Urinary Total NNAL Research Articles

Secondhand smoke exposure in school children in Malta assessed through urinary biomarkers

School children may be exposed to secondhand smoke (SHS) either at home, in transit or in social gatherings permitting smoking in their presence. Questionnaires about SHS often underestimate prevalence and extent of exposure. A more accurate tool is the use of biomarkers such as cotinine (COT) and trans-3′-hydrocycotinine (3HC) as biomarkers of SHS exposure, alongside 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), a reduction product in the body of the tobacco-specific nitrosamine, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), both potent carcinogens. We measured urinary COT, 3HC and total NNAL using sensitive and specific high-performance LC-MS/MS methods. The limit of quantification (LOQ) for each assay were 0.05 ng/mL, 0.1 ng/mL and 0.25 pg/mL respectively. The aim of this study was to evaluate the exposure to SHS of school children (9–11 years), from five public schools in the island of Malta, from questionnaire information about smoking at home and verify it by urinary biomarker data of COT, 3HC and NNAL. These biomarkers were measurable in 99.4%, 95.4% and 98.3% of the participating children respectively. From the children reporting smoking at home, 11% had a history of asthma and had COT, 3HC and NNAL geometric mean concentrations double compared to the non-asthmatic group. In has been confirmed that non-smokers exposed to SHS and THS have a higher NNAL/COT ratio than the group identified as smokers according to specific and defined COT threshold levels (despite the fact that a priori, the entire study group was composed of non-smokers). The implication of high measured levels of urinary NNAL in children should be of concern given its potency. A main effects multifactor ANOVA model was developed and the children's house and school locations and the smoking frequency were statistically significant to predict the levels of the three metabolites. For 3HC only, the status of the employment of the mother was also an important predictor.

Effect of an electronic nicotine delivery system with 0, 8, or 36 mg/mL liquid nicotine versus a cigarette substitute on tobacco-related toxicant exposure: a four-arm, parallel-group, randomised, controlled trial

Electronic nicotine delivery systems (ENDSs) are used by some smokers to reduce cigarette consumption, but their effectiveness is uncertain. We aimed to examine the extent to which ENDSs or a non-nicotine cigarette substitute influence tobacco-related toxicant exposure and cigarette consumption in smokers interested in smoking reduction. We did a four-arm, parallel-group, randomised controlled trial at two sites in the USA (Penn State University, Hershey, PA, and Virginia Commonwealth University, Richmond, VA). We enrolled adults aged 21-65 years who smoked more than nine cigarettes per day (for at least the past year), with exhaled CO of more than 9 parts per million at screening, who were not currently using an ENDS, and who were interested in reducing smoking but not quitting. Participants were randomised (site-specific with allocation concealment; 1:1:1:1) to receive either a cartomiser-based, pen-style ENDS (eGo-style) paired with 0, 8, or 36 mg/mL liquid nicotine (participants and researchers masked to concentration) or a non-ENDS cigarette-shaped plastic tube that delivered no nicotine or aerosol (cigarette substitute; unmasked) for 24 weeks. Conditions were chosen to reflect a range of nicotine delivery including none (cigarette substitute and 0 mg/mL ENDS), low (8 mg/mL), and cigarette-like (36 mg/mL), and all conditions were paired with smoking reduction instructions. The primary outcome was concentration of the tobacco-specific carcinogen metabolite 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL; urinary total) collected at randomisation and at 4, 12, and 24 weeks. Multiple imputation with and without covariate adjustment was used in addition to sensitivity analyses. This trial is registered with ClinicalTrials.gov, NCT02342795. Between July 22, 2015, and Nov 16, 2017, 684 individuals were screened and 520 (76%) were enrolled and randomised. 188 (36%) of 520 participants were lost to follow-up by week 24; attrition did not differ by study group (39 [30%] of 130 in the cigarette substitute group, 56 [43%] of 130 in the ENDS with 0 mg/mL nicotine group, 49 [38%] of 130 in the ENDS 8 mg/mL group, and 44 [34%] of 130 in the ENDS 36 mg/mL group). Urinary total NNAL at 24 weeks in the ENDS with 36 mg/mL nicotine group was 210·80 pg/mg creatinine (95% CI 163·03-274·42) compared with 346·09 pg/mg creatinine (265·00-455·32) in the cigarette substitute group (p=0·0061). No other significant differences between groups were observed for any time point for urinary total NNAL. Serious adverse event frequency was similar across groups (12 events in 12 participants [9%] in the ENDS with 36 mg/mL nicotine group, seven events in six participants [5%] in the 8 mg/mL group, 11 events in ten participants [8%] in the 0 mg/mL group, and 13 events in 13 participants [10%] in the cigarette substitute group), and all of these were deemed unrelated or unlikely to be related to study product use. There was one death between randomisation and 24 weeks (suicide; in the ENDS with 0 mg/mL nicotine group). Use of an ENDS with cigarette-like nicotine delivery can reduce exposure to a major pulmonary carcinogen, NNAL, even with concurrent smoking. Future ENDS trials should involve products with well characterised nicotine delivery, including those with nicotine delivery approaching that of a cigarette. National Institutes of Health, US Food and Drug Administration.

