Changes in Biomarkers of Tobacco Exposure among Cigarette Smokers Transitioning to ENDS Use: The Population Assessment of Tobacco and Health Study, 2013-2015.

Limited data are available for how biomarkers of tobacco exposure (BOE) change when cigarette smokers transition to using electronic nicotine delivery systems (ENDS). Using biomarker data from Waves 1 (2013-2014) and 2 (2014-2015) of the PATH Study, we examined how mean BOE concentrations, including metabolites of nicotine, tobacco-specific nitrosamines (TSNA), polycyclic aromatic hydrocarbons (PAH), and volatile organic compounds (VOC) and metals, changed when 2475 adult smokers transitioned to using ENDS or quit tobacco products. Exclusive smokers who transitioned to dual use had a significant decrease in NNAL (4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol), but not nicotine metabolites, most PAHs, metals, or VOCs. Exclusive smokers who became dual users had significant reductions in total nicotine equivalents, NNAL, and 2CyEMA (acrylonitrile metabolite), but only in those who reduced cigarettes per day (CPD) by >=50%. Smokers who transitioned to exclusive ENDS use had significant reductions in most TSNAs, PAHs, and VOCs; however, nicotine metabolites did not decrease in dual users who became exclusive ENDS users. Smokers who quit tobacco use had significant decreases in nicotine metabolites, all TSNAs, most PAHs, and most VOCs. Cigarette smokers who became dual users did not experience significant reductions in most BOEs. Reductions were impacted by changes in CPD. However, transitioning from smoking to no tobacco or exclusive ENDS use was associated with reduced exposure to most BOEs measured. Future analyses could incorporate additional waves of PATH data and examine changes in biomarker exposure by ENDS device type and CPD.

Modes of delivery in concurrent nicotine and cannabis use ("co-use") among youth: Findings from the International Tobacco Control (ITC) Survey.

BACKGROUND: Cannabis use is more common among nicotine users than non-users. This study characterized concurrent use of nicotine and cannabis ("co-use") among 12,064 youth aged 16-19 years residing in Canada, the United States, and England in 2017. Methods: Data were from the ITC Youth Tobacco & Vaping Survey (Wave 1). Seven modes of cannabis delivery (MOD) were characterized by country of residence and past 30-day use of combusted tobacco and electronic cigarettes. Weighted multivariable regression models were fitted to assess the correlates of co-use and each cannabis MOD. Results: Seventy percent of cannabis users reported nicotine use. Co-users exhibited behavioral and demographic differences compared to exclusive users of either substance. "Smoking cannabis without tobacco" was the most popular form of use (78%). Use of nicotine-containing e-cigarettes was associated with "using an e-cigarette to vape cannabis oil/liquid" (aOR: 4.96, 95%CI: 2.23-11.06). Combustible tobacco use was associated with "smoking cannabis with tobacco in a joint/blunt" (aOR: 2.93, 95%CI: 1.89-4.56). Country-level differences were detected. Conclusions: Nicotine use is substantial among cannabis users, and associations exist between modes of delivery for both drugs. Results underscore the importance of studying cannabis and nicotine use concurrently, and the need to address the use of both substances in developing interventions for youth users.

Emerging Chemicals of Health Concern in Electronic Nicotine Delivery Systems.

