Mainstream Smoke Research Articles

A Method to Amend Cigarettes With Menthol for Use in Research.

Studies have evaluated the role of menthol cigarettes on various addiction-related outcomes; however, the effect of varying menthol content on these outcomes has not been evaluated. We developed a method to amend non-menthol SPECTRUM Research Cigarettes to contain menthol at four different levels. SPECTRUM Research Cigarettes, NRC 600 (0.8 mg nicotine; 10 mg tar), were modified to contain target menthol amounts at 3, 6, and 12 mg/cigarette by injecting 25 µL ethanol/triacetin/menthol solutions of varying concentrations (120 mg menthol/mL, 240 mg/mL, and 480 mg/mL) into four distinct locations in the filter and tobacco rod. Menthol content was tested in triplicate in the whole cigarette and in the tobacco rod and filter at 1, 24, 48, and 72 hours for each target menthol level using an extraction solution of quinoline in methyl-tert-butyl ether and measured using gas chromatography with flame ionization detection. Injections into the filter and tobacco rod (12.5 µL each) yielded equal menthol distribution up to 72 hours. However, total menthol content decreased from an average of 90.3% of the target menthol concentration at 1 hour to 80.7% at 72 hours in cigarettes stored individually in glass tubes at room temperature. Analysis of urinary menthol glucuronide confirmed that amended cigarettes used within 24 hours of injection delivered dose-related menthol levels to participants in a clinical laboratory setting. This method can be used to modify cigarettes with a range of reliable menthol levels in both filter and tobacco rod for use in laboratory and clinical research. This study presents a technique for modifying cigarettes with different levels of menthol that can reliably deliver dose-related menthol levels to participants when smoked in a clinical study. The technique can be used to quickly amend cigarettes to examine the independent effects of varying flavor and additive levels on smoking behavior, nicotine pharmacokinetics, mainstream smoke emissions, and other laboratory or clinical research outcomes.

Carbonyl Yields in Cigars under Three Smoking Regimens Using a Linear Smoking Machine.

This study used standard linear smoking machines and puffing protocols to generate data on carbonyl yields in mainstream smoke from 11 unfiltered sheet-wrapped cigars (SWC), seven leaf-wrapped cigars (LWC), and two Kentucky reference cigarettes (3R4F, 1R6F). Carbonyl yields in cigar and cigarette products were determined using three different smoking regimens: International Organization for Standardization (ISO), Canadian Intense (CI), and Cooperation Centre for Scientific Research Relative to Tobacco (CORESTA) Recommended Method (CRM) No. 64 (CRM64, Routine Analytical Cigar-Smoking Machine─Specifications, Definitions and Standard Conditions). Mainstream tobacco smoke was collected using a smoking machine fitted with an impinger containing 2,4-dinitrophenylhydrazine (DNPH) and carbonyl compounds quantified using liquid chromatography with an ultraviolet detector. Commercial SWC and LWC generated comparable formaldehyde yields (SWC, 9.4-28 μg/cigar [ISO], 8.2-43 μg/cigar [CI], 8.6-13 μg/cigar [CRM64]; LWC, 11-13 μg/cigar [ISO], 11-22 μg/cigar [CI], 16-21 μg/cigar [CRM64]) and acrolein yields; however, LWC generated higher acetaldehyde yields compared to SWC, using CI and CRM64 regimens. Reference cigarettes using standard puffing regimens generated carbonyl yields within reported ranges and 5-10% RSDs, whereas the CRM64 regimen generated lower carbonyl yields and 12-14% RSDs. Reference cigarettes generated higher formaldehyde yields using cigarette smoking regimens (21-28 μg/cigarette under ISO, 76-96 μg/cigarette under CI) but comparable formaldehyde yields under CRM64 (12-14 μg/cigarette). In addition, this study evaluated physical parameters (e.g., tobacco weight, length, diameter, circumference, tobacco rod density) that show the correlation between tobacco weight, tobacco rod density, and acetaldehyde yields under the three smoking regimens. Carbonyl yields in the mainstream smoke of cigar products using the three smoking regimens were highly variable; however, the CI smoking regimen may provide meaningful analytical information regarding cigar smoke constituents, with lower likelihood of self-extinguishment due to the short puffing intervals.

Cytotoxic effect of gas phase extract of mainstream smoke derived from heated tobacco products on vascular smooth muscle cells.