Relationship Between Urinary 4-(Methylnitrosamino)-1-(3-Pyridyl)-1-Butanol and Lung Cancer Risk in the General Population: A Community-Based Prospective Cohort Study.

No published studies have prospectively evaluated the association between urinary 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) levels and lung cancer risk in the general population. Here, we conducted a prospective community-based cohort study in the Republic of Korea to evaluate the relationship between urinary NNAL levels and lung cancer risk using prediagnostic urine samples. This prospective cohort study was based on the Korean National Cancer Center Community Cohort. During the follow-up period, 173 primary lung cancer cases were identified. Total urinary NNAL levels were measured by liquid chromatography-tandem mass spectrometry, and data were analyzed using multivariable Cox proportional hazards regression models. The risk of lung cancer was significantly increased per unit of natural log-transformed urinary NNAL (HR, 1.27; 95% CI, 1.09–1.48), after adjusting for age, region, entry year into the cohort, education achievement, alcohol consumption status, BMI, smoking status, and urinary cotinine levels. Cox proportional-hazards models with NNAL quartiles also showed positive dose-response relationships with risk of lung cancer. A significantly increased risk of lung cancer was found in the fourth quartile of urinary NNAL levels (HR, 3.27; 95% CI, 1.37–7.79, P for trend < 0.01). After stratification with sex, the significant association remained in only men. Urinary NNAL levels are associated with the risk of lung cancer in the general population, and this association is independent from the quantification of cigarette smoking and nicotine uptake.

Abstract 3333: Potential of kava in reducing lung cancer risk, tobacco use, and associated disparities

Abstract Tobacco use is the primary risk factor for lung cancer, the leading cause of cancer deaths in the US. Quitting, however, is challenging due to the addictive nature of nicotine. Thus, preventing lung carcinogenesis in conjunction with smoking cessation may be necessary. Racial disparities also exist - African American (AA) smokers, with the same level of tobacco consumption, are at the highest risk of developing lung cancer. 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) is one key carcinogen in tobacco for human lung cancer risk and its differential uptake in AA and CA smokers may contribute to the disparity. AA smokers also have the lowest success rate in quitting. Kava is a daily beverage to help people relax and improve the quality of sleep. It is commercially available in US as a dietary supplement. Epidemiological data suggest that kava consumption may reduce cancer risk. Its relaxing property may help reduce tobacco dependence/use. Our preclinical data showed that kava completely blocked NNK induced tumorigenesis in A/J mice with enhancing NNK detoxification and reducing DNA damage as the potential mechanism. Building upon these, a pilot pre- and post- one-week kava trial was performed among smokers (n = 21). The results showed that kava was well-tolerated with high compliance and there were no signs of adverse effects when rigorous safety measures were implemented. Excitingly, the results suggested that one-week kava intake helped smokers 1) reduce tobacco use; 2) reduce the amounts of urinary DNA adducts; and 3) increase urinary excretion of NNAL. Specifically, urinary Total Nicotine Equivalent (TNE), the sum of nicotine N-oxide, total nicotine, cotinine, and 3-HO-cotinine, was used to estimate tobacco use and one-week kava use led to a 29.8% reduction in TNE (p &amp;lt; 0.001). Urinary 3-methyladenine (3-mA), a NNK-dependent DNA adduct biomarker, was used to estimate NNK-induced DNA damage and one week kava used led to a 33.6% reduction (p = 0.014). Lastly, urinary total NNAL was used as a biomarker for NNK excretion and one-week kava use led to a 95% increase (p = 0.002). Plasma cortisol of smokers, a stress biomarker, decreased by &amp;gt; 40% (p&amp;lt;0.01) after kava use, which may contribute to the reduction in tobacco use. The 21 participants include 13 CA, 7 AA and one American Indian. The impact of kava was also analyzed between AA and CA smokers. The reductions in urinary TNE and 3-mA were more pronounced in AA smokers while the increase in urinary NNAL was higher in AA smokers as well. Consistently, the reduction in plasma cortisol was greater in AA than CA smokers. The results from this pilot trial indicate kava’s potential to reduce lung cancer risk via reducing tobacco use, enhancing NNK/NNAL detoxification, and reducing DNA damage. The results also strongly suggest kava’s unique potential to address the associated racial disparities. Future rigorous trial is needed to confirm these observations. Note: This abstract was not presented at the meeting. Citation Format: Chengguo Xing, Yi Wang, Naomi Fujioka, Sreekanth Narayanapillai, Junxuan Lu. Potential of kava in reducing lung cancer risk, tobacco use, and associated disparities [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3333.