Electronic nicotine delivery systems (ENDS), by virtue of their highly engineered construction (plastics, glass, e-liquids), may contain a number of emerging chemicals of concern (ECCs), including phthalates, phenolic compounds, and flame retardants. Current knowledge regarding the safety of ENDS may underestimate the health risks from ECCs. In this study, we examined the types and levels of those three groups of chemicals in the components and parts of ENDS devices, including refill liquids, tanks/cartridges, atomizers, drip tips/mouthpieces, and sealing materials. Our results suggest that phthalates were the most prevalent chemicals in all tested samples, followed by parabens and organophosphate flame retardants (OPFRs). Particularly, all measured chemicals had significantly higher detection rates in cartridges/tanks, drip tips/mouthpieces, and sealing materials in contrast to e-liquids and coil samples. Among all those three types of ENDS components, phthalates generally had the highest concentrations (0.279-3790 ng/unit) in the drip tip/mouthpiece samples, followed by the sealing materials (0.380-508.8 ng/unit) and the empty tank/cartridge samples (up to 761.7 ng/unit). For parabens, highest concentrations were observed in drip tip/mouthpiece samples (1.152-130.1 ng/unit), followed by sealing materials (0.220-30.08 ng/unit) and the tank/cartridge samples (1.794-34.24 ng/unit). For OPFRs, tris(1,3-dichloro-2-propyl) phosphate had the highest concentrations (39.40-774.1 ng/unit) in all component samples. High concentrations (20.25-260.4 ng/unit) were also observed for several OPFRs in sealing materials and drip tip/mouthpiece samples. These findings will contribute to addressing the information gaps pertinent to the presence of ECCs in ENDS and will warrant further studies for understanding the potential negative health effects and to what extent those chemicals may cause potential negative health effects when using the ENDS. The findings will also contribute to developing evidence-based standards for the regulatory control of the types and levels of ECCs in ENDS products.

Use of Electronic Cigarettes in Smoke-Free Spaces by Smokers: Results from the 2014-2015 Population Assessment on Tobacco and Health Study.

Background: Smoke-free air policies exist to protect users and nonusers from exposure to tobacco smoke. Although electronic nicotine delivery systems (ENDS) may expose passerby to nicotine and particulate matter, few US states regulate indoor use of ENDS. The purpose of this study was to investigate reported rationales for ENDS use and reported ENDS use in public smoke-free places by dual cigarette/ENDS users. Methods: A population of ENDS/cigarette co-users (n = 2051) was drawn from Wave 2 of the Population Assessment of Tobacco and Health (PATH) dataset (2014-2015). Harm reduction beliefs and cessation behavior of co-users were investigated as predictors of ENDS use in public smoke-free places using logistic regression. Results: Fifty-eight percent of dual users reported past 30-day ENDS use in public smoke-free places. Reported use of ENDS to cut down on cigarette smoking (OR: 2.38, 95% CI: 1.86, 3.05), as an alternative to quitting tobacco (OR: 1.71, 95% CI: 1.37, 2.13), or because of belief that ENDS help people to quit cigarettes (OR: 1.52, 95% CI: 1.20, 1.92) were significantly associated with increased odds of ENDS use in smoke-free places. Conclusions: Beliefs that ENDS were useful as cessation tools or posed modified risk to users and nonusers were associated with elevated odds of use ENDS in locations where conventional tobacco is prohibited. Due to limitations in the survey instrument, in-home ENDS use could not be directly assessed in this analysis. However, these self-reported findings suggest that use of ENDS in public places where cigarette use is prohibited is prevalent enough to be of concern for future regulation and enforcement efforts.

Association between Urinary Metabolite Levels of Organophosphorus Flame Retardants and Serum Sex Hormone Levels Measured in a Reference Sample of the US General Population.

Use of organophosphorus flame retardants (OPFRs) in consumer materials have led to widespread human exposure.Research is needed to examine the health effects attributable to the general population's exposure to OPFRs. Using the data from the National Health and Nutrition Examination Surveys (NHANES) (2013-2014), multiple regression analyses were performed to compare the adjusted geometric means (aGMs) of serum sex hormone by quartiles of urinary metabolites of OPFRs, including diphenyl phosphate (DPhP), bis(1,3-dichloro-2-propyl) phosphate (BDCPP), bis(2-chloroethyl) phosphate (BCEP) and dibutyl phosphate(DBuP), in children (6 - 9 years old), adolescents (10 - 19 years old) and adults(≥ 20 years old), while accounting for potential confounding factors. The aGMs of sex hormone-binding globulin increased by 36% (95% CI: 6.1 - 56.7%) in female children (p = 0.03), 44% (95%CI: 16 - 63%) in female adolescents (p = 0.010), and 22% (95%CI: 3.51 - 37%) in female adults (p = 0.025), from the 1st to 4th quartiles of the levels of DPhP, BDCPP, DBUP, respectively. The aGMs of estradiol (EST) decreased by 64% and 77% from the 1st to 4th quartiles of the DBUP levels in female children (p = 0.015) and female adolescent (p = 0.020), respectively. The aGMs of EST increased by 31% (95%CI: 3.8 - 51%) from the 1st to 4th quartiles of the DBUP levels in female adults (p = 0.031). These findings suggest that exposure to certain OPFRs is associated with the altered sex hormone levels in this sample of US population. More studies are needed to examine the mechanisms responsible for these observations.

