Michèle Bouchard

Urinary 1-hydroxypyrene as a biomarker of exposure to polycyclic aromatic hydrocarbons: biological monitoring strategies and methodology for determining biological exposure indices for various work environments

This article reviews the published studies on urinary 1-hydroxypyrene (1-OHP) as a biomarker of exposure to polycyclic aromatic hydrocarbons (PAHs) in work environments. Sampling and analysis strategies as well as a methodology for determining biological exposure indices (BEIs) of 1-OHP in urine for different work environments are proposed for the biological monitoring of occupational exposure to PAHs. Owing to the kinetics of absorption of pyrene by different exposure routes and excretion of 1-OHP in urine, in general, 1-OHP urinary excretion levels increase during the course of a workday, reaching maximum values 3-9 h after the end of work. When the contribution of dermal exposure is important, post-shift 1-OHP excretion can however be lower than pre-shift levels in the case where a worker has been exposed occupationally to PAHs on the day prior to sampling. In addition, 1-OHP excretion levels in either pre-shift, post-shift or evening samples increase during the course of a work-week, levelling off after three consecutive days of work. Consequently, ideally, for a first characterization of a work environment and for an indication of the major exposure route, considering a 5-day work-week (Monday to Friday), the best sampling strategy would be to collect all micturitions over 24 h starting on Monday morning. Alternatively, collection of pre-shift, post-shift and evening urine samples on the first day of the work-week and at the end of the work-week is recommended. For routine monitoring, pre-shift samples on Monday and post-shift samples on Friday should be collected when pulmonary exposure is the main route of exposure. On the other hand, pre-shift samples on Monday and Friday should be collected when the contribution of skin uptake is important. The difference between beginning and end of work-week excretion will give an indication of the average exposure over the workweek. Pre-shift samples on the first day of the work-week will indicate background values, and, hence, reflect general environment exposure and body burden of pyrene and/or its metabolites. On the other hand, since PAH profile can vary substantially in different work sites, a single BEI cannot apply to all workplaces. A simple equation was therefore developed to establish BEIs for workers exposed to PAHs in different work environments by using a BEI already established for a given work environment and by introducing a correction factor corresponding to the ratio of the airborne concentration of the sum of benzo(a)pyrene (BaP) equivalent to that of pyrene. The sum of BaP equivalent concentrations represents the sum of carcinogenic PAH concentrations expressed as BaP using toxic equivalent factors. Based on a previously estimated BEI of 2.3 μmol 1-OHP mol(-1) creatinine for coke-oven workers, BEIs of 4.4, 8.0 and 9.8 μmol 1-OHP mol(-1) creatinine were respectively calculated for vertical pin Söderberg workers, anode workers and pre-bake workers of aluminium plants and a BEI of 1.2 μmol 1-OHP mol(-1) creatinine was estimated for iron foundry workers. This approach will allow the potential risk of cancer in individuals occupationally exposed to PAHs to be assessed better.

Biological monitoring of exposure to pyrethrins and pyrethroids in a metropolitan population of the Province of Quebec, Canada.