Children are particularly vulnerable to environmental tobacco smoke exposure: Evidence from biomarkers of tobacco-specific nitrosamines, and oxidative stress

BackgroundWorldwide, smoking is a major public health problem, with exposure to environmental tobacco smoke (ETS) affecting both smokers, and passive smokers, including children. Despite ETS also describing secondhand, and thirdhand smoke (SHS, and THS respectively), the health effects of exposure to passive smoking via these sources are not fully understood, particularly in children. Although cotinine, the primary proximate metabolite of nicotine, has been widely used as a biomarker of ETS exposure, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), the metabolite of the tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), provides a uniquely important contribution, both as a biomarker of exposure, and as a specific risk indicator for pulmonary carcinogenesis. MethodsWe used LC-MS/MS to study NNK metabolites, cotinine, and 8-oxo-7,8-dihydro-2′-deoxyguanosine (a biomarker of oxidative stress), in the urine of 110 non-smoking adults (age range: 23–62) and 101 children (age range: 9–11), exposed to ETS. ResultsIn our study of passive smoking adults, and children exposed to ETS, we showed that although the children had a similar urinary level of cotinine compared to the adults, the children had approximately two times higher levels of urinary total NNAL (P = 0.002), and free NNAL (P = 0.01), than adults. The children also had three times lower ability to detoxify NNK than adults (P < 0.001). Furthermore, the children showed 1.5 times higher ratio of total NNAL/cotinine than adults (P = 0.01), implying that THS is another important source of ETS in this population. Furthermore, ETS exposure in children appeared to lead to an increase in levels of oxidative stress. ConclusionsTaken together, our results demonstrate that, in children, THS may play an important role in the ETS exposure, and that children are at particular risk of ETS-induced health effects.

Carcinogen Exposure among Canadian Tobacco Users: Changes in NNK Exposure from 2007-2009 through 2012-2013.

Background: Tobacco-specific nitrosamines (TSNAs) are a class of carcinogens found in tobacco products, whose levels can vary considerably depending on tobacco blends and manufacturing processes. The current study examined whether recent increases in levels of the TSNA NNK [4-(methylnitrosamino-1-(3-pyridyl)-1-butanone] in Canadian cigarettes translated into differences in exposure among Canadian tobacco users.Methods: Nationally representative data from the Canadian Health Measures Survey (CHMS) were used to measure levels of total urinary NNAL [4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol], a metabolite of the TSNA NNK, among tobacco users. Data from CHMS Cycle 3 (2012-13) were used to examine NNAL, and linear regression was used to examine predictors. Data from CHMS Cycle 1 (2007-09) and Cycle 3 (2012-13) were used to examine changes in NNAL over time.Results: From 2007-2009 through 2012-2013, levels of creatinine-corrected NNAL increased by 64% (P < 0.0001). Levels of NNAL in 2012-2013 were higher among older respondents (P = 0.04), among females (P = 0.03), among respondents identifying as "white" and "Aboriginal" (P < 0.0001), and among those with greater daily cigarette consumption (P < 0.001), as well as greater levels of urinary free cotinine (P < 0.0001) and urinary creatinine (P < 0.0001).Conclusions: The findings indicate that exposure to the TSNA NNK among Canadian tobacco users has increased considerably from 2007-2009 through 2012-2013, in parallel to changes in TSNA levels in Canadian cigarettes. In the absence of epidemiologic data, it is unclear whether this change translates into increased risk.Impact: The study findings have potential implications for tobacco manufacturers, who bear a responsibility to reduce levels of tobacco carcinogens to the full extent possible. Cancer Epidemiol Biomarkers Prev; 27(3); 262-7. ©2018 AACR.

