Aromatic and polycyclic hydrocarbons in air and their urinary metabolites in coke plant workers

This study describes the exposure of coke plant workers to hydrocarbons. Aromatic hydrocarbons (AHs) and polycyclic aromatic hydrocarbons (PAHs) in the breathing zone air and their oxygenated metabolites in the urine of coke plant workers are qualitatively and quantitatively determined. Concentrations of benzene, toluene, naphthalene, m+p-xylene, o-xylene and 14 different PAHs were measured at the different workplaces by personal air sampling. O-cresol, 1- and 2-naphthol, methylhippuric acid, and 1-hydroxypyrene were determined in hydrolyzed urine of workers collected after the work shift. The gas chromatography–mass spectrometry (GC/MS) method was applied to identify AHs in air and in urine samples. Time-weighted values of exposure to aromatic hydrocarbons at a coke plant were: benzene (0.06–9.82 mg/m³), toluene (0.05–4.71 mg/m³), naphthalene (0.01–3.28 mg/m³), o-xylene (0.01–1.76 mg/m³) and m + p-xylene (0.01–2.62 mg/m³). At the coke batteries, the total concentration of PAHs ranged from 7.27 to 21.92 μg/m³. At the sorting department, the total concentration of PAHs were about half this value. Concentration of the urinary metabolites (naphthols and methylhippuric acid) detected in workers at the tar distillation department are three times higher than those for the coke batteries and sorting department workers. A correlation between inhaled toluene, naphthalene, xylene, and urinary excretion of metabolites has been found. Time-weighted average concentrations of AHs in the breathing zone air show that exposure levels of the workers are rather low in comparison to exposure limits. The 1-hydroxypyrene concentration is below 24.75 μmol/mol creatinine. The GC/MS analysis reveals the presence of AHs, mainly benzene and naphthalene homologues. It has been found that coke plant workers are simultaneously exposed to the mixture of aromatic and polycyclic hydrocarbons present in the breathing zone air of a coke plant. Exposure levels are significantly influenced by job categories. Compounds identified in the urine appear to be the products of the hydroxylation of AHs present in the air as well as unmetabolized hydrocarbons. Am. J. Ind. Med. 34:445–454, 1998. © 1998 Wiley-Liss, Inc.     

Influence of time and mode of exposure on biotransformation of naphthalene by Juvenile starry flounder (Platichthys stellatus) and rock sole (Lepidopsetta bilineata)

Juvenile starry flounder (Platichthys stellatus) and rock sole (Lepidopsetta bilineata) were force-fed 56 Ci each of 1-³H-naphthalene dissolved in salmon oil. Values for radioactivity associated with naphthalene and the metabolite fraction were determined for various tissues and body fluids. Results show that these pleuronectids extensively metabolize dietary naphthalene. The rates of decline in naphthalene concentrations (expressed as disintegrations per minute per milligram of dry tissue) were greater than the rates of decline in metabolite concentrations (dpm/mg) in liver, blood, and skin; therefore, relative proportion of metabolites to naphthalene increased with time and at 168 hr after the initiation of the naphthalene-exposure, more than half of the total radioactivity in both species of fish was associated with the metabolites.Profiles of metabolites in liver, skin, and bile were obtained using thin-layer chromatography. 1,2-Dihydro-1,2-dihydroxynaphthalene constituted 38.7 and 39.7%, respectively, of the total extracted metabolites in livers of the naphthalene-exposed rock sole and starry flounder at 24 hr, whereas the bile from both species contained primarily (>90%) conjugates. From 24 to 168 hr, a significant (P < 0.05) decrease in the proportion of the dihydrodiol derivative and a concomitant increase in the proportion of conjugates—specifically, sulfate/glucoside fraction-were observed with livers of both rock sole and starry flounder. No significant change occurred in the spectrum of biliary metabolites with time. The pattern of metabolites in skin of both species was qualitatively similar to that in liver; however, the proportion of the dihydrodiol was greater in skin than in liver at 24 hr.When naphthalene (56Ci) dissolved in salmon oil was administered to starry flounder via intraperitoneal injection, the extent of biotransformation was less than after dietary exposure. Moreover, metabolites in the livers of the fish in the injection study were predominantly (76.7% of total extracted metabolites) non-conjugates at 24 hr. Once again, from 24 to 168 hr, an increase in the proportion of the sulfate/glucoside fraction and a concomitant decrease in the proportion of the dihydrodiol was observed with liver.These studies demonstrate that the extent of biotransformation of naphthalene and the types of metabolites remaining in tissues (e.g., liver) of flatfish are greatly influenced by both the mode of exposure and the time elapsed after the exposure is initiated. It appears therefore, that different exposures (e.g., in water, food, or sediment) of pleuronectids to polycyclic aromatic hydrocarbons may result in different degrees of alteration in genetic material because of variability in accumulation of non-conjugated metabolites, some of which are implicated in covalent binding with DNA in terrestrial mammals.     

