Simple and reliable method to simultaneously determine urinary 1- and 2-naphthol using in situ derivatization and gas chromatography-mass spectrometry for biological monitoring of naphthalene exposure in occupational health practice.

OBJECTIVES: The aim of this study was to develop and validate a simple and reliable gas chromatography-mass spectrometry (GC-MS) method to simultaneously determine urinary 1-naphthol (1-NAP) and 2-naphthol (2-NAP) for biological monitoring of occupational exposure to naphthalene. METHODS: NAPs were derivatized in situ with acetic anhydride after enzymatic hydrolysis, extracted with n-hexane, and analyzed using GC-MS. Validation of the proposed method was conducted in accordance with US Food and Drug Administration guidance. A final validation was performed by analyzing a ClinChek® -Control for phenolic compounds. RESULTS: The linearity of calibration curves was indicated by a high correlation coefficient (>0.999) in the concentration range 1-100 µg/L for each NAP. The limits of detection and quantification for each NAP were 0.30 and 1.00 µg/L, respectively. The recovery was 90.8%-98.1%. The intraday and interday accuracies, expressed as the deviation from the nominal value, were 92.2%-99.9% and 93.4%-99.9%, respectively. The intraday and interday precision, expressed as the relative standard deviation, was 0.3%-3.9% and 0.4%-4.1%, respectively. The ClinChek® values obtained using our method were sufficiently accurate. CONCLUSIONS: The proposed method is simple, reliable, and appropriate for routine analyses, and is useful for biological monitoring of naphthalene exposure in occupational health practice.

Production of a specific monoclonal antibody for 1-naphthol based on novel hapten strategy and development of an easy-to-use ELISA in urine samples.

1-naphthol (1-NAP) is the main metabolite of pesticide carbaryl and naphthalene, and is also a genotoxic and carcinogenic intermediate in the synthesis of organic compound, dyes, pigment and pharmaceutical industry. In this work, two novel haptens were designed and synthesized for developing a competitive indirect enzyme-linked immunosorbent assay (ciELISA) method for 1-NAP in urine samples. The assay showed a limit of detection of 2.21 ng/mL and working range from 4.02 ng/mL to 31.25 ng/mL for 1-NAP in optimized working buffer. The matrix effect of samples was eliminated via 15-fold dilution of optimized working buffer. Good average recoveries (102.4%-123.4%) with a coefficient of variation from 11.7% to 14.7% was obtained for spiked urine samples. Subsequent instrument verification test showed good correlation between the results of ciELISA and high-performance liquid chromatography. The developed ciELISA is a high-throughput tool to monitor 1-NAP in urine, which can provide technical support for the establishment of biological exposure level for the exposure to carbaryl, naphthalene and other related pollutants.

Alterations in platelet indices link polycyclic aromatic hydrocarbons toxicity to low-grade inflammation in preschool children.

BACKGROUND: Environmental exposure to carcinogenic polycyclic aromatic hydrocarbons (PAHs) can disturb the immune response. However, the effect of PAHs on low-grade inflammation related to platelets in humans is unknown. OBJECTIVES: We investigated the association of PAH exposure with low-grade inflammation and platelet parameters in healthy preschoolers. METHODS: The present study recruited 239 participants, aged 2-7 years, from an electronic-waste (e-waste)-exposed (n = 118) and a reference (n = 121) area. We measured ten urinary PAH metabolites, four types of immune cells and cytokines, and seven platelet parameters, and compared their differences between children from the two groups. Spearman correlation analysis was performed to explore the potential risk factors for PAH exposure and the associations between urinary monohydroxylated PAHs (OH-PAHs) and biological parameters. Associations between urinary PAH metabolites and platelet indices were analyzed using quantile regression models. Mediation analysis was used to understand the relationship between urinary total hydroxynaphthalene (ΣOHNa) and interleukin (IL)-1ß through seven platelet indices, as mediator variables. RESULTS: We found higher urinary monohydroxylated PAH (OH-PAH) concentrations, especially 1-hydroxynaphthalene (1-OHNa) and 2-hydroxynaphthalene (2-OHNa), in children from the e-waste-exposed group than in the reference group. These were closely associated with child personal habits and family environment. A decreased lymphocyte ratio and increased pro-inflammatory cytokines, such as gamma interferon-inducible protein (IP)-10 and IL-1ß, were found in the e-waste-exposed children. After adjustment for confounding factors, significantly negative correlations were found between levels of mean platelet volume (MPV), platelet distribution width (PDW), platelet-large cell ratio (P-LCR) and ratio of mean platelet volume to platelet count (MPVP) and OH-PAHs. In addition, ΣOHNa was positively associated with IL-1ß mediated through MPV, PDW, P-LCR, and ratio of platelet count to lymphocyte count (PLR). CONCLUSIONS: Platelet indices were significantly associated with the changes in urinary OH-PAH levels, which may can be regarded as effective biomarkers of low-grade inflammation resulting from low PAH exposure in healthy children.

