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A method for the measurement of 24 hydroxylated polycyclic aromatic hydrocarbon metabolites (OH-PAHs) in urine has been developed. The method is based on enzymatic deconjugation, automated liquid--liquid extraction, and gas chromatography/isotope dilution high-resolution mass spectrometry after derivatization of the OH-PAHs to the trimethylsilylated derivatives. The metabolites included in the current method are formed from eight different parent compounds. The limits of detection were below 7 pg/mL when using a sample size of 2 mL of urine, except for 1- and 2-naphthols (18 and 12 pg/mL, respectively). The enzymatic deconjugation efficiency, verified by deconjugation of urine samples spiked with [alpha]-naphthyl [beta]-D-glucuronide sodium salt (1-NAP-GLU) and pyrene-1-sulfate potassium salt (1-PYR-SULF), was determined to be 97% for 1-NAP-GLU conjugate and 84% for 1-PYR-SULF. The overall coefficients of variance for six batches of quality control samples (n = 42), was 2.9-11%. Mean method recoveries of the [sup.13]C-labeled internal standards were 66-72%, except for [sup.13][C.sub.6-1-naphthol (46%). The throughput of this method has been determined to be 40 samples per day per analyst. This method is currently applied to epidemiological studies, such as the National Exposure and Nutrition Examination Survey (NHANES), to measure human exposure to PAHs.

The ability to estimate low-dose human exposure to commonly used pesticides often is requested in epidemiologic studies. Therefore, fast and robust methods are necessary that can measure many analytes in the same sample. We have developed a method for high-throughput analysis of 19 markers of commonly used pesticides in human urine. The analytes were seven specific metabolites of organophosphorus pesticides, five metabolites of synthetic pyrethroids, six herbicides or their metabolites, and one insect repellant. Human urine (2 mL) was spiked with stable isotopically labeled analogues of the analytes, enzymatically hydrolyzed, extracted using solid-phase extraction, concentrated, and analyzed using high-performance liquid chromatography-tandem mass spectrometry. The sample was divided into two portions and analyzed on two different mass spectrometers, one using atmospheric pressure chemical ionization (APCI) and the other using turbo ion spray atmospheric pressure ionization (TIS). All analytes except the pyrethroid metabolites were analyzed using APCI. The detection limits for all analytes ranged from 0.1 to 1.5 ng/mL of urine, with the majority (17) below 0.5 ng/mL. The analytical precision for the different analytes, estimated as both the within-day and between-day variation, was 3-14 and 4-19%, respectively. The extraction recoveries of the analytes ranged from 68 to 114%. The throughput, including calibration standards and quality control samples, is ~50 samples a day. However, the analysis time with the TIS application is much shorter, and if only pyrethroid metabolite data are of interest, the throughput can be increased to 100-150 samples/day.

Abstract Background Urban air pollution is an increasing health problem, particularly in Asia, where the combustion of fossil fuels has increased rapidly as a result of industrialization and socio-economic development. The adverse health impacts of urban air pollution are well established, but less is known about effective intervention strategies. In this demonstration study we set out to establish methods to assess whether wearing an R95 activated carbon respirator could reduce intake of polycyclic aromatic hydrocarbons (PAH) in street workers in Hanoi, Vietnam. Methods In this demonstration study we performed a cross-over study in which non-smoking participants that worked at least 4 hours per day on the street in Hanoi were randomly allocated to specific respirator wearing sequences for a duration of 2 weeks. Urines were collected after each period, i.e. twice per week, at the end of the working day to measure hydroxy PAHs (OH-PAH) using gas chromatography/high resolution mass spectrometry. The primary endpoint was the urinary concentration of 1-hydroxypyrene (1-OHP). Results Forty-four participants (54.5% male, median age 40 years) were enrolled with the majority being motorbike taxi drivers (38.6%) or street vendors (34.1%). The baseline creatinine corrected urinary level for 1-OHP was much higher than other international comparisons: 1020 ng/g creatinine (IQR: 604–1551). Wearing a R95 mask had no significant effect on 1-OHP levels: estimated multiplicative effect 1.0 (95% CI: 0.92-1.09) or other OH-PAHs, except 1-hydroxynaphthalene (1-OHN): 0.86 (95% CI: 0.11-0.96). Conclusions High levels of urine OH-PAHs were found in Hanoi street workers. No effect was seen on urine OH-PAH levels by wearing R95 particulate respirators in an area of high urban air pollution, except for 1-OHN. A lack of effect may be de to gaseous phase PAHs that were not filtered efficiently by the respirator. The high levels of urinary OH-PAHs found, urges for effective interventions. Trial registration ISRCTN74390617 (date of assignation: 04/08/2009).