Genotoxic risk assessment in white blood cells of occupationally exposed workers before and after alteration of the polycyclic aromatic hydrocarbon (PAH) profile in the production material: comparison with PAH air and urinary metabolite levels

Objective: Workers in various industries can be exposed to polycyclic aromatic hydrocarbons (PAHs). The relationship between biomarkers of genotoxic risk, PAH compounds in air (ambient monitoring) and PAH metabolites in urine (internal exposure) were studied in 17 workers exposed to PAHs in a fireproof-material producing plant before and 3 months after the PAH profile was altered in the binding pitch. Methods: Two biomarkers of exposure, specific DNA adducts of (±)-r-7,t-8-dihydroxy-t-9,10-oxy-7,8,9,10-tetrahydrobenzo[a]pyrene (anti-BPDE) and non-specific DNA adduct of 8-oxo-7,8-dihydro-2-deoxyguanosine (8-oxodGuo) were determined in white blood cells (WBCs). In addition, DNA strand breaks were analysed in lymphocytes by single-cell gel electrophoresis in a genotoxic risk assessment. Sixteen PAH compounds in air were determined by personal air sampling, and hydroxylated metabolites of phenanthrene, pyrene and naphthalene were determined in urine. Results: After substitution of the binding pitch the concentrations of benzo[a]pyrene in air decreased (P<0.01). No changes could be observed for pyrene, while levels of phenanthrene (P=0.0013) and naphthalene (P=0.0346) in air increased. Consequently, median DNA adduct rates of anti-BPDE decreased after alteration of the production material (from 0.9 to <0.5 adducts/10⁸ nucleotides). No changes in the excretion of 1-hydroxypyrene in urine could be determined, whereas increased levels of 1-, 2+9-, 3- and 4-hydroxyphenanthrene (P<0.0001) and 1-naphthol and 2-naphthol (P=0.0072) were found in urine. In addition, a statistically significant increase in DNA strand break frequencies (P<0.01) and elevated 8-oxodGuo adduct levels (P=0.7819, not statistically significant) were found in the WBCs of exposed workers 3 months after the PAH profile in the binding pitch had been altered. Conclusion: The results presented here show that the increased concentration of naphthalene and/or phenanthrene in the air at the work place could induce the formation of DNA strand breaks and alkali-labile sites in WBCs of exposed workers.     

Urinary 1-naphthol and 1-pyrenol as indicators of exposure to coal tar products

Chemical exposure of assemblers handling creosote-impregnated wood and of a single worker chiselling coal tar pitch layer was assessed by measuring airborne naphthalene and various polycyclic aromatic hydrocarbons (PAHs), and by measurement of urinary excretion of 1-naphthol and 1-pyrenol. The sum concentration of PAHs and of 4–6 aromatic ring-containing PAHs were high, 440 g/m³ and 290 g/m³, respectively, when chiselling. In the assembler's workplace, the PAH concentrations were about 1/50 of this value. Regarding airborne naphthalene concentrations the situation was reversed (assemblers, 1000 g/m³; chiseller, 160 g/m³). Correspondingly, the assemblers' urinary 1-napthol concentrations were 15–20 times higher than those of the chiseller. The urinary 1-pyrenol concentration of the chiseller was 2–4 times higher than among the assemblers. As the estimated pyrene inhalation doses among the assemblers could account for only about 2%–25% of the 24-h pyrenol excretion in urine, the skin was presumably the main route of uptake. For an assessment of the exposure to PAHs, air measurements, monitoring of metabolites in urine and preferably also data on the composition of the skin-contaminating product are needed.     

