Urinary phenyl mercapturic acid (PhMA) is a specific biomarker of benzene exposure that has been widely used in biomonitoring of the general population and in occupational exposure studies. However, previous research has identified significant interlaboratory variation in urinary PhMA concentrations due to differences in the acidity of the sample treatment conditions. This variation arises from the need to convert the benzene's precursor metabolite 6-hydroxy-2,4-cyclohexadienyl mercapturic acid (pre-PhMA) to PhMA. In this study, we systematically examined the influence of sample treatment pH on this reaction across various acidic treatment conditions representative of the reported PhMA assays. The resulting pre-PhMA and PhMA levels were quantified using an established liquid chromatography-tandem mass spectrometry assay. PhMA levels increased with more acidic treatment conditions until pH -0.6 when PhMA formation was the greatest at 53.1% formation of the total pre-PhMA. The formation of PhMA was dependent on sample treatment pH. Thus, a quadratic regression was modeled on PhMA formation vs pH across all acid types. The resulting regression model (y = 0.874 × pH2 - 12.146 × pH + 41.99, R2 = 0.978) can be used to determine the extent of PhMA formation for a specific treatment pH, improving the ability to compare PhMA results among studies with different analytical methods or to absolute health-based cutoffs such as the biological exposure index. To explain the incomplete PhMA formation, we utilized gas chromatography-mass spectrometry to identify and quantify the formation of benzene as a major byproduct of the acid-derived dehydration of pre-PhMA. Further, base-derived dehydration of pre-PhMA to PhMA was performed to validate the benzene formation mechanism observed with acid dehydration.

Exposure assessment of hazardous volatile organic compounds (VOCs) requires accurate quantification of internal dose when establishing limits or identifying significant differences within and among populations. Even though accurate internal dose can be directly measured in blood, it is not always practical or possible to collect a suitable blood specimen. This work studies the relationship between blood and urine levels for certain smoke biomarkers (e.g., tobacco, marijuana) measured in self-reported cigarette smokers. Urine and blood specimens were collected as matched pairs from individuals at the same time. We used our latest specimen collection and VOC analysis protocols to minimize sample collection, handling, and analysis biases. From these analyses, unmetabolized urine benzene, furan, 2,5-dimethylfuran, isobutyronitrile, and benzonitrile levels were found to trend with blood levels. In addition, we measured urine creatinine levels, which were found to be significantly associated with all blood analyte concentrations (p-value ranging from <0.0063 to <0.0001) except for isobutyronitrile (p = 0.3347). For the analytes that were associated with urine creatinine levels, the ratios of urine-to-blood concentrations were substantially higher than those predicted from the urine/blood partition coefficients (Kurine/blood), which should occur if VOCs can freely equilibrate (i.e., passive diffusion) between the blood and urine. The urine isobutyronitrile concentration, which was the only analyte that was not associated with the urine creatinine level, had a urine-to-blood ratio similar to Kurine/blood. These results suggest either that urine VOC levels for certain VOCs do not equilibrate with blood levels in the urinary tract or that there is a conversion of conjugated to free forms, increasing urine VOC levels. Nevertheless, these deviations from partition theory (e.g., Henry's Law) are analyte-specific and require characterization to establish a relationship between blood and urine levels.

Methylcarbamoyl mercapturic acid (MCAMA, N-acetyl-S-(N-methylcarbamoyl)-L-cysteine) is a urinary metabolite of N,N-dimethylformamide and methyl isocyanate, which are volatile organic compounds that are harmful to humans. N,N-dimethylformamide exposure causes liver damage, and methyl isocyanate inhalation damages the lining of the respiratory tract, which can increase risk of chronic obstructive pulmonary disease and asthma. This study characterizes urinary MCAMA levels in the US population and explores associations of MCAMA concentrations with select demographic and environmental factors. We used liquid chromatography tandem mass spectrometry to measure MCAMA in urine collected from study participants ≥ 12 years old (N = 8272) as part of the National Health and Nutrition Examination Survey 2005–2006 and 2011–2016. We produced multiple regression models with MCAMA concentrations as the dependent variable and sex, age, fasting time, race/ethnicity, diet, and cigarette smoking as independent variables. Cigarette smokers and nonsmokers had median urinary MCAMA concentrations of 517 μg/g creatinine and 127 μg/g creatinine, respectively. Sample-weighted multiple regression analysis showed that MCAMA was positively associated with serum cotinine (p < 0.0001). Compared to non-exposed participants (serum cotinine ≤ 0.015 ng/mL), presumptive exposure to second-hand tobacco smoke (serum cotinine > 0.015–≤ 10 ng/mL and 0 cigarettes smoked per day) was associated with 20% higher MCAMA (p < 0.0001). Additionally, smoking 1–10 cigarettes per day was associated with 261% higher MCAMA (p < 0.0001), smoking 11–20 cigarettes per day was associated with 357% higher MCAMA (p < 0.0001), and smoking > 20 cigarettes per day was associated with 416% higher MCAMA (p < 0.0001). These findings underscore the strong association of tobacco smoke exposure with urinary MCAMA biomarker levels.

