Evaluation of occupational exposure: comparison of biological and environmental variabilities using physiologically based toxicokinetic modeling.

PURPOSE: Few studies compare the variabilities that characterize environmental (EM) and biological monitoring (BM) data. Indeed, comparing their respective variabilities can help to identify the best strategy for evaluating occupational exposure. The objective of this study is to quantify the biological variability associated with 18 bio-indicators currently used in work environments. METHOD: Intra-individual (BV(intra)), inter-individual (BV(inter)), and total biological variability (BV(total)) were quantified using validated physiologically based toxicokinetic (PBTK) models coupled with Monte Carlo simulations. Two environmental exposure profiles with different levels of variability were considered (GSD of 1.5 and 2.0). RESULTS: PBTK models coupled with Monte Carlo simulations were successfully used to predict the biological variability of biological exposure indicators. The predicted values follow a lognormal distribution, characterized by GSD ranging from 1.1 to 2.3. Our results show that there is a link between biological variability and the half-life of bio-indicators, since BV(intra) and BV(total) both decrease as the biological indicator half-lives increase. BV(intra) is always lower than the variability in the air concentrations. On an individual basis, this means that the variability associated with the measurement of biological indicators is always lower than the variability characterizing airborne levels of contaminants. For a group of workers, BM is less variable than EM for bio-indicators with half-lives longer than 10-15 h. CONCLUSION: The variability data obtained in the present study can be useful in the development of BM strategies for exposure assessment and can be used to calculate the number of samples required for guiding industrial hygienists or medical doctors in decision-making.

Impact of biological and environmental variabilities on biological monitoring--an approach using toxicokinetic models.

Biological monitoring of occupational exposure is characterized by important variability, due both to variability in the environment and to biological differences between workers. A quantitative description and understanding of this variability is important for a dependable application of biological monitoring. This work describes this variability, using a toxicokinetic model, for a large range of chemicals for which reference biological reference values exist. A toxicokinetic compartmental model describing both the parent compound and its metabolites was used. For each chemical, compartments were given physiological meaning. Models were elaborated based on physiological, physicochemical, and biochemical data when available, and on half-lives and central compartment concentrations when not available. Fourteen chemicals were studied (arsenic, cadmium, carbon monoxide, chromium, cobalt, ethylbenzene, ethyleneglycol monomethylether, fluorides, lead, mercury, methyl isobutyl ketone, penthachlorophenol, phenol, and toluene), representing 20 biological indicators. Occupational exposures were simulated using Monte Carlo techniques with realistic distributions of both individual physiological parameters and exposure conditions. Resulting biological indicator levels were then analyzed to identify the contribution of environmental and biological variability to total variability. Comparison of predicted biological indicator levels with biological exposure limits showed a high correlation with the model for 19 out of 20 indicators. Variability associated with changes in exposure levels (GSD of 1.5 and 2.0) is shown to be mainly influenced by the kinetics of the biological indicator. Thus, with regard to variability, we can conclude that, for the 14 chemicals modeled, biological monitoring would be preferable to air monitoring. For short half-lives (less than 7 hr), this is very similar to the environmental variability. However, for longer half-lives, estimated variability decreased.

Toxicokinetics of p-tert-octylphenol in male and female Sprague-Dawley rats after intravenous, oral, or subcutaneous exposures.