Pyrethroid and pyrethrins are neurotoxic insecticides widely used to control agricultural and domestic insect pests. The general population is potentially chronically exposed through food consumption, but the actual exposure is poorly documented in Canada. This study aimed at obtaining an indication of the absorption of those insecticides in residents of Montreal Island, the largest metropolitan area of the Province of Quebec, Canada. We randomly recruited 120 adults and 120 children aged 18-64 and 6-12 years old, of which 81 adults and 89 children completed the study. The absorption of pyrethroids and pyrethrins was assessed through measurements of six urinary metabolites: chrysanthemum dicarboxylic acid (CDCA), cis- and trans-2,2-(dichlorovinyl)-2,2-dimethylcyclopropane carboxylic acids (cDCCA and tDCCA), cis-2,2-(dibromovinyl)-2,2-dimethylcyclopropane carboxylic acid (DBCA), 3-phenoxybenzoic acid (PBA) and 4-fluoro-3-phenoxybenzoic acid (FPBA). Metabolites were determined in 12-h urine collections for children and 2-consecutive 12-h collections for adults, and were analyzed by gas-chromatography/mass spectrometry. In both adults and children, the relative distribution of the various metabolites was as follows: tDCCA>PBA>cDCCA>CDCA>DBCA>FPBA. In adults, median (95th percentiles) cumulative amounts of these metabolites were 12.0 (231.1), 8.2 (177.9), 5.0 (110.1), 0.3 (8.2), 0.1 (4.7) and 0.1 (0.5)pmol/kg bw, respectively, in nighttime 12-h urine collections. Corresponding values in children were 12.6 (207.7), 10.2 (73.2), 5.1 (59.6), 2.1 (14.2), 0.1 (4.9) and 0.1 (0.8)pmol/kg bw. The main metabolites observed are indicative of exposure mainly to permethrin and cypermethrin and amounts absorbed are in the same range in adults and children. The distribution levels of the main metabolites in our sample also appeared similar to those reported in the US population.

Exposure to phthalates, bisphenol A and metals in pregnancy and the association with impaired glucose tolerance and gestational diabetes mellitus: The MIREC study.

BACKGROUND Studies from several countries report increases in rates of gestational diabetes mellitus (GDM) over recent decades. Exposure to environmental chemicals could contribute to this trend. OBJECTIVES To determine the associations between plasticisers and metals measured in early pregnancy with impaired glucose tolerance (IGT) and GDM in a Canadian pregnancy cohort. METHODS Women enrolled in the Maternal-Infant Research on Environmental Chemicals (MIREC) Study were included if they had a singleton delivery and did not have pre-existing diabetes. Eleven phthalate metabolites and total bisphenol A (BPA) were measured in first-trimester urine samples, and four metals (lead, cadmium, mercury and arsenic) were measured in first-trimester blood samples. IGT and GDM were assessed in accordance with standard guidelines by chart review. Chemical concentrations were grouped by quartiles, and associations with outcomes were examined using logistic regression with adjustment for maternal age, race, pre-pregnancy BMI, and education. Restricted cubic spline analysis was performed to help assess linearity and nature of any dose-response relationships. RESULTS Of 2001 women recruited into the MIREC cohort, 1274 met the inclusion criteria and had outcome data and biomonitoring data measured for at least one of the chemicals we examined. Elevated odds of GDM were observed in the highest quartile of arsenic exposure (OR = 3.7, 95% CI = 1.4-9.6) in the adjusted analyses. A significant dose-response relationship was observed in a cubic spline model between arsenic and odds of GDM (p < 0.01). No statistically significant associations were observed between phthalates or BPA or other metals with IGT or GDM. CONCLUSIONS Our findings add to the growing body of evidence supporting the role of maternal arsenic exposure as a risk factor for gestational diabetes.

The toxicokinetics of pyrene and its metabolites in rats

Five experiments were conducted in male Sprague-Dawley rats regarding the kinetic of urinary excretion of 1-hydroxypyrene (1-OHP) following i.v., oral and dermal exposure to 0.5-50 micromol/kg pyrene either as a single substance or as mixture of various polycyclic aromatic hydrocarbons (PAH). Frequent urine collections over 48 h after exposure and a tissue versus time distribution experiment using [14C]pyrene allowed to define the kinetic profile of both pyrene and 1-OHP. For all exposure routes, there is a linear relationship over two orders of magnitude between the dose of pyrene and the urinary excretion of 1-OHP. Differences in biliary/urinary 1-OHP excretion ratio in canulated rats (3) versus faecal/urinary 1-OHP excretion ratio in non-canulated rats (0.6) indicate major enterohepatic recirculation of the metabolite. Half-lives of both pyrene and 1-OHP in all measured tissues were all comprised between 3.1 and 5.4 h, and 5.2-6.7 h, respectively, so that no long term accumulation would be predicted from these values for any tissue. Binary and ternary mixtures involving naphthalene and benzo(a)pyrene in addition to pyrene has no influence on the urinary excretion profile of 1-OHP. All these observations led to the proposal of a dynamic compartment model of pyrene and metabolite flows indicating that following rapid initial distribution to fatty tissues, pyrene is rapidly biotransformed into various metabolites and undergoes major enterohepatic recycling. Part of the initially formed and part of the recirculated 1-OHP eventually undergoes urinary excretion such that close to 60% of pyrene is eliminated as metabolites in urine by 24 h after injection while 20% is excreted in the faeces over the same period.