Urinary hydroxy-metabolites of naphthalene,phenanthrene and pyrene as markers of exposure to diesel exhaust

Objective The objective of this study was to assess the exposure of bus-garage and waste-collection workers to polycyclic aromatic hydrocarbons (PAHs) derived from diesel exhaust by the measurement of levels of seven urinary PAH metabolites: 2-naphthol, 1-hydroxyphenanthrene, 2-hydroxyphenanthrene, 3-hydroxyphenanthrene, 1+9-hydroxyphenanthrene, 4-hydroxyphenanthrene and 1-hydroxypyrene.Subjects and methods One urine sample from each of 46 control persons, and one pre-shift and two post-shift spot urine samples from 32 exposed workers were obtained in winter and in summer. The metabolites were analysed after enzymatic hydrolysis by high performance liquid chromatography (HPLC) with fluorescence detection.Results The sum of seven PAH metabolites (mean 3.94±3.40 and 5.60±6.37 mol/mol creatinine in winter and summer, respectively) was higher [P=0.01, degrees of freedom (df) =61.2 and P=0.01, df=67.6 in winter and summer, respectively] in the exposed group than in the control group (mean 3.18±3.99 and 3.03±2.01 mol/mol creatinine in winter and summer, respectively). The mean concentrations of 2-naphthol among exposed and controls ranged between 3.34 and 4.85 mol/mol creatinine and 2.51 and 2.58 mol/mol creatinine, respectively (P<0.01 in winter, P<0.03 in summer). The mean level of the hydroxyphenanthrenes in the samples of exposed workers was between 0.40 and 0.70 mol/mol creatinine and in the control samples 0.40–0.60 mol/mol creatinine. The concentration of 1-hydroxypyrene was higher among exposed workers in both pre-shift and post-shift samples (mean 0.10–0.15 mol/mol creatinine) than in control group (mean 0.05–0.06 mol/mol creatinine) in winter (P=0.002, df=78) and in summer (P<0.001, df=68).Conclusions The urinary hydroxy-metabolites of naphthalene, phenanthrene and pyrene showed low exposure to diesel-derived PAHs; however, it was higher in exposed workers than in control group. Urinary PAH monohydroxy-metabolites measured in this study did not correlate with the PAHs in the air samples, reported earlier, in 2002 and 2003.     

Accumulation of14C-naphthalene in the tissues of redhead ducks fed oil-contaminated crayfish

Crayfish, artificially contaminated with¹⁴C-naphthalene-5% water-soluble fraction of No. 2 fuel oil, were force-fed to one-year-old redhead ducks to determine the accumulation of petroleum hydrocarbons. The relative distribution of carbon-14 activity in the gall bladder containing bile, and fat were similar, and significantly greater (P < 0.05) than the activity in the blood, brain, liver, and kidney. There was a significant increase (P < 0.05) in the disintegrations per minute per gram (dpm/g) in the blood, brain, kidney, and liver between days 1 and 3 of feeding, indicating a progressive accumulation of carbon-14 activity (naphthalene and presumably its metabolites). There was no significant effect of sex or the interaction of the duration of feeding and sex on carbon-14 activity in any of the tissues. The low daily dose of petroleum hydrocarbons (a total of approximately 1.25 mg/day) received by the ducks from the crayfish and the relatively short feeding regimen did not cause any overt signs of toxicity in the ducks.     

occupational exposure to aromatic hydrocarbons at a coke plant: Part II. Exposure assessment of volatile organic compounds

The objective of the study is to assess the external and internal exposures to aromatic hydrocarbons in the tar and oil naphthalene distillation processes at a coke plant. 69 workers engaged as operators in tar and oil naphthalene distillation processes and 25 non-exposed subjects were examined. Personal analyses of the benzene, toluene, xylene isomers, ethylbenzene, naphthalene, indan, indene and acenaphthene in the breathing zone air allowed us to determine the time weighted average exposure levels to the aromatic hydrocarbons listed above. The internal exposure was investigated by measurement of the urinary excretion of naphthols, 2-methylphenol and dimethylphenol isomers by means of gas chromatography with a flame ionization detection (GC/FID). Urine metabolites were extracted after enzymatic hydrolysis by solid-phase extraction with styrene-divinylbenzene resin. The time-weighted average concentrations of the hydrocarbons detected in the breathing zone air shows that the exposure levels of the workers are relatively low in comparison to the exposure limits. Statistically significant differences between average concentrations of aromatic hydrocarbons (benzene, toluene, xylene isomers) determined at the workplaces in the tar distillation department have been found. Concentrations of the naphthalene and acenaphthene detected in workers from the oil distillation department are higher that those from the tar distillation department. Concentrations of naphthols, 2-methoxyphenol and dimethylphenol isomers in the urine of occupationally exposed workers were significantly higher than those of non-exposed subjects. Concentrations of the 2-methoxyphenol and dimethylphenol isomers in urine were significantly higher for the tar distillation workers, whereas concentrations of naphthols were higher for the oil naphthalene distillation workers. Operators at the tar and naphthalene oil distillation processes are simultaneously exposed to a mixture of different hydrocarbons, mainly benzene and naphthalene homologues.     