Validity of different biomonitoring parameters in human urine for the assessment of occupational exposure to naphthalene.

Up to date, information on the validity of human biomonitoring (HBM) parameters of naphthalene exposure is poor. This study was performed to reveal the relation between occupational exposure to naphthalene and biological exposure markers. Therefore, ten lowly and highly exposed workers from the abrasives industry were selected to characterise a broad exposure range. Naphthalene in air was determined by personal air monitoring during one shift. For biological monitoring, pre- and post-shift urine samples collected on 2 days of a working week were analysed for 1,2-dihydroxynaphthalene (1,2-DHN), 1- and 2-naphthol, 1- and 2-naphthylmercapturic acid (NMA). The naphthalene concentration in air was in the range of 0.5 to 11.6 mg/m3. The biomarkers in urine showed post-shift concentration in the range of 114-51,809 µg/L for 1,2-DHN, 0.8-666 µg/L for 1-NMA, 2-2698 µg/L for 1-naphthol and 4-1135 µg/L for 2-naphthol, respectively. 2-NMA was not detected. The urinary levels increased significantly from pre- to post-shift for all analysed parameters and an accumulation over the working week was observed. Significant positive correlations were observed between 1,2-DHN, 1-NMA, 1- and 2-naphthol in post-shift urine samples and personal exposure to naphthalene in the air. 1-NMA and 1,2-DHN, 1- and 2-naphthol have been demonstrated as suitable biomarkers for naphthalene exposure monitoring. Of the determined biomarkers, 1,2-DHN is by far the metabolite with the highest concentration in the urine samples.

Human Semen Quality, Sperm DNA Damage, and the Level of Urinary Concentrations of 1N and TCPY, the Biomarkers of Nonpersistent Insecticides.

The aim of the study was to evaluate the association between environmental exposure to nonpersistent insecticides and semen quality (concentration, motility, morphology, computer-aided semen analysis [CASA] parameters, and sperm DNA damage). Urine samples ( n = 315) collected from men who attended the infertility clinic with normal semen concentration of 15 to 300 mln/ml and age under 45 years were analyzed for two metabolites (1-naphthol [1N] and 3,5,6-trichloro-2-pyridinol [TCPY]) of nonpersistent insecticides. Participants provided semen, blood, and saliva samples; additionally, men filled a detailed questionnaire. The results identified that urinary TCPY concentration was significantly associated with a decrease in motility; also there was a positive association between TCPY and DNA fragmentation index (DFI). 1N concentration was negatively associated with a percentage of sperm with normal morphology and positively with one of the CASA parameters (curvilinear velocity [VCL]). The results suggest that environmental exposure to nonpersistent insecticides may have an impact on semen quality parameters and sperm DNA damage.

Maternal urinary 2-hydroxynaphthalene and birth outcomes in Taiyuan, China.