Introduction: Since 2004, several jurisdictions have mandated that cigarettes show reduced ignition propensity (RIP) in laboratory testing. RIP cigarettes may limit fires caused by smoldering cigarettes, reducing fire-related deaths and injury. However, some evidence suggests that RIP cigarettes emit more carbon monoxide and polycyclic aromatic hydrocarbons, and smokers may alter their smoking patterns in response to RIP cigarettes. Both of these could increase smokers’ exposures to harmful constituents in cigarettes. Methods: An 18-day switching study with a comparison group was conducted in Boston, MA (N = 77), and Buffalo, NY (N = 83), in 2006–2007. Current daily smokers completed 4 laboratory visits and two 48-hr field data collections. After a 4-day baseline, Boston participants switched to RIP cigarettes for 14 days, whereas Buffalo participants smoked RIP cigarettes throughout. Outcome measures included cigarettes smoked per day; smoking topography; salivary cotinine; breath CO; and hydroxylated metabolites of pyrene, naphthalene, phenanthrene, and fluorene. Because the groups differed demographically, analyses adjusted for race, age, and sex. Results: We observed no significant changes in smoking topography or CO exposure among participants who switched to RIP cigarettes. Cigarette use decreased significantly in the switched group (37.7 cigarettes/48 hr vs. 32.6 cigarettes/48 hr, p = .031), while hydroxyphenanthrenes increased significantly (555 ng/g creatinine vs. 669 ng/g creatinine, p = .007). No other biomarkers were significantly affected. Discussion: Small increases in exposure to phenanthrene among smokers who switched to RIP versions were observed, while other exposures and smoking topography were not significantly affected. Toxicological implications of these findings are unclear. These findings should be weighed against the potential public health benefits of adopting RIP design standards for cigarette products. [ABSTRACT FROM PUBLISHER]

Urinary mono-hydroxy polycyclic aromatic hydrocarbons (OH-PAHs) are commonly used in biomonitoring to assess exposure to polycyclic aromatic hydrocarbons (PAHs). Similar to other biologically non-persistent chemicals, OH-PAHs have relatively short biological half-lives (4.4–35 h). Little information is available on their variability in urinary concentrations over time in non-occupationally exposed subjects. This study was designed to (i) examine the variability of nine urinary OH-PAH metabolite concentrations over time and (ii) calculate sample size requirements for future epidemiological studies on the basis of spot urine, first-morning void, and 24-h void sampling. Individual urine samples (n=427) were collected during 1 week from 8 non-occupationally exposed adults. We recorded the time and volume of each urine excretion, dietary details, and driving activities of the participants. Within subjects, the coefficients of variation (CVs) for the wet-weight concentration of OH-PAHs in all samples ranged from 45% to 297%; creatinine adjustment reduced the CV to 19–288% (P<0.001; paired t-test). The simulated 24-h void concentrations were the least variable measure, with CVs ranging from 13% to 182% for the 9 OH-PAHs. Within-day variability contributed on average 84%, and between-day variability accounted for 16% of the total variance of 1-hydroxypyrene (1-PYR). Intraclass correlation coefficients of 1-PYR levels were 0.55 for spot urine samples, 0.60 for first-morning voids, and 0.76 for 24-h voids, indicating a high degree of correlation between urine measurements collected from the same subject over time. Sample size calculations were performed to estimate the number of subjects required for detecting differences in the geometric mean at a statistical power of 80% for spot urine, first-morning, and 24-h void sampling. These data will aid in the design of future studies of PAHs and possibly other biologically non-persistent chemicals and in the interpretation of their analytical results. [ABSTRACT FROM AUTHOR]

We are reporting a method for measuring 43 polycyclic aromatic hydrocarbons (PAH) and their methylated derivatives (Me-PAHs) in air particulate matter (PM) samples using isotope dilution gas chromatography/high-resolution mass spectrometry (GC/HRMS). In this method, PM samples were spiked with internal standards, loaded into solid phase extraction cartridges, and eluted by dichloromethane. The extracts were concentrated, spiked with a recovery standard, and analyzed by GC/HRMS at 10,000 resolution. Sixteen 13C-labeled PAHs and two deuterated Me-PAHs were used as internal standards to account for instrument variability and losses during sample preparation. Recovery of labeled internal standards was in the range of 86–115%. The proposed method is less time-consuming than commonly used extraction methods, such as sonication and accelerated solvent extraction (ASE), and it eliminates the need for a filtration step required after the sonication extraction method. Limits of detection ranged from 41 to 332 pg/sample for the 43 analytes. This method was used to analyze reference materials from the National Institute of Standards and Technology. The results were consistent with those from ASE and sonication extraction, and these results were also in good agreement with the certified or reference concentrations. The proposed method was then used to measure PAHs on PM2.5 samples collected at three sites (urban, suburban, and rural) in Atlanta, GA. The results showed distinct seasonal and spatial variation and were consistent with an earlier study measuring PM2.5 samples using an ASE method, further demonstrating the compatibility of this method and the commonly used ASE method. [Figure not available: see fulltext.] [ABSTRACT FROM AUTHOR]

