Solanum pseudocapsicum vs Capsicum annum; comparative phenolics profiling using green ultrasonic extraction and UHPLC analysis.

BACKGROUND: Solanum pseudocapsicum (PC) and Capsicum annum (CA) belongs to the family of Solanaceae. CA have been reported a rich source of phenolics whereas, the phenolics content of GA (gallic acid), SC (scopoletin), RA (rosmarinic acid), and RV (resveratrol) are yet to be reported for the PC-fruit. This study comparatively evaluates the phenolics profile for different parts (seeds and skin) and colors (green and red) of the PC- and CA-fruits using the green solvents of ethanol (ET), acetone (AC), water (H2O), and different combinations of these solvents. METHODOLOGY: Ultrasonics extraction (US) and UHPLC analysis were employed for phenolics evaluation. RESULTS: The USMD (method development) revealed the highest extract yield of 62 mg/100 mg for the PC-skin in ET:AC (70:30) solvent whereas, more phenolics (ppm) were observed for PC-seeds in ET:AC (50:50) solvent, particularly the SC (29.46) and GA (16.92). The UHPLCMDMV exhibited significant accuracies (100.70-114.14 %) with r2-values (0.9993-0.9997) in the linearity range of 1-200 ppm. The USMV (method validation) in PC- and CA-fruit parts and colors revealed more extract yields for the red skin part of the PC- (180.5 mg) and CA-fruit (126.2 mg). The phenolics were seen more in the green seeds of the PC-fruit (ppm); SC (276), GA (147.36), RV (28.54), and RA (23.87) followed by the green PC-skin, and red/green CA-seeds. The statistical models of mean differences, ANOVA, and Pearson's correlation showed significant differences for the PC-fruit parts (seeds and skin) and colors (red and green) vs extract yield and phenolics content (P = 0.05). CONCLUSION: PC-and CA-fruits were successfully evaluated where the seeds for the green fruits exhibited more phenolics amount.

Evaluation of the in vitro schistosomicidal, leishmanicidal, and trypanocidal activities of the capsaicin metabolite, Capsicum frutescens, and Capsicum baccatum extracts and of their analysis of the main constituents by HPLC/UV and CG/MS.

Neglected tropical diseases are significant causes of death and temporary or permanent disability for millions living in developing countries. Unfortunately, there is no effective treatment for these diseases. Thus, this work aimed to conduct a chemical analysis using HPLC/UV and GC/MS to identify the major constituents of the hydroalcoholic extracts of Capsicum frutescens and Capsicum baccatum fruits, evaluating these extracts and their constituents' schistosomicidal, leishmanicidal and trypanocidal activities. The results obtained for the extracts of C. frutescens are better when compared to those obtained for C. baccatum, which can be related to the different concentrations of capsaicin (1) present in the extracts. The lysis of trypomastigote forms results for capsaicin (1) led to a significant value of IC50 = 6.23 µM. Thus, the results point to capsaicin (1) as a possible active constituent in these extracts.

Menthol and Other Flavor Chemicals in Cigarettes from Vietnam and the Philippines.

INTRODUCTION: Tobacco product flavors can increase product appeal, adolescent initiation and experimentation, and difficulty quitting. Flavored tobacco products are not restricted in Vietnam or the Philippines despite the high smoking prevalence among those 15 years of age and older (24% and 23%, respectively). There are no published reports to our knowledge on the levels of flavor chemicals in the cigarettes sold in these two countries. METHODS: Cigarettes were purchased in Vietnam (32 brand variants) and the Philippines (19 brand variants) during 2020. Chemical analyses gave the mg/filter, mg/rod, and mg/stick (= mg/(filter + rod)) values for 180 individual flavor chemicals. Values were calculated for menthol, clove-related compounds, and "other flavor chemicals" (OFCs). RESULTS: Five flavor groupings were found among the brand variants purchased in Vietnam: menthol + OFCs (n = 15), OFCs only (n = 8), nonflavored (n = 7), menthol + OFCs with a clove flavorant (n = 1) and menthol only (n = 1). Three flavor groupings were found among the brand variants purchased in the Philippines: menthol + OFCs (n = 10), nonflavored (n = 5), and menthol only (n = 4). CONCLUSIONS: A range of flavored cigarette products are being offered by tobacco companies in Vietnam and the Philippines, presumably to maximize cigarette sales. Regulation of flavor chemicals should be considered in these two countries. IMPLICATIONS: Article 9 of the WHO Framework Convention on Tobacco Control (FCTC), ratified by both Vietnam and the Philippines, states that "there is no justification for permitting the use of ingredients, such as flavoring agents, which help make tobacco products attractive." Flavors increase product appeal, adolescent initiation and experimentation, and difficulty quitting. These analyses found that cigarettes purchased in Vietnam and the Philippines contained menthol and other flavor chemicals. Tobacco companies are offering multiple flavor chemical profiles and nominally nonflavored versions in these countries; regulation of flavor chemicals should be considered in these two countries.