BACKGROUND: Naphthalene is the simplest polycyclic aromatic hydrocarbon (PAH). It is easily emitted into the atmosphere, posing a significant risk to human health. However, limited studies have described the impact of naphthalene exposure on birth outcomes. In this study, we investigated the association between the maternal urinary metabolites of naphthalene, 2-hydroxynaphthalene (2-OH NAP), and birth outcomes. METHOD: In the present study, four urinary PAH metabolites were measured in 263 pregnant women during late pregnancy. Multiple linear regression analysis was used to analyze the relationship between the concentrations of 2-OH NAP and birth outcomes, and restricted cubic spline models were further used to examine the shapes of the dose-response association. RESULT: General linear models showed that prenatal urinary 2-OH NAP was associated with lower birth weight (BW) (- 4.38% for the high vs. low exposure group of 2-OH NAP; p for trend = 0.049) and higher cephalization index (CI) (4.30% for the high vs. low exposure group of 2-OH NAP; p for trend = 0.038). These associations were linear and significant when 2-OH NAP was modeled as a continuous variable in restricted cubic spline models (P linear = 0.0293 for 2-OH NAP and BW; P linear = 0.0326 for 2-OH NAP and CI). Multiple linear regression data indicated that each 1 ln-unit increase in 2-OH NAP was significantly associated with a 2.09 g/cm increase in the CI. The associations among 2-OH NAP, BW, and CI were also observed in a subset of participants residing close to arterial traffic. CONCLUSION: Our data indicated that prenatal exposure to naphthalene had an adverse effect on fetal birth outcomes, especially the brain development index. Reduced exposure to naphthalene may improve newborn health outcomes. In Taiyuan, naphthalene may result from traffic pollution.

2-Naphthol Levels and Allergic Disorders in Children.

BACKGROUND: The measurement of polycyclic aromatic hydrocarbons (PAH) in ambient air is quite difficult to perform. Using urine biomarkers of PAH such as 2-naphthol is one approach to this problem. This study explored the association between urine 2-naphthol levels and allergic diseases. The associations between 2-naphthol levels and oxidative stress biomarkers for the possible disease pathogenesis were also investigated. METHOD: A total of 453 kindergarten children from the (Childhood Environment and Allergic Diseases Study) CEAS cohort with urine samples were recruited. Urine 2-naphthol levels were measured by high-performance liquid chromatography mass spectrometry (HPLC-MS/MS) and markers of oxidative stress (8OHdG) were measured by enzyme-linked immunosorbent assays (ELISA). Information on environmental risk factors and allergic diseases were also collected. The association between 2-naphthol levels, 8OHdG levels, IgE, and allergic diseases were evaluated by multivariate linear regression and logistic regression. RESULTS: Levels of 2-naphthol were positively correlated with 8OHdG levels. A one ln-unit increase in the 2-naphthol level was positively associated to 8OHdG levels (per ln-unit: ß = 100.61, p < 0.001). When dividing 2-naphthol levels into quartiles, asthma was significantly associated with 2-naphthol levels at a concentration of >1.60 ng/mL (adjusted OR: 3.14, 95% CI 1.34⁻7.35). CONCLUSION: Urine 2-naphthol levels are associated with markers of oxidative stress and the risk of allergic diseases in young children.

Suitability of several naphthalene metabolites for their application in biomonitoring studies.

Naphthalene occurs together with polycyclic aromatic hydrocarbons (PAHs) at industrial workplaces and is ubiquitous in the environment. For biological monitoring of naphthalene exposures, up to now mainly 1- and 2-naphthol in urine have been used. Recently, we proposed 1,2-dihydroxynaphthalene (1,2-DHN) and the 1- and 2-naphthylmercapturic acid (1- and 2-NMA) as new urinary biomarkers to characterise a naphthalene exposure. In this study, in a collective of nine occupationally exposed workers handling with creosote the naphthalene metabolites 1,2-DHN, 1- and 2-NMA as well as 1- and 2-naphthol were analysed in order to evaluate the suitability of the different parameters for their application in biomonitoring studies. Additionally, air sampling was conducted to characterise the exposure in task related exposure situations at different workplaces. In the analysed 51 urine samples, 1,2-DHN was the main metabolite with concentrations ranging from 2.3 to 886 µg/g creatinine (crea) (median 34 µg/g crea). For the sum of 1- and 2-naphthol, concentrations in the range of 2.6-174 µg/g crea (median 15 µg/g crea) were observed. 1-NMA concentrations were in the range of < LOD-2.4 µg/g crea (61% > LOD), while 2-NMA was not detected in the analysed urine samples. The biomarkers 1,2-DHN, 1- and 2-naphthol as well as 1-NMA showed significant correlations, which pointed out to naphthalene as the common exposure source. The poor correlations between naphthalene in the air and the biomarkers in urine may be a result of the varying exposure situations and may indicate not solely inhalative, but additional dermal uptake. 1,2-DHN was the most sensitive and, together with 1-NMA, the most specific parameter of the biological monitoring of naphthalene exposure at workplaces. Further studies with this parameter are needed for individuals at different workplaces as well as for persons of the general population without occupational PAH exposure to characterise 1,2-DHN levels as well as to establish their relationship with the naphthalene exposure.