Tobacco Product Use Patterns, and Nicotine and Tobacco-Specific Nitrosamine Exposure: NHANES 1999-2012.

Background: Few studies have examined differences in product consumption patterns and nicotine and tobacco-specific nitrosamines (TSNA) exposure between single versus dual- and poly-tobacco users. We applied the Tobacco Product Use Patterns (T-PUPs) model to fill this gap in the literature.Methods: Data from adults (age ≥18 years) who used any tobacco products during the 5 days prior to participating in the 1999-2012 National Health and Nutrition Examination Survey (NHANES) were analyzed. Participants were classified into seven T-PUPs: (1) cigarettes only, (2) noncigarette combustibles only, (3) noncombustibles only, (4) dual noncigarette combustibles and noncombustibles, (5) dual cigarettes and noncombustibles, (6) dual cigarettes and noncigarette combustibles, and (7) poly-tobacco use. Weighted regression models were used to compare product consumption, serum cotinine, and urinary total 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (i.e., NNAL) levels between single-, dual-, and poly-tobacco T-PUPs.Results: Dual- and poly-tobacco T-PUPs were associated with lower product consumption compared with single-product T-PUPs only in some cases (e.g., dual cigarette and noncombustible users smoked cigarettes on 0.6 fewer days in the past 5 days compared with cigarette-only users; P < 0.05). Dual- and poly-tobacco T-PUPs had either nondistinguishable or higher levels of serum cotinine and urinary total NNAL than corresponding single-product T-PUPs.Conclusions: Product consumption, and nicotine and TSNAs exposure of dual- and poly-tobacco product category users somewhat differ from those of single-product category users as defined by the T-TUPs model.Impact: Higher levels of cotinine and NNAL among dual- and poly-tobacco T-TUPs users compared with the single-product T-TUPs users may indicate health concerns. Cancer Epidemiol Biomarkers Prev; 26(10); 1525-30. ©2017 AACR.

A two-site, two-arm, 34-week, double-blind, parallel-group, randomized controlled trial of reduced nicotine cigarettes in smokers with mood and/or anxiety disorders: trial design and protocol

BackgroundThe U.S. Food and Drug Administration can set standards for cigarettes that could include reducing their nicotine content. Such a standard should improve public health without causing unintended serious consequences for sub-populations. This study evaluates the effect of progressive nicotine reduction in cigarettes on smoking behavior, toxicant exposure, and psychiatric symptoms in smokers with comorbid mood and/or anxiety disorders using a two-site, two-arm, double-blind, parallel group, randomized controlled trial (RCT) in four phases over 34 weeks.MethodsAdult smokers (N = 200) of 5 or more cigarettes per day will be randomized across two sites (Penn State and Massachusetts General). Participants must have not had a quit attempt in the prior month, nor be planning to quit in the next 6 months, meet criteria for a current or lifetime unipolar mood and/or anxiety disorder based on the structured Mini-International Neuropsychiatric Interview, and must not have an unstable medical or psychiatric condition. After a week of smoking their own cigarettes, participants receive two weeks of Spectrum research cigarettes with usual nicotine content (11.6 mg). After this baseline period, participants will be randomly assigned to continue smoking Spectrum research cigarettes that contain either (a) Usual Nicotine Content (11.6 mg); or (b) Reduced Nicotine Content: the nicotine content per cigarette is progressively reduced from approximately 11.6 mg to 0.2 mg in five steps over 18 weeks. At the end of the randomization phase, participants will be offered the choice to either (a) quit smoking with assistance, (b) continue smoking free research cigarettes, or (c) return to purchasing their own cigarettes, for the final 12 weeks of the study. The primary outcome measure is blood cotinine; key secondary outcomes are: exhaled carbon monoxide, urinary total NNAL- 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol and 1-hydroxypyrene, oxidative stress biomarkers including 8-isoprostanes, measures of psychiatric symptoms (e.g., depression, anxiety), smoking behavior and dependence (e.g., cigarette consumption, quit attempts), and health effects (e.g., blood pressure, respiratory symptoms).DiscussionResults from this study will inform FDA on the potential effects of regulating the nicotine content of cigarettes and help determine whether smokers with mood and/or anxiety disorders can safely transition to significantly reduced nicotine content cigarettes.Trial registrationTRN: NCT01928758, registered August 21, 2013.