Nicotine and Toxicant Exposure Among Concurrent Users (Co-Users) of Tobacco and Cannabis.

BACKGROUND: Smoking cannabis may potentially increase exposure to numerous toxic chemicals that are commonly associated with tobacco use. There is a paucity of data related to toxicant exposures among concurrent users of tobacco and cannabis (co-users). METHODS: Data are from the Population Assessment of Tobacco and Health Study Wave 1 Biomarker Restricted-Use Files. Analyses focused on adults who provided urine samples (N = 5859). Urine samples were analyzed for biomarkers of exposure to nicotine, tobacco-specific nitrosamines, polycyclic aromatic hydrocarbons, and volatile organic compounds. Using weighted linear regression, we compared adjusted geometric mean concentrations of 15 biomarkers between user groups of various tobacco product types according to their self-reported past 30-day cannabis use. RESULTS: Past 30-day cannabis use was similar across various types of tobacco product use subgroups (range: 13%-23%) and significantly more common compared to non-tobacco users (1.0%; p < .001). Across all groups of tobacco users, those who co-used cannabis exhibited significantly higher concentrations of the biomarker of exposure to acrylonitrile (CYMA) compared to non-cannabis users (by 39%-464%). Tobacco-cannabis co-users also showed significantly elevated levels of the biomarker of exposure to acrylamide (AAMA) compared to exclusive tobacco users, and significantly higher exposure to many polycyclic aromatic hydrocarbons (including fluorene and pyrene). CONCLUSIONS: Co-users exhibited higher concentrations for biomarkers of exposure to many combustion byproducts, compared to exclusive tobacco users. More robust measurements of cannabis use can address potential confounding in assessments of exposures to tobacco-related constituents, and potential health effects resulting from co-use. IMPLICATIONS: With disproportionately greater rates of cannabis use occurring among tobacco users, it is critical to consider how concurrent cannabis use may influence health-related outcomes among smokers. Our findings suggest potential additive toxicant exposures among co-users of tobacco and cannabis. Lack of consideration and measurement of cannabis use in assessing tobacco-related exposures may confound estimates thought to be attributable to tobacco, particularly for non-specific biomarkers. Assessing tobacco and cannabis use in tandem will allow for more precise measurement of outcomes related to one or both substances, and can provide additional information on potential health effects related to co-use.

Concurrent Quantification of Emerging Chemicals of Health Concern in e-Cigarette Liquids by High-Performance Liquid Chromatography-Tandem Mass Spectrometry.

Emerging chemicals of concern (ECCs), including phthalate plasticizers, flame retardants, and phenolic compounds, are likely present in electronic nicotine delivery system (ENDS) replacement solutions (e-liquids) which are often packaged, stored in, and/or can contact with, plastic, glass, and metal materials. Developing and validating an efficient analytical method for concurrent quantification of ECCs in e-liquids are thus needed to inform evidence-based safety evaluation of ENDS products. In this study, we developed and validated a "dilute-and-shoot" method using high-performance liquid chromatography coupled with tandem mass spectrometry to simultaneously measure organophosphate flame retardants (OPFRs), phthalate plasticizers, and tetrabromobisphenol A (TBBPA) in e-liquids. We analyzed samples in positive electrospray ionization mode (ESI+) for OPFRs and phthalates and negative ESI- for TBBPA. The method has a total runtime of 10 min. The optimized procedure was able to deliver broad dynamic linearity ranges with coefficients of determination (R 2) above 0.995, limits of detection ranging from 0.020 to 10 ng/mL, average accuracy within ±15%, and imprecision ≤ 15.0% for all analytes. To our knowledge, this is the first multianalysis method for measuring ECCs in e-liquid samples, and the validation results show that it is sensitive, accurate, precise, and efficient.