Environmental and biological monitoring of occupational exposure to polynuclear aromatic hydrocarbons during highway pavement construction in Italy.

OBJECTIVES: We performed a cross-sectional study with the main aim of evaluating occupational exposure to polycyclic aromatic hydrocarbons (PAHs) in workers involved in the pavement construction of a new highway in Northern Italy, where modified bitumen was used as binder for Hot Mix Asphalt. METHODS: We applied a combined approach of air and biological monitoring. Both the aerosol and vapour phases of bitumen were collected applying the NIOSH 5506 method. The 16 PAHs listed as high priority by EPA were determined by HPLC-UV. End-of-shift urine samples were collected from 144 workers to determine 1-hydroxypyrene (1-OHP) and 2-naphthol (2-NAP) concentrations after enzyme digestion and HPLC-UV analysis. Socio-demographic and lifestyle information was collected by a questionnaire. RESULTS: Paving workers were actually exposed to PAHs, including carcinogenic compounds, that were measurable only in the aerosol phase. Higher exposure as well as dose levels were measured for the paver group. Biological monitoring confirmed that 1-OHP was less affected by smoking habits as compared to 2-NAP and showed a higher association with occupational exposure. CONCLUSION: Carcinogenic PAH compounds were detectable only in the aerosol phase and this must be taken into account in the adoption of preventive measures. Biomonitoring supported the superiority of 1-OHP as compared to 2-NAP in assessing the internal dose in such workers.

S phase arrest in lymphocytes induced by urinary 1-hydroxypyrene and alcohol drinking in coke oven workers.

Arrest of the cell cycle after DNA damage is believed to promote DNA repair. We aim to investigate the main factors affecting cell cycle arrest of lymphocytes in coke oven workers. A total of 600 workers were included in this study, and their urinary levels of four polycyclic aromatic hydrocarbons (PAH) metabolites, 8-hydroxydeoxyguanosine (8-OHdG), and cell cycle distribution were determined. Urinary PAH metabolites were significantly increased in coke oven workers ( p < 0.01). It was found that only urinary 2-hydroxynaphthalene and 1-hydroxypyrene showed significant positive linear dose-response effects on 8-OHdG in this study population ( ptrend = 0.025 and 0.017, respectively). The dose-response effect was also observed for smoking and drinking on 8-OHdG ( ptrend < 0.001 and 0.034, respectively). Multivariate logistic regression analysis revealed that high levels of urinary 1-hydroxypyrene were associated with a significantly increased risk of S phase arrest (odds ratio (OR) = 1.32, p = 0.03), so as heavy alcohol drinking (OR = 1.31, p = 0.02). Drinking can significantly modify the effects of urinary 1-hydroxypyrene on S phase arrest, during co-exposure to both heavy drinking and median or high 1-hydroxypyrene levels (OR = 3.31, 95% confidence interval (CI) = 1.21-7.63 and OR = 2.56, 95% CI = 1.08-6.06, respectively). Our findings demonstrate that coke oven workers with heavy drinking will cause S phase arrest so as to repair more serious DNA damage.

Prenatal urinary polycyclic aromatic hydrocarbon metabolites, global DNA methylation in cord blood, and birth outcomes: A cohort study in China.

