Sex differences in urinary levels of several biological indicators of exposure: a simulation study using a compartmental-based toxicokinetic model.

Toxicokinetic modeling is a useful tool to describe or predict the behavior of a chemical agent in the human or animal organism. A general model based on four compartments was developed in a previous study to quantify the effect of human variability on a wide range of biological exposure indicators. The aim of this study was to adapt this existing general toxicokinetic model to three organic solvents--methyl ethyl ketone, 1-methoxy-2-propanol, and 1,1,1,-trichloroethane--and to take into account sex differences. In a previous human volunteer study we assessed the impact of sex on different biomarkers of exposure corresponding to the three organic solvents mentioned above. Results from that study suggested that not only physiological differences between men and women but also differences due to sex hormones levels could influence the toxicokinetics of the solvents. In fact the use of hormonal contraceptive had an effect on the urinary levels of several biomarkers, suggesting that exogenous sex hormones could influence CYP2E1 enzyme activity. These experimental data were used to calibrate the toxicokinetic models developed in this study. Our results showed that it was possible to use an existing general toxicokinetic model for other compounds. In fact, most of the simulation results showed good agreement with the experimental data obtained for the studied solvents, with a percentage of model predictions that lies within the 95% confidence interval varying from 44.4 to 90%. Results pointed out that for same exposure conditions, men and women can show important differences in urinary levels of biological indicators of exposure. Moreover, when running the models by simulating industrial working conditions, these differences could be even more pronounced. A general and simple toxicokinetic model, adapted for three well-known organic solvents, allowed us to show that metabolic parameters can have an important impact on the urinary levels of the corresponding biomarkers. These observations give evidence of an interindividual variability, an aspect that should have its place in the approaches for setting limits of occupational exposure.

Sex differences in urinary levels of several biological indicators of exposure: a human volunteer study.

The aim of the study was to quantify the variability on biological indicators of exposure between men and women for three well known solvents: methyl ethyl ketone, 1-methoxy-2-propanol and 1,1,1-trichloroethane. Another purpose was to explore the effect of selected CYP2E1 polymorphisms on the toxicokinetic profile. Controlled human exposures were carried out in a 12 m³ exposure chamber for each solvent separately, during 6h and at half of the threshold limit value. The human volunteers groups were composed of ten young men and fifteen young women, including ten women using hormonal contraceptive. An analysis of variance mainly showed an effect on the urinary levels of several biomarkers of exposure among women due to the use of hormonal contraceptive, with an increase of more than 50% in metabolites concentrations and a decrease of up to 50% in unchanged substances concentrations, suggesting an increase in their metabolism rate. The results also showed a difference due to the genotype CYP2E1*6, when exposed to methyl ethyl ketone, with a tendency to increase CYP2E1 activity when volunteers were carriers of the mutant allele. Our study suggests that not only physiological differences between men and women but also differences due to sex hormones levels can have an impact on urinary concentrations of several biomarkers of exposure. The observed variability due to sex among biological exposure indices can lead to misinterpretation of biomonitoring results. This aspect should have its place in the approaches for setting limits of occupational exposure.

Evaluation of dynamic headspace with gas chromatography/mass spectrometry for the determination of 1,1,1-trichloroethane, trichloroethanol, and trichloroacetic acid in biological samples.

A sensitive and reproducible method is described for the analysis of trichloroacetic acid in urine and 1,1,1-trichloroethane in blood using dynamic headspace GC/MS. Samples were analyzed using the soil module of a modified purge and trap autosampler to facilitate the use of disposable purging vessels. Coefficients of variation were below 3.5% for both analytes, and response was linear in the range of 0.01-7.0 microg/ml for trichloroacetic acid and 0.9 ng/ml-2.2 microg/ml for 1,1,1-trichloroethane. Attempts at using dynamic headspace for the analysis of trichloroethanol in urine were unsuccessful.

The importance of measured end-points in demonstrating the occurrence of interactions: a case study with methylchloroform and m-xylene.