A method to achieve accurate measurement of unmetabolized volatile organic compounds (VOCs) in urine was developed and characterized. The method incorporates a novel preanalytical approach of adding isotopically labeled internal standard (ISTD) analogues directly to the collection container at the point of collection to compensate for analyte loss to the headspace and the collection container surfaces. Using this approach, 45 toxic VOCs ranging in water solubility and boiling point were evaluated and analyzed by headspace solid-phase microextraction/gas chromatography–mass spectrometry. Results show that urine VOCs could be equally lost to the container headspace as to the container surface suggesting similarity of these two regions as partition phases. Surface adsorption loss was found to trend with compound water solubility. In particular, with no headspace, more nonpolar VOCs experienced substantial losses (e.g., 48% for hexane) in a standard 120 mL urine cup at concentrations in the low- and sub-ppb range. The most polar VOCs evaluated (e.g., tetrahydrofuran) showed no significant loss. Other commonly practiced methods for urine sample collection and analysis such as aliquoting, specimen freezing, and use of surrogate ISTD were found to significantly bias results. With this method, we achieved errors ranging from −8.0 to 4.8% of spiked urine specimens. Paired urine and blood specimens from cigarette smokers were compared to assess this method.

RationaleOver 2800 e‐cigarette, or vaping, product use‐associated lung injury (EVALI) cases were reported to the Centers for Disease Control and Prevention (CDC) during August 2019 to February 2020. Bronchoalveolar lavage (BAL) fluid samples from 51 EVALI and 99 non‐EVALI cases were analyzed for toxicants including terpenes. We describe a novel method to measure selected terpenes in BAL fluid by gas chromatography/tandem mass spectrometry (GC/MS/MS).Methodsα‐Pinene, β‐pinene, β‐myrcene, 3‐carene, and limonene were measured in BAL fluid specimens by headspace solid‐phase microextraction/gas chromatography/tandem mass spectrometry. We created and characterized BAL fluid pools from non‐EVALI individuals to determine assay accuracy, precision, linearity, limits of detection, and analytical specificity. All measurements were conducted in accordance with the CDC's Division of Laboratory Sciences rigorous method validation procedures.ResultsMatrix validation experiments showed that calibration curves in BAL fluid and saline had similar slopes, with differences of less than 7%. The assay precision ranged from 2.52% to 5.30%. In addition, the limits of detection for the five analytes ranged from 1.80 to 16.8 ng/L, and the linearity was confirmed with R2 values >0.99.ConclusionsWe developed and validated a method to quantify selected terpenes in BAL fluid specimens using GC/MS/MS. The assay provided accurate and precise analyses of EVALI and non‐EVALI BAL fluid specimens in support of CDC's EVALI response. This method is applicable to the determination of a broad range of terpenes in BAL fluid specimens.