What is the impact of e-cigarettes on periodontal stem cells as revealed by transcriptomic analyses?

DESIGN: The research used an in vitro cell exposure model and multi-omics integration of transcriptome and epigenome profiling to compare the molecular effects of e-cigarettes and tobacco smoke on dental stem cells. AIM: The study aimed to compare the effects of e-cigarette and tobacco smoke on periodontal stem cells using a multi-omics approach to understand gene regulation. METHODS: This research studied primary human gingival mesenchymal stem cells (GMSCs) and periodontal ligament stem cells (PDLSCs) obtained from healthy donors. The cells were subjected to tobacco smoke, e-cigarette aerosol (both tobacco and menthol flavors), e-cigarette liquid (both tobacco and menthol flavors), or untreated conditions using an in vitro exposure system. RNA sequencing and bioinformatics analysis were used to profile the transcriptome and identify differential gene expression. Additionally, chromatin immunoprecipitation sequencing (ChIP-seq) was used to conduct genome-wide histone modification mapping for H3K27me3. Transcriptome profiling was combined with histone modification characterization to understand gene regulatory mechanisms. The study compared the effects of smoke versus e-cigarette, aerosol versus liquid exposure, and tobacco versus menthol flavor on gene expression and epigenetic landscapes in the two oral stem cell populations. RESULTS: The use of tobacco smoke caused damage to the DNA and nucleus in GMSCs, as well as mitochondrial dysfunction in PDLSCs. Regarding e-cigarettes, the aerosol and liquid affected non-coding RNA expression differently. The chemokine CXCL2 was found to be downregulated by aerosol but upregulated by liquid in GMSCs. An integrative analysis revealed that the upregulation of CXCL2 caused by e-liquid involved reduced H3K27me3 and activation of distal enhancers. On the other hand, aerosol exposure maintained H3K27me3 levels, while direct e-liquid exposure resulted in genome-wide reductions in H3K27me3, particularly in enhancer regions. Overall, the specific delivery methods and components of e-cigarettes caused unique changes in the transcriptome and epigenome of oral stem cells. CONCLUSIONS: E-cigarettes affect oral stem cells differently than tobacco smoke. Their aerosol and liquid have varying impacts on gene expression and regulatory landscapes in oral cells. Multi-omics approaches are important to understanding the molecular changes caused by e-cigarette components. This can help with toxicological assessments and determine their impact on periodontal health. Transcriptome and epigenome profiling are powerful tools to examine the unique molecular mechanisms involved in cellular responses to e-cigarettes.

Detection of the Synthetic Coolant Menthone 1,2-Glycerol Ketal in an e-Liquid and in Electronic Waterpipe Aerosols Therefrom.

The presence and toxicological risks of synthetic coolants in electronic nicotine delivery systems (ENDS) have not been thoroughly studied. We identified the synthetic coolant menthone 1,2-glycerol ketal (MGK) in a menthol-flavored e-liquid at a concentration of ∼170 µg/mL. We also detected MGK in aerosols resulting from heating the e-liquid with an electronic waterpipe. MGK was initially detected in the e-liquid by two-dimensional gas chromatography-time-of-flight mass spectrometry. To avoid potential analytical artifacts that could result from heating samples in the injection port of the gas chromatograph, quantitation of MGK in the e-liquid was accomplished using a liquid chromatography-tandem mass spectrometry method. Following recent reports identifying other synthetic coolants in e-liquids, these results add knowledge about inhalation exposures from ENDS use and suggest the importance of future research to study the potential inhalation toxicity related to the use of MGK-containing e-liquids in ENDS devices. Furthermore, the results demonstrate the ability to quantify ketals in e-liquids using liquid chromatography methods.

The mutation of CaCKI1 causes seedless fruits in chili pepper (Capsicum annuum).