A toxicokinetic study to elucidate 3-hydroxybenzo(a)pyrene atypical urinary excretion profile following intravenous injection of benzo(a)pyrene in rats.

The toxicokinetics of benzo(a)pyrene (BaP) and 3‐hydroxybenzo(a)pyrene (3‐OHBaP) were assessed in 36 male Sprague–Dawley rats injected intravenously with 40 µmol kg1 of BaP to explain the reported atypical urinary excretion profile of 3‐OHBaP. Blood, liver, kidney, lung, adipose tissue, skin, urine and feces were collected at t = 2, 4, 8, 16, 24, 33, 48, 72 h post‐dosing. BaP and 3‐OHBaP were measured by high‐performance liquid chromatography/fluorescence. A biexponential elimination of BaP was observed in blood, liver, skin and kidney (t½ of 4.2–6.1 h and 12.3–14.9 h for initial and terminal phases, respectively), while a monoexponential elimination was found in adipose tissue and lung (t½ of 31.2 and 31.5 h, respectively). A biexponential elimination of 3‐OHBaP was apparent in blood, liver and skin (t½ of 7.3–11.7 h and 15.6–17.8 h for initial and terminal phases, respectively), contrary to adipose tissue, lung and kidney. In adipose tissue and lung, a monophasic elimination of 3‐OHBaP was observed (t½ of 27.0 h and 24.1 h, respectively). In kidney, 3‐OHBaP kinetics showed a distinct pattern with an initial buildup during the first 8 h post‐dosing followed by a gradual elimination (t½ of 15.6 h). In the 72‐h post‐treatment, 0.21 ± 0.09% (mean ± SD) of dose was excreted as 3‐OHBaP in urine and 12.9 ± 1.0% in feces while total BaP in feces represented 0.40 ± 0.16% of dose. This study allowed the identification of the kidney as a retention compartment governing 3‐OHBaP atypical urinary excretion. Copyright

Reconstruction of methylmercury intakes in indigenous populations from biomarker data

Significant amounts of methylmercury (MeHg) can bioaccumulate in fish and sea mammals. To monitor MeHg exposure in individuals, organic and inorganic mercury are often measured in blood samples or in hair strands, the latter being by far the best integrator of past exposure. With knowledge of the MeHg kinetics in humans, the levels of both biomarkers can be related to MeHg body burden and intakes. In the present study, we use the toxicokinetic model of Carrier et al. (2001) describing the distribution and excretion of MeHg in humans, to reconstruct the history of MeHg intakes of indigenous women of the Inuvik region in Canada starting from total mercury concentrations in hair segments. From these reconstructed MeHg intakes, the corresponding simulated mercury blood concentrations are found to be good predictors of the concentrations actually measured in blood samples. An important conclusion of this study is that, for almost all subjects, the reconstructed history of their MeHg intakes provides much lower intake values than intakes estimated from questionnaires on food consumption and estimated MeHg levels in these foods; the mean value of the reconstructed MeHg intakes is 0.03 μg/kg/day compared with the mean value of 0.20 μg/kg/day obtained from questionnaires. The model was also used to back-calculate the MeHg intakes from concentrations in hair strands collected from aboriginals of the Amazon region in Brazil, a population significantly more exposed than the population of the Inuvik region.