Cytotoxic effect of gas phase extract of mainstream smoke derived from heated tobacco products on vascular smooth muscle cells.

Conventional tobacco products harbor unique and heterogenous microbiomes

While an increasing number of studies have evaluated tobacco microbiomes, comparative microbiome analyses across diverse tobacco products are non-existent. Moreover, to our knowledge, no previous studies have characterized the metabolically-active (live) fraction of tobacco bacterial communities and compared them across products. To address these knowledge gaps, we compared bacterial communities across four commercial products (cigarettes, little cigars, cigarillos and hookah) and one research cigarette product. After total DNA extraction (n = 414) from all samples, the V3V4 region of the 16S rRNA gene was sequenced on the Illumina HiSeq platform. To identify metabolically-active bacterial communities within these products, we applied a coupled 5-bromo-2′-deoxyuridine labeling and sequencing approach to a subset of samples (n = 56). Each tobacco product was characterized by its signature microbiome, along with a shared microbiome across all tobacco products consisting of Pseudomonas aeruginosa, P. putida, P. alcaligenes, Bacillus subtilis, and Klebsiella pneumoniae. Comparing across products (using Linear discriminant analysis Effect Size (LEfSe)), a significantly higher (p < 0.05) relative abundance of Klebsiella and Acinetobacter was observed in commercial cigarettes, while a higher relative abundance of Pseudomonas and Pantoea was observed in research cigarettes. Methylorubrum and Paenibacillus were higher in hookah, and Brevibacillus, Lactobacillus, Bacillus, Lysinibacillus, and Staphylococcus were higher in little cigars and cigarillos. Across all products, the majority of the metabolically-active bacterial communities belonged to the genus Pseudomonas, followed by several genera within the Firmicutes phylum (Bacillus, Terribacillus, and Oceanobacillus). Identification of some metabolically-active pathogens such as Bacillus cereus and Haemophilus parainfluenzae in commercial products is of concern because of the potential for these microorganisms to be transferred to users' respiratory tracts via mainstream smoke. Future work is warranted to evaluate the potential impact of these tobacco bacterial communities on users' oral and lung microbiomes, which play such an important role on the spectrum from health to disease.

Sidestream Smoke Extracts from Harm-Reduction and Conventional Camel Cigarettes Inhibit Osteogenic Differentiation via Oxidative Stress and Differential Activation of intrinsic Apoptotic Pathways.

Epidemiological studies suggest cigarette smoking as a probable environmental factor for a variety of congenital anomalies, including low bone mass, increased fracture risk and poor skeletal health. Human and animal in vitro models have confirmed hypomineralization of differentiating cell lines with sidestream smoke being more harmful to developing cells than mainstream smoke. Furthermore, first reports are emerging to suggest a differential impact of conventional versus harm-reduction tobacco products on bone tissue as it develops in the embryo or in vitro. To gather first insight into the molecular mechanism of such differences, we assessed the effect of sidestream smoke solutions from Camel (conventional) and Camel Blue (harm-reduction) cigarettes using a human embryonic stem cell osteogenic differentiation model. Sidestream smoke from the conventional Camel cigarettes concentration-dependently inhibited in vitro calcification triggered by high levels of mitochondrially generated oxidative stress, loss of mitochondrial membrane potential, and reduced ATP production. Camel sidestream smoke also induced DNA damage and caspase 9-dependent apoptosis. Camel Blue-exposed cells, in contrast, invoked only intermediate levels of reactive oxygen species insufficient to activate caspase 3/7. Despite the absence of apoptotic gene activation, damage to the mitochondrial phenotype was still noted concomitant with activation of an anti-inflammatory gene signature and inhibited mineralization. Collectively, the presented findings in differentiating pluripotent stem cells imply that embryos may exhibit low bone mineral density if exposed to environmental smoke during development.