ED01.02 Tobacco Carcinogens and Lung Cancer Susceptibility

ED01.02 Tobacco Carcinogens and Lung Cancer Susceptibility

P1.01-012 Kava Effects on the Metabolism of Tobacco-Specific Carcinogen 4-(Methylnitrosamino)-1-(3-Pyridyl)-1-Butanone (NNK) in Humans

Kava is extracted from the roots of piper methysticum and is consumed by South Pacific Islanders as a relaxing beverage. Epidemiologic evidence points to a protective effect of kava against tobacco-induced lung cancer. NNK is a potent tobacco-specific carcinogen indisputably linked to lung cancer formation. Kava reduced NNK-induced lung adenoma formation in the A/J mouse model. Data also suggest that enhanced NNAL detoxification may be a potential mechanism by which kava exerts a chemopreventive effect. In humans, urinary NNAL is a validated biomarker of NNK uptake. We conducted a clinical trial in smokers to assess the effect of kava on NNK metabolism. The primary objective was to compare urinary total NNAL before and after kava administration. Secondary objectives included comparing the NNAL-gluc/NNAL-free ratio, determining the safety of kava, and quantifying O6-methylguanine adducts. The hypothesis was that kava administration would result in increased levels of NNAL in the urine (and increased NNAL-gluc/NNAL-free ratio), reflecting increased elimination and/or increased detoxification of NNK. Additionally, we hypothesized that kava could reduce O6-methylguanine adducts. We conducted a single-arm, open-label clinical trial in adult healthy smokers, in which subjects took a commercial kava supplement three times daily for seven days. Twenty-four hour urine collections were collected at baseline, days 4-5, and days 6-7 of the kava intervention for NNAL quantification. Blood samples were collected at baseline, day 4, and day 7 of the kava intervention for safety monitoring and for DNA adduct analysis. Subjects also completed a detailed tobacco questionnaire, food diary, smoking diary, and cigarette evaluation scale (CES) questionnaire. To date, 17 subjects (goal = 18) have completed the study. The results and statistics are being finalized. Short-term kava administration was safe with no evidence of hepatotoxicity. Subjects experienced less of the reinforcing effects of smoking after short-term kava administration as determined by the CES scores. The total CES score decreased on average by 4.47, from 45.53 to 41.06 (p=0.053, 95% CI -0.06-9.01). Notably, the smoking “satisfaction” scores decreased by 0.607 (p=0.024, 95% CI 0.09-1.12). This is the first study investigating the effect of kava on NNK metabolism in humans and is the first step to gain a more sophisticated mechanistic understanding of kava’s role in potentially modulating tobacco-related lung cancer risk. Short-term kava administration is safe in healthy adult smokers. Kava holds potential as a possible chemopreventive agent for smokers or tobacco cessation aide.

Association between Secondhand Smoke in Hospitality Venues and Urinary 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol Concentrations in Non-Smoking Staff.

The purpose of this study was to determine the relationship between urinary cotinine and total 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) concentrations in non-smoking staff and the indoor levels of fine particles (PM2.5) in hospitality venues that allow smoking, with respect to demographic and indoor environmental factors. We evaluated 62 hospitality venues that allowed smoking in Seoul, Korea. A real-time aerosol monitor was used to measure indoor PM2.5 concentrations. Field technicians recorded indoor environmental characteristics. One non-smoking staff member in each hospitality venue was tested for urinary cotinine and total NNAL concentrations. Demographic characteristics were obtained from self-reported staff questionnaires. Natural-log (ln)-transformed PM2.5 concentrations were significantly correlated with the ln-transformed cotinine (r = 0.31) and the total NNAL concentrations (r = 0.32). In multivariable regression analysis, the urinary cotinine concentrations of the staff members were significantly correlated with indoor PM2.5 concentrations; those with the highest concentrations were more likely to be women or staff members that worked in venues with a volume <375 m3. Total NNAL concentrations were significantly correlated only with indoor PM2.5 concentrations. Indoor PM2.5 may be used as an indicator for urinary cotinine and total NNAL concentrations in non-smoking staff members in hospitality venues that allow smoking.

Review of the state of the art of human biomonitoring for chemical substances and its application to human exposure assessment for food safety

Review of the state of the art of human biomonitoring for chemical substances and its application to human exposure assessment for food safety

Simple high-throughput analytical method using ultra-performance liquid chromatography coupled with tandem mass spectrometry to quantify total 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol in urine.