BACKGROUND: Prenatal exposure to polycyclic aromatic hydrocarbons (PAHs) is a potential risk factor for adverse birth outcomes. Epigenetic mechanisms may play a key role in which PAHs exert its effects. OBJECTIVE: Our study aimed to examine whether prenatal PAH exposure was associated with adverse birth outcomes and altered DNA methylation and to explore potential mediating roles of DNA methylation. METHODS: Ten urinary PAH metabolites were measured from 106 pregnant women during late pregnancy in a Chinese cohort study. Cord blood DNA methylation in long interspersed nucleotide element-1 (LINE-1) and Alu repetitive elements as surrogates of global DNA methylation was analyzed by bisulfite pyrosequencing. Multivariable linear regression was used to estimate the associations of urinary PAH metabolites with birth outcomes and DNA methylation, and a mediation analysis was also conducted. RESULTS: Prenatal urinary 2-hydroxynaphthalene (2-OHNa), ∑OHNa (sum of 1- and 2-OHNa), and sum of monohydroxy-PAH (∑OH-PAHs) were associated with lower birth length (e.g., -0.80%, 95% CI: -1.39%, -0.20% for the third vs. first tertile of 2-OHNa; p for trend = 0.01). Prenatal urinary 2-OHNa and 1-hydroxyphenanthrene (1-OHPh) were associated with lower Alu and LINE-1 methylation (e.g., -1.88%, 95% CI: -3.73%, -0.10% for the third vs. first tertile tertile of 2-OHNa in Alu methylation; p for trend = 0.04). Mediation analysis failed to show a mediator effect of global DNA methylation in the association between prenatal urinary OH-PAHs and birth outcomes. CONCLUSIONS: Prenatal specific PAH exposures are associated with decreased birth length and global DNA methylation. However, global DNA methylation does not mediate the associations of prenatal PAH exposure with birth outcomes. Further studies are needed to confirm the results.

Reliable quantification of 1,2-dihydroxynaphthalene in urine using a conjugated reference compound for calibration.

After environmental and occupational exposure to naphthalene, 1,2-dihydroxynaphthalene (1,2-DHN) was shown to be one major metabolite in human naphthalene metabolism. However, the instability of free 1,2-DHN complicates the reliable determination of this promising biomarker in urine. To solve this stability problem, glucuronide conjugates of 1,2-DHN and the corresponding isotopically labelled D6-1,2-dihydroxynaphthalene (D6-1,2-DHN) were synthesised and applied as reference material and internal standard in a gas chromatographic-tandem mass spectrometric (GC-MS/MS) method. The determination of 1- and 2-naphthol (1-MHN, 2-MHN) was included in the procedure to enable a comprehensive assessment of naphthalene metabolism and exposure. The results of the validation showed a high reliability and sensitivity of the method. The detection limits range from 0.05 to 0.16 µg/L. Precision and repeatability were determined to range from 1.4 to 6.6% for all parameters. The simultaneous determination of 1- and 2-MHN as additional parameters besides 1,2-DHN enables the application of the method for further metabolism and kinetic studies on naphthalene. The use of glucuronide-derivative reference substances and the application of structurally matched isotopic-labelled internal standards for each substance guarantee a reliable quantification of the main naphthalene metabolites 1,2-DHN and 1- and 2-MHN. Graphical abstract Reliable quantification of 1,2-dihydroxynaphthalene in urine using a conjugated reference compound for calibration.

JP8 exposure and neurocognitive performance among US Air Force personnel.