Background The causative agents for the current national outbreak of electronic-cigarette, or vaping, product use-associated lung injury (EVALI) have not been established. Detection of toxicants in bronchoalveolar-lavage (BAL) fluid from patients with EVALI can provide direct information on exposure within the lung. Methods BAL fluids were collected from 51 patients with EVALI in 16 states and from 99 healthy participants who were part of an ongoing study of smoking involving nonsmokers, exclusive users of e-cigarettes or vaping products, and exclusive cigarette smokers that was initiated in 2015. Using the BAL fluid, we performed isotope dilution mass spectrometry to measure several priority toxicants: vitamin E acetate, plant oils, medium-chain triglyceride oil, coconut oil, petroleum distillates, and diluent terpenes. Results State and local health departments assigned EVALI case status as confirmed for 25 patients and as probable for 26 patients. Vitamin E acetate was identified in BAL fluid obtained from 48 of 51 case patients (94%) in 16 states but not in such fluid obtained from the healthy comparator group. No other priority toxicants were found in BAL fluid from the case patients or the comparator group, except for coconut oil and limonene, which were found in 1 patient each. Among the case patients for whom laboratory or epidemiologic data were available, 47 of 50 (94%) had detectable tetrahydrocannabinol (THC) or its metabolites in BAL fluid or had reported vaping THC products in the 90 days before the onset of illness. Nicotine or its metabolites were detected in 30 of 47 of the case patients (64%). Conclusions Vitamin E acetate was associated with EVALI in a convenience sample of 51 patients in 16 states across the United States. (Funded by the National Cancer Institute and others.) In a study involving 51 patients with electronic-cigarette, or vaping, product use-associated lung injury in 16 states across the United States, vitamin E acetate was detected in samples of bronchoalveolar-lavage fluid from 94% of the patients but not in samples from a healthy comparator group.

CDC, the Food and Drug Administration (FDA), state and local health departments, and multiple public health and clinical partners are investigating a national outbreak of e-cigarette, or vaping, product use-associated lung injury (EVALI). Based on data collected as of October 15, 2019, 86% of 867 EVALI patients reported using tetrahydrocannabinol (THC)-containing products in the 3 months preceding symptom onset (1). Analyses of THC-containing product samples by FDA and state public health laboratories have identified potentially harmful constituents in these products, such as vitamin E acetate, medium chain triglyceride oil (MCT oil), and other lipids (2,3) (personal communication, D.T. Heitkemper, FDA Forensic Chemistry Center, November 2019). Vitamin E acetate, in particular, might be used as an additive in the production of e-cigarette, or vaping, products; it also can be used as a thickening agent in THC products (4). Inhalation of vitamin E acetate might impair lung function (5-7).

Nail technicians are exposed to volatile organic compounds (VOCs) from nail products, but no studies have previously measured VOC biomarkers for these workers. This study of 10 nail technicians aimed to identify VOCs in nail salons and explore relationships between air concentrations and biomarkers. Personal and area air samples were collected using thermal desorption tubes during a work shift and analyzed using gas chromatography/mass spectrometry (GC/MS) for 71 VOCs. Whole blood samples were collected pre-shift and post-shift, and analyzed using GC/MS for 43 VOCs. Ventilation rates were determined using continuous CO2 measurements. Predominant air VOC levels were ethyl methacrylate (median 240 µg/m3 ), methyl methacrylate (median 205 µg/m3 ), toluene (median 100 µg/m3 ), and ethyl acetate (median 639 µg/m3 ). Blood levels were significantly higher post-shift than pre-shift for toluene (median pre-shift 0.158 µg/L and post-shift 0.360 µg/L) and ethyl acetate (median pre-shift <0.158 µg/L and post-shift 0.510 µg/L); methacrylates were not measured in blood because of their instability. Based on VOCs measured in these seven nail salons, we estimated that emissions from Greater Boston area nail salons may contribute to ambient VOCs. Ventilation rates did not always meet the ASHRAE guideline for nail salons. There is a need for changes in nail product formulation and better ventilation to reduce VOC occupational exposures.

Studies of exposure to petroleum (crude oil/fuel) often involve monitoring benzene, toluene, ethylbenzene, xylenes (BTEX), and styrene (BTEXS) because of their toxicity and gas-phase prevalence, where exposure is typically by inhalation. However, BTEXS levels in the general U.S. population are primarily from exposure to tobacco smoke, where smokers have blood levels on average up to eight times higher than nonsmokers. This work describes a method using partition theory and artificial neural network (ANN) pattern recognition to classify exposure source based on relative BTEXS and 2,5-dimethylfuran blood levels. A method using surrogate signatures to train the ANN was validated by comparing blood levels among cigarette smokers from the National Health and Nutrition Examination Survey (NHANES) with BTEXS and 2,5-dimethylfuran signatures derived from the smoke of machine-smoked cigarettes. Classification agreement for an ANN model trained with relative VOC levels was up to 99.8% for nonsmokers and 100.0% for smokers. As such, because there is limited blood level data on individuals exposed to crude oil/fuel, only surrogate signatures derived from crude oil and fuel were used for training the ANN. For the 2007-2008 NHANES data, the ANN model assigned 7 out of 1998 specimens (0.35%) and for the 2013-2014 NHANES data 12 out of 2906 specimens (0.41%) to the crude oil/fuel signature category.