Since the urinary concentration of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) is a reliable biomarker of exposure to tobacco smoke, we developed a relatively simple high-throughput chromatographic method to quantify total urinary NNAL concentrations in the general population. The high-throughput analytical method was developed using ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS) to identify and quantify total urinary NNAL concentrations in 10 non-smokers and 15 otherwise healthy smokers. Loss of nitric oxide at m/z 30 was found to be the predominant mass transitioned, and therefore was used as the SIM transition to quantify both NNAL and NNAL-methyl-d3 in urine. The analytical method did not require sample derivatization. Standard curves for total NNAL concentrations were linear between 20 and 1500 pg/mL, with coefficients of determination >0.95. Precision and accuracy ranged from 2.2% to 8.6% (CV) and from -5.6% to 10.9% (percent error), respectively. The lowest limit of quantification was 6.7 pg/mL, and 2.0 pg/mL the lowest limit of detection (LLOD). Total urinary NNAL concentrations in non-smoker subjects were <LLOD, whereas in smokers varied between <LLOD to 112.1 pg/mL. An UPLC-MS/MS analytical method to quantify total urinary NNAL concentrations in smokers that does not require sample derivatization is presented herein. The method could be useful in clarifying the toxicities associated with human exposure to cigarette smoking. However, quantification might be adversely affected by co-eluting interfering compounds or selective ion suppression or enhancement as a result of having only one ion transition to monitor NNAL and NNAL-methyl-d3 in urine.

TSNA Exposure: Levels of NNAL Among Canadian Tobacco Users.

Tobacco smoke contains more than 7,000 chemicals, including known carcinogens, such as tobacco-specific nitrosamines (TSNAs). TSNA levels in cigarettes vary considerably within and across markets; however, the extent to which these different TSNA levels translate into differences in human exposure and risk remains unclear. The current study sought to examine TSNA exposure among Canadian tobacco users. Data from the 2007-2009 Canadian Health Measures Survey were used to measure levels of urinary NNAL [4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol], a metabolite of the TSNA NNK [4-(methylnitrosamino-1-(3-pyridyl)-1-butanone], among tobacco users (n = 507). Geometric mean concentrations of total urinary NNAL and creatinine-corrected total urinary NNAL were calculated. A linear regression model was used to examine predictors of urinary levels of NNAL. The mean population level of total urinary NNAL and creatinine-corrected total urinary NNAL was 71.2 pg/ml and 82.0 pg/mg creatinine, respectively. NNAL levels were higher among older respondents (p = .02), among females (p = .04), and among those with greater daily cigarette consumption (p < .0001), greater levels of urinary free cotinine (p < .0001), and greater levels of urinary creatinine (p < .0001). Overall, the mean level of urinary total NNAL among Canadian tobacco users was approximately one fourth that of their U.S. counterparts. The study findings provide the first nationally representative characterization of TSNA exposure among Canadian tobacco users. Although the findings indicate marked differences in TSNA exposure between Canadian and American populations of tobacco users, it is not known whether these differences in exposure translate into differences in risk.

Second-Hand Tobacco Smoke Exposure in Various US Occupations (NHANES 2007–08): Assessment Using Serum Cotinine and Urinary Total NNAL

Second-Hand Tobacco Smoke Exposure in Various US Occupations (NHANES 2007–08): Assessment Using Serum Cotinine and Urinary Total NNALAbstract Number:2127 Binnian Wei*, John T. Bernert, Benjamin C. Blount, Connie S. Sosnoff, and Lanqing Wang Binnian Wei* Centers for Disease Control and Prevention, United States, E-mail Address: [email protected] Search for more papers by this author , John T. Bernert Centers for Disease Control and Prevention, United States, E-mail Address: [email protected] Search for more papers by this author , Benjamin C. Blount Centers for Disease Control and Prevention, United States, E-mail Address: [email protected] Search for more papers by this author , Connie S. Sosnoff Centers for Disease Control and Prevention, United States, E-mail Address: [email protected] Search for more papers by this author , and Lanqing Wang Centers for Disease Control and Prevention, United States, E-mail Address: [email protected] Search for more papers by this author AbstractBackground: Our previous study using serum cotinine has shown that second-hand smoke (SHS) exposure varied significantly among different occupations. Urinary 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), a major metabolite of the tobacco-specific carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), has an excretion half-life of 10–18 days. Thus urinary NNAL could reflect a relatively longer SHS exposure interval compared to serum cotinine which has an half-life of only about 16 hours.Objectives: The objective of this study is to describe the total urinary NNAL concentrations among US occupations.Methods: Samples came from the National Health and Nutrition Examination Survey (NHANES) 2007-2008. Nonsmokers were those participants with serum cotinine <=10 ng/mL, aged 18 and above.Results: For total urinary NNAL, non-smoking workers in legal and scientific occupations had the lowest geometric mean (GM) values of 0.56 and 0.57 ng/g creatinine, respectively, while the food preparation/serving occupation had the highest GM of 1.47 ng/g creatinine, followed by installation/maintenance/repair workers (1.24). Those two occupations were 79% and 51%, respectively, higher than the GM (0.82 ng/g creatinine) for all occupation groups. For serum cotinine, the lowest GM values were found in legal (0.018 ng/mL) and education/training/library (0.018 ng/mL) occupations, while the highest GM values were again identified in food preparation/serving (0.096 ng/mL) and installation/maintenance/repair (0.081 ng/mL) occupations. Those two occupations had 148% and 109% higher serum cotinine, respectively, compared to GM for all occupation groups (0.039 ng/mL).Conclusions: Occupations that had the highest total urinary levels of the tobacco-specific carcinogen NNAL were consistent with those found to have the highest serum cotinine levels. Targeted efforts may be needed to decrease SHS for those occupations, especially within food preparation/service workplaces.