Mixed exposures may result in significant changes in one biomarker of exposure without altering another biomarker, and this may have unknown significance in terms of exposure assessment and overall toxicity of the mixture. Results from a previous investigation showed that human exposure to methylchloroform (MC, 400 ppm) and m-xylene (XYL, 200 ppm) during 4 h did not result in any significant effect on blood concentrations of these solvents, suggesting the absence of interaction between MC and XYL. Those results were adequately described by conducting a physiologically-based toxicokinetic (PBTK) modeling of the MC-XYL interaction in humans; however, the model suggested that urinary excretion of MC metabolites would be reduced as a result of combined exposure, whereas that of XYL metabolites would not be modified. An experimental verification of this model prediction was then undertaken with rats. In this study, Sprague-Dawley rats (n, 5) were exposed during 4 h to MC (400 ppm) or XYL (200 ppm), alone or as a mixture. Results showed that combined exposure did not affect the blood concentration of MC whereas that of XYL was increased throughout the 2-h blood collection period following exposure. The excretion of MC metabolites during a period of 48 h following the onset of exposure, i.e., trichloroethanol (TCE: -71%) and trichloroacetic acid (TCA: -73%), were significantly reduced. Methylhippuric acid (MHA) was not affected by co-exposure to MC as expected from the PBTK model forecasts. These results exemplify the use of a priori PBPK modeling for designing interaction studies and choosing appropriate/sensitive end-points for demonstrating the occurrence of potential interactions.

Effect of various exposure scenarios on the biological monitoring of organic solvents in alveolar air. II. 1,1,1-Trichloroethane and trichloroethylene.

The purpose of the present study was to investigate the influence of different exposure scenarios on the elimination of trichloroethylene (TRI) and 1,1,1-trichloroethane (1,1,1-TRI) in alveolar air and other biological fluids in human volunteers. In addition, it was sought to establish an interactive process between experimental data gathering and simulation modeling in an attempt to predict the influence of the different scenarios of exposure to TRI and 1,1,1-TRI on their respective biological monitoring indices and thus to establish the flexibility and validity of simulation models. Two adult male and two adult female Caucasian volunteers were exposed by inhalation, in a dynamic controlled exposure chamber, to various concentrations of TRI (12.5-25 ppm) or 1,1,1-TRI (87.5-175 ppm) in order to establish the influence of exposure concentration, duration of exposure, variation of concentration within day, and work load on biological exposure indices. The concentrations of unchanged solvents in end-exhaled air and in blood as well as the urinary excretion of trichloroethanol (TCE) and trichloroacetic acid (TCA) were determined. The results show that doubling the exposure concentration for both solvents led to a proportional increase in the concentrations of unchanged solvents in alveolar air and blood at the end of a 7-h exposure period; this proportionality was still observable in 1,1,1-TRI expired air samples 16 h after the end of the third exposure day. In the case of urinary excretion of TCE and TCA, the proportionality between excretion and exposure concentration was not as good. It was once again observed that the slow excretion of both metabolites leads to progressive cumulation and that their urinary determination is subject to considerable interindividual variations. After adjustment (lowering) of the exposure concentration to account for a prolongation of the duration of exposure (from 8 to 12 h) it was observed that the concentrations of TRI or 1,1,1-TRI towards the end of both exposure periods are more a reflection of the actual exposure concentration than of the exposure duration. Despite important interindividual variations, these adjusted and nonadjusted exposures led to almost identical average total urinary excretion over 24 h) of TCE and TCA after exposure to 1,1,1-TRI, as was also the case for TCE but not for TCA after exposure to TRI. Induced within-day variations in the exposure concentration led to corresponding but not proportional changes in alveolar air concentrations for both solvents. After exposure to peak concentrations there was a lag period before alveolar air concentrations returned to prepeak levels. At the end of repeated 10-min periods of physical exercise at 50 W, alveolar air concentrations of TRI were increased by 50% while those of 1,1,1-TRI increased by only 12%. After optimization of the physiologically based toxicokinetic model parameters with experimental data collected during the first exposure scenario, results pertaining to the three other scenarios were adequately simulated by the optimized models. Overall, the results of the present study suggest that alveolar air solvent concentration is a reliable index of exposure to both TRI and 1,1,1-TRI under various experimental exposure scenarios. These results also suggest that urinary excretion of TCE and TCA must be interpreted with caution when assessing exposure to either solvents. For exposure situations likely to be encountered in the workplace, physiologically based toxicokinetic modeling appears to be a useful tool both for developing strategies of biological monitoring of exposure to volatile organic solvents and for predicting alveolar air concentrations under a given set of exposure conditions.

Urinary methylchloroform rather than urinary metabolites as an indicator of occupational exposure to methylchloroform.