Marijuana and Tobacco Coexposure in Hospitalized Children.

BACKGROUND: The impact of secondhand marijuana smoke exposure on children is unknown. New methods allow for the detection of marijuana smoke exposure in children. METHODS: We studied children who were hospitalized in Colorado and had a parent participating in a smoking cessation study; all children had urine samples remaining from the original study as well as consent for future research. Parents completed a survey and urine samples were analyzed for cotinine and marijuana metabolites, including 11-hydroxy-Δ9-tetrahydrocannabinol (COOH-THC), by using liquid chromatography-tandem mass spectrometry. RESULTS: The median age of the children was 6.0 years (range 0-17 years); 57% were boys. Half (55%) were white, 12% were African American, and 33% were of another race; 39% identified as Hispanic. Approximately 46% had detectable COOH-THC, and 11% had detectable THC. Of those with detectable THC, 3 were teenagers, and 6 were <8 years of age. There were no significant differences in urinary COOH-THC concentrations by age, sex, race and/or ethnicity, or socioeconomic status. Children with positive results for COOH-THC were more likely to have parents who use marijuana daily, smoke marijuana versus other forms of use, use daily in the home, and smoke marijuana in another room if the children are around compared with smoking outside. CONCLUSIONS: Approximately half of the children who qualified for our study had biological evidence of exposure to marijuana. Researchers in studies such as this provide valuable data on secondhand exposure to children from parents using tobacco and marijuana and can inform public health policies to reduce harm.

Examining the Association between Body Burdens of Harmful Chemicals and Heaviness of Marijuana Smoking.

Marijuana is seeing increased use both globally and domestically. However, the scientific community has not fully reached a consensus about what negative health effects and to what extent marijuana smoking may cause. In this study, we evaluated the associations between the body burdens of polycyclic aromatic hydrocarbons (PAHs) among marijuana smokers and the smoking heaviness. We observed clear, increasing trends of PAH body burdens as marijuana smoking increased. The findings highlight the importance of capturing the heaviness of marijuana smoking in future studies and support the need for well-designed toxicological and epidemiological studies for understanding the health effects of marijuana use in a changing environment.

Brief Report: Lead Levels in Selected Electronic Cigarettes from Canada and the United States.

Few published studies have investigated the presence of lead in the e-liquid of electronic nicotine delivery systems (ENDS). Lead inhalation is associated with increased risk of stroke, heart disease, and other diseases. This study used a novel application of graphite furnace technology to compare the concentration of lead between e-liquids of different packaging and product designs using e-liquids that are or were commercially available in the United States and Canada. Eleven nicotine-free disposable ENDS devices and 12 bottled refill solutions that contained nicotine were purchased from retailers in Canada and the United States between 2015 and 2017. E-liquids extracted from the disposable products and individual containers were analyzed for lead content by graphite furnace using atomic absorption detection. The lead concentration of open-wick ENDS devices ranged from 25.2 ppb to 838.4 ppb, with a standard deviation of 187.4 ppb. None of the bottled e-liquids contained quantifiable levels of lead. This study found that quantifiable levels of lead are present in certain disposable e-cigarette devices, and there is evidence from this study that the design of ENDS devices may contribute to lead exposure. These findings suggest that lead testing should be incorporated into future chemical analyses of ENDS devices.

Urinary Metabolite Levels of Flame Retardants in Electronic Cigarette Users: A Study Using the Data from NHANES 2013-2014.