Development of cigarette filter with biomass-based activated carbon for naphthalene removal from mainstream smoke

Development of cigarette filter with biomass-based activated carbon for naphthalene removal from mainstream smoke

Modulation of mouse laryngeal inflammatory and immune cell responses by low and high doses of mainstream cigarette smoke

Cigarette smoking is a major risk factor for laryngeal diseases. Despite well-documented cigarette smoke (CS) induced laryngeal histopathological changes, the underlying immunopathological mechanisms remain largely unexplored. The goal of this study was to evaluate inflammatory and immune cell responses in a CS-exposed larynx. Specifically, we used a 4-week subacute whole-body CS inhalation mouse model to assess these responses in the laryngeal mucosa upon exposure to low (LD; 1 h/day) and high dose (HD; 4 h/day) CS. Laryngeal tissues were harvested and evaluated using a 254-plex NanoString inflammation panel and neutrophil/macrophage/T-cell immunohistochemistry (IHC). NanoString global and differential gene expression analysis revealed a unique expression profile only in the HD group, with 26 significant differentially expressed genes (DEGs). StringDB KEGG pathway enrichment analysis revealed the involvement of these DEGs with pro-inflammatory pathways including TNF/TNFα and IL-17. Furthermore, inflammatory responses remained inhibited in conjunction with predicted activated states of anti-inflammatory regulators like PPARγ and NFE2L2 upon Ingenuity Pathway Analysis (IPA). Subglottic T-cell levels remained significantly inhibited as corroborated by IPA predictions. Overall, our key findings are consistent with HD exposures being anti-inflammatory and immunosuppressive. Furthermore, the identification of important regulatory genes and enriched pathways may help improve clinical interventions for CS-induced laryngeal diseases.

Aerosol Formation and Transfer in Open- and Closed-Ended Heated Tobacco Products

Summary A lit cigarette forms a positive and negative pressure zone by a puff with respect to the position of its paper burn line. Smoke aerosols generated from the two zones are then pulled through the rod under the puff to form the mainstream smoke. This phenomenon is fundamental to the thermophysics and the resultant chemical composition of the mainstream smoke. In this study, we created two different airflow pathways inside a heated tobacco rod by a puff, and investigated the differences in aerosol formation and its chemical compositions. The two different pressure-induced conditions, one through an open-ended tobacco rod (marked as HNB, a label of a designed airflow pathway of commercial heated tobacco products called heat-not-burn prior), and the other through a closed-ended tobacco rod (marked as NSC, a label of a novel-designed airflow pathway of heated tobacco products), were compared for their aerosol collected mass (ACM), the contents of nicotine, water and added aerosol agents such as propylene glycol (PG) and glycerol (VG), as well as selected aldehydes and ketones in the mainstream aerosol. Aerosol particle distribution and the heated temperature along different rod locations were also compared during a puff. The results indicated marked differences in the aerosol formation processes between the two HNB and NSC systems. The transfer ratios of the main aerosol components were significantly higher for the NSC; the levels of formaldehyde and acetaldehyde were significantly lower under the NSC than the HNB condition. There were also significant differences for the aerosol particle number concentration (APNC) and count median diameter (CMD) for the two systems. The lack of convective heat transfer in the aerosol formation under the NSC condition resulted in a relatively stable thermal aerosol generation zone, reflected by the temperature difference between the two systems in the selected locations. The NSC mode of tobacco heating thus offers a novel and enhanced aerosol generation for heat tobacco product designs.

Accuracy improvement of determination of seven minor tobacco alkaloids in mainstream cigarette smoke using analyte protectants by gas chromatography-mass spectrometry

Tobacco alkaloids are important precursors of carcinogenic tobacco-specific nitrosamines. Therefore, accurate quantification of tobacco alkaloids is highly important. This study investigates the compensation effects of novel analyte protectants (APs) for matrix effects (MEs) to determine seven minor tobacco alkaloids (nornicotine, myosmine, anabasine, anatabine, nicotyrine, 2,3′-bipyridine, and cotinine) in mainstream cigarette smoke with high accuracy and robustness. By comparing the heights and shapes of the chromatographic peaks before and after the addition of APs to standard solutions prepared in dichloromethane and cigarette smoke solutions, the compensation effects of 12 APs and their combinations on the MEs of seven minor tobacco alkaloids were evaluated, and the best combination of 2-pyridylethylamine (2 mg/mL)+1,2-decanediol (1 mg/mL) was identified. This AP combination could effectively improve the shapes and increase the heights (by 7–371%) of chromatographic peaks for standard solutions prepared in dichloromethane and cigarette smoke solutions. Before the addition of this AP combination, the slope ratios of the calibration curves for two types of standard solutions of the seven target chemicals were 71.4–159.8%, while they were 87.4–105.6% after the addition, indicating that this AP combination reduced the matrix difference between pure solvent and cigarette smoke solution. After adding the AP combination, the standard curves of solutions prepared in dichloromethane showed good linearity (r2 ≥ 0.999), the spiked recoveries were between 80.9% and 119.6%, and the inter- and intraday precisions were between 1.5–9.5% and 3.1–8.5%, respectively. Three commercial cigarette samples and one mixed standard solution were also tested under four different instrument working conditions to verify the long-term accuracy and ruggedness of the approach in routine real-world analysis of the method. The results showed that the RSD values were higher (3.5–25.4%) without the AP combination than that (<6.7%) with the AP combination. Because of its high accuracy, precision, and robustness, this method has good practical prospects.