This study was undertaken to characterize the toxicokinetics of p-tert-octylphenol (OP), a weak estrogenic compound, in male and female rats. Male and female Sprague-Dawley rats were given a single dose of OP either by oral gavage (50, 125 or 250 mg/kg), by intravenous (iv) injection (2, 4, or 8 mg/kg), or by subcutaneous (sc) injection (125 mg/kg). In a repeated dosing experiment, rats were given OP (oral) daily (25, 50, or 125 mg/kg) for 35 d (female) or 60 d (male). Blood and tissue samples were collected and analyzed for OP content using gas chromatography with detection by mass spectrometry. Blood OP concentrations were generally higher in female than male rats following a single oral or sc administration but were similar following a single iv injection. Tissue OP concentrations were also higher in female than male rats following oral exposure, consistent with the faster metabolism of OP observed in male rat liver microsomes. After subchronic administration, blood OP concentrations were higher at the end of exposure for female (33 d) (2.26-fold, not significant) and male (57 d) (3.47-fold) rats than single dosing but there was no change in the tissue OP concentrations. Gender differences in tissue OP concentrations may contribute, in part, to gender differences in the toxicity of OP in rats. The fact that OP was found in all reproductive tissues confirms its potential for direct endocrine-like effects.

Determination of p-tert-octylphenol in blood and tissues by gas chromatography coupled with mass spectrometry.

A sensitive and reproducible procedure using gas chromatography coupled with mass spectrometry is described for the determination of p-tert-octylphenol (OP), a persistent degradation product of alkylphenol ethoxylates that binds to the estrogen receptor in blood and tissues. The first step involved the extraction of blood (200 microL) or tissue homogenate (400 microL) with methyl tert-butyl ether, including p-tert-butylphenol (BP) as internal standard. After extraction, the sample was evaporated to dryness with a gentle stream of nitrogen at 45 degrees C, and OP and BP were derivatized with an acetylation reaction involving acetic anhydride and catalyzed by pyridine. Samples were then analyzed by a gas chromatograph equipped with a mass spectrometer (single ion monitoring) with a Varian VF-5ms capillary column. The limit of detection and the limit of quantification of the method in blood were 4.6 and 15.5 ng/mL, respectively. The linearity and reproducibility of the method were acceptable, with coefficients of variation of approximately 10% for blood and ranging between 9% and 27% for tissues. This method was applied to the determination of unchanged OP in blood and tissues obtained from Sprague-Dawley rats after oral and IV OP administration.

Physiologically based modeling of the inhalation pharmacokinetics of ethylbenzene in B6C3F1 mice.

A physiologically based pharmacokinetic (PBPK) model was developed for inhaled ethylbenzene (EB) in B6C3F1 mice. The mouse physiological parameters were obtained from the literature, but the blood:air and tissue:air partition coefficients were determined by vial equilibration technique. The maximal velocity for hepatic metabolism (Vmax) obtained from a previously published rat study was increased by a factor of approximately 3 to account for enzyme induction during repeated exposures. The Michaelis affinity constant (Km) for hepatic metabolism of EB, obtained from a previously published rat PBPK modeling study, was kept unchanged during single and repeated exposure scenarios. Hepatic metabolism alone could not adequately describe the clearance of EB from mouse blood. Additional metabolism was assumed to be localized in the lung. The parameters for pulmonary metabolism were obtained by optimization of PBPK model fits to kinetic data collected following exposures to 75-1000 ppm. The PBPK model successfully predicted all available blood and tissue concentration data in mice exposed to 75 or 750 ppm EB. Overall, the results indicate that the rate of EB clearance is markedly higher in B6C3F1 mice than rats or humans and exceeds the hepatic metabolism capacity. Available biochemical evidence is consistent with a significant role for pulmonary metabolism; however, the extent to which the extrahepatic metabolism is localized in the lung is unclear. Overall, the PBPK model developed for the mouse adequately simulated the blood and tissue kinetics of EB by accounting for its high rate of clearance.

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.

Influence of oral administration of a quaternary mixture of trihalomethanes on their blood kinetics in the rat.