Current external and internal exposure to naphthalene of workers occupationally exposed to polycyclic aromatic hydrocarbons in different industries

Objectives: External and internal exposure to naphthalene was examined in the most important industries that are typically concerned with polycyclic aromatic hydrocarbon (PAH)-induced diseases (cancer). Furthermore, a control collective from the general population was investigated. Methods: External naphthalene was determined by personal air sampling (n=205). The internal exposure was examined by urinary metabolites 1-naphthol and 2-naphthol (n=277). Results: Highest median concentrations of naphthalene in air were found in converter infeed (93.2 g/m³) and coal-tar distillation (35.8 g/m³). Moderate and low levels were determined in coking plants (14.5 g/m³) and in the production of refractories (6.1 g/m³) and graphite electrodes (0.7 g/m³). Biological monitoring revealed concentrations of the sum of both metabolites [(1+2)-NOL] in smokers to be increased by 1.6–6.4 times compared with that in non-smokers at the same workplaces. Among non-smokers we found high median (1+2)-NOL levels in converter bricklayers (120.1 g/l), in coal-tar distillation workers (56.0 g/l) and in coking plant workers (29.5 g/l). (1+2)-NOL concentrations around 10 g/l were found in the production of refractories and graphite electrodes. There was a rough coherency between external and internal naphthalene exposure. In the controls, median (1+2)-NOL concentrations were 10.9 g/l in non-smokers urine and 40.3 g/l in smokers urine samples. Conclusions: Actual conditions of occupational hygiene at the workplaces investigated in this comprehensive study are better than those that current limit values of 50,000 g/m³ (TLV, TRK) seem to induce. It has become obvious that tobacco smoking is a crucial confounding factor in biological monitoring of naphthalene-exposed humans, making interpretation of occupationally increased naphthol excretions very difficult at low exposure levels.     

Effects of naphthalene and benzene on fathead minnows and rainbow trout

Fathead minnows (FHM) and rainbow trout (RBT) were used in flow-through bioassays to determine the acute toxicity of benzene and naphthalene, and to determine the embryo-larval effects of naphthalene on FHM. On an acute basis, naphthalene was more toxic than benzene (naphthalene LC₅₀ values were 1.6 mg/L for RBT and 7.9 mg/L for FHM; benzene LC₅₀ values were 5.3 mg/L for RBT and >15.1 mg/L for FHM). In the embryo-larval test naphthalene significantly (= 0.05) reduced FHM growth at concentrations as low as 0.85 mg/L. The highest concentration producing no effect was 0.45 mg/L naphthalene, which was 5.7% of the FHM 96-hr LC₅₀. Based upon long-term no-effects naphthalene concentration, the best estimate of the maximum acceptable toxicant concentration (MATC) was >0.45 to <0.85 mg/L naphthalene.Work funded under an Interagency Agreement between the U.S. Department of Energy and the U.S. Environmental Protection Agency under Contract No. DE-AS20-79 LC 01761 to the Rocky Mountain Institute of Energy and Environment, University of Wyoming     

Concentration and profile of 22 urinary polycyclic aromatic hydrocarbon metabolites in the US population

Urinary monohydroxy polycyclic aromatic hydrocarbons (OH-PAHs) are a class of PAH metabolites used as biomarkers for assessing human exposure to PAHs. The Centers for Disease Control and Prevention's National Health and Nutrition Examination Survey (NHANES) uses OH-PAHs to establish reference range concentrations for the US population, and to set benchmarks for future epidemiologic and biomonitoring studies. For the years 2001 and 2002, 22 OH-PAH metabolites were measured in urine specimens from 2748 NHANES participants. Percentages of samples with detectable levels ranged from nearly 100% for metabolites of naphthalene, fluorene, phenanthrene, and pyrene, to less than 5% for metabolites from parent compounds with higher molecular weight such as chrysene, benzo[c]phenanthrene, and benz[a]anthracene. The geometric mean for 1-hydroxypyrene (1-PYR)—the most commonly used biomarker for PAH exposure—was 49.6 ng/L urine, or 46.4 ng/g creatinine. Children (ages 6–11) generally had higher levels than did adolescents (ages 12–19) or adults (ages 20 and older). Model-adjusted, least-square geometric means for 1-PYR were 87, 53 and 43 ng/L for children, adolescents (ages 12–19) and adults (ages 20 years and older), respectively. Log-transformed concentrations for major detectable OH-PAHs were significantly correlated with each other. The correlation coefficients between 1-PYR and other metabolites ranging from 0.17 to 0.63 support the use of 1-PYR as a useful surrogate representing PAH exposure.     