Evaluating the safety of e-cigarettes and making informed judgement about developing potential standards require sufficient scientific evidence. Since e-cigarettes are highly engineered products containing plastic, glass and metal parts, and e-liquids are largely different matrices, many toxic compounds which are not typical hazards for the users of combustible tobacco products (e.g., cigarettes), could exist in e-liquids, and consequently, posing potential health risk to e-cigarette users. We combined the measurements of urinary metabolites of organophosphate flame retardants (OPFRs) with questionnaire data collected in the National Health and Nutrition Examination Surveys (NHANES) from 2013 to 2014, and we compared adjusted geometric means (GM) for each biomarker in e-cigarette users with levels in non-users and users of various tobacco products using multiple regression analyses to adjust for potential confounders. We found diphenyl phosphate (DPhP), bis(1,3-dichloro-2-propyl) phosphate (BDCPP), bis(2-chloroethyl) phosphate (BCEP), and dibutyl phosphate (DBUP) were detected in all e-cigarette users. The adjusted GM of BCEP, the metabolite of tris(2-chloroethyl) phosphate (TCEP), was 81% higher than nonusers (p = 0.0124) and significantly higher than those for both cigarette and cigar users (p < 0.05). The findings in this pilot study suggest that certain OPFRs may present in e-cigarettes as contaminants, and consequently, resulting in higher exposure levels in e-cigarette users compared to nonusers. As we only identified 14 e-cigarette users in the survey, the findings in this study need to be confirmed in future study at a larger scale. A better examination of the types and levels of FRs and their potential contamination sources in e-cigarettes is also needed.

Detecting biomarkers of secondhand marijuana smoke in young children.

BACKGROUND: The impact of secondhand marijuana smoke exposure on children is unknown. New methods allow detection of secondhand marijuana smoke in children. METHODS: We studied children ages 1 mo to 2 y hospitalized with bronchiolitis in Colorado from 2013 to 2015. Parents completed a survey, and urine samples were analyzed for cotinine using LC/MS/MS (limits of detection 0.03 ng/ml) and marijuana metabolites including COOH-THC (limits of detection 0.015 ng/ml). RESULTS: A total of 43 subjects had urine samples available for analysis. Most (77%) of the subjects were male, and 52% were less than 1 y of age. COOH-THC was detectable in 16% of the samples analyzed (THC+); the range in COOH-THC concentration was 0.03-1.5 ng/ml. Two subjects had levels >1 ng/ml. Exposure did not differ by gender or age. Non-white children had more exposure than white children (44 vs. 9%; P < 0.05). 56% of children with cotinine >2.0 ng/ml were THC+, compared with 7% of those with lower cotinine (P < 0.01). CONCLUSION: Metabolites of marijuana smoke can be detected in children; in this cohort, 16% were exposed. Detectable COOH-THC is more common in children with tobacco smoke exposure. More research is needed to assess the health impacts of marijuana smoke exposure on children and inform public health policy.

Temporal Trends of Secondhand Smoke Exposure: Nonsmoking Workers in the United States (NHANES 2001-2010).

BACKGROUND: The workplace is one of the major locations outside of the home for nonsmokers' exposure to secondhand smoke (SHS). New policies in many U.S. states and localities restrict or prohibit smoking in the workplace, and information on current trends in the exposure of nonsmokers to SHS across various occupational groups is therefore needed. OBJECTIVE: We evaluated temporal trends in SHS exposure among nonsmoking workers in the United States and identified those occupations with workers with the highest levels of SHS exposure. METHODS: We combined serum cotinine (sCOT) measurements and questionnaire data from five survey cycles of the National Health and Nutrition Examination Survey (NHANES: 2001-2010). Trends in SHS exposure by occupations were determined from percent changes and least-squares geometric means (LSGMs) of sCOT concentrations computed using sample-weighted multiple regression models. RESULTS: Between NHANES 2001-2002 and NHANES 2009-2010, LSGMs of sCOT levels had changed -25% (95% CI: -39, -7%) in nonsmoking workers. The largest decrease was identified among food preparation workers [-54% (95% CI: -74, -19%)], followed by white-collar [-40%, (95% CI: -56, -19%)] and blue-collar workers (-32%, 95% CI: -51, -5%). LSGMs of sCOT remained highest in food preparation workers in all survey cycles, but the gap between occupations narrowed in the latest survey cycle (2009-2010). For example, the gap in LSGMs of sCOT between food preparation and science/education workers dropped > 70% during 2000 to 2010. CONCLUSIONS: During the period from 2001 to 2010, the overall SHS exposure in nonsmoking workers declined with substantial drops in food preparation/service and blue-collar workers. Although disparities persist in SHS exposure, the gaps among occupations have narrowed. CITATION: Wei B, Bernert JT, Blount BC, Sosnoff CS, Wang L, Richter P, Pirkle JL. 2016. Temporal trends of secondhand smoke exposure: nonsmoking workers in the United States (NHANES 2001-2010). Environ Health Perspect 124:1568-1574; http://dx.doi.org/10.1289/EHP165.