KEY MESSAGE: The seedless mutant tn-1 in chili pepper is caused by a mutation in CaCKI1 (CA12g21620), which encodes histidine kinase involving female gametophyte development. An amino acid insertion in the receiver domain of CaCKI1 may be the mutation responsible for tn-1. Seedlessness is a desirable trait in fruit crops because the removal of seeds is a troublesome step for consumers and processing industries. However, little knowledge is available to develop seedless chili peppers. In a previous study, a chili pepper mutant tn-1, which stably produces seedless fruits, was isolated. In this study, we report characterization of tn-1 and identification of the causative gene. Although pollen germination was normal, confocal laser microscopy observations revealed deficiency in embryo sac development in tn-1. By marker analysis, the tn-1 locus was narrowed down to a 313 kb region on chromosome 12. Further analysis combined with mapping-by-sequencing identified CA12g21620, which encodes histidine kinase as a candidate gene. Phylogenetic analysis revealed CA12g21620 was the homolog of Arabidopsis CKI1 (Cytokinin Independent 1), which plays an important role in female gametophyte development, and CA12g21620 was designated as CaCKI1. Sequence analysis revealed that tn-1 has a 3-bp insertion in the 6th exon resulting in one lysine (K) residue insertion in receiver domain of CaCKI1, and the sequence nearby the insertion is widely conserved among CKI1 orthologs in various plants. This suggested that one K residue insertion may reduce the phosphorylation relay downstream of CaCKI1 and impair normal development of female gametophyte, resulting in seedless fruits production in tn-1. Furthermore, we demonstrated that virus-induced gene silencing of CaCKI1 reduced normally developed female gametophyte in chili pepper. This study describes the significant role of CaCKI1 in seed development in chili pepper and the possibility of developing seedless cultivars using its mutation.

Protective effects of Capsicum fruits and their constituents on damage in TNF-α-stimulated human dermal fibroblasts.

BACKGROUND: Antioxidant and anti-inflammatory effects of natural products on skin cells have been proved to be effective in improving skin damage. Capsicum species contain capsaicinoids that have antioxidant and anti-inflammatory properties, and various subspecies are cultivated. In this study, the effects of four Capsicum fruits and major constituents on oxidative stress and inflammatory reactions were measured using human dermal fibroblasts (HDFs) to verify their effects on skin damage. RESULTS: The inhibitory effects of nitric oxide (NO), reactive oxygen species (ROS), and prostaglandin E2 (PGE2 ) by cucumber hot pepper, red pepper (RDP), Shishito pepper (SSP), and Cheongyang pepper were determined in HDFs. RDP and SSP inhibited the production of NO, ROS, and PGE2 in tumor necrosis factor-alpha-stimulated HDFs. Additionally, SSP seeds restored tumor necrosis factor-alpha-induced increase in matrix metalloproteinase-1 and decreased procollagen I α1 (COLIA1). In high-performance liquid chromatography analysis of the capsaicinoids capsaicin (CAP) and dihydrocapsaicin (DHC), CAP was detected at a higher level than DHC in the peel and seeds of all four types of Capsicum fruits, and the total amount of capsaicinoids was the highest in SSP. CAP and DHC, which are major constituents of Capsicum fruits, also inhibited NO, ROS, and PGE2 and restored matrix metalloproteinase-1 and procollagen I α1. CONCLUSION: RDP and SSP were shown to have a significant protective effect on skin damage, including oxidative stress, inflammatory reactions, and reduction of collagens. Capsaicinoids CAP and DHC were proved as active constituents. This research may provide basic data for developing Capsicum fruits as ingredients to improve skin damage, such as inflammation and skin aging. © 2022 Society of Chemical Industry.

Separation of Nonivamide and Capsaicin by Ultrahigh-Performance Liquid Chromatography for Routine Analysis of Capsaicinoids in Capsicum Products.