A method for the measurement of 24 hydroxylated polycyclic aromatic hydrocarbon metabolites (OH-PAHs) in urine has been developed. The method is based on enzymatic deconjugation, automated liquid-liquid extraction, and gas chromatography/isotope dilution high-resolution mass spectrometry after derivatization of the OH-PAHs to the trimethylsilylated derivatives. The metabolites included in the current method are formed from eight different parent compounds. The limits of detection were below 7 pg/mL when using a sample size of 2 mL of urine, except for 1- and 2-naphthols (18 and 12 pg/mL, respectively). The enzymatic deconjugation efficiency, verified by deconjugation of urine samples spiked with α-naphthyl β-D-glucuronide sodium salt (1-NAP-GLU) and pyrene-1-sulfate potassium salt (1-PYR-SULF), was determined to be 97% for 1-NAP-GLU conjugate and 84% for 1-PYR-SULF. The overall coefficients of variance for six batches of quality control samples (n = 42), was 2.9–11%. Mean method recoveries of the 13C-labeled internal standards were 66–72%, except for 13C6-1-naphthol (46%). The throughput of this method has been determined to be 40 samples per day per analyst. This method is currently applied to epidemiological studies, such as the National Exposure and Nutrition Examination Survey (NHANES), to measure human exposure to PAHs. [ABSTRACT FROM AUTHOR]

Public transport vehicle drivers, especially in highly polluted or trafficked areas, are exposed to high levels of air pollutants. In this study, we assessed the influence of traffic on levels of hydroxy polycyclic aromatic hydrocarbons (OH-PAHs) in commercial bus drivers in Trujillo, Peru, by measuring the within-shift changes in the urinary whole weight and creatinine-corrected concentrations of the PAH metabolites. We measured personal PM(2.5) as a proxy of exposure to traffic emission. Urine samples were collected daily from two bus drivers and three minivan drivers in Trujillo, pre-, mid-, post-work shift and on days when the drivers were off work (total n = 144). Ten OH-PAH metabolites were measured in the urine samples. Drivers were also monitored for exposure to PM(2.5) (n = 41). Daily work shift (mean = 13.1 ± 1.3 hr) integrated PM(2.5) was measured in the breathing zones of the drivers for an average of 10.5 days per driver. The differences across shift in OH-PAH concentrations were not statistically significant except for urinary 2-hydroxyfluorene (2-FLU) (p = 0.04) and 4-hydroxyphenanthrene (4-PHE) (p = 0.01) and creatinine-corrected 4-hydroxyphenanthrene (p = 0.01). Correlation between pairs of hydroxy-PAHs (ρ = 0.50 to 0.93) were highest for mid-shift samples. Concentrations of PM(2.5) (geometric mean = 64 μg/m(3); 95% confidence limits = 52 μg/m(3), 78 μg/m(3)) is similar to those measured in many other studies of traffic exposure. There was significant change across work shift for concentrations of only two of the OH-PAHs (2-FLU and 4-PHE). Results indicate that the drivers may have had limited time for clearance of PAH exposure from the body between work shifts. Comparisons of the concentrations of creatinine-corrected hydroxy-PAH to those reported in other studies indicate that exposure of public transport drivers to PAH could be similar. By following the subjects over multiple days, this study gives an indication of appropriate exposure situations for the use of hydroxy-PAHs and will be beneficial in designing future occupational studies of PAH exposure.

A method for the measurement of 24 hydroxylated polycyclic aromatic hydrocarbon metabolites (OH-PAHs) in urine has been developed. The method is based on enzymatic deconjugation, automated liquid-liquid extraction, and gas chromatography/isotope dilution high-resolution mass spectrometry after derivatization of the OH-PAHs to the trimethylsilylated derivatives. The metabolites included in the current method are formed from eight different parent compounds. The limits of detection were below 7 pg/mL when using a sample size of 2 mL of urine, except for 1- and 2-naphthols (18 and 12 pg/mL, respectively). The enzymatic deconjugation efficiency, verified by deconjugation of urine samples spiked with alpha-naphthyl beta-d-glucuronide sodium salt (1-NAP-GLU) and pyrene-1-sulfate potassium salt (1-PYR-SULF), was determined to be 97% for 1-NAP-GLU conjugate and 84% for 1-PYR-SULF. The overall coefficients of variance for six batches of quality control samples (n = 42), was 2.9-11%. Mean method recoveries of the 13C-labeled internal standards were 66-72%, except for 13C6-1-naphthol (46%). The throughput of this method has been determined to be 40 samples per day per analyst. This method is currently applied to epidemiological studies, such as the National Exposure and Nutrition Examination Survey (NHANES), to measure human exposure to PAHs.