Concentrations versus amounts of biomarkers in urine: a comparison of approaches to assess pyrethroid exposure.

BackgroundAssessment of human exposure to non-persistent pesticides such as pyrethroids is often based on urinary biomarker measurements. Urinary metabolite levels of these pesticides are usually reported in volume-weighted concentrations or creatinine-adjusted concentrations measured in spot urine samples. It is known that these units are subject to intra- and inter-individual variations. This research aimed at studying the impact of these variations on the assessment of pyrethroid absorbed doses at individual and population levels.MethodsUsing data obtained from various adult and infantile populations, the intra and inter-individual variability in the urinary flow rate and creatinine excretion rate was first estimated. Individual absorbed doses were then calculated using volume-weighted or creatinine-adjusted concentrations according to published approaches and compared to those estimated from the amounts of biomarkers excreted in 15- or 24-h urine collections, the latter serving as a benchmark unit. The effect of the units of measurements (volume-weighted or creatinine adjusted concentrations or 24-h amounts) on results of the comparison of pyrethroid biomarker levels between two populations was also evaluated.ResultsEstimation of daily absorbed doses of permethrin from volume-weighted or creatinine-adjusted concentrations of biomarkers was found to potentially lead to substantial under or overestimation when compared to doses reconstructed directly from amounts excreted in urine during a given period of time (-70 to +573% and -83 to +167%, respectively). It was also shown that the variability in creatinine excretion rate and urinary flow rate may introduce a bias in the case of between population comparisons.ConclusionThe unit chosen to express biomonitoring data may influence the validity of estimated individual absorbed dose as well as the outcome of between population comparisons.

Urinary and biliary excretion kinetics of 1-hydroxypyrene following intravenous and oral administration of pyrene in rats

The urinary and biliary excretion kinetics of 1-hydroxypyrene (1-OHP) were compared in male Sprague-Dawley rats exposed intravenously and orally to 1.5, 5, 15, 50 and 100 micromol/kg pyrene. Urine and bile samples were collected at 6-h intervals for up to 24 h. Results showed that the kinetics of 1-OHP were similar for both biliary and urinary excretion whatever the administered dose or exposure route. Furthermore, the time course of 1-OHP excretion in either bile or urine following intravenous dosing resembled that observed after oral administration. In addition, the exposure route and dose had no significant effect on the fraction of dose recovered in urine and bile as 1-OHP after 6, 12, 18 and 24 h post-dosing. Following intravenous injection of 1.5, 5, 15, 50 and 100 micromol/kg pyrene, the mean cumulative percent of dose excreted as 1-OHP in urine over 24 h ranged from 1.7 to 3.2%, while biliary values ranged from 6.5 to 9.5%. Correspondingly, after oral administration, on average, 2.6-3.3% of dose was excreted as 1-OHP in urine and 7.9-10.9% was recovered in bile. Overall, the linear dose-excretion relationship following either exposure routes supports the usefulness of 1-OHP in urine as a bioindicator of polycyclic aromatic hydrocarbon (PAH) exposure. Results further suggest that tissue uptake and distribution of intravenously and orally administered pyrene proceeds similarly. By comparing these data with predicted values from a previously published physiologically based pharmacokinetic model for pyrene in the rat, it also appears that a small fraction of pyrene dose (12%) remains in the body after 24 h and that metabolites other than 1-OHP as measured in the current study are present in significant proportions in urine and feces.

Urinary excretion of benzo[a]pyrene metabolites following intravenous, oral, and cutaneous benzo[a]pyrene administration.