BACKGROUND: Baseline separation of nonivamide (NON) and capsaicin (CAP) has not been achieved by using the existing liquid chromatography (LC) methods for the capsaicinoid analysis. This could lead to large errors in the determination of capsaicinoids for capsicum products. OBJECTIVE: The development of an ultrahigh-performance liquid chromatography (UHPLC) method that simultaneously separates NON and CAP as well as other capsaicinoids for the routine analysis of capsaicinoids in capsicum products. METHOD: Capsaicinoids were separated on a Waters CORTECSTM T3 Column (2.1 mm i.d. × 150 mm, 1.6 µm particle size) that was maintained at 45°C on a UHPLC system with a 3-step gradient elution using a binary mobile phase system consisting of water and acetonitrile. Florescence detection was set at 280 nm excitation wavelength and 325 nm emission wavelength. RESULTS: The UHPLC method was able to simultaneously separate NON and CAP, with a minimum resolution of 1.5, as well as other seven capsaicinoids with a total run time of 27 min. Method selectivity, robustness, accuracy, and precision were evaluated, and excellent performance was achieved. CONCLUSIONS: The UHPLC method for NON and CAP and other seven capsaicinoids has been successfully developed and found suitable for the routine analysis of capsaicinoids. HIGHLIGHTS: For the first time, NON and CAP are well separated (Rs >1.5) in a 27 min LC separation. This UHPLC method offers a suitable solution for the determination of nine capsaicinoids in QC labs.

Computational modeling method to estimate secondhand exposure potential from exhalations during e-vapor product use under various real-world scenarios.

Potential secondhand exposure of exhaled constituents from e-vapor product (EVP) use is a public health concern. We present a computational modeling method to predict air levels of exhaled constituents from EVP use. We measured select constituent levels in exhaled breath from adult e-vapor product users, then used a validated computational model to predict constituent levels under three scenarios (car, office, and restaurant) to estimate likely secondhand exposure to non-users. The model was based on physical/thermodynamic interactions between air, vapor, and particulate phase of the aerosol. Input variables included space setting, ventilation rate, total aerosol amount exhaled, and aerosol composition. Exhaled breath samples were analyzed after the use of four different e-liquids in a cartridge-based EVP. Nicotine, propylene glycol, glycerin, menthol, formaldehyde, acetaldehyde, and acrolein levels were measured and reported based on a linear mixed model for analysis of covariance. The ranges of nicotine, propylene glycol, glycerin, and formaldehyde in exhaled breath were 89.44-195.70 µg, 1199.7-3354.5 µg, 5366.8-6484.7 µg, and 0.25-0.34 µg, respectively. Acetaldehyde and acrolein were below detectable limits; thus, no estimated exposure to non-EVP users is reported. The model predicted that nicotine and formaldehyde exposure to non-users was substantially lower during EVPs use compared to cigarettes. The model also predicted that exposure to propylene glycol, glycerin, nicotine and formaldehyde among non-users was below permissible exposure limits.

Composition of aerosols from thermal degradation of flavors used in ENDS and tobacco products.

Aim: The cardiovascular toxicity of unheated and heated flavorants and their products as commonly present in electronic cigarette liquids (e-liquids) was evaluated previously in vitro. Based on the results of in vitro assays, cinnamaldehyde, eugenol, menthol, and vanillin were selected to conduct a detailed chemical analysis of the aerosol generated following heating of each compound both at 250 and 750 °C. Materials and Methods: Each flavoring was heated in a drop-tube furnace within a quartz tube. The combustion atmosphere was captured using different methods to enable analysis of 308 formed compounds. Volatile organic compounds (VOCs) were captured with an evacuated Summa canister and assayed via gas chromatography interfaced with mass spectrometry (GC-MS). Carbonyls (aldehydes and ketones) were captured using a 2,4-dinitrophenylhydrazine (DNPH) cartridge and assayed via a high-performance liquid chromatography-ultra-violet (HPLC-UV) assay. Polyaromatic hydrocarbons (PAHs) were captured using an XAD cartridge and filter, and extracts were assayed using GC-MS/MS. Polar compounds were assayed after derivatization of the XAD/filter extracts and analyzed via GC-MS. Conclusion: At higher temperature, both cinnamaldehyde and menthol combustion significantly increased formaldehyde and acetaldehyde levels. At higher temperature, cinnamaldehyde, eugenol, and menthol resulted in increased benzene concentrations. At low temperature, all four compounds led to higher levels of benzoic acid. These data show that products of thermal degradation of common flavorant compounds vary by flavorant and by temperature and include a wide variety of harmful and potentially harmful constituents (HPHCs).

Comparison of Carcinogen Biomarkers in Smokers of Menthol and Nonmenthol Cigarettes: The 2015-2016 National Health and Nutrition Examination Survey Special Sample.