For over three decades our laboratory has been developing and improving methods for quantifying toxic volatile organic compounds (VOCs) in blood in support of numerous national and regional studies, including nine National Health and Nutrition Examination Survey (NHANES) cycles. Absorption of VOCs most commonly occurs through inhalation, as 100% of blood circulates to the lungs for exchange with alveolar gas. The high blood:gas partition ratios of most VOCs favor preconcentration in the blood, but blood VOC equilibration with tissues and organs is what influences VOC elimination half-life. As such, VOC analysis in blood is best-suited for compounds that are stable in the body by offering a direct measure of VOC burden experienced by tissues and organs.The current analysis method uses headspace solid-phase microextraction (HS-SPME) and gas chromatography-mass spectrometry (GC-MS). Important to the success of this method is the use of isotopically labeled analogs specific to every compound to compensate for competition effects in the headspace and SPME fiber, as well as adsorption and volatilization losses. The combination of these techniques has enabled us to simultaneously quantify a broad array of VOCs (boiling points from 32 to 204 °C) in the low parts-per-trillion (ng/L) range from a 3-mL blood sample.This presentation will include an overview of the blood VOC method and describe recent improvements to achieve accuracy and precision of < 15% for nonpolar compounds (e.g., alkanes) and within 5% for most of the other VOCs. We will also describe noteworthy blood VOC trends in the United States and reveal new analytes that are to be included in future studies and NHANES cycles. In addition to individual VOC trends, we will describe recent work comparing relative VOC levels among participants using artificial neural networks as a means to objectively distinguish exposure between different VOC sources within a large population.

This work describes a quantitative high-throughput analytical method for the simultaneous measurement of small aliphatic nitrogenous biomarkers, i.e., 1,6-hexamethylenediamine (HDA), isophoronediamine (IPDA), β-methylamino-l-alanine (BMAA), and trimethylamine N-oxide (TMAO), in human urine. Urinary aliphatic diamines, HDA and IPDA, are potential biomarkers of environmental exposure to their corresponding diisocyanates. Urinary BMAA forms as a result of human exposure to blue-green algae contaminated food. And, TMAO is excreted in urine due to the consumption of carnitine- and choline-rich diets. These urinary biomarkers represent classes of small aliphatic nitrogen-containing compounds (N-compounds) that have a high aqueous solubility, low logP, and/or high basic pKa. Because of the highly polar characteristics, analysis of these compounds in complex sample matrices is often challenging. We report on the development of ion-pairing chemistry based ultra-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UPLC-ESI-MS/MS) method for the simultaneous measurement of these biomarkers in human urine. Chromatographic separation was optimized using heptafluorobutyric acid-(HFBA-) based mobile phase and a reversed-phase C18 column. All four analytes were baseline separated within 2.6 min with an overall run time of 5 min per sample injection. Sample preparation involved 4 h of acid hydrolysis followed by automated solid phase extraction (SPE) performed using strong cation exchange sorbent bed with 7 N ammonia solution in methanol as eluent. Limits of detection ranged from 0.05 ng/mL to 1.60 ng/mL. The inter-day and intra-day accuracy were within 10%, and reproducibility within 15%. The method is accurate, fast, and well-suited for biomonitoring studies within targeted groups, as well as larger population-based studies such as the U. S. National Health and Nutrition Examination Survey (NHANES).

To address concerns among Gulf Coast residents about ongoing exposures to volatile organic compounds, including benzene, toluene, ethylbenzene, o -xylene, and m -xylene/ p -xylene (BTEX), we characterized current blood levels and identified predictors of BTEX among Gulf state residents. We collected questionnaire data on recent exposures and measured blood BTEX levels in a convenience sample of 718 Gulf residents. Because BTEX is rapidly cleared from the body, blood levels represent recent exposures in the past 24 h. We compared participants’ levels of blood BTEX to a nationally representative sample. Among nonsmokers we assessed predictors of blood BTEX levels using linear regression, and predicted the risk of elevated BTEX levels using modified Poisson regression. Blood BTEX levels in Gulf residents were similar to national levels. Among nonsmokers, sex and reporting recent smoky/chemical odors predicted blood BTEX. The change in log benzene was −0.26 (95% CI: −0.47, −0.04) and 0.72 (0.02, 1.42) for women and those who reported odors, respectively. Season, time spent away from home, and self-reported residential proximity to Superfund sites (within a half mile) were statistically associated with benzene only, however mean concentration was nearly an order of magnitude below that of cigarette smokers. Among these Gulf residents, smoking was the primary contributor to blood BTEX levels, but other factors were also relevant.