The effect of the administration route, dose, and sampling time on the total urinary excretion of four major benzo[a]pyrene (BaP) metabolites, 3-hydroxyBaP (3-OHBaP), 9-hydroxyBaP 9-hydroxyBaP (9-OHBaP), trans-4,5-dihydrodiolBaP (4,5-diolBaP), and trans-9,10-dihydrodiolBaP (9,10-diolBaP), was studied in male Sprague-Dawley rats exposed to a single intravenous, oral, and cutaneous dose of 2, 6, 20, and 60 mumol BaP/kg. Urine samples were collected at 24-h intervals following treatment. Over the 0-72 h period and for a given dose, amounts of BaP metabolites were 3-OHBaP > 4,5-diolBaP > > 9-OHBaP following intravenous and oral dosing, and 3-OHBaP > > 9-OHBaP > or = 4,5-diolBaP after cutaneous treatment. 9,10-diolBaP was barely detected. On the other hand, amounts of 3-OHBaP and 4,5-diolBaP excreted in urine over the 0-72 h period and for a given dose appeared in the following order: intravenous approximately oral > or = cutaneous. Amounts of 9-OHBaP excreted varied as follows: oral > or = cutaneous > intravenous. For all routes of administration, excretion of 4,5-diolBaP was almost complete over the 0-24 h period in contrast with 3-OHBaP and 9-OHBaP. Peak excretion of 3-OHBaP and 9-OHBaP was reached in the 0-24 h period following intravenous and oral treatment and in the 24-48 h period following cutaneous application. Overall, for a given administration route and dose, there were variations in the time profiles between metabolites. In general, there was nonetheless a good correlation between the BaP dose and urinary excretion of 3-OHBaP, 9-OHBaP, and 4,5-diolBaP. Furthermore, total urinary excretion of a specific metabolite, its time profile, and the relative proportion of the metabolites studied depended on the administration route. Data also suggest that a measure of the concentration ratio of the different metabolites could reflect the time and main route of exposure.

Assessment of exposure to pyrethroids and pyrethrins in a rural population of the Montérégie area, Quebec, Canada.

Pesticide use remains a preoccupation of the population and public health authorities given its possible impact on health. Pyrethroids can be listed among the widely used pesticides. The general population is potentially chronically exposed to pyrethroids mainly through food intake, but acute or sporadic exposures can also occur by other routes. Although pyrethroids are considered among the least toxic pesticides, their neurotoxic properties can affect humans, but current exposure levels in the population of Quebec are not known. The study thus aimed at assessing pyrethroid exposure in a rural, agricultural population during summer through measurements of urinary biomarkers. A total of 163 volunteers, 49 children and 114 adults, living in the Montérégie area of Quebec, participated in the study, which took place from June to August 2006, the period of intensive application of pesticides. Participants were asked to collect all their micturitions from 6 p.m. until the next morning, including first morning void, and to fill out a questionnaire to document factors that could potentially contribute to exposure. A gas-chromatography mass-spectrometry method was used to quantify six urinary metabolites resulting from pyrethroid biotransformation: cis- and trans-2,2-(dichlorovinyl)-2,2-dimethylcyclopropane carboxylic acid (cDCCA and tDCCA), 3-phenoxybenzoic acid (PBA), chrysanthemum dicarboxylic acid (CDCA), cis-2,2-(dibromovinyl)-2,2-dimethylcyclopropane carboxylic acid (DBCA) and 4-fluoro-3-phenoxybenzoic acid (FPBA). Distributions of amounts of the six metabolites excreted per unit of body weight, during a standardized 12-hr period, followed the same decreasing pattern in adults and in children: tDCCA > PBA > cDCCA > CDCA > DBCA > FPBA. No statistically significant difference was observed between the two groups, but amounts of metabolites varied greatly among individuals, suggesting important interindividual variations in the absorbed doses of these compounds. No consistent associations were observed between the excretion of correlated metabolites and the various factors documented by questionnaire (personal factors, life habits, sources of exposure). Comparison of the current data with those observed in an urban population of the same province during the summer of 2005 suggests a greater summertime exposure to some pyrethroids in the rural population.