Petroleum-based fuels such as jet propellant (JP) 4, JP5, JP8, and jet A1 (JetA) are among the most common occupational chemical exposures encountered by military and civilian workforces. Although acute toxicity following high-level exposures to JP8 and similar chemical mixtures has been reported, the relationship between persistent low-level occupational exposures to jet fuels and both acute and longer-term central nervous system (CNS) function has been comparatively less well characterized. This paper describes results of neurocognitive assessments acquired repeatedly across a work week study design (Friday to Friday) as part of the Occupational JP8 Exposure Neuroepidemiology Study (OJENES) involving U.S. Air Force (AF) personnel with varying levels of exposure to jet fuel (JP8). JP8 exposure levels were quantified using both personal air monitoring and urinary biomarkers of exposure. Neurocognitive performance was evaluated using an objective, standardized battery of tests. No significant associations with neurocognitive performances were observed between individuals having regular contact and those with minimal/no direct contact with JP8 (measured by average work week levels of personal breathing zone exposure). Also, no significant findings were noted between repeated measures of absorbed dose (multi-day pre-shift urinary 1- and 2-naphthol) and reduced proficiency on neurocognitive tasks across the work week. Results suggest that occupational exposure to lower (than regulated standards) levels of JP8 do not appear to be associated with acute, measurable differences or changes in neurocognitive performance.

Association of atmospheric concentrations of polycyclic aromatic hydrocarbons with their urinary metabolites in children and adolescents.

This study aims to determine the atmospheric concentrations of particulate matter 2.5 (PM2.5)-bounded polycyclic aromatic hydrocarbons (PAHs) and their association with their urinary metabolites in children and adolescents. This study was conducted from October 2014 to March 2016 in Isfahan, Iran. We measured 16 species of PAHs bounded to PM2.5 by gas chromatography mass spectrometry (GC/MS) from 7 parts of the city. Moreover, PAH urinary metabolites were measured in 186 children and adolescents, randomly selected from households. Urinary metabolites consisted of 1-hydroxy naphthalene (1-naphthol), 2-hydroxy naphthalene (2-naphthol), 9-hydroxy phenanthrene (9-phenanthrol), and 1-hydroxy pyrene using GC/MS. Considering the short half-lives of PAHs, we measured the metabolites twice with 4 to 6 months of time interval. We found that the ambient concentrations of PAHs were significantly associated with their urinary metabolites. 1-hydroxy naphthalene and 2-hydroxy naphthalene concentrations showed an increase of 1.049 (95% CI: 1.030, 1.069) and 1.047 (95% CI: 1.025, 1.066) for each unit increase (1 ng/m3) in ambient naphthalene. Similarly, 1-hydroxy pyrene showed an increase of 1.009 (95% CI: 1.006-1.011) for each unit increase (1 ng/m3) in ambient pyrene concentration after adjustment for body mass index, physical activity level, urinary creatinine, age, and sex. The association of urinary 9-hydroxyphenanthrene and ambient phenantherene was significant in the crude model; however after adjustment for the abovementioned covariates, it was no more significant. We found significant correlations between exposure to ambient PM2.5-bounded PAHs and their urinary excretion. Considering the adverse health effects of PAHs in the pediatric age group, biomonitoring of PAHs should be underscored; preventive measures need to be intensified.

Quantification of 1-hydroxypyrene, 1- and 2-hydroxynaphthalene, 3-hydroxybenzo[a]pyrene and 6-hydroxynitropyrene by HPLC-MS/MS in human urine as exposure biomarkers for environmental and occupational surveys.

CONTEXT: Several urinary PAHs metabolites can be detected by HPLC-MS/MS for individual exposure assessment. OBJECTIVE: Quantitation of urinary metabolites of four PAHs, selected on the basis of their significance, with reduced costs and high sensitivity. MATERIALS AND METHODS: HPLC-MS/MS was used and pure standards and isotope-labeled internal analogs of the analytes. Two hundred samples were tested after enzymatic hydrolysis. RESULTS: Accuracy was higher than 90% and variability lower than 19%; LODs permit to measure 1-hydroxypyrene, 1 and 2-hydroxynaphthalene in all subjects, 6-hydroxynitropyrene in the 65% and 3-hydroxybenzo[a]pyrene in the 70%. DISCUSSION AND CONCLUSION: The method is suitable both for occupational and for environmental studies. This is the first paper reporting urinary levels of 6-hydroxynitropyrene in European subjects, nonoccupationally exposed to nitro-PAHs.

Biomonitoring Human Exposure to Household Air Pollution and Association with Self-reported Health Symptoms - A Stove Intervention Study in Peru.