Tobacco-specific N-nitrosamine exposures and cancer risk in the Shanghai Cohort Study: remarkable coherence with rat tumor sites.

The tobacco-specific nitrosamines N'-nitrosonornicotine (NNN) and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) are potent carcinogens for the rat esophagus and lung, respectively. Consistent with the animal carcinogenicity data, we previously reported a remarkably strong association between prospectively measured urinary total NNN, a biomarker of human NNN intake, and the risk of developing esophageal cancer among smokers in the Shanghai Cohort Study. We also demonstrated that urinary total 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), a biomarker of exposure to the lung carcinogen NNK, is strongly associated with the risk of lung, but not esophageal cancer in smokers. In this study, we investigated the potential relationship between NNN intake and lung cancer risk in the same cohort. The prospectively collected urine samples from lung cancer cases and matching controls selected for this study, all current smokers, have been previously analyzed for total NNAL, cotinine (a biomarker of nicotine intake) and phenanthrene tetraol (PheT) (a biomarker of exposure to polycyclic aromatic hydrocarbons). Urinary levels of total NNN were not associated with the risk of lung cancer: odds ratios (95% confidence intervals) associated with the second and third tertiles of total NNN, relative to the lowest tertile, were 0.82 (0.36-1.88) and 1.02 (0.39-2.89), respectively (p for trend = 0.959), after adjustment for self-reported smoking history, urinary cotinine and PheT. The results of this study reaffirm the previously reported specificity of urinary total NNN and total NNAL as predictors of esophageal and lung cancer risks, respectively, in smokers, and demonstrate remarkable coherence between rat target tissues of these carcinogens and susceptibility to cancer in smokers.

Cigarette Rod Length and Its Impact on Serum Cotinine and Urinary Total NNAL Levels, NHANES 2007-2010

Research suggests that smokers of slim, long, or ultralong cigarettes may have a perception of reduced harm from their own brand. This study compared serum cotinine and urinary total 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanonol (NNAL) levels among smokers of regular-sized (68-72mm), king-sized (79-88mm), and long (94-101mm) or ultralong (110-121mm) cigarettes. Nationally representative data from the 2007-2008 and 2009-2010 National Health and Nutrition Examination Survey were analyzed for 3,699 current smokers aged ≥20 years. Biomarker levels were summarized using geometric means (GMs). Multivariate linear regression analyses were performed to assess the effect of cigarette rod length on log-transformed serum cotinine and creatinine-adjusted urinary total NNAL levels. The GM of serum cotinine level was higher among smokers of long/ultralong cigarettes (263.15ng/ml) compared with smokers of regular-sized (173.13ng/ml) or king-sized (213.79ng/ml) cigarettes. Similarly, the GM of creatinine-adjusted NNAL levels was higher among smokers of long/ultralong cigarettes (0.48ng NNAL/mg of creatinine) compared with smokers of regular-sized (0.34ng NNAL/mg of creatinine) or king-sized (0.33ng NNAL/mg of creatinine) cigarettes. After adjusting for potential confounders, mean cotinine and NNAL levels were both significantly higher among smokers of long/ultralong cigarettes compared with levels observed in smokers of either regular-sized or king-sized cigarettes. However, no significant differences were observed between smokers of regular-sized and king-sized cigarettes in mean levels of cotinine or NNAL. Significantly elevated tobacco biomarker levels were observed among smokers of long/ultralong cigarettes compared with smokers of regular-sized or king-sized cigarettes. This underscores the need to educate the public about the dangers of all tobacco products.