BACKGROUND: The US FDA announced its commitment to prohibiting menthol as a characterizing flavor in tobacco. The relationship between cigarette menthol and exposure to toxic substances in mainstream tobacco smoke is not well characterized. METHODS: Data from the National Health and Nutrition Examination Survey (NHANES) 2015 to 2016 special sample were used to study markers of 26 harmful and potentially harmful constituents (HPHC) in tobacco smoke. These include urine metabolites of polycyclic aromatic hydrocarbons (PAH), volatile organic compounds (VOC), and heavy metals in exclusive menthol (n = 162) and nonmenthol (n = 189) cigarette smokers. Urine metabolites of 7 PAHs, 15 VOCs, and 4 heavy metal biomarkers were compared by menthol status. Multivariable analyses were conducted on creatinine-adjusted concentrations. RESULTS: There were no significant differences in cotinine levels or in 22 of 26 HPHCs. Among the urine metabolites of PAHs, the levels of 1-hydroxyphenanthrene were about 16% lower in menthol smokers. Among the urine metabolites of VOCs, menthol cigarette smokers presented significantly lower concentrations of acrylamide, N,N-dimethylformamide, and acrylonitrile. Menthol and nonmenthol smokers presented similar levels of heavy metals. Menthol did not affect the levels of cotinine and the nicotine metabolite ratio in urine. CONCLUSIONS: Menthol and nonmenthol cigarettes deliver similar levels of most HPHCs. IMPACT: Findings on toxicity are similar for menthol and nonmenthol cigarettes.

'Menthol-Plus': a major category of cigarette found among 'concept' descriptor cigarettes from Mexico.

BACKGROUND: Tobacco companies are offering cigarettes with 'concept' descriptor names that suggest sensation and/or flavour properties (eg, Marlboro 'Velvet Fusion'). Little has been known about the identities and levels of flavour chemicals in such cigarettes. METHODS: Thirty-three filter cigarette variants from 27 packs (including two sampler packs with four variations each) from Canada and Mexico were analysed (rod + filter) for 177 flavour chemicals plus triacetin, a filter plasticiser and possible flavourant. Five brands of US mentholated filter cigarettes were also analysed. RESULTS: Twenty-seven of the 33 cigarettes (all were Mexican variants) were categorised as 'menthol-plus': significant menthol (3.0-11.9 mg/cigarette), plus varying amounts (0.32-3.4 mg/cigarette) of total other flavour chemicals (TOFCs) (excludes triacetin). For 10 of the 27, TOFCs >1.0 mg/cigarette. For 7 of the 27, the TOFCs profile was categorised as containing total fruit flavour compounds (TFFCs) >1.0 mg/cigarette. One Mexican variant was categorised as 'menthol-only' (TOFCs ≤0.15 mg/cigarette). All menthol-plus and menthol-only cigarettes contained one or two optional-crush capsules in their filters (crushed prior to analysis). All five Canadian brand variants were 'non-flavoured'. All five US brand variants were 'menthol-only'. CONCLUSIONS: All but one of the 'concept' descriptor cigarettes from Mexico were 'menthol-plus'. While the Canadian cigarettes complied with Canada's flavour chemical ban, concept descriptors on the packs may increase appeal. Given the scale of the problem posed by menthol alone, health officials seeking to decrease the appeal of smoked tobacco should examine the extent to which 'concept descriptor' cigarettes using 'menthol-plus' flavour profiling together with artful descriptors are furthering the problem of smoked tobacco.

Minty aroma compounds in red wine: Development of a novel automated HS-SPME-arrow and gas chromatography-tandem mass spectrometry quantification method.

A novel automated method was developed for the quantitative determination of nine terpenoids that could contribute to the minty notes of red wine bouquet. The method couples headspace SPME-Arrow extraction with GC-MS/MS analysis. PDMS/DVB fiber was chosen for the extraction and an ionization energy of 30 eV permitted to optimize the analyte detection. The optimal sample preparation consists of a two-fold dilution of the wine sample with addition of 4 g of sodium chloride while the most suitable extraction conditions take place at 50 °C for 1 h. The method shows good linearity, intraday variations between 2 and 25%, interday variations between 7 and 23% and recoveries between 80 and 119%. The method exhibits the required low detection (between 3 and 60 ng/L) and quantification (between 6 ng/L and 200 ng/L) limits. These limits have permitted the quantification of the pool of minty terpenoids in fourteen red Bordeaux wines.

Health Care Utilization of Menthol and Non-menthol Cigarette Smokers.