Decreasing urinary PAH metabolites and 7-methylguanine after smoking cessation

Objective: Humans are exposed to various carcinogens by smoking. Urinary metabolites of polycyclic aromatic hydrocarbons (PAH), one of the major carcinogens in cigarette smoke, were measured as the environmental carcinogen exposure marker for humans. We evaluated urinary exposure markers for smoking cessation. Method: In this study, we measured cigarette smoke exposure markers, such as urinary cotinine, PAH exposure markers, such as urinary 1-hydroxypyrene (1-OHP), 2-naphthol (2-NP) and 1-naphthol (1-NP), as well as a methylating chemical exposure marker, 7-methylguanine (7-MeG). The before smoking cessation levels of these markers, and the after smoking cessation levels were then compared. Eighteen subjects participated in this smoking cessation program. Results: Levels of all of four markers were found to have decreased by 19–54% after smoking cessation. Urinary cotinine, 1-OHP, 2-NP and 7-MeG levels were found to have significantly decreased after smoking cessation. There were positive correlations between cotinine and three urinary PAH markers and between 1-OHP, 2-NP and 7-MeG. Conclusion: PAH metabolites were better biomarkers of smoking cessation than 7-MeG. Analyzing urinary metabolites or urinary DNA adducts is suitable for epidemiological studies.     

Identification by physical means of organic moieties of conjugates produced from carbaryl by tobacco cells in suspension culture

Carbaryl (1-naphthyl methylcarbamate), labeled with¹⁴C in the C₁-naphthyl, carbonyl, orN-methyl position, was introduced into the culture medium of tobacco cells in suspension culture. Following incubation, cells were homogenized in water, centrifuged, and supernatants hydrolyzed with-glucosidase or HCl. Organic moieties (moieties) were characterized by two-dimensional thin-layer chromatography (TLC), and many were subsequently identified by infrared and mass spectrometry. On the basis of the data obtained with¹⁴C₁-naphthyl-labeled carbaryl, it appeared that 18.4% of the total characterized metabolites represented unconjugatedN-CH₂OH- carbaryl [1-naphthylN-(hydroxymethyl)carbamate], excreted by the cells into the culture medium. The metabolites found in the cells primarily consisted of conjugates of 1-naphthol (73.6% of the total characterized metabolites) andN-CH₂OH-carbaryl (2.5%). Conjugates of 7-hydroxycarbaryl (7-hydroxy-1-naphthyl methylcarbamate), 4-hydroxycarbaryl (4-hydroxy-1-naphthyl methylcarbamate), and 5-hydroxycarbaryl (5-hydroxy-1-naphthyl methylcarbamate) were also detected in small amounts. Of five unknown¹⁴C₁-naphthyl-labeled carbaryl metabolites, three were tentatively characterized as:O-1-naphthylcholesterol (cholest-5-en-3-yl-1-naphthol; 3.0%); an unconjugated hydroxylated 1,4-dihydro-1,4-epiperoxynaphthalene (1.4%); and an acidlabile,-glucosidase-resistant conjugate of acis-dihydrodiol of 1-naphthol (0.3%; other than thetrans-5,6-dihydrodiol). The cholesterol derivative may represent a new detoxification mechanism in plants; the epiperoxide may help to elucidate plant oxidation mechanisms. A new TLC procedure was developed which successfully separated the acetate derivative ofN-hydroxycarbaryl (1-naphthylN-hydroxy-N-methylcarbamate) from 12 other common moieties of carbaryl metabolites and their acetate derivatives. A new two-dimensional TLC system was developed for the separation of underivatizedN-hydroxycarbaryl from 14 other moieties of carbaryl metabolites; two additional two-dimensional TLC systems were utilized for moiety separations. With these TLC procedures, no conjugated or unconjugatedN-hydroxycarbaryl could be detected in any tobacco cell culture fraction after incubation of cells in medium containing radiolabeled carbaryl. Authentic¹⁴C₁-naphthyllabeledN-CH₂OH-carbaryl was shown to be converted to desmethylcarbaryl (1-naphthylcarbamate) (97%) and 1-naphthol (3%) by 0.1N HCl hydrolysis.Deceased.     