BACKGROUND: Household air pollution (HAP) from indoor biomass stoves contains harmful pollutants, such as polycyclic aromatic hydrocarbons (PAHs), and is a leading risk factor for global disease burden. We used biomonitoring to assess HAP exposure and association with self-reported symptoms in 334 non-smoking Peruvian women to evaluate the efficacy of a stove intervention program. METHODS: We conducted a cross-sectional study within the framework of a community randomized control trial. Using urinary PAH metabolites (OH-PAHs) as the exposure biomarkers, we investigated whether the intervention group (n=155, with new chimney-equipped stoves) were less exposed to HAP compared to the control group (n=179, with mostly open-fire stoves). We also estimated associations between the exposure biomarkers, risk factors, and self-reported health symptoms, such as recent eye conditions, respiratory conditions, and headache. RESULTS: We observed reduced headache and ocular symptoms in the intervention group than the control group. Urinary 2-naphthol, a suggested biomarker for inhalation PAH exposure, was significantly lower in the intervention group (GM with 95% CI: 13.4 [12.3, 14.6] µg/g creatinine) compared to control group (16.5 [15.0, 18.0] µg/g creatinine). Stove type and/or 2-naphthol was associated with a number of self-reported symptoms, such as red eye (adjusted OR with 95% CI: 3.80 [1.32, 10.9]) in the past 48h. CONCLUSIONS: Even with the improved stoves, the biomarker concentrations in this study far exceeded those of the general populations and were higher than a no-observed-genotoxic-effect-level, indicating high exposure and a potential for increased cancer risk in the population.

Urinary concentrations of organophosphate and carbamate pesticides in residents of a vegetarian community.

Few population studies have measured urinary levels of pesticides in individuals with vegan, vegetarian, or organic diets. The objectives of this study were to evaluate whether a vegan/vegetarian diet was associated with increased exposure to organophosphate and carbamate pesticides, and to evaluate the impact of organic consumption on pesticide exposure in vegans and vegetarians. In the current pilot study conducted in 2013-2014, we collected spot urine samples and detailed 24h recall dietary data in 42 adult residents of Amirim, a vegetarian community in Northern Israel. We measured urinary levels of non-specific organophosphate pesticide metabolites (dialkylphosphates, (DAPs)) and specific metabolites of the current-use pesticides chlorpyrifos (3,5,6-trichloro-2-pyridinol (TCPy)), propoxur (-isopropoxyphenol (IPPX)), and carbaryl (1-naphthol). Six DAP metabolites were detected in between 67 and 100% of urine samples, with highest geometric mean concentrations for dimethylphosphate (19.2µg/g). Creatinine-adjusted median concentrations of total DAPs and of TCPy were significantly higher in Amirim residents compared to the general Jewish population in Israel (0.29µmol/g compared to 0.16, p<0.05 for DAPs and 4.32µg/g compared to 2.34µg/g, p<0.05 for TCPy). Within Amirim residents, we observed a positive association between vegetable intake and urinary TCPy levels (rho=0.47, p<0.05) and lower median total dimethyl phosphate levels in individuals reporting that >25% of the produce they consume is organic (0.065µmol/L compared to 0.22, p<0.05). Results from this pilot study indicate relatively high levels of urinary organophosphate pesticide metabolite concentrations in residents of a vegetarian community, a positive association between vegetable intake and urinary levels of a chlorpyrifos specific metabolite, and lower levels of total dimethyl phosphate in individuals reporting higher intake of organic produce. Results suggest that consumption of organic produce may offer some protection from increased exposure to organophosphate pesticide residues in vegetarians.

Capillary electrophoresis chemiluminescence assay of naphthol isomers in human urine and river water using Ni(IV) complex-luminol system.