Proposed Cutoff for Identifying Adult Smokeless Tobacco Users With Urinary Total 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanonol: An Aggregated Analysis of NHANES 2007–2010 Data

NNAL [4-(methylnitrosamino)-1-(3-pyridyl)-1-butanonol] is a valid biomarker of tobacco use. However; no study has assessed its use in distinguishing current smokeless tobacco (SLT) users from nonusers. Therefore, this study used aggregated data from the 2007-2008 and 2009-2010 waves of the National Health and Nutrition Examination Survey to determine an optimal threshold for identifying SLT users with NNAL. Optimal urinary total NNAL concentrations for discriminating SLT-only users from nonusers of any tobacco were determined using receiver operating characteristic analysis. Percentage agreement between self-reported SLT use status and NNAL levels was calculated overall and by sociodemographic characteristics. All analyses were weighted and performed with Stata, Version 11, and MedCalc for Windows, Version 9.5.0.0. In total, 264 individuals reported exclusively using SLT (and no other combustible tobacco product) within the past 5 days, whereas 14,824 were self-reported nonusers of any combustible or smokeless tobacco product. The optimal NNAL cutoff point was 34.0 pg/ml, which was associated with a high sensitivity (95.2%), specificity (93.4%), and overall correct classification rate (93.5%). The area under the curve was 98.3% and the corresponding Youden's Index was 88.7%. There was high agreement between the proposed NNAL cutoff point and self-reported SLT-only use (95.6%) and self-reported SLT nonuse (93.9%). The proposed cutoff point of 34.0 pg/ml had high sensitivity and specificity and may be used by clinicians and researchers to verify or detect recent SLT use. This study also indicated that self-reported SLT use among adults is a reliable measure and has high agreement with biochemical assessment.

TSNA exposure from cigarette smoking: 18 Years of urinary NNAL excretion data

The objective of this work was to characterize trends over time in urinary excretion of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) among cigarette smokers in the US. We identified 35 studies presenting data that either reported, or could be converted to, common units of total urinary NNAL excretion as pmol/mg creatinine. The studies spanned 18years, reported urinary NNAL excretion estimates for 61 defined populations, and included a combined total of 3941 study participants. Analyses show that urinary NNAL excretion trends downward with study publication year, and the trend is statistically significant. The trend does not appear to be accounted for by a reduction in cigarettes smoked per day by study participants over the same time period. This trend is consistent with reductions in tobacco specific nitrosamine (TSNA) levels in both cigarette tobacco filler and mainstream cigarette smoke observed over the past decade and with efforts by the tobacco industry and the agricultural community to reduce levels of TSNAs in tobacco and cigarette smoke.

Joint Effect of Arsenic Methylation Profile and NNK Metabolites on Urothelial Carcinoma

Cigarette smoking interacts with the urinary arsenic profile to modify the risk of urothelial carcinoma. NNK (4-[methylnitrosamino]-1-[3-pyridyl]-1-butanone) and its metabolite NNAL (4-[methylnitrosamino]-1-[3-pyridyl]-1-butanol) are biomarkers for cigarette smoking exposure. We explored the joint effects of urinary NNK metabolites and arsenic species on urothelial carcinoma risk. We recruited 137 pairs of urothelial carcinoma cases and matched healthy participants for a hospital based case-control study. Participants completed questionnaires and provided 50 ml urine samples. Urine samples were analyzed for free NNAL and NNAL-glucuronides using liquid chromatography-tandem mass spectrometry. Samples were analyzed for arsenic species using high performance liquid chromatography hydride generator atomic absorption spectrometry. Overall, subjects with high urinary total NNAL and high total arsenic had a greater urothelial carcinoma risk than those with a low total NNAL and low total arsenic. Subjects with a lower ratio of NNAL-glucuronides-to-free NNAL and higher total arsenic had a greater urothelial carcinoma risk than those with a higher NNAL-glucuronides-to-free NNAL ratio and lower total arsenic. This is the first study to our knowledge to demonstrate a significant trend of progressively increased risk of urothelial carcinoma in subjects who had none, one or both of the factors of urinary total arsenic and total NNAL or urinary total arsenic and the ratio of NNAL-glucuronides-to-free NNAL.