Urinary Polycyclic Aromatic Hydrocarbons and Childhood Obesity: NHANES (2001–2006)

Background: Polycyclic aromatic hydrocarbons (PAHs) are known carcinogens and suspected endocrine disruptors. Prenatal exposure to PAHs has been associated with obesity in early childhood.Objective: We examined the association of urinary PAH metabolites with adiposity outcomes [body mass index (BMI) z-score, waist circumference (WC), and rate of obesity] in children and adolescents.Methods: We performed whole-sample analyses of 3,189 individuals 6–19 years of age who participated in the 2001–2006 National Health and Nutrition Examination Survey. We performed multivariate linear and logistic regression to analyze the association of BMI z-score, WC, and obesity with concentrations of single urinary PAH compounds and the sum of PAHs. Furthermore, the analyses were stratified by developmental stage [i.e., children (6–11 years) and adolescents (12–19 years)].Results: BMI z-score, WC, and obesity were positively associated with the molecular mass sum of the PAHs and the total sum of naphthalene metabolites. Most associations increased monotonically with increasing quartiles of exposure among children 6–11 years of age, whereas dose–response trends were less consistent for adolescents (12–19 years of age). Neither total PAHs nor total naphthalene metabolites were associated with overweight in either age group, and there was little evidence of associations between the outcomes and individual PAHs.Conclusions: Total urinary PAH metabolites and naphthalene metabolites were associated with higher BMI, WC, and obesity in children 6–11 years of age, with positive but less consistent associations among adolescents.Citation: Scinicariello F, Buser MC. 2014. Urinary polycyclic aromatic hydrocarbons and childhood obesity: NHANES (2001–2006). Environ Health Perspect 122:299–303; http://dx.doi.org/10.1289/ehp.1307234     

萘代谢产物-白蛋白加合物作为焦炉工接触生物标志物的初步探索

目的探讨萘代谢产物的白蛋白加合物作为焦炉工接触生物标志物的可行性。方法随机选取某焦化厂28名焦炉作业工人(接触组)和22名对照个体(对照组),工人在连续工作4d的班后采集静脉血和尿样,并使用调查表收集个人信息。采用气相色谱质谱联机方法测定血浆中白蛋白加合物。结果在接触组及对照组中均检测出1,2萘醌及1,4萘醌与白蛋白的加合物(1,2NPQAlb和1,4NPQAlb)。接触组1,2NPQAlb水平(766pmol/g)高于对照组(449pmol/g),差异有统计学意义;1,4NPQAlb水平在接触组(486pmol/g)和对照组(442pmol/g)间差异无统计学意义。接触组1,2NPQAlb水平(766pmol/g)高于1,4NPQAlb(486pmol/g),差异有显著性。尿中萘、1萘酚、2萘酚及1羟基芘浓度与1,2NPQAlb水平间呈显著相关性,而与1,4NPQAlb之间未见明显的相关关系。结论1,2NPQAlb可有效地反映个体中长期暴露于气态多环芳烃的内剂量水平,有可能作为生物标志物用于焦炉工的生物监测。     

Effect of environmental temperature on naphthalene metabolism by Juvenile Starry flounder (Platichthys stellatus)

Juvenile starry flounder (Platichthys stellatus) maintained at 4° or 12°C were forced-fed³H-1-naphthalene. At 24 hr, after the initiation of exposure, significantly (p < 0.05) higher concentrations (2 to 15 times) of naphthalene were present in tissues of starry flounder at 4°C than those present in fish held at 12°C. The influence of lowering of water temperature on naphthalene retention was even more marked after one week. At this time, muscle and liver of fish at 4°C contained 26 and 34 times, respectively, more naphthalene than did muscle and liver of fish at 12°C. Concentrations of total metabolites, in most tissues were not substantially higher at the lower temperature either 24 or 168 hr after the naphthalene-exposure.Thin-layer Chromatographic separation of the metabolites revealed that at 24 hr, 1,2-dihydro-1,2-dihydroxynaphthalene (dihydrodiol) was the major component in liver (40 to 50% of extracted metabolites) and muscle (-80% of extracted metabolites) regardless of the temperature. Bile contained, primarily, conjugates (e.g., glucuronides), which yielded the dihydrodiol as the principal metabolite on enzymatic hydrolysis. From 24 to 168 hr, the concentrations of each metabolite class did not vary directly with the concentrations of total metabolites. Accordingly, at 168 hr, the ratio of total metabolite concentrations in liver of fish at 4°C compared to 12°C was 1.6, whereas the ratios for the dihydrodiol, sulfate/glucoside conjugates and glucuronide conjugates were 4.5, 0.6 and 3.8 respectively.Generally, lowered water temperature increased tissue concentrations of the parent hydrocarbon and its metabolites. However, the magnitude of the increase was dependent upon the compound, the tissue, and the time after the initiation of the exposure. The results emphasize the importance of determining concentrations of individual metabolites together with parent hydrocarbons in tissues of fish when assessing effects of environmental parameters on xenobiotic toxicity.     