Urinary concentrations of PAH and VOC metabolites in marijuana users.

BACKGROUND: Marijuana is seeing increased therapeutic use, and is the world's third most-popular recreational drug following alcohol and tobacco. This widening use poses increased exposure to potentially toxic combustion by-products from marijuana smoke and the potential for public health concerns. OBJECTIVES: To compare urinary metabolites of polycyclic aromatic hydrocarbons (PAHs) and volatile organic compounds (VOCs) among self-reported recent marijuana users and nonusers, while accounting for tobacco smoke exposure. METHODS: Measurements of PAH and VOC metabolites in urine samples were combined with questionnaire data collected from participants in the National Health and Nutrition Examination Surveys (NHANES) from 2005 to 2012 in order to categorize participants (≥18years) into exclusive recent marijuana users and nonusers. Adjusted geometric means (GMs) of urinary concentrations were computed for these groups using multiple regression analyses to adjust for potential confounders. RESULTS: Adjusted GMs of many individual monohydroxy PAHs (OH-PAHs) were significantly higher in recent marijuana users than in nonusers (p<0.05). Urinary thiocyanate (p<0.001) and urinary concentrations of many VOC metabolites, including metabolites of acrylonitrile (p<0.001) and acrylamide (p<0.001), were significantly higher in recent marijuana users than in nonusers. CONCLUSIONS: We found elevated levels of biomarkers for potentially harmful chemicals among self-identified, recent marijuana users compared with nonusers. These findings suggest that further studies are needed to evaluate the potential health risks to humans from the exposure to these agents when smoking marijuana.

Assessing exposure to tobacco-specific carcinogen NNK using its urinary metabolite NNAL measured in US population: 2011-2012.

Carcinogenic tobacco-specific nitrosamines (TSNAs) such as 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) are found only in tobacco and derived products. Food and Drug Administration of the United States (US FDA) lists NNK as one of the 93 harmful and potentially harmful constituents (HPHCs) found in tobacco products and tobacco smoke. The aim of this study was to use the urinary concentration of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), a major metabolite of NNK, to quantitatively estimate exposure to NNK in the US general population. In 2011-2012, the Centers for Disease Control and Prevention's National Health and Nutrition Examination Survey (NHANES) collected urine and serum samples from a representative sample of US residents. We used a serum cotinine cutoff of 10 ng/ml with combination of questionnaire data to select non-users from cigarette users and used self-reported data to determine different tobacco product user groups. We estimated the absorbed total daily dose of NNK using a probabilistic method based on a two-compartment model. The geometric mean (GM) for the daily dose of NNK among smokers aged 12-16 years was significantly higher than that for non-users at the same age stage exposed to second-hand smoke (SHS) (P<0.001). Among those exposed to SHS, the GM for daily dose of NNK in young children (6-11 years) was nearly three times of those for adults in the age range 21-59 years. Among cigarette users, non-Hispanic Whites had the highest NNK daily dose and Mexican Americans had the lowest levels. Exclusive snuff or chewing product users had significantly higher daily dose of NNK than did cigarette smokers. Our study found that the maximum daily dose of NNK for children aged from 6 to 11 years and that for a significant percentage of cigarette users, chewing product and snuff users were higher than an estimated provisional "reference" risk level.

Sensitive Quantification of Cannabinoids in Milk by Alkaline Saponification-Solid Phase Extraction Combined with Isotope Dilution UPLC-MS/MS.