In order to compare methylchloroform (MC, or 1,1,1-trichloroethane) per se and its metabolites in urine as indicators of occupational exposure to this solvent, 50 male solvent workers were studied in the second half of a working week to evaluate the exposure-excretion relationship. The time-weighted average intensity of solvent exposure of individuals during an 8-h shift was monitored by personal diffusive sampling. Urine samples were collected near the end of the shift and were analyzed for MC and its metabolites [i.e., trichloroacetic acid (TCA), trichloroethanol (TCE) and total trichloro-compounds (TTC; the sum of TCA and TCE)] by head-space gas chromatography. MC per se, TCA, TCE, and TTC in urine correlated significantly (P < 0.01) with MC in ambient air, and among the four the correlation coefficient was highest for MC. The same result were obtained by multiple regression analysis in which ambient air MC was taken as the dependent variable and either the three indicators urinary MC, TCA, and TCE or the two indicators urinary MC and TTC were taken as independent variables. Taking the specificity and selectivity of the analyte as well as the simple and hazardous chemical-free procedure of analysis into consideration, it is concluded that MC is the analyte of choice as an indicator of occupational exposure to MC, when urine is selected as a specimen available by noninvasive sampling.

Determinacao espectrofotometrica e cromatografica em fase gasosa de tricloretanol em urina / Spectrophotometric and gas chromatographic determination of trichlorethanol in urine

O tricloretanol (TCE), produto de biotransformacao dos solventes 1,1,1-tricloretano (1,1,1-TRI) e tricloretileno, e usado como indicador biologico de dose interna na vigilancia de individuos expostos ocupacionalmente a estes compostos. Neste trabalho foi realizada a otimizacao de metodo espectrofotometrico e cromatografico em fase gasosa para a determinacao do TCE, bem como se estudou a aplicacao destes metodos a analise de urina de trabalhadores expostos ao 1,1,1-TRI. A analise cromatografica com detector de captura eletronica foi efetuada em coluna SE-30 - 15//, usando-se o diclorobenzeno como padrao externo. Ambos os metodos mostraram precisao e recuperacao adequados, respectivamente de 2,2//de CV e 95,6//para o espectrofotometrico e de 2,3//de CV e 95,1//para o cromatografico. O teor medio de tricloretanol em urina, expresso em mg/g de creatinina, foi de 4,4 quando usada a espectrofotometria e de 2,4 pela cromatografia, para o grupo de individuos expostos ao 1,1,1-tricloretano. Os resultados obtidos mostraram a maior sensibilidade e especificidade da cromatografia em fase gasosa na determinacao do tricloretanol em urina

Exposure of 1,1,1-trichloroethane and dose-related excretion of metabolites in urine of printing workers.

The quantitative relationship between the time-weighted average exposure by inhalation to 1,1,1-trichloroethane and excretion of metabolites in urine at the end of a work shift was investigated in 48 male printing workers exposed to the solvent at levels below 65 ppm. Male control subjects, 10 in all, were also investigated. Statistical analysis showed that there is a linear relationship between the two exposure indicators, suggesting that occupational exposure to 1,1,1-trichloroethane can be bio-monitored by means of urinalysis for either total trichloro-compounds or trichloroacetic acid. The correlation coefficient was higher for total trichloro-compounds (r = 0.86-0.89 depending on correction for urine density) than trichloroacetic acid (r = 0.50-0.71), which suggests that the former is a better indicator of exposure to 1,1,1-trichloroethane than the latter. The calculations based on the present study suggest that, at the end of the shift, only less than 2% of 1,1,1-trichloroethane absorbed will be excreted as metabolites in urine.

1,1,1-Trichloroethane (methyl chloroform) in urine as biological index of exposure.