Determination and Difference Analysis of Phenolic Compounds in Smokers' Saliva and Mainstream Smoke.

To study the differences in phenolic compounds between tobacco smokers' saliva and mainstream smoke, a method was developed for the analysis of 12 phenolic compounds in saliva and mainstream smoke based on ultrahigh-performance liquid chromatography with fluorescence detection (UPLC-FLD). The contents and distributions of phenolic compounds in tobacco smokers' saliva and mainstream smoke were compared. The results were as follows: (1) Phenolic compounds were quantitatively analyzed by the internal standard method using 4-fluorophenol as an internal standard. For smokers' saliva samples, the limits of quantification (LOQs) ranged from 2.2 to 19.1 μg/L, and the recoveries were from 80.2% to 119.2% at the three spiked levels. For mainstream smoke samples, the LOQs ranged from 0.03 to 0.26 μg/cig, and the recoveries ranged from 84.9% to 107.0% at the three spiked levels. (2) The contents of phenolic compounds from 14 cigarettes in mainstream smoke and smokers' saliva were determined. In mainstream smoking, the main phenolic compounds were hydroquinone, catechol, phenol, meta- and para-Cresol, and o-methylhydroquinone. In smokers' saliva, the main phenolic compounds were phenol and meta- and para-Cresol and the contents of phenolic compounds in smokers' saliva from different cigarettes were significantly different. (3) The content distribution patterns of phenolic compounds in smokers' saliva differed from those in mainstream smoke. The predominant phenolic compound in mainstream smoke was dihydroxybenzene, while monophenols predominated in smokers' saliva. (4) The contents of phenolic compounds from five kinds of cigarettes were analyzed in the saliva of different smokers using principal component analysis, which indicated that cigarettes with different sensory effects were clearly distinguished by differences in the contents of phenolic compounds in saliva.

The influencing factors of health hazards of benzo[a]pyrene in cigarette mainstream smoke: The example of one brand in Beijing.

INTRODUCTIONThe study focused on the influence of tar concentrations, smoking regimen, and smoking behavior, on benzo[a]pyrene (B[a]P) emission from cigarette mainstream smoke and related health hazards to determine the key factors influencing B[a]P reduction and protection of the smoker’s health.METHODSA locally popular brand of cigarettes in Beijing was selected with tar concentrations of 1, 3, 5, 8, 10, and 11 mg/cigarette. Two different machine smoking regimens, the Canada Intense (HCI) regimen and the International Organization for Standardization (ISO) regimen, were adopted to collect the cigarette mainstream smoke. The B[a]P emission concentrations were then measured by gas chromatography and mass spectrum.RESULTSThe average B[a]P emission was 8.14–17.6 ng/cigarette for the HCI regimen and 0.92–3.46 ng/cigarette for the ISO regimen. As expected, the tar concentrations and B[a]P emissions exhibited a positive relation in both the HCI and ISO regimens, the cancer risk and non-cancer risk increased with an increase in tar concentrations for both the ISO and HCI regimens, and the smoking behavior also affected the B[a]P emissions with a tendency of VB (ventilation blocking) > HVB (half ventilation blocking) > DP (deep puff), under the same smoking regimen. Under the same conditions, the cancer risk and non-cancer risk in men were 1.19 and 1.11 times, respectively, higher than in women.CONCLUSIONSThe smoking regimen influences the B[a]P emission relatively more than the cigarette tar concentration and smoking behavior. The cancer risk and non-cancer risk are higher in men than in women that possibly due to longer smoking duration and greater smoking intensity.