Maternal exposure to marijuana during the lactation period-either active or passive-has prompted concerns about transmission of cannabinoids to breastfed infants and possible subsequent adverse health consequences. Assessing these health risks requires a sensitive analytical approach that is able to quantitatively measure trace-level cannabinoids in breast milk. Here, we describe a saponification-solid phase extraction approach combined with ultra-high-pressure liquid chromatography-tandem mass spectrometry for simultaneously quantifying Δ9-tetrahydrocannabinol (THC), cannabidiol (CBD), and cannabinol (CBN) in breast milk. We demonstrate for the first time that constraints on sensitivity can be overcome by utilizing alkaline saponification of the milk samples. After extensively optimizing the saponification procedure, the validated method exhibited limits of detections of 13, 4, and 66 pg/mL for THC, CBN, and CBD, respectively. Notably, the sensitivity achieved was significantly improved, for instance, the limits of detection for THC is at least 100-fold more sensitive compared to that previously reported in the literature. This is essential for monitoring cannabinoids in breast milk resulting from passive or nonrecent active maternal exposure. Furthermore, we simultaneously acquired multiple reaction monitoring transitions for 12C- and 13C-analyte isotopes. This combined analysis largely facilitated data acquisition by reducing the repetitive analysis rate for samples exceeding the linear limits of 12C-analytes. In addition to high sensitivity and broad quantitation range, this method delivers excellent accuracy (relative error within ±10%), precision (relative standard deviation <10%), and efficient analysis. In future studies, we expect this method to play a critical role in assessing infant exposure to cannabinoids through breastfeeding.

Analysis of Cannabinoids and Their Metabolites in Human Urine.

Biologically monitoring marijuana exposure from active and passive use requires both a wide linear range and sensitive detection. We have developed and validated a multifunctional method using ultrahigh performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS) for analysis of urinary Δ9-tetrahydrocannabinol (THC), cannabidiol and cannabinol, and two major metabolites of THC, 11-nor-9-carboxy-THC and 11-hydroxy-THC, in active users and particularly in people exposed to secondhand marijuana smoke (SHMS). The method used positive electrospray ionization (ESI) mode to reach the sensitivity needed to detect trace SHMS exposure with limits of detection (LOD) ranging from 0.002 to 0.008 nanograms per milliliter (ng/mL) and 0.005 to 0.017 ng/mL for "free" (unconjugated forms) and "total" (unconjugated plus conjugated forms) measurements, respectively. These LODs were approximately 10-100 times more sensitive than those reported in the literature. To reduce or avoid time-consuming repetitive sample preparation and analysis, the method simultaneously monitored multiple reaction monitoring transitions in negative ESI mode to quantify high analyte levels typically found in the urine of active marijuana users (linear dynamic range of 12.5-800 ng/mL). The validation results indicated this method was accurate (average inter/intra-day bias, <10%), precise (inter/intra-day imprecision, <10%), and fast (6 min run time). In addition, sample preparation throughput was greatly improved using an automation liquid-handling system, meeting the needs for potential large-scale population studies.

Validation of a LC-MS/MS method for quantifying urinary nicotine, six nicotine metabolites and the minor tobacco alkaloids--anatabine and anabasine--in smokers' urine.

