Crispin H. Pierce

Toluene metabolites as biological indicators of exposure

The measurement of exhaled and excreted xenobiotics and their metabolites can provide accurate, non-invasive, and time-flexible measurements of internal dose. We analyzed rates of exhaled (2)H(8)-toluene and excreted urinary metabolites from 33 exposures of men to 50 ppm of (2)H(8)-toluene for 2 h at rest. The total dose was distributed as follows: exhaled (2)H(8)-toluene, 13 +/- 6.2%; (2)H(5)-hippuric acid, 75 +/- 6.4%; (2)H(7)-o-cresol, 0.31 +/- 0.22%; (2)H(7)-m-cresol, 0.53 +/- 0.44%; and (2)H(7)-p-cresol, 11 +/- 3.8%. Interindividual variability was assessed using the coefficients of variation for peak exhalation or excretion rates, and fractions of dose excreted: (2)H(8)-toluene, c.v.=60, 47%; (2)H(5)-hippuric acid, 29, 8.6%; (2)H(7)-o-cresol, 80, 73%; (2)H(7)-m-cresol, 37, 83%; and (2)H(7)-p-cresol, 38, 34%. Excretion rates of the cresols were stable over the first 5 h post-exposure, and o-cresol was determined to be the best urinary indicator of exposure, given the lower background levels of this isomer. The hippuric acid/cresol rate ratios for the first 5 h post-exposure could be described by single exponential terms, and thus provided a means for estimating time since exposure for any finite toluene duration/exposure combination.

Modeling the acute neurotoxicity of styrene

Styrene is a widely used industrial solvent associated with acute neurotoxicity. To investigate the relationships between exposure, blood concentrations, and the appearance of neurotoxic effects, four healthy males were exposed to styrene concentrations of 5-200 ppm in four different exposure-time profiles. A digit recognition test and P300 event-related evoked potential were used to measure neurologic function. A physiologically based kinetic (PBK) model generated close predictions of measured styrene blood concentrations, in the range of 0.01-12 mg/L, from this and 21 previous studies. Simulated peak brain concentration, durationXaverage exposure, and peak exposure level were predictive of toxicity. Central nervous system effects were expected at a blood concentration near 2.4 mg/L. A standard of 20 ppm was expected to protect styrene-exposed workers from acute central nervous system toxicity under light work conditions.

Exponential Modeling, Washout Curve Reconstruction, and Estimation of Half-Life of Toluene and its Metabolites

Health risks from ostensible occupational and environmental toxicant exposure are difficult to quantify. Maximal use of limited biological measurements of xenobiotic or metabolite concentration in the body is therefore essential. Elimination rates of exhaled [2H8]toluene and urinary metabolites were analyzed from 33 exposures of males to 50 ppm [2H8]toluene for 2 h at rest. It was hypothesized that the shapes from our decay curves would be applicable to any occupational or environmental toluene exposure. Except for a rapid decline in toluene blood and breath levels in the 0–0.1 h period, this “curve reconstruction” method successfully fit data from published studies. Urinary hippuric acid concentrations were not well fit due to substantial background levels, whereas o-cresol levels were accurately described. Our approach was able to reconstruct data from studies where exposure duration ranged from 10 min to 7 h, and where activity level ranged from rest to 150 W (strenuous exercise). Using this approach, limited biological data following toluene exposure could be back-extrapolated to immediate postexposure concentrations, which in turn could be compared to biological indicators of exposure to determine risk.