Trihalomethanes (THMs; chloroform, bromoform, bromodichloromethane, dibromochloromethane), formed as by-products of chlorine disinfection, are found to occur in combination in drinking water supplies. THMs are metabolized by cytochromes P-450 and are likely substrates of CYP2E1. Therefore, it is possible that mixed exposure results in toxicokinetic interactions among THMs. The toxicokinetics of THMs during mixed exposures has not been investigated previously. The purpose of this study was to characterize the blood kinetics of the four THMs administered singly or in combination in the rat. A single dose of 0.25 mmol/kg or 0.5 mmol/kg b.w., of each THM alone, or of a quaternary mixture containing 0.25 mmol/kg of each THM (total dose of 1.0 mmol/kg) was administered by gavage. The venous blood concentrations of the THMs were measured by headspace gas chromatography (GC) at 20, 40, 60, 120, 180, 270 and 360 min post-administration. Results showed a nonlinear relationship between the area under the blood concentration versus time curves (AUCs) and administered doses of THMs, suggesting that metabolism is saturated in this dose range. The venous blood concentrations of THMs following administration of the quaternary mixture were significantly higher compared to single exposures. The altered kinetics of THMs during combined exposures is consistent with the occurrence of mutual inhibition of their hepatic metabolism. Simulation exercises conducted with physiologically based toxicokinetic models support metabolic inhibition as the possible mechanism of the interaction among THMs. The data reported in this study provide the starting point for evaluating the significance of interactions among THMs in the risk assessment process.

Physiologically based modeling of the maximal effect of metabolic interactions on the kinetics of components of complex chemical mixtures.

The objective of this study was to predict and validate the theoretically possible, maximal impact of metabolic interactions on the blood concentration profile of each component in mixtures of volatile organic chemicals (VOCs) [dichloromethane (DCM), benzene (BEN), trichloroethylene (TCE), toluene (TOL), tetrachloroethylene (PER), ethylbenzene (EBZ), styrene (STY), as well as para, ortho-, and meta-xylene (p-XYL, o-XYL, m-XYL)] in the rat. The methodology consisted of: (1) obtaining the validated, physiologically based toxicokinetic (PBTK) model for each of the mixture components from the literature, (2) substituting the Michaelis-Menten description of metabolism with an equation based on the hepatic extraction ratio (E) for simulating the maximal impact of metabolic interactions (i.e., by setting E to 0 or 1 for simulating maximal inhibition or induction, respectively), and (3) validating the PBTK model simulations by comparing the predicted boundaries of venous blood concentrations with the experimental data obtained following exposure to various mixtures of VOCs. All experimental venous blood concentration data for 9 of the 10 chemicals investigated in the present study (PER excepted) fell within the boundaries of the maximal impact of metabolic inhibition and induction predicted by the PBTK model. The modeling approach validated in this study represents a potentially useful tool for screening/identifying the chemicals for which metabolic interactions are likely to be important in the context of mixed exposures and mixture risk assessment.

Evaluation of the pharmacokinetic interactions between orally administered trihalomethanes in the rat.

The blood kinetics of trihalomethanes has recently been reported to differ between an oral administration of any single trihalomethane (0.25 mmol/kg) [THMs: chloroform, bromoform, bromodichloromethane (BDCM), dibromochloromethane (DBCM)] and a combined administration of 0.25 mmol/kg of each of the 4 THMs. The significant increase in blood concentrations of THMs could be a consequence of pharmacokinetic interactions between two or more of the THMs present simultaneously. The objective of the present study was to characterize the blood kinetics of THMs following oral administration singly or as binary mixtures in order to assess the relative contribution of each THM to the kinetic interferences observed with the quaternary mixture. A single dose of each THM (0.5 mmol/kg) alone or of a binary mixture containing 0.5 mmol/kg of each THM was administered by gavage to male Sprague-Dawley rats. The venous blood concentrations of unchanged THMs were measured for up to 720 min postadministration by headspace gas chromatography. Results showed that, compared to single administration, each binary mixture caused a significant increase in the blood concentrations of both THMs present and this effect increased with time. The impact, however, was not similar for each mixture, especially during the first hour following administration of the compounds (bromoform and DBCM). Among the four THMs, bromoform and DBCM kinetics appeared to be more sensitive to the mixture effect and to exert the greatest impact on the kinetics of the second THM present in the mixture. Simulation exercises conducted with physiologically based toxicokinetic models suggest metabolic inhibition as the possible mechanism of the interaction between THMs. In conclusion, the results of this study show that, at the dose level investigated, every binary combination of THMs, when orally administered, resulted in a significant modulation of their pharmacokinetics and suggest that this is probably the consequence of a mutual metabolic inhibition between the THMs.