Tobacco use is a major contributor to premature morbidity and mortality. The measurement of nicotine and its metabolites in urine is a valuable tool for evaluating nicotine exposure and for nicotine metabolic profiling--i.e., metabolite ratios. In addition, the minor tobacco alkaloids--anabasine and anatabine--can be useful for monitoring compliance in smoking cessation programs that use nicotine replacement therapy. Because of an increasing demand for the measurement of urinary nicotine metabolites, we developed a rapid, low-cost method that uses isotope dilution liquid chromatography-tandem mass spectrometry (LC-MS/MS) for simultaneously quantifying nicotine, six nicotine metabolites, and two minor tobacco alkaloids in smokers' urine. This method enzymatically hydrolyzes conjugated nicotine (primarily glucuronides) and its metabolites. We then use acetone pretreatment to precipitate matrix components (endogenous proteins, salts, phospholipids, and exogenous enzyme) that may interfere with LC-MS/MS analysis. Subsequently, analytes (nicotine, cotinine, hydroxycotinine, norcotinine, nornicotine, cotinine N-oxide, nicotine 1'-N-oxide, anatabine, and anabasine) are chromatographically resolved within a cycle time of 13.5 minutes. The optimized assay produces linear responses across the analyte concentrations typically found in urine collected from daily smokers. Because matrix ion suppression may influence accuracy, we include a discussion of conventions employed in this procedure to minimize matrix interferences. Simplicity, low cost, low maintenance combined with high mean metabolite recovery (76-99%), specificity, accuracy (0-10% bias) and reproducibility (2-9% C.V.) make this method ideal for large high through-put studies.

A high-throughput robotic sample preparation system and HPLC-MS/MS for measuring urinary anatabine, anabasine, nicotine and major nicotine metabolites.

BACKGROUND: Most sample preparation methods characteristically involve intensive and repetitive labor, which is inefficient when preparing large numbers of samples from population-scale studies. METHODS: This study presents a robotic system designed to meet the sampling requirements for large population-scale studies. Using this robotic system, we developed and validated a method to simultaneously measure urinary anatabine, anabasine, nicotine and seven major nicotine metabolites: 4-Hydroxy-4-(3-pyridyl)butanoic acid, cotinine-N-oxide, nicotine-N-oxide, trans-3'-hydroxycotinine, norcotinine, cotinine and nornicotine. We analyzed robotically prepared samples using high-performance liquid chromatography (HPLC) coupled with triple quadrupole mass spectrometry in positive electrospray ionization mode using scheduled multiple reaction monitoring (sMRM) with a total runtime of 8.5 min. RESULTS: The optimized procedure was able to deliver linear analyte responses over a broad range of concentrations. Responses of urine-based calibrators delivered coefficients of determination (R(2)) of >0.995. Sample preparation recovery was generally higher than 80%. The robotic system was able to prepare four 96-well plate (384 urine samples) per day, and the overall method afforded an accuracy range of 92-115%, and an imprecision of <15.0% on average. CONCLUSIONS: The validation results demonstrate that the method is accurate, precise, sensitive, robust, and most significantly labor-saving for sample preparation, making it efficient and practical for routine measurements in large population-scale studies such as the National Health and Nutrition Examination Survey (NHANES) and the Population Assessment of Tobacco and Health (PATH) study.

Studying permethrin exposure in flight attendants using a physiologically based pharmacokinetic model.

Assessment of potential health risks to flight attendants from exposure to pyrethroid insecticides, used for aircraft disinsection, is limited because of (a) lack of information on exposures to these insecticides, and (b) lack of tools for linking these exposures to biomarker data. We developed and evaluated a physiologically based pharmacokinetic (PBPK) model to assess the exposure of flight attendants to the pyrethroid insecticide permethrin attributable to aircraft disinsection. The permethrin PBPK model was developed by adapting previous models for pyrethroids, and was parameterized using currently available metabolic parameters for permethrin. The human permethrin model was first evaluated with data from published human studies. Then, it was used to estimate urinary metabolite concentrations of permethrin in flight attendants who worked in aircrafts, which underwent residual and pre-flight spray treatments. The human model was also applied to analyze the toxicokinetics following permethrin exposures attributable to other aircraft disinsection scenarios. Predicted levels of urinary 3-phenoxybenzoic acid (3-PBA), a metabolite of permethrin, following residual disinsection treatment were comparable to the measurements made for flight attendants. Simulations showed that the median contributions of the dermal, oral and inhalation routes to permethrin exposure in flight attendants were 83.5%, 16.1% and 0.4% under residual treatment scenario, respectively, and were 5.3%, 5.0% and 89.7% under pre-flight spray scenario, respectively. The PBPK model provides the capability to simulate the toxicokinetic profiles of permethrin, and can be used in the studies on human exposure to permethrin.