The presence of 1-naphthol in the urine of industrial workers exposed to naphthalene.

1-Naphthol at concentrations ranging from 0.4 to 34.6 mg/l was found in urine collected directly after the end of the work shift from a group of industrial workers employed in distillation of naphthalene oil. The maximum excretion was found one hour after the end of the shift and the mean excretion rate was 0.57 mg/h. Coke plant workers exposed to naphthalene and other aromatic and polycyclic hydrocarbons also had 1-naphthol in their urine. Mean values were 0.89 mg/l (working with new technology) and 4.86 mg/l (working with old technology) and the excretion rates were 0.19 and 0.31 mg/h respectively. The maximum excretion was shifted to two to three hours after the end of the exposure. For non-exposed subjects the mean urinary 1-naphthol concentration was 120 micrograms/l and the excretion rate was 7.0 micrograms/h.     

In vivo effects of the soluble fraction of light cycle oil on immune functions in the European sea bass, Dicentrarchus labrax (Linné)

Hydrocarbons are major contaminants that may affect biota at various trophic levels in estuaries and coastal ecosystems. The effects of accidental pollution by light cycle oil (LCO), a refined product of heavy fuel oil, on bioaccumulation, depuration processes and immune-related parameters in the European sea bass, Dicentrarchus labrax, were investigated in the laboratory after 7 days of exposure and a 2-week recovery period. Exposure of fish to the soluble fraction of LCO (1600 ng L⁻¹) for 7 days led to the bioaccumulation of some polycyclic aromatic hydrocarbons (PAHs) in muscles: naphthalene, acenaphthene, fluorene, phenanthrene and anthracene. After 7 days of recovery period, half-elimination of naphthalene was reported in fish muscles due to facilitated diffusive loss by the epithelium and a faster elimination rate proven by the presence of a high level of naphthalene biliary metabolites. The other bioaccumulated molecules displayed a slower depuration rate due to their elimination by the formation of hydrophobic metabolites excreted through bile or urine. Three days after the beginning of the recovery period, each contaminated fish showed severe external lesions (tissue necrosis, suppurative exudates, haemorrhagic area). The hypothesis of a possible link with inflammatory phenomenon was supported by (i) an inversion of the leucocyte sub-population percentage, (ii) a significant up-expression in the spleen of the tumour necrosis factor alpha gene, (iii) a significant increase in ACH₅₀. Moreover, the lack of C3 gene regulation in the spleen suggested a non-renewal of this component. The reduction of phagocytic activity and lysozyme concentration reflected immune suppression. Finally, LCO toxicity in this fish was clearly demonstrated to be related to inflammatory reaction and immune depletion.     

A Study for the Proper Application of Urinary Naphthols, New Biomarkers for Airborne Polycyclic Aromatic Hydrocarbons