Fifteen human volunteers were exposed to 1,1,1-trichloroethane (methyl chloroform) vapor at 72-495 mg/m3 for a period of 2 to 4 hours at rest (ten cases) and during light physical exercise (five cases). Subsequently 60 workers occupationally exposed to 1,1,1-trichloroethane in a refrigerator manufacturing plant were studied (median value: 178 mg/m3; geometrical standard deviation: 2.19 mg/m3). As expected, the relative uptake (R) of 1,1,1-trichloroethane decreased in the course of exposure at rest (R = 0.44 after 20 minutes of exposure; R = 0.26 after 240 minutes of exposure). Both in the experimentally exposed subjects and in the occupationally exposed workers, the urinary concentration of 1,1,1-trichloroethane showed a linear relationship to the corresponding environmental time-weighted average concentration. The correlation coefficients (r) were 0.95 in occupationally exposed subjects and more than 0.90 in experimentally exposed groups. A linear equation also existed between urinary concentration and amount of 1,1,1-trichloroethane absorbed (r = 0.88). The findings indicate that the urinary concentration of 1,1,1-trichloroethane can be used as an appropriate biological exposure indicator. In occupationally exposed subjects performing moderate work, the urinary 1,1,1-trichloroethane concentration corresponding to the time-weighted average of the threshold limit value was found to be 860 micrograms/L and its 95% lower confidence limit (biological threshold) 805 micrograms/L.

[Use of gas chromatography with flame ionization (GC-FID) in the measurement of solvents in the urine]. / Impiego della gas cromatografia a ionizzazione di fiamma (GC-FID) per la misura dei solventi tal quali nelle urine.

The urinary concentration of some solvents (acetone, cyclohexane, 1,2 dichloropropane, n-hexane, methyl ethyl ketone, perchloroethylene, styrene, toluene, 1,1,1, trichloroethane) was measured by means of a gas chromatography Hewlett-Packard 5890 supplied with a flame ionization detector (GC-FID, DANI HS 3950). The coefficient of variation of the method was lower than 5%. The sensitivity of the GC-FID was very similar to what of mass spectrometer detector (GC-MSD, HP 5970 A).

Metabolites of chlorinated solvents in blood and urine of subjects exposed at environmental level.

After a two-level selection, 141 blood donors living in Milan, Italy, were analyzed for their content of plasma and urinary trichloroacetic acid (TCA) and trichloroethanol (TCE). Environmental levels of exposure to trichloroethylene (TRI) and tetrachloroethylene (PER) through drinking water and air were also measured. The plasma TCA levels were in the range of previously found concentrations. Relationships among plasma and urinary metabolites were found and discussed.

[Necessity for total trichloride compound measurement in screening tests for workers exposed to trichloroethylene or 1,1,1,-trichloroethane].

In 9 series of physical examinations legally required for trichloroethylene (TCE) workers, levels of urinary total trichloride compounds (TTC) were revealed to be generally high in 323 specimens, including 250 male ones, collected from a medium sized company. Sixty four specimens (19.8%), including 56 male ones (22.4%), exceeded 300 mg/l which are regarded to correspond to the threshold limit for TCE in the air of the work site. Four specimens had over 900 mg/l. In all 323 specimens, however, no abnormalities were found with respect to specific gravity of blood, hemoglobin concentration and urinary tests. TTC concentration values increased in specimens in the last 4 physical checkups, compared with the values measured in the first 5. The cause of this increase seemed to be related to increases in the number of products and the hours of overtime work. Symptoms felt by these workers gradually subsided as they were continuously exposed to TCE for more than 2 or 3 years. Some workers of long service had a tendency toward addiction to TCE. In the legally required environmental monitoring of TCE, and other harmful organic solvents, the 8 workplaces fell into either classification 1 or 2, indicating that the environmental conditions are not harmful. More importantly, the above evaluations did not take into account long hours of overtime put in by those workers. Turning to the checkups of 1,1,1-trichloroethane workers, the results of legally required physical checkups and environmental monitoring were almost the same as those for TCE.(ABSTRACT TRUNCATED AT 250 WORDS)

1,1,1-Trichloroethane exposure, biologic monitoring by breath and urine analyses.

Absorption and excretion of 1,1,1-trichloroethane, as well as the kinetics of formation and elimination of trichloroethanol (TCE) and trichloroacetic acid (TCA) were simulated by a mathematical model. The results of this model were compared with experimental one on pulmonary elimination of the solvent and urinary excretion of the metabolites. The influences of duration and repetition of exposure on the pulmonary and urinary eliminations were studied. A tentative method of biologic monitoring is proposed. Theoretically, the most suitable method of biologic monitoring is proposed. Theoretically, the most suitable method to estimate the exposure is by two determinations, before and after a work shift. Following this procedure, analysis of TCE in the urine is more sensitive than determination of 1,1,1-trichloroethane in the breath. As an indicator of exposure risk, TCA is not considered sensitive enough if variations in the inspired concentration occur.