational Safety and Health Administration (OSHA) permissible workplace exposure levels, use of US Environmental Protection Agency (EPA) toxicity criteria supplemented with California EPA criteria, and use of the Human Exposure -Rodent Potential methodology and database developed by AMES et al. when data are available. There appears to be a wide divergence in the permissible exposures allowable in the workplace and those advocated by environmental regulators.

Analysis of α- and β-carbolines in mainstream smoke of reference cigarettes by gas chromatography–mass spectrometry

Analysis of α- and β-carbolines in mainstream smoke of reference cigarettes by gas chromatography–mass spectrometry

Synthesis and application of a molecularly imprinted polymer as a filter to reduce polycyclic aromatic hydrocarbon levels in mainstream cigarette smoke

Tobacco-specific nitrosamines are one of the most important groups of carcinogens in tobacco products. Using adsorbents as filter additives is an effective way to reduce tobacco-specific nitrosamines in cigarette smoke. Molecularly imprinted polymers (MIPs) using nicotinamide as template were grafted on the silica gel surface to obtain MIP@SiO2 and employed as filter additives to absorb tobacco-specific nitrosamines in mainstream cigarette smoke. Four milligrams of MIP@SiO2 per cigarette was added to the interface between filter and tobacco rod to prepare a binary filter system. The mainstream smoke was collected on an industry-standard Cambridge filter pad and extracted with ammonium acetate aqueous solution before analysis. Compared to the cigarette smoke of the control group, the levels of tobacco-specific nitrosamines with silica gel and with MIP@SiO2 were both reduced, and the adsorption rates of N-nitrosonornicotine, N-nitrosoanabasine, N-nitrosoanatabine, and 4-(methylnitrosamino)-1-(3-pyridine)-1-butanone with silica gel and with MIP@SiO2 were 20.76, 15.32, 18.79, and 18.01%, and 41.33, 34.04, 37.86, and 35.53%, respectively. Furthermore the content of total particle materials in cigarette smoke with silica gel was decreased evidently but showed no observable change with MIP@SiO2 . It indicated MIP@SiO2 could selectively reduce tobacco-specific nitrosamines in the mainstream cigarette smoke with no change to the cigarette flavor.

Inder order to determine Benzopyrene (BaP) in Mainstream and Sidestream Smoke, a Solid phase extraction (SPE) cartridge was used to isolate the BaP fraction from the total particulate matter of mainstream cigarette smoke and sidestream cigarette smoke and the BaP were measured by RP-HPLC with ultraviolet detection. The result showed that the recovery rate of BaP extracted is 94.5%, compared with the relative standard of 6.95%. The yields of BaP in mainstream smoke for Virginia cigarette are higher than those for blended cigarettes. The yields of BaP in sidestream smoke are much higher than those in mainstream smoke. The proposed method involves an solid phase extract and HPLC-UV analysis procedure. With this method, the BaP yields in mainstream and sidestream cigarette smoke can be measured fast, easily and precisely using readily available apparatus and instruments.

N-nitrosonornicotine (NNN), 4-( methylnitrosamino )-1-(3-pyridyl )-1-butanone (NNK), N-nitrosoanatabine (NAT) and N-nitrosoanabasine (NAB) are the most abundant carcinogens identified in tobacco and tobacco smoke. The accurate quantifications of NNN, NNK, NAT and NAB are necessary to evaluate its impact on the public health. A liquid chromatography-electrospray tandem mass spectrometry (LC-ESI MS/MS) method was developed to simultaneously determine NNN, NNK, NAT and NAB in mainstream cigarette smoke. Mainstream smoke was collected in a Cambridge filter pad and then was extracted by 10 mL 100 mmol/L ammonium acetate after 100 microL of mixed deuterated internal standards was added. Then the extract was detected by using positive electrospray ionization on a tandem mass spectrometer in multiple reaction monitoring (MRM) mode. NNN, NNK, NAT and NAB were separated on a Zorbax Eclipse XDB-C18 column with the gradient elution using mobile phase A (0.1% acetic acid in water) and mobile phase B (0.1% acetic acid in methanol). The detection limits for NNN, NNK, NAT and NAB were 0.019, 0.002, 0.008 and 0.007 microg/L, respectively. The recoveries were varied from 84.9% to 104.5% for Chinese Virginia cigarettes and the relative standard deviations (n = 8) ranged from 2.96% to 6.65%. This proposed approach, which provides a higher sensitivity and specificity, is suitable for the determination of NNN, NNK, NAT and NAB in mainstream cigarette smoke.

A method for differentiation of gas- and particulate-phase mercury in mainstream cigarette smoke was developed using electrostatic precipitation (EP) as the trap for the particulate phase and impingers containing acidic potassium permanganate solution as the trap for the gas-phase portion. The mercury collected from the gas phase was analyzed by conventional cold vapor atomic absorption spectrometry (CVAAS) and the particulate phase was analyzed by gold amalgamation CVAAS. Cigarettes were smoked under two smoking regimes, FTC (35-mL puff volume, 2 s puff duration and one puff every 60 s) and an alternative (45-mL puff volume, 2 s puff duration, one puff every 30 s and 50% of any ventilation holes blocked) currently recommended by the Massachusetts Department of Health. For the 1R4F reference cigarette smoked under the FTC smoking regime, the mercury found in the particulate phase was less than 0.2 ng/cig, compared with 4.9 ng/cig in the gas phase. By changing smoking parameters, the mercury concentration in mainstream smoke was found to change proportional to the delivery of cigarette smoke condensate (CSC) for the same type of cigarette. However, the mercury level for different types of cigarettes smoked under the same smoking parameters had no linear relationship with CSC delivery. Spiked recovery was 98% AA± 8% for gas-phase mercury and 97% AA± 2% for the particulate phase. These results indicate that the analytical method developed is suitable for the determination of mercury in mainstream smoke. For routine analytical work in a smoking laboratory, only the gas phase needs to be analyzed for determination of mercury in mainstream smoke because the amount of mercury in the particulate phase is negligible.

The purpose of this study was to test the hypothesis that components in mainstream (MS) and sidestream (SS) cigarette smoke inhibit growth and angiogenesis using the chick chorioallantoic membrane (CAM). Varying doses of whole or gas-phase MS and SS smoke solutions were placed on day 5 CAMs, and their effects on angiogenesis were evaluated on day 6. All parameters evaluated (CAM area, major blood vessel area, major blood vessel diameter, blood vessel pattern formation, and capillary plexus formation) were inhibited to different degrees in a dose-dependent manner by both MS and SS smoke treatment. Inhibition of growth and vessel development was correlated with inhibition of cell proliferation. Inhibition of capillary plexus formation was caused by failure of mesodermal blood vessels to migrate to the ectoderm. SS smoke solution was more inhibitory than MS smoke solution in all assays, except for capillary plexus formation. In all assays, the toxicants in SS smoke partitioned mainly with the gas phase, whereas those in MS smoke were deduced to be mainly in the particulate phase in the proliferation-dependent assays (CAM area, blood vessel area, blood vessel diameter) and in both the gas and particulate phase in the pattern formation and plexus formation assays. Some of the inhibitory doses of MS and SS smoke solutions had nicotine concentrations within the range found in human smokers. Taken together, these data demonstrate that exposure to complex mixtures of chemicals in MS and SS cigarette smoke adversely affect growth, vessel development, vessel migration, and cell proliferation.