INTRODUCTION: To study the association between health care utilization and menthol cigarette use and whether the association differed between African American (AA) and non-AA smokers. METHODS: We analyzed the three most recent 2005, 2010, and 2015 National Health Interview Survey Cancer Control Supplements. After incorporating propensity score weights adjusting for observed differences between menthol and non-menthol users, we estimated Zero-Inflated Poisson models on hospital nights, emergency department visits, doctor visits, and home visits as a function of menthol use status and other covariates separately for current cigarette smokers and recent quitters (former smokers quitting cigarette smoking ≤ 4 years). RESULTS: Although current menthol smokers smoked fewer cigarettes per day than current non-menthol smokers, they did not differ from current non-menthol smokers in health care utilization. Among recent quitters, those who used to smoke menthol cigarettes had higher odds of having hospital nights than those who used to smoke non-menthol cigarettes. However, we did not find any significant association between menthol use and other health care utilization-emergency department visits, doctor visits, and home visits-among recent quitters. Moreover, compared with non-AA recent quitters, AA recent quitters had higher odds of having home visits, but fewer home visits, if they used to smoke menthol cigarettes. CONCLUSION: Menthol use was associated with greater hospitalization among recent quitters, and the association between home visits and menthol use differed between AA and non-AA recent quitters. IMPLICATIONS: This is the first study that used econometric models to study the association between health care utilization and menthol cigarette use and examine whether the association differed between AA and non-AA smokers. Our study found health care utilization did not differ by menthol use status for current smokers, although current menthol smokers smoked fewer cigarettes per day than current non-menthol smokers. However, we found menthol use was associated with higher odds of having hospital nights for recent quitters. We also found AA recent quitters had a different association between home visits and menthol use compared with non-AA recent quitters.

Real-time breath analysis of exhaled compounds upon peppermint oil ingestion by secondary electrospray ionization-high resolution mass spectrometry: technical aspects.

Breath analysis by secondary electrospray ionization high-resolution mass spectrometry (SESI-HRMS) has potential for clinical diagnosis and drug monitoring. However, there is still a lack of benchmarking data that shows the capability of this technique and allows comparability with other breath analysis techniques. In this regard, the goal of this study was the identification of volatile compounds upon ingestion of a specific peppermint oil capsule to get benchmark data for real-time breath analysis with SESI-HRMS. This was done in the framework of a consortium set up by the International Association of Breath Research (IABR), aimed at comparing several analytical instruments for breath analysis. Breath temporal profiles of two subjects were analyzed with SESI-HRMS before and after ingestion of a peppermint oil capsule. The measurements were performed at two different locations using identical SESI-HRMS platforms to allow for comparability and benchmarking. Remarkably, along with the four major compounds (monoterpenes/cineole, menthone, menthofuran and menthol) reported by other members of the consortium, we detected 57 additional features significantly associated (ρ > 0.8) with the peppermint oil capsule, suggesting that this relatively simple intervention might trigger a more complex metabolic cascade than initially expected. This observation was made on both sites. Additional replicate experiments for one of the subjects suggested that a core of 35-40 unique molecules are consistently detected in exhaled breath upon ingestion of the capsule. In addition, we illustrate the analytical capabilities of real-time SESI-HRMS/MS to assist in the identification of unknown compounds. The results outlined herein showcase the performance of SESI-HRMS and enable comparison with other breath analysis techniques. Along with that, they strengthen the potential of this analytical technique for non-invasive drug monitoring and clinical diagnostic purposes.

Green HPTLC and stability-indicating RP-HPLC for the assay of dextromethorphan hydrobromide and menthol in their lozenges.

Two chromatographic methods were developed for the assay of the FDA approved lozenges containing dextromethorphan hydrobromide (DXT) and menthol (MNT). The first was a green HPTLC method which uses a mobile phase of methanol-ammonia (10:0.1, v/v). The densitometric measurements of the spots which were retained at 0.28±0.01 for DXT and 0.76±0.02 for MNT was done at 210nm. The other method was RP-HPLC method with stability indicating merits at which a mixture of 20mM phosphate buffer pH 3 and acetonitrile as mobile phase in isocratic mode was used. The cited drugs were resolved in RP-HPLC method using isocratic elution using 20mM phosphate buffer: acetonitrile (65:35 v/v) with retention times of 2.21 and 3.47min for MNT and DXT, respectively and quantified using 215nm. Both methods were entirely validated and both methods were successfully able to analyze both drugs in presence of lozenges inactive ingredients. HPLC method had the advantage of being stability indicating at which resolution of the drugs from their forced degradation products was successfully attained. For HPTLC method, both drugs showed reasonable RF values when compared to rapidly eluted MNT in RP-HPLC; also it was more environmentally friendly than RP-HPLC as it used solvents which are less toxic and greener.