Urinary naphthols, 1- and 2-naphthol, recently have been suggested as route-specific biomarkers for exposure to airborne polycyclic aromatic hydrocarbons. For the proper application of urinary naphthols as biomarkers, we studied effects of lifestyle on urinary naphthols levels in 119 Japanese male workers. After improving the detection limit of urinary naphthols up to 0.27 μg/L by high-resolution capillary gas chromatography/mass spectrometry/selected ion monitoring, urinary naphthols were detectable in approximately 90% of the subjects. Among detectable samples, the geometrical mean (GM) of urinary 1-naphthol concentration was 5.13 μg/L (geometrical standard deviation, GSD, 4.90), while the GM of urinary 2-naphthol concentration was 3.16 μg/L (GSD, 5.61). We observed that urinary 1- and 2-naphthol level were three- and sevenfold higher, respectively, among smokers than among nonsmokers (p < 0.01). The ratios of urinary 2-naphthol to 1-naphthol were significantly higher among smokers than nonsmokers (p < 0.05). The number of cigarettes smoked and urinary cotinine levels were also positively related to the concentration of urinary naphthols (p < 0.01), while other lifestyle factors, i.e., age and consumption of alcohol, greasy or salty food, sweets, fruits, vegetables, meat, or fish, were not. We also studied whether genetic polymorphisms of enzymes, which were involved in naphthalene metabolism, affected urinary naphthols levels. The cytochrome P450 (CYP) 1A1 exon 7 genetic polymorphism was not related to urinary naphthol levels. Among smokers, the subjects with c1/c2 or c2/c2 type of CYP2E1, which was determined by CYP2E1 RsaI polymorphism in 5′-flaking region, showed higher concentrations of urinary 2-naphthol than the subjects with c1/c1 type regardless of creatinine-correction (p < 0.05) and the subjects with glutathione S-transferase (GST) M1 deficient type showed higher concentrations of both urinary 1- and 2-naphthol than those with GSTM1 normal type but only without creatinine-correction (p < 0.05). Thus, when urinary naphthols are used as biomarkers, smoking and the genetic polymorphisms of CYP2E1 and GSTM1 should be considered. Received: 6 January 1998/Accepted: 10 July 1998     

Analysis of urinary metabolites of polycyclic aromatic hydrocarbons in incineration workers

Incineration workers are exposed to various pyrolysis products of organic materials, heavy metals and polycyclic aromatic hydrocarbons (PAHs). In this study, the exposure of incineration workers to PAHs was evaluated by measuring urinary metabolites of pyrene and naphthalene. The concentrations of urinary 1-hydroxypyrene (1OHP), a metabolite of pyrene, and 2-naphthol (2NP), a metabolite of naphthalene, were measured among 100 workers in 4 different types of incinerators, both before and after their work shifts. These incinerators were two old types, one modern type and one outdoors. The medians of urinary 1OHP of before and after the work shifts obtained from all workers were 0.067 and 0.044 mug/gCr, respectively; and the medians of urinary 2NP were 7.5 and 10.0 mug/gCr, respectively. A significant increase of 2NP after the work shift was found at one old incinerator. A significant decrease of metabolites was found at the other old incinerator. Significant correlations were found between urinary metabolites and cigarettes smoked per day. The effect of smoking on urinary metabolite levels was also important. Significant correlations were found between urinary 1OHP and 2NP levels in all workers. In multiple regression analysis smoking habit and incinerator type were found as significant factors. The improvement of the work environment, through decreasing exposure to both tobacco smoke and hazardous work shift-related substances, should be an occupational health aim.     

trans,trans-Muconic acid,a reliable biological indicator for the detection of individual benzene exposure down to the ppm level

Summary trans,trans-Muconic acid (2,4-hexadienedioic acid) (t,t-MA) is a minor benzene metabolite which can be used as a biological indicator for benzene exposure. The purpose of the study was to evaluate the limits of use of t,t-MA for detection and quantification of occupational exposures to benzene, particularly on an individual scale, phenol being used as the metabolite of reference. A simple and sensitve method previously described by the authors was carried out to analyse t,t-MA in 105 end-of-shift urinary samples from 23 workers exposed to benzene used as an extraction solvent for concretes recovery in the perfume industry. Good correlations were found between atmospheric benzene and both metabolites (uncorrected or corrected for creatinine) or between the metabolites themselves, with correlation coefficients from 0.81 to 0.91 (P < 0.0001). Correlation-coefficients were not improved after correction for creatinine. The overall individual benzene exposure range, median, and arithmetic mean were respectively 0.1–75, 4.5, and 9.0 ppm with corresponding t,t-MA excretion of 0.1–47.9, 5.2 and 8.9 mg/l (uncorrected) and phenol excretion of 1.4–298, 30.9, and 42.2 mg/l (uncorrected). In the control group (145 determinations for t,t-MA and 76 for phenol from 79 individuals) the range, median, and arithmetic mean were respectively < 0.04–0.66, 0.08, and 0.13 mg/l (uncorrected t,t-MA) and 1.5–42.0, 9.85 and 11.3 mg/l (uncorrected phenol). t,t-MA was far more specific than phenol and could be easily and practically used to estimate with a given probability the upper or lower corresponding benzene concentrations down to around the ppm level. Biological exposure indices for benzene exposure to 10, 5, or 1 ppm could be set at 10, 5, or 1 mg t,t-MA/l (uncorrected).