A comparison of 1H8-and 2H8-toluene toxicokinetics in men

1. To examine the bioequivalence of an isotope-labelled tracer to study toxicant disposition, we conducted 33 controlled human exposures to a mixture of 50 ppm 1H8-toluene and 50 ppm 2H8-toluene for 2 h, and measured concentrations in blood and breath, and metabolite levels in urine for 100 h post-exposure. 2. A physiologically based kinetic (PBK) model found that compared with 1H8-toluene, 2H8-toluene had a 6.4+/-13% (mean+/-SD) lower AUC, a 6.5+/-13% higher systemic clearance (1.46+/-0.27 versus 1.38+/-0.25 l/h-kg), a 17+/-22% larger terminal volume of distribution (66.4+/-14 versus 57.2+/-10 l/kg) and a 9.7+/-26% longer terminal half-life (38+/-12 versus 34+/-10 h) (p < 0.05 for all comparisons). 3. The higher 2H8-toluene clearance may have been due to an increased rate of ring oxidation, consistent with the 17% higher observed fraction of 2H5- versus 1H5-cresol metabolites in urine. 4. The larger terminal volume and half-lives for 2H8-toluene suggested a higher adipose tissue/blood partition coefficient. 5. Observed isotope differences were small compared with interindividual differences in 1H8-toluene kinetics from previous studies. 6. The PBK model allowed us to ascribe observed isotope differences in solvent toxicokinetics to underlying physiologic mechanisms.

Estimation of Background Exposure to Toluene Using a Physiologically-Based Kinetic Model

Estimation of Background Exposure to Toluene Using a Physiologically‐Based Kinetic Model: Crispin H. Pierce, et al. Department of Environmental Health, University of Washington—Estimation of environmental exposure to toxicants has generally been limited to concentration measurements in air, water, and foods. Measurement of background levels of toxicants in biological tissues for this purpose has been limited by analytical detection. After developing a sensitive headspace GC‐MS method, we conducted 33 controlled human exposures of 50 ppm 1Hg8‐toluene and 50 ppm 2H8‐toluene for 2 h, and measured concentrations in blood and breath for 100 h post‐exposure. Blood and breath samples from a separate cohort of 9 men exposed to 2H8‐toluene only were also measured for background 1H ‐toluene levels. A physiologically‐based kinetic (PBK) model, previously constructed and validated in an analysis of the 2H8‐toluene data, was used to predict the level of ambient 1H8‐toluene exposure that produced the observed breath levels. The model‐derived estimate of mean background 1H8‐toluene exposure was 47 ± 44 ppb (mean ± s.d.), which is consistent with indoor air measurements from this and previous studies of 3‐27 ppb and outdoor measurements of 2‐43 ppb. According to the PBK model, background exposure was expected to produce an average blood concentration of 5.9 nmol/l, which was within a measured range of 3‐16 nmol/l, and a corresponding alveolar air concentration of 310 nmol/m3, within a range of 138‐764 nmol/m3. This work extends the use of physiologic modeling to back‐predict environmental dose, and found that significant differences in inter‐individual 1H8‐toluene background exposures exist.

Individual prior information in a physiological model of 2H8-toluene kinetics

Physiologically-based toxicokinetic (PBTK) models are widely used to quantify whole-body kinetics of various substances. However, since they attempt to reproduce anatomical structures and physiological events, they have a high number of parameters. Their identification from kinetic data alone is often impossible, and other information about the parameters is needed to render the model identifiable. The most commonly used approach consists of independently measuring, or taking from literature sources, some of the parameters, fixing them in the kinetic model, and then performing model identification on a reduced number of less certain parameters. This results in a substantial reduction of the degrees of freedom of the model. In this study, we show that this method results in final estimates of the free parameters whose precision is overestimated. We then compared this approach with an empirical Bayes approach, which takes into account not only the mean value, but also the error associated with the independently determined parameters. Blood and breath 2H8-toluene washout curves, obtained in 17 subjects, were analyzed with a previously presented PBTK model suitable for person-specific dosimetry. Model parameters with the greatest effect on predicted levels were alveolar ventilation rate QPC, fat tissue fraction VFC, blood-air partition coefficient Kb, fraction of cardiac output to fat Qa/co and rate of extrahepatic metabolism Vmax-p. Differences in the measured and Bayesian-fitted values of QPC, VFC and Kb were significant (p < 0.05), and the precision of the fitted values Vmax-p and Qa/co went from 11 ± 5% to 75 ± 170% (NS) and from 8 ± 2% to 9 ± 2% (p < 0.05) respectively. The empirical Bayes approach did not result in less reliable parameter estimates: rather, it pointed out that the precision of parameter estimates can be overly optimistic when other parameters in the model, either directly measured or taken from literature sources, are treated as known without error. In conclusion, an empirical Bayes approach to parameter estimation resulted in a better model fit, different final parameter estimates, and more realistic parameter precisions.