The burning and pollutant formation processes during cigarette smoking under various inhalation frequency

In current study, particle number (PN), CO, HC and NOx in both main and side stream smoke were measured, the pyrolysis of tobacco leaf was measured by differential scanning calorimeter and the morphology of leaf was observed by laser scanning confocal microscope. Main findings are: the diameter of the particulate matter was changed between 10 nm to 80 nm; pollutants in main stream smoke decrease with interval time; PN and HC in side stream smoke increase with interval time. But for CO and NOx exposure in side stream smoke, the highest amount appeared at interval 3 s followed by 4 s and then 9 s. So, the common smoking habits did not achieve the best trade-off between inhaled and exposure amount. The test uncertainty was analyzed in detail. So the present study is to provide a good demonstration for smoking research and implications for improvement of smoking habit.

Effects of tobacco nitrate content on free radical levels in mainstream smoke.

Tobacco smoke free radicals play an important role in the development of smoking related adverse health effects. We previously reported that gas phase (GP) radicals vary greatly by cigarette brand and tobacco variety and are highly correlated with levels of NNK in smoke. Since NNK production in tobacco is dependent on nitrate, we proposed that GP radical production may also be associated with tobacco nitrate content. To test this, we examined the relationship between intrinsic nitrate levels in 15 individual tobacco types and the levels of free radicals delivered in mainstream smoke from cigarettes produced from these tobaccos. Intrinsic nitrate levels varied >250-fold among the tobacco types, ranging from <0.1 mg/g tobacco in the Bright Leaf types to 24.1 ± 0.4 mg/g in Light Fire Cured Virginia tobacco. Among the tobacco types tested, GP radicals were highly correlated with nitrate levels (r = 0.96, p < 0.0001). To investigate nitrate-specific changes to free radical production during smoking, different concentrations of exogenous sodium nitrate were added to unsmoked shredded leaves of 4 different tobacco types (Bright Leaf Sweet Virginia, American Virginia, Semi-Oriental 456, and reconstituted). Nitrate addition resulted in dose-dependent increases in GP radicals in the corresponding smoke, supporting our hypothesis that intrinsic nitrate levels are responsible for GP radical production in cigarette smoke. We also observed increases in NNK levels as a function of added nitrate that varied significantly among the 4 tobacco types tested, implying that other tobacco-type related factors may be impacting nicotine nitrosation during pyrolysis. Altogether, these findings have identified tobacco nitrate as a key factor in the production of GP radicals, but to a lesser extent with PP radicals, as well as NNK during combustion and highlight its potential implication as a target for regulation.

Development of cigarette filter with biomass-based activated carbon for naphthalene removal from mainstream smoke

Naphthalene is one of the concentrated polycyclic aromatic hydrocarbons (PAHs) in cigarette mainstream smoke. Accumulated exposure to naphthalene causes various health problems such as irritation, gastrointestinal problems, neurological symptoms, renal problems, hematological disorders, and cancer. This study aimed to develop a biomass-based activated carbon from dried hemp (Cannabis sativa) stem for the removal of naphthalene from cigarette mainstream smoke. Activated carbon was prepared at an activation temperature of 350 to 550 °C using phosphoric acid as the activating agent. The elemental composition, porous texture, and surface functional groups of the activated carbon were determined by elemental analysis, nitrogen adsorption measurements, and temperature-programmed desorption (TPD). We found that the specific surface area and oxygen-containing functional groups of the carbon facilitate naphthalene adsorption where 77–96% of the naphthalene was removed from cigarette mainstream smoke. In addition, a computational investigation with a semiempirical PM7 calculation has provided the adsorption energies of naphthalene on oxygen-containing functional groups. As expected, the carboxyl group showed high efficiency in naphthalene adsorption, which is consistent with the experimental results.

Effects of smoking on the tissue regeneration-associated functions of human endometrial stem cells via a novel target gene SERPINB2