Evaluation of the health risk associated with exposure to chloroform in indoor swimming pools.

The exposure of swimmers to chloroform (CHCl3) was investigated in indoor swimming pools of the Quebec City region along with the associated carcinogenic risk. Six training sessions involving 52 competition swimmers (11 to 20 yr old) were conducted in 3 different pools, while 12 adult leisure swimmers attended 5 sessions, each held in a different pool. For each session, water and ambient air CHCl3 concentrations were measured and CHCl3 levels in alveolar air samples (CHCl3 ALV) collected from swimmers prior to entering the swimming pool premises and after 15, 35, and 60 min of swimming. Mean water concentrations varied from 18 microg/L to 80 microg/L, while those in air ranged from 78 microg/m3 to 329 microg/m3. Multiple linear regression analyses revealed that CHCl3 ALV values in competition swimmers were strongly correlated to ambient air and water levels, and to a lesser degree to the intensity of training. Only ambient air concentration was positively correlated to CHCl3 ALV in the leisure group. Concentrations of CHCl3 metabolites bound to hepatic and renal macromolecules, estimated using a physiologically based pharmacokinetic (PBPK) model, were 1.6 and 1.9 times higher for the competition swimmers than for the leisure swimmers, respectively. The highest hepatic concentration predicted in competition swimmers, 0.22 microg CHCl3 equivalents/kg of tissue, was at least 10,000 times lower than the smallest no observed effect level for liver tumors in animals. Data indicate that the safety margin is therefore very large, for competitive swimmers as well as for leisure swimmers.

Blood:air partition coefficients of individual and mixtures of trihalomethanes.

The objective of the present study was to determine the rat blood:air partition coefficients (P(b:a)) of chloroform, bromodichloromethane, dibromochloromethane and bromoform present in vitro individually or as mixtures. The experimentally determined P(b:a) of chloroform, bromodichloromethane, dibromochloromethane and bromoform present individually corresponded to (mean +/- SD, n = 8) 21.3 +/- 1.8, 41.8 +/- 6.2, 97.5 +/- 4.1, and 187 +/- 7.4, respectively. The P(b:a) of these trihalomethanes (THMs) showed a decreasing trend during mixed in vitro exposures to 0.138 +/- 0.002 or 0.273 +/- 0.002 micromol of each of the four THMs. In general, the P(b:a) determined during mixed exposures differed by < or = 15% of the average P(b:a) determined for THMs present individually. The results of this study suggest that an alteration of P(b:a) of the individual THMs is unlikely to occur at the blood concentrations of THMs observed during mixed exposures in rats.

Inhalation pharmacokinetics of ethylbenzene in B6C3F1 mice.