OBJECTIVEOur objective was to improve understanding of the differences in use behavior and exposure when smoking menthol and non-menthol cigarettes using a 2-part cross-over design.METHODSAdult daily smokers were assigned randomly to alternate between 2 weeks of exclusively smoking a menthol test cigarette or a non-menthol test cigarette. Urine and saliva were collected for biomarker measurements; carbon monoxide (CO) was measured, and participants smoked test cigarettes through a CreSS® smoking topography device during 3 clinic visits. Participants turned in their cigarette butts from the test periods for determination of mouth level nicotine and completed subjective questionnaires related to the test cigarettes.RESULTSRegardless of cigarette preference, participants had higher salivary cotinine when smoking the non-menthol test cigarette, but there were no significant differences detected in urine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol between the 2 test cigarettes. Mouth level nicotine, puff volume, and puff duration were significantly higher when smoking the menthol brand. Both menthol and non-menthol smokers reported significantly lower enjoyment and satisfaction scores for test cigarettes compared with their brand of choice.CONCLUSIONSOur results suggest that mentholation has an effect on measures of smoking behavior and that mouth level nicotine is a useful indicator of between-brand smoke exposure.

To accurately measure menthol levels in human urine, we developed a method using gas chromatography/electron ionization mass spectrometry with menthol-d4 stable isotope internal standardization. We used solid phase microextraction (SPME) headspace sampling for collection, preconcentration and automation. Conjugated forms of menthol were released using β-glucuronidase/sulfatase to allow for measuring total menthol. Additionally, we processed the specimens without using β-glucuronidase/sulfatase to quantify the levels of unconjugated (free) menthol in urine. This method was developed to verify mentholated cigarette smoking status to study the influence of menthol on smoking behaviour and exposure. This objective was accomplished with this method, which has no carryover or memory from the SPME fiber assembly, a method detection limit of 0.0017 μg/mL, a broad linear range of 0.002–0.5 μg/mL for free menthol and 0.01–10 μg/mL for total menthol, a 7.6% precision and 88.5% accuracy, and an analysis runtime of 17 min. We applied this method in analysis of urine specimens collected from cigarette smokers who smoke either mentholated or non-mentholated cigarettes. Among these smokers, the average total urinary menthol levels was three-fold higher (p < 0.001) among mentholated cigarette smokers compared with non-mentholated cigarette smokers.

INTRODUCTION A significant portion of the increased risk of cancer and respiratory disease from exposure to cigarette smoke is attributed to volatile organic compounds (VOCs). In this study, 21 VOCs were quantified in mainstream cigarette smoke from 50U.S. domestic brand varieties that included high market share brands and 2 Kentucky research cigarettes (3R4F and 1R5F). METHODS Mainstream smoke was generated under ISO 3308 and Canadian Intense (CI) smoking protocols with linear smoking machines with a gas sampling bag collection followed by solid phase microextraction/gas chromatography/mass spectrometry (SPME/GC/MS) analysis. RESULTS For both protocols, mainstream smoke VOC amounts among the different brand varieties were strongly correlated between the majority of the analytes. Overall, Pearson correlation (r) ranged from 0.68 to 0.99 for ISO and 0.36 to 0.95 for CI. However, monoaromatic compounds were found to increase disproportionately compared to unsaturated, nitro, and carbonyl compounds under the CI smoking protocol where filter ventilation is blocked. CONCLUSIONS Overall, machine generated "vapor phase" amounts (µg/cigarette) are primarily attributed to smoking protocol (e.g., blocking of vent holes, puff volume, and puff duration) and filter ventilation. A possible cause for the disproportionate increase in monoaromatic compounds could be increased pyrolysis under low oxygen conditions associated with the CI protocol. IMPLICATIONS This is the most comprehensive assessment of volatile organic compounds (VOCs) in cigarette smoke to date, encompassing 21 toxic VOCs, 50 different cigarette brand varieties, and 2 different machine smoking protocols (ISO and CI). For most analytes relative proportions remain consistent among U.S. cigarette brand varieties regardless of smoking protocol, however the CI smoking protocol did cause up to a factor of 6 increase in the proportion of monoaromatic compounds. This study serves as a basis to assess VOC exposure as cigarette smoke is a principle source of overall population-level VOC exposure in the United States.