BackgroundSmokers directly inhale mainstream cigarette smoke, which contains numerous known and potential toxic substances, and thus, smoking is expected to have broad harmful effects that cause tissue injury and dysfunction. Interestingly, many studies have suggested that the recent decline in female fertility and increased rate of spontaneous abortion could be associated with increased smoking rates. Indeed, women that smoked for 10 years or more were reported to have a ~ 20% higher infertility rate than women that had never smoked. However, the reasons for the underlying harmful aspects of smoking on female fertility remain a matter of debate. Importantly, a previous study revealed that resident endometrial stem cell deficiency significantly limits the cyclic regeneration potential of endometrium, which, in turn, decreases successful pregnancy outcomes. In this context, we postulated that exposure to mainstream cigarette smoke extracts might decrease female fertility by inhibiting the functions of resident endometrial stem cells.MethodsWe investigated whether cigarette mainstream smoke exposure directly inhibits various tissue regeneration-associated functions of endometrial stem cells, such as self-renewal, migration, pluripotency, and differentiation capacity in vitro. Next, we determined whether SERPINB2 mediates cigarette smoke-induced suppressive effects on various tissue regeneration-associated functions by depleting SERPINB2 expression with specific shRNA targeting SERPINB2. Mice were injected intraperitoneally with low (0.5 mg/kg) or high (1 mg/kg) doses of cigarette smoke extract (10 times for two weeks), and endometrial stem cells were then isolated from mice uterine tissues.ResultsWe found that exposure to cigarette smoke extracts remarkably suppressed various tissue regeneration-associated functions of endometrial stem cells, such as self-renewal, migration, multilineage differentiation ability, and pluripotency in vitro and in vivo by activating the SERPINB2 gene. Indeed, cigarette smoke-induced inhibitory effects on various endometrial stem cell functions were significantly abolished by SERPINB2 knockdown.ConclusionsThese findings provide valuable information on the harmful effects of cigarette smoking on resident endometrial stem cells and hopefully will facilitate the developments of promising therapeutic strategies for subfertile or infertile women that smoke cigarettes.

A Bioprinted Bruch's Membrane for Modeling Smoke-Induced Retinal Pigment Epithelium Degeneration via Hybrid Membrane Printing Technology.

The retinal pigment epithelium (RPE) not only forms the outer blood-retinal barrier (oBRB) but also plays a multifunctional role in the ocular system. The loss of this epithelium leads to serious diseases resulting in vision impairment. No effective treatment is available for the repair of RPE damage. A functional in vitro RPE model that allows the recapitulation of oBRB-related pathophysiological responses is lacking. Here, a hybrid membrane printing technology is developed to fabricate cellular monolayers on the basement membrane to mimic human Bruch's membrane (BM). Using this technology, in vitro oBRB model containing the RPE monolayer on the printed BM with stable mechanical properties and fibril diameter similar to that of natural BM is developed. Compared to traditional collagen bioink, BM-based bioink significantly promotes RPE functions in vitro. Finally, smoking-like conditions are exposed to the model to recapitulate the absorption of mainstream cigarette smoke which is known as one of the risk factors for the disease progression. RPE function is damaged due to oxidative stress. Furthermore, the versatility of the model as a drug-testing platform is confirmed by the suppression of oxidative stress via antioxidants. This technology shows potential for fabricating a functional oBRB model that reflects patient conditions.

Exposure to Cigarette Smoke Impedes Human Osteoblast Differentiation Independently of Nicotine.

Tobacco smoking has been implicated in an array of adverse health outcomes, including those that affect adult bone. However, little is known about the impact of tobacco products on developing bone tissue as it develops in the embryo. Here, human embryonic stem cells were differentiated into osteoblasts in vitro and concomitantly exposed to various concentrations of smoke solutions from two conventional, one additive-free and two harm-reduction brands of cigarettes. Differentiation inhibition was determined by calcium assays that quantified matrix mineralization and compared to the cytotoxicity of the tobacco product. Exposure to mainstream smoke from conventional and additive-free cigarettes caused no inhibition of cell viability or mineralization, while sidestream smoke (SS) concentration-dependently produced cell death. In contrast, mineralization was inhibited only by the highest mainstream concentration of harm-reduction smoke solution. Additionally, sidestream smoke solution from the harm-reduction cigarettes impeded calcification at concentrations lower than those determined to be cytotoxic for conventional products. Sidestream smoke impaired in vitro osteogenesis at subtoxic concentrations. In addition, though often perceived as safer, smoke from harm-reduction cigarettes was more potent in inhibiting in vitro osteogenesis than smoke from conventional cigarettes. This study adds to a growing list of adverse outcomes associated with pre-natal tobacco exposure. Specifically, in vitro exposure to tobacco products interfered with osteogenic differentiation of human embryonic stem cells, a well-established surrogate model for human embryonic bone development. Contrasting a diverse array of tobacco products unveiled that sidestream smoke was generally more developmentally osteotoxic than mainstream smoke and that harm-reduction products may not be less harmful than conventional products, adverse effects that were seemingly independent of nicotine.