The objective of the present study was to characterize the inhalation pharmacokinetics of ethylbenzene (EB) in male and female B6C3F1 mice following single and repeated exposures. Initially, groups of 28 male and female mice were exposed for 4 h to 75, 200, 500, or 1000 ppm in order to determine potential non-linearity in the kinetics of EB. Then, groups of male and female mice were exposed for 6 h to 75 ppm and 750 ppm (corresponding to the NTP exposures) for 1 or 7 consecutive days, to evaluate whether EB kinetics was altered during repeated exposures, The maximal blood concentration (Cmax; mean+/-SD, n=4) observed in female mice at the end of a 4-h exposure to 75, 200, 500, and 1000 ppm was 0.53+/-0.18, 2.26+/-0.38, 19.17+/-2.74, and 82.36+/-16.66 mg/L, respectively. The areas under the concentration vs. time curve (AUCs) following 4-h exposure to 75, 200, 500, and 1000 ppm were 88.5, 414.0, 3612.2, and 19,104.1 mg/L/min, respectively, in female mice, and 116.7, 425.7, 3148.3, and 16,039.1 mg/L/min in male mice. The comparison of Cmax and the kinetic profile of EB in mice exposed to 75 ppm suggests that they are similar between 1-day and 7-day exposures. However, at 750 ppm, the rate of EB elimination would appear to be greater after repeated exposures than single exposure, the pattern being evident in both male and female mice. Overall, the single and repeated exposure pharmacokinetic data collected in the present study suggest that EB kinetics is saturable at exposure concentrations exceeding 500 ppm (and therefore at 750 ppm used in the NTP mouse cancer bioassay) but is in the linear range at the lower concentration used in the bioassay (75 ppm). These data suggest that consideration of the nature and magnitude of non-linear kinetics and induction of metabolism during repeated exposures is essential for the conduct of a scientifically sound analysis of EB cancer dose-response data collected in B6C3F1 mice.

Physiologically based modeling of n-hexane kinetics in humans following inhalation exposure at rest and under physical exertion: impact on free 2,5-hexanedione in urine and on n-hexane in alveolar air.

We used a modified physiologically based pharmacokinetic (PBPK) to describe/predict n-hexane (HEX) alveolar air concentrations and free 2,5-HD urinary concentrations in humans exposed to n-HEX by inhalation during a typical workweek. The effect of an increase in workload intensity on these two exposure indicators was assessed and, using Monte Carlo simulation, the impact of biological variability was investigated. The model predicted HEX alveolar air concentrations at rest of 19.0 ppm (25 ppm exposure) and 38.7 ppm (50 ppm exposure) at the end of the last working day (day 5), while free 2,5-HD urinary concentrations of 3.4 micromol/L (25 ppm) and 6.3 micromol/L (50 ppm) were predicted for the same period (last 4.5 hours of Day 5). Monte Carlo simulations showed that the range of values expected to occur in a group of 1000 individuals exposed to 50 ppm of HEX (95% confidence interval) for free 2,5-HD (1.7-14.7 micromol/L) is much higher compared with alveolar air HEX (33.4-46 ppm). Simulations of exposure at 50 ppm with different workloads predicted that an increase in workload intensity would not greatly affect both indicators studied. However, the alveolar air HEX concentration is more sensitive to modifications of workload intensity and time of sampling, after the end of exposure, compared with 2,5-HD. The PBPK model successfully described the HEX alveolar air concentrations and free 2,5-HD urinary concentrations measured in human volunteers and is the first, to our knowledge, to describe the excretion kinetics of free 2,5-HD in humans over a 5-day period.

Validation of a physiological modeling framework for simulating the toxicokinetics of chemicals in mixtures.

The objective of this study was to investigate the usefulness of a physiologically based toxicokinetic (PBTK) modeling framework for simulating the kinetics of chemicals in mixtures of varying complexities and composition. The approach involved the simulation of the kinetics of components in two situations: (i) when one of the mixture components was substituted with another (i.e., benzene in the benzene (B)-toluene (T)-ethyl benzene (E)-m-xylene (X) mixture was substituted with dichloromethane (D)), and (ii) when another chemical was added to the existing four-chemical mixture model (i.e., when D was added to the existing BTEX mixture model). In both cases, differing compositions of mixtures were used to obtain simulations and to generate experimental data on kinetics for validation purposes. Since the quantitative and qualitative mechanisms of interaction among B, T, E, and X have already been established, the mechanisms of binary interactions between D and the BTEX components (e.g., competitive, noncompetitive, or uncompetitive metabolic inhibition) were investigated in the present study. The analysis of rat blood kinetic data (4-h inhalation exposures, 50-200 ppm each) to all binary combinations (D-B, D-T, D-E, and D-X) investigated in the present study was suggestive of competitive metabolic inhibition as the plausible interaction mechanism. By incorporating the newly estimated values of metabolic inhibition constant (K(i)) for each of these binary combinations within the five-chemical PBTK model (i.e., for the DBTEX mixture), the model adequately predicted the venous blood kinetics of chemicals in rats following a 4-h inhalation exposure to various mixtures (mixture 1:100 ppm of D and 50 ppm each of T, E, and X; mixture 2: 100 ppm each of D, T, E, and X; mixture 3: 100 ppm of D and 50 ppm each of B, T, E, and X; mixture 4: 100 ppm each of D, B, T, E, and X). The results of the present study suggest that the PBTK model framework is useful for conducting extrapolations of the kinetics of chemicals from one mixture to another differing in complexity and composition, based on mechanistic considerations of interactions elucidated at the binary level.