Urinary diamines are biomarkers of diisocyanate exposure. Diisocyanates are considered as skin and respiratory sensitizers and are the most frequently reported cause of occupational asthma. Herein we report on the development and validation of an ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the measurement of five aromatic diamines, 4,4'-methylenedianiline (MDA), 2,4-toluenediamine (4TDA), 2,6-toluenediamine (6TDA), 1,5-naphthalenediamine (NDA), and p-phenylenediamine (PPDA) in human urine. The method incorporates sample preparation steps, which include a 4 h acid hydrolysis followed by high-throughput solid-phase extraction prior to chromatographic separation. Chromatographic separation was achieved using a C18 reversed phase column with gradient elution of basic mobile phases (pH 9.2). The duty cycle of the method was less than 5 min, including both the column equilibration and autosampler movement. Analytical detection was performed using positive ion atmospheric pressure chemical ionization tandem mass spectrometry (APCI-MS/MS) in scheduled multiple reaction monitoring (sMRM) mode. Excellent linearity was observed over standard calibration curve concentration ranges of 3 orders of magnitude with method detection limit ranging from 10 to 100 pg/mL. The interday and intraday reproducibility and accuracy were within ±15%. This method is fast, accurate, and reproducible and is suitable for assessment of exposure to the most common aromatic diisocyanates within targeted groups as well as larger population studies such as the National Health and Nutrition Examination Survey (NHANES).

Objective: This study aimed to evaluate blood volatile organic compound (VOC) levels as biomarkers of occupational jet propulsion fuel 8 (JP-8) exposure while controlling for smoking. Methods: Among 69 Air Force personnel, post-shift blood samples were analyzed for components of JP-8, including ethylbenzene, toluene, o-xylene, and m/p-xylene, and for the smoking biomarker, 2,5-dimethylfuran. JP-8 exposure was characterized based on self-report and measured work shift levels of total hydrocarbons in personal air. Multivariate regression was used to evaluate the relationship between JP-8 exposure and post-shift blood VOCs while controlling for potential confounding from smoking. Results: Blood VOC concentrations were higher among US Air Force personnel who reported JP-8 exposure and work shift smoking. Breathing zone total hydrocarbons was a significant predictor of VOC blood levels, after controlling for smoking. Conclusions: These findings support the use of blood VOCs as a biomarker of occupational JP-8 exposure.

Quantifying volatile organic compounds (VOCs) in cigarette smoke is necessary to establish smoke-related exposure estimates and evaluate emerging products and potential reduced-exposure products. In response to this need, we developed an automated, multi-VOC quantification method for machine-generated, mainstream cigarette smoke using solid-phase microextraction gas chromatography-mass spectrometry (SPME-GC-MS). This method was developed to simultaneously quantify a broad range of smoke VOCs (i.e., carbonyls and volatiles, which historically have been measured by separate assays) for large exposure assessment studies. Our approach collects and maintains vapor-phase smoke in a gas sampling bag, where it is homogenized with isotopically labeled analogue internal standards and sampled using gas-phase SPME. High throughput is achieved by SPME automation using a CTC Analytics platform and custom bag tray. This method has successfully quantified 22 structurally diverse VOCs (e.g., benzene and associated monoaromatics, aldehydes and ketones, furans, acrylonitrile, 1,3-butadiene, vinyl chloride, and nitromethane) in the microgram range in mainstream smoke from 1R5F and 3R4F research cigarettes smoked under ISO (Cambridge Filter or FTC) and Intense (Health Canada or Canadian Intense) conditions. Our results are comparable to previous studies with few exceptions. Method accuracy was evaluated with third-party reference samples (≤15% error). Short-term diffusion losses from the gas sampling bag were minimal, with a 10% decrease in absolute response after 24 h. For most analytes, research cigarette inter- and intrarun precisions were ≤20% relative standard deviation (RSD). This method provides an accurate and robust means to quantify VOCs in cigarette smoke spanning a range of yields that is sufficient to characterize smoke exposure estimates.