Differences in Smoking Topography and Subjective Responses to Smoking Among African American and White Menthol and Non-Menthol Smokers.

INTRODUCTION: It has been suggested that menthol increases exposure to harmful elements of smoking and makes smoking more rewarding, easier to initiate, and harder to quit. Isolating the direct effects of menthol is challenging as African American (AA) race and menthol preference are highly overlapping. This study evaluated smoking behavior and subjective responses among a balanced sample of AA and white menthol and non-menthol smokers. In addition, smoking topography (ST) was compared to naturalistic smoking (NS) and interactions with menthol and race were explored. AIMS AND METHODS: Smokers (N = 100) smoked and rated their preferred brand of cigarettes via ST or NS during two laboratory visits (counterbalanced). RESULTS: Controlling for baseline differences among the groups (eg, nicotine dependence), menthol smokers took shorter and smaller puffs and AA smokers took longer puffs, but there were no differences in total puff volume, carbon monoxide, or other ST parameters. Menthol smokers reported greater urge reduction and lower sensory stimulation. The smoking method (ST vs. NS) had no effects on smoking behavior or exposure. Cigarettes smoked via ST were rated stronger. Differences in satisfaction based on the smoking method interacted with race and menthol status. Ratings of aversion differed by race and menthol status. CONCLUSIONS: Menthol was not associated with increased smoke exposure or reward (except for urge reduction). ST caused minimal experimental reactivity relative to NS. Additional research that isolates the effects of menthol and examines potential interactive effects with race and other variables is needed to better understand its role in smoking-related health disparities. IMPLICATIONS: Menthol and non-menthol smokers differed on some demographic variables and menthol preference was associated with greater nicotine dependence and greater urge reduction after smoking. Menthol was not associated with greater smoke exposure. Future research that investigates the unique risks associated with menthol and examines potential interactive effects with race and other related variables is warranted to better understand the role of menthol in smoking-related health disparities.

Mentholation triggers brand-specific shifts in the bacterial microbiota of commercial cigarette products.

Bacterial communities are integral constituents of tobacco products. They originate from tobacco plants and are acquired during manufacturing processes, where they play a role in the production of tobacco-specific nitrosamines. In addition, tobacco bacterial constituents may play an important role in the development of infectious and chronic diseases among users. Nevertheless, tobacco bacterial communities have been largely unexplored, and the influence of tobacco flavor additives such as menthol (a natural antimicrobial) on tobacco bacterial communities is unclear. To bridge this knowledge gap, time series experiments including 5 mentholated and non-mentholated commercially available cigarettes-Marlboro red (non-menthol), Marlboro menthol, Newport menthol box, Newport menthol gold, and Newport non-menthol-were conducted. Each brand was stored under three different temperature and relative humidity conditions. To characterize bacterial communities, total DNA was extracted on days 0 and 14. Resulting DNA was purified and subjected to PCR of the V3V4 region of the 16S rRNA gene, followed by sequencing on the Illumina HiSeq platform and analysis using the QIIME, phyloseq, metagenomeSeq, and DESeq software packages. Ordination analyses showed that the bacterial community composition of Marlboro cigarettes was different from that of Newport cigarettes. Additionally, bacterial profiles significantly differed between mentholated and non-mentholated Newports. Independently of storage conditions, tobacco brands were dominated by Proteobacteria, with the most dominant bacterial genera being Pseudomonas, unclassified Enterobacteriaceae, Bacillus, Erwinia, Sphingomonas, Acinetobacter, Agrobacterium, Staphylococcus, and Terribacillus. These data suggest that the bacterial communities of tobacco products differ across brands and that mentholation of tobacco can alter bacterial community composition of select brands. KEY POINTS: • Bacterial composition differed between the two brands of cigarettes. • Mentholation impacts cigarette microbiota. • Pseudomonas and Bacillus dominated the commercial cigarettes. Graphical abstract.

Trends and Associations of Menthol Cigarette Smoking Among US Middle and High School Students-National Youth Tobacco Survey, 2011-2018.