Physiologically based pharmacokinetic modeling of a ternary mixture of alkyl benzenes in rats and humans.

The objective of the present study was to develop a physiologically based pharmacokinetic (PBPK) model for a ternary mixture of alkyl benzenes [toluene (TOL), m-xylene (XYL), and ethylbenzene (EBZ)] in rats and humans. The approach involved the development of the mixture PBPK model in the rat and extrapolation to humans by substituting rat physiological parameters and blood:air partition coefficients in the model with those of humans, scaling maximal velocity for metabolism on the basis of body weight0.75 and keeping all other model parameters species-invariant. The development of the PBPK model for the ternary mixture in the rat was accomplished by initially validating or refining the existing PBPK models for TOL, XYL, and EBZ and linking the individual chemical models via the hepatic metabolism term. Accordingly, the Michaelis-Menten equation for each solvent was modified to test four possible mechanisms of metabolic interaction (i.e., no interaction, competitive inhibition, noncompetitive inhibition, and uncompetitive inhibition). The metabolic inhibition constant (Ki) for each binary pair of alkyl benzenes was estimated by fitting the binary chemical PBPK model simulations to previously published data on blood concentrations of TOL, XYL, and EBZ in rats exposed for 4 hr to a binary combination of 100 or 200 ppm of each of these solvents. Competitive metabolic inhibition appeared to be the most plausible mechanism of interaction at relevant exposure concentrations for all binary mixtures of alkyl benzenes in the rat (Ki,TOL-XYL = 0.17; Ki,TOL-EBZ = 0.79; Ki,XYL-TOL = 0.77; Ki,XYL-EBZ = 1.50; Ki,EBZ-TOL = 0.33; Ki,EBZ-XYL = 0.23 mg/L). Incorporating the Ki values obtained with the binary chemical mixtures, the PBPK model for the ternary mixture simulated adequately the time course of the venous blood concentrations of TOL, XYL, and EBZ in rats exposed to a mixture containing 100 ppm each of these solvents. Following the validation of the ternary mixture model in the rat, it was scaled to predict the kinetics of TOL, XYL, and EBZ in blood and alveolar air of human volunteers exposed for 7 hr to a combination of 17, 33, and 33 ppm, respectively, of these solvents. Model simulations and experimental data obtained in humans indicated that exposure to atmospheric concentrations of TOL, XYL, and EBZ that remain within the permissible concentrations for a mixture would not result in biologically significant modifications of their pharmacokinetics. Overall, this study demonstrates the utility of PBPK models in the prediction of the kinetics of components of chemical mixtures, by accounting for mechanisms of binary chemical interactions.

Comparison of the influence of binary mixtures versus a ternary mixture of inhaled aromatic hydrocarbons on their blood kinetics in the rat.