A capillary electrophoresis method involving online indirect chemiluminescence (CL) detection was used to determine naphthol (NAP) isomers. The method was based on the quenching effect of 1- and 2-NAP on a new CL reaction of luminol with Ni(IV) complex in an alkaline medium. Separation was conducted with a 25.0 mM sodium borate buffer containing 0.8 mmol/L luminol. Under optimized conditions, 1- and 2-NAP were baseline separated and detected in less than 8 min. The limits of detection of 1- and 2-NAP were 3.1 and 2.7 µg/L, respectively (S/N = 3), with a linear range of 4.0-80.0 µg/L (r > 0.995). Analysis of real samples demonstrated that the spiked recoveries were in the range of 89.2-107.5% (n = 3). The proposed method was successfully used to determine 1- and 2-NAP contents in three environmental water samples and 14 human urine samples. No derivatization or tedious pretreatment was required in the analysis. The proposed method is a potential approach for routine tests of naphthol isomers in a facile CE-CL system.

Genome-Wide Analysis of DNA Methylation and Cigarette Smoking in a Chinese Population.

BACKGROUND: Smoking is a risk factor for many human diseases. DNA methylation has been related to smoking, but genome-wide methylation data for smoking in Chinese populations is limited. OBJECTIVES: We aimed to investigate epigenome-wide methylation in relation to smoking in a Chinese population. METHODS: We measured the methylation levels at > 485,000 CpG sites (CpGs) in DNA from leukocytes using a methylation array and conducted a genome-wide meta-analysis of DNA methylation and smoking in a total of 596 Chinese participants. We further evaluated the associations of smoking-related CpGs with internal polycyclic aromatic hydrocarbon (PAH) biomarkers and their correlations with the expression of corresponding genes. RESULTS: We identified 318 CpGs whose methylation levels were associated with smoking at a genome-wide significance level (false discovery rate < 0.05), among which 161 CpGs annotated to 123 genes were not associated with smoking in recent studies of Europeans and African Americans. Of these smoking-related CpGs, methylation levels at 80 CpGs showed significant correlations with the expression of corresponding genes (including RUNX3, IL6R, PTAFR, ANKRD11, CEP135 and CDH23), and methylation at 15 CpGs was significantly associated with urinary 2-hydroxynaphthalene, the most representative internal monohydroxy-PAH biomarker for smoking. CONCLUSION: We identified DNA methylation markers associated with smoking in a Chinese population, including some markers that were also correlated with gene expression. Exposure to naphthalene, a byproduct of tobacco smoke, may contribute to smoking-related methylation. CITATION: Zhu X, Li J, Deng S, Yu K, Liu X, Deng Q, Sun H, Zhang X, He M, Guo H, Chen W, Yuan J, Zhang B, Kuang D, He X, Bai Y, Han X, Liu B, Li X, Yang L, Jiang H, Zhang Y, Hu J, Cheng L, Luo X, Mei W, Zhou Z, Sun S, Zhang L, Liu C, Guo Y, Zhang Z, Hu FB, Liang L, Wu T. 2016. Genome-wide analysis of DNA methylation and cigarette smoking in Chinese. Environ Health Perspect 124:966-973; http://dx.doi.org/10.1289/ehp.1509834.

Monitoring exposure to polycyclic aromatic hydrocarbons in an Australian population using pooled urine samples.

Integrated exposure to polycyclic aromatic hydrocarbons (PAHs) can be assessed through monitoring of urinary mono-hydroxylated PAHs (OH-PAHs). The aim of this study was to provide the first assessment of exposure to PAHs in a large sample of the population in Queensland, Australia including exposure to infant (0-4years). De-identified urine specimens, obtained from a pathology laboratory, were stratified by age and sex, and pooled (n=24 pools of 100) and OH-PAHs were measured by gas chromatography-isotope dilution-tandem mass spectrometry. Geometric mean (GM) concentrations ranged from 30ng/L (4-hydroxyphenanthrene) to 9221ng/L (1-naphthol). GM of 1-hydroxypyrene, the most commonly used PAH exposure biomarker, was 142ng/L. The concentrations of OH-PAHs found in this study are consistent with those in developed countries and lower than those in developing countries. We observed no association between sex and OH-PAH concentrations. However, we observed lower urinary concentrations of all OH-PAHs in samples from infants (0-4years), children (5-14years) and the elderly (>60year old) compared with samples from other age groups (15-29, 30-44 and 45-59years) which may be attributed to age-dependent behaviour-specific exposure sources.