Trends in Analytical Methods for Analysis of Tobacco Products: An Overview

Abstract. Currently, due to the strong trends in policies to reduce smoking, several concerns related to quality control and regulation of tobacco products have been raised, as a great variety of products are available to consumers in the market. Considering that development of robust and selective analytical methods for tobacco and derivative products has been an important and necessary task over the years, however laborious, due to the complexity of the matrices. This work presents an overview of advances, innovation, and trends in analytical methods, focused on the chromatographic analysis of nicotine, tobacco-specific nitrosamines (TSNAs), humectants, pesticides, polycyclic aromatic hydrocarbons (PAHs) and sugars in different tobacco products. Gas chromatography (GC) coupled with different detectors is widely used to analyze nicotine, humectants and TSNAs in tobacco leaves and derivative products, while liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been frequently used to analyze TSNAs in mainstream cigarette smoke and smokeless tobacco products. Pesticides were mainly analyzed in tobacco using GC or LC coupled with mass detector (MS), while PAHs and sugars were generally analyzed in tobacco and smoke using LC-MS and GC-MS techniques, respectively. In addition to already established methods and despite the lack of a worldwide standardization of methods, significant efforts have been made to develop analytical procedures for a wide variety of tobacco products, with a broad range of innovative chromatographic methods available. In this sense, a potential trend is the possibility of simultaneous determination of multiple components aiming to reduce the analysis time. The present study examined the main works that developed or improved analytical methods for identifying and quantifying priority compounds in different tobacco matrices and aims to contribute to future research with this objective, in addition, to promoting standardization of technical terms used in this analytical area. It is noteworthy that some of the methods mentioned here have not been validated and further studies are needed in order to obtain reproducible analytical methods for regulatory purposes.&#x0D; Keywords: chromatography analyses; tobacco leaves; cigarette; cigarette smoke; smokeless tobacco products.&#x0D;

210Po and 210Pb content in the smoke of Heated Tobacco Products versus Conventional Cigarette smoking

210Po is a radioactive component of conventional cigarette tobacco smoke and is a recognized carcinogen. Despite the expanding market of heated tobacco products, no data are available on the activity of 210Po in the smoke of IQOS Heets cigarette. We determined the 210Po activity in the mainstream smoke of thirteen cigarette brands available on the Swiss market using a smoking machine and compared the results to the 210Po activity measured in the mainstream smoke of the IQOS system. In addition, we measured the 210Po and 210Pb loss on heating after uniform heating from 50 to 600 °C for several cigarette brands and the Heets cigarettes. 13.6 ± 4.1% of 210Po activity was found in the mainstream smoke in conventional cigarette smoking (7% for 210Pb). This dropped to 1.8 ± 0.3% in the mainstream smoke of IQOS Heets. Conversely, when the tobacco was heated uniformly at 330 °C, a loss of 210Po of more than 80% was observed for all type of cigarettes. Apparently, IQOS significantly reduced the 210Po and 210Pb activities in the mainstream smoke. However, our results show that only 15% of the Heets tobacco reaches 330 °C with IQOS. While IQOS reduces the 210Po and 210Pb activities in the mainstream smoke compared to conventional cigarettes, it only heats a marginal fraction of the tobacco present in the Heets cigarette. Because smoking is an addiction (mostly due to nicotine), IQOS could possibly deliver an unsatisfactory dose of nicotine to a Heets cigarette smoker, as most of the tobacco is left unaltered.

A Comprehensive Analysis of Inorganic Ions and Their Selective Removal from the Reconstituted Tobacco Extract Using Electrodialysis.

Many tobacco stalks, dust, and fines are discharged in the tobacco industry, rich in inorganic minerals ions and nicotine salts. The high salinity and nicotine salts are challenging to be addressed by traditional treatment and are a severe threat that ought to be overcome. Thus, proper techniques can regenerate the tobacco stalks into reconstituted tobacco flakes used as cigarette filler. The electrodialysis process has been a viable approach to removing the inorganic ingredients in wastewater. We studied concentration, pH, and co-related influences with the nicotine and sugar/nicotine contents on the desalination performance. The results show that the inorganic ions such as Cl−, K+, Ca2+, and Mg2+ ions were successfully removed. When the feed concentration ranges from 3 to 15%, the removal ratio of the K+ ions is higher than Ca2+ and Mg2+ ions. As we reported previously, the K+ and Ca2+ ions are unfavorable for the total particulate matter emission but beneficial to decreasing the HCN delivery in mainstream cigarette smoke. Selective ED is a robust technology to reduce the harmful component delivery in cigarette smoke.