The objective of the present study was to compare the influence of various binary mixtures containing ethylbenzene (EBZ), toluene (TOL) or xylene (XYL) administered by inhalation, with the influence exerted by a ternary mixture, on the kinetics of these solvents in blood. Groups of four rats were exposed for 4 h to TOL, XYL and EBZ, singly or in combination. The concentration of TOL, XYL and EBZ in blood was measured at various times (5, 30, 60, 90 and 120 min) following the end of exposure and the areas under the blood concentration curves (AUC) were calculated. Results showed that exposures to binary and ternary mixtures resulted in significantly higher (P < 0.05) blood concentrations of unchanged solvents as a result of metabolic interaction between these solvents. When the comparison was based on individual solvents, there was no difference between effect exerted by the ternary mixture and the binary mixtures, except for one. However, a comparison based on the total concentration of unchanged solvents disclosed that exposure to the ternary mixture resulted in greater interactive effects (3.17-fold increase) than exposures to binary mixtures (1.97-fold increase), whereas four out of six binary mixtures produced higher total levels of unchanged solvents in blood compared to the ternary mixture. This study shows that the greater risk of toxicity often thought to be associated with exposures to complex mixtures should not only be related to the magnitude of interactive effects among components (i.e., degree of mutual metabolic interaction) resulting from combined exposures, but also should take into account, as is universally recognized, the internal total dose of toxic chemicals in target organs/tissues.

Physiological modeling of the toxicokinetic interactions in a quaternary mixture of aromatic hydrocarbons.

The available data on binary interactions are yet to be considered within the context of mixture risk assessments because of our inability to predict the effect of a third or fourth chemical in the mixture on the interacting binary pairs. Physiologically based toxicokinetic (PBTK) models represent a framework that can be potentially used for predicting the impact of multiple interactions on component kinetics at any level of complexity. The objective of this study was to develop and validate an interaction-based PBTK model for simulating the toxicokinetics of the components of a quaternary mixture of aromatic hydrocarbons [benzene (B), toluene (T), ethylbenzene (E), m-xylene (X)] in the rat. The methodology consisted of: (1) obtaining and refining the validated individual chemical PBTK models from the literature, (2) interconnecting all individual chemical PBTK models at the level of liver on the basis of the mechanism of binary chemical interactions (e.g., competitive, noncompetitive, or uncompetitive metabolic inhibition), and (3) comparing the a priori predictions of the interaction-based model to corresponding experimental data on venous blood concentrations of B, T, E, and X during mixture exposures. The analysis of blood kinetics data from inhalation exposures (4 h, 50-200 ppm each) of rats to all binary combinations of B, T, E, and X was suggestive of competitive metabolic inhibition as the plausible interaction mechanism. The metabolic inhibition constant (K(i)) for each binary combination was quantified and incorporated within the mixture PBTK model. The binary interaction-based PBTK model predicted adequately the inhalation toxicokinetics of all four components in rats following exposure to mixtures of BTEX (50 ppm each of B, T, E, and X, 4 h; 100 ppm each of B, T, E and X, 4 h; 100 ppm B + 50 ppm each of T, E, and X, 4 h). The results of the present study suggest that data on interactions at the binary level alone are required and sufficient for predicting the kinetics of components in complex mixtures.

Physiologically-based pharmacokinetic modeling of a mixture of toluene and xylene in humans.

A physiologically-based pharmacokinetic (PBPK) model for a mixture of toluene (TOL) and xylene (XYL), developed and validated in the rat, was used to predict the uptake and disposition kinetics of TOL/XYL mixture in humans. This was accomplished by substituting the rat physiological parameters and the blood:air partition coefficient with those of humans, scaling the maximal velocity for hepatic metabolism on the basis of body weight0.75, and keeping all other model parameters species-invariant. The human TOL/XYL mixture PBPK model, developed based on the quantitative biochemical mechanism of interaction elucidated in the rat (i.e., competitive metabolic inhibition), simulated adequately the kinetics of TOL and XYL during combined exposures in humans. The simulations with this PBPK model indicate that an eight hour co-exposure to concentrations that remain within the current threshold limit values of TOL (50 ppm) and XYL (100 ppm) would not result in significant pharmacokinetic interferences, thus implying that data on biological monitoring of worker exposure to these solvents would be unaffected during co-exposures.