INTRODUCTION: Youth cigarette smoking decreased significantly over the last two decades in the United States. This study provides estimates and trends from 2011 to 2018 and factors associated with youth menthol and non-menthol smoking from 2016 to 2018. METHODS: Using data from the 2011-2018 National Youth Tobacco Surveys, past 30-day (current) menthol and non-menthol cigarette smoking were estimated for all youth (prevalence) and youth smokers (proportions). Trends were examined using Joinpoint regression, calculating the annual percent change (APC). Multivariate logistic regression analyses identified factors associated with menthol smoking. RESULTS: From 2011 to 2018, menthol cigarette smoking among current youth cigarette smokers significantly decreased from 57.3% to 45.7% (APC: -3.0%), while non-menthol (38.2% to 47.3% [APC: 2.9%]) and unknown menthol status (not sure\missing) (4.5% to 7.0% [APC: 7.1%]) significantly increased. Menthol cigarette smoking among high school, male, female, and non-Hispanic white current cigarette smokers decreased, but remained unchanged among middle school, non-Hispanic black, and Hispanic smokers. Significantly higher proportions of menthol cigarette smokers smoked on ≥20 days, ≥2 cigarettes per day, and ≥100 cigarettes in their lifetime compared to non-menthol smokers. Among current cigarette smokers, non-Hispanic blacks, Hispanics, flavored non-cigarette tobacco users, frequent smokers (≥20 days), those smoking 2-5 cigarettes per day, and those living with someone who uses tobacco had higher odds of menthol cigarette smoking. CONCLUSIONS: In 2018, nearly half of current youth cigarette smokers smoked menthol cigarettes. While menthol cigarette smoking declined from 2011 to 2018 among all youth and among youth smokers, there was no change in menthol cigarette smoking among non-Hispanic black, Hispanic, and middle school cigarette smokers. IMPLICATIONS: This study finds that overall cigarette and menthol cigarette smoking declined in youth from 2011 to 2018. However, menthol cigarette smoking among non-Hispanic black, Hispanic, and middle school youth cigarette smokers did not change. Information from this study can help inform efforts to reduce menthol cigarette smoking among US youth, particularly racial/ethnic minority populations.

High Rates of Menthol Cigarette Use Among Pregnant Smokers: Preliminary Findings and Call for Future Research.

INTRODUCTION: Smoking during pregnancy remains widespread and is causally associated with infant morbidity and mortality. Despite links between menthol cigarette use and decreased smoking cessation, little is known regarding rates or characteristics of pregnant menthol cigarette smokers. METHODS: Participants were drawn from two low-income, racially/ethnically diverse cohorts of pregnant smokers recruited from 2006 to 2015 (N = 166, Mage = 25 ± 5). Demographics, menthol cigarette use, daily cigarette use, quit status, and consecutive weeks quit were assessed by prospective interviews. Nicotine dependence was assessed using the Fagerström Test of Nicotine Dependence. Nicotine levels were assessed via saliva cotinine. RESULTS: High rates of menthol use were found in both cohorts (85% and 87%). Across both cohorts, menthol smokers were more likely to identify as racial/ethnic minorities, were less educated, and reported lower income than non-menthol smokers (ps < .03). Menthol smokers also reported fewer continuous weeks quit (8.4 vs. 14.5 weeks quit; p < .03) and a tendency toward decreased likelihood of quitting smoking over pregnancy (29% vs. 48%; p < .08) in unadjusted but not in covariate-adjusted analyses. No differences emerged in cigarettes per day, nicotine dependence or nicotine exposure. CONCLUSIONS: We found very high rates of menthol cigarette use in pregnant smokers-particularly among racial/ethnic minority and low socioeconomic status smokers-and some evidence for associations with reduced smoking cessation in pregnancy. Consideration of pregnant smokers as a uniquely vulnerable population is warranted in evaluating regulation of menthol in cigarettes. Further research is needed regarding the impact of menthol on smoking persistence in pregnancy and on maternal and infant health outcomes. IMPLICATIONS: This study highlights high rates of menthol cigarette use in pregnant women in the Northeast, United States, with evidence for higher rates of menthol use among racial/ethnic minority, less educated and low-income pregnant smokers, and preliminary evidence for associations between menthol cigarette use and reduced smoking cessation. Consideration of the effects of menthol on smoking persistence in pregnant women and on the health of their offspring is warranted in the development of regulations regarding menthol in cigarettes.