A toxicokinetic (TK) model was developed to describe the inhalation exposure in humans to methyl formate (MF), a catalyst used in foundries, and to discuss biological monitoring. The TK model consisted of four compartments: MF, the metabolites – methanol (MeOH) and formic acid (FA) – and, in addition, a urinary compartment describing the saturable reabsorption of FA. Levels of MeOH and FA in urine, from an experimental study (100 ppm MF, 8 h at rest), validated the present model. The TK model describes well the general behaviour of MeOH and FA in urine after MF exposure. A nonlinear and a linear relationship respectively, was predicted between MF exposure and FA or MeOH excretion in urine, and this has previously been seen after occupational MF exposure. The present model has been modified to simulate MeOH exposure as well. Generally low exposures (concentration or exercise) produce only marginal increases in FA urinary excretions, but when exposure is elevated, urinary FA excretion increases because of saturation in the mechanism of reabsorption. Using FA urinary excretion as the critical indicator, because of its link to health effects, an occupational exposure limit value for MF of no greater than 50 ppm should be selected (based on predictions with the TK model). MeOH in urine can be considered as a biomarker for MF at low exposure, because of lower background values and of a linear relationship with exposure. At higher exposures, however, FA could be used as a biomarker as it becomes progressively more sensitive. But the use of biological monitoring for MF is difficult because of individual variations in background values. Under the present state of knowledge both FA and MeOH should be used to estimate only group exposures, rather than individual exposures.

A physiologically based toxicokinetic (PBTK) model was developed for evaluation of inhalation exposure in humans to the gasoline additive, ethyl tertiary-butyl ether (ETBE). PBTK models are useful tools to relate external exposure to internal doses and biological markers of exposure in humans. To describe the kinetics of ETBE, the following compartments were used: lungs (including arterial blood), liver, fat, rapidly perfused tissues, resting muscles, and working muscles. The same set of compartments and, in addition, a urinary excretion compartment were used for the metabolite tertiary-butyl alcohol (TBA). First order metabolism was assumed in the model, since linear kinetics has been shown experimentally in humans after inhalation exposure up to 50 ppm ETBE. Organ volumes and blood flows were calculated from individual body composition based on published equations, and tissue/blood partition coefficients were calculated from liquid/air partition coefficients and tissue composition. Estimates of individual metabolite parameters of 8 subjects were obtained by fitting the PBTK model to experimental data from humans (5, 25, 50 ppm ETBE, 2-h exposure; Nihlén et al., Toxicol. Sci., 1998; 46, 1-10). The PBTK model was then used to predict levels of the biomarkers ETBE and TBA in blood, urine, and exhaled air after various scenarios, such as prolonged exposure, fluctuating exposure, and exposure during physical activity. In addition, the interindividual variability in biomarker levels was predicted, in the eight experimentally exposed subjects after a working week. According to the model, raising the work load from rest to heavy exercise increases all biomarker levels by approximately 2-fold at the end of the work shift, and by 3-fold the next morning. A small accumulation of all biomarkers was seen during one week of simulated exposure. Further predictions suggested that the interindividual variability in biomarker levels would be higher the next morning than at the end of the work shift, and higher for TBA than for ETBE. Monte Carlo simulations were used to describe fluctuating exposure scenarios. These simulations suggest that ETBE levels in blood and exhaled air at the end of the working day are highly sensitive to exposure fluctuations, whereas ETBE levels the next morning and TBA in urine and blood are less sensitive. Considering these simulations, data from the previous toxicokinetic study and practical issues, we suggest that TBA in urine is a suitable biomarker for exposure to ETBE and gasoline vapor.

Ethers or other oxygen-containing compounds are used as replacements for lead in gasoline and to ensure complete combustion. Methyl tertiary-butyl ether (MTBE) is already in use world-wide and ethyl tertiary-butyl ether (ETBE) may be used increasingly in the future. The aims of the present thesis were to study the uptake and disposition (toxicokinetics) of MTBE and ETBE in humans, to address the issue of biological monitoring and to measure acute effects (assessed by questionnaire, lung function, nasal and ocular measurements). Healthy male volunteers were experimentally exposed during 2 h to vaporized MTBE (5, 25 and 50 ppm, n=10) or ETBE (0, 5, 25 and 50 ppm, n=8) in a chamber (whole-body) and 13C-labeled MTBE (50 ppm, n=4) via a face-mask. The toxicokinetics of MTBE and ETBE in humans were quite similar, judging from breath, blood and urine profiles. However, the respiratory uptake of MTBE was higher (45% MTBE and 33% ETBE) and respiratory exhalation of unmetabolized MTBE was lower (38% MTBE and 47% ETBE) compared to ETBE. Linear kinetics was seen for the ethers and TBA up to 50 ppm. Low urinary excretion of unmetabolized ether (tertiary-butyl alcohol (TBA) (13C-labeled MTBE exposure two urinary metabolites, 2-methyl-1,2-propanediol and (x-hydroxyisobutyric acid, were characterized by 13C-NMR. These metabolites were excreted in markedly higher amounts than TBA. A physiologically based toxicokinetic model was developed for ETBE using the determined partition coefficients and data from the chamber exposure. The model indicates that workload has a profound influence on internal dose. TBA excretion seemed to be less sensitive to fluctuations in exposure levels compared to the ether, especially in samples taken at the end of a workshift. Only a small accumulation of ether and TBA in the body was seen throughout a week of repeated exposure. MTBE vapor had very small or no effects up to 50 ppm. After ETBE exposure, elevated ratings of irritation in the throat and airways (50 ppm), and, in addition, minor pulmonary function impairments were seen (25 and 50 ppm ETBE, lung measures not performed for MTBE). This effect as such is not of clinical concern in healthy individuals. However, it cannot be excluded that other subjects may show more severe reactions. In conclusion, the toxicokinetic results as well as considering practical issues (i.e. sampling and analysis), implies that the concentration of TBA in urine is a good biological exposure marker for ether and gasoline vapor. However, further studies of other metabolites that are excreted in higher amounts in the urine (e.g., [alpha]-hydroxyisobutyric acid), may identify even more suitable candidates as biomarkers. The knowledge about the toxicokinetics and acute effects of MTBE and ETBE in humans presented in this thesis is relevant in e.g., risk assessments.

After exposure to methyl tert-butyl ether (MTBE), a gasoline additive, only one metabolite [tert-butyl alcohol (TBA), <1% of dose] has been identified in human urine [Nihlén, A., et al. (1998) Toxicol. Appl. Pharmacol. 148, 274-280]. In the study presented here, metabolites of MTBE were characterized by (1)H-decoupled (13)C NMR spectroscopy in urine obtained from four volunteers experimentally exposed to 50 ppm (13)C-labeled MTBE ([1,2-(13)C(2)]MTBE) vapor (facemask) for 2 h during a light physical work load (50 W). Chemical shifts for the two adjacent (13)C-labeled carbons in [1, 2-(13)C(2)]MTBE-derived metabolites were consistent with the shifts obtained for spiked standards of alpha-hydroxyisobutyric acid (HBA) and 2-methyl-1,2-propanediol (MPD). NMR signals were not detected for labeled MTBE, TBA, or possible MTBE-derived conjugates. Quantification of HBA and MPD was performed by NMR for two urine samples (collected 20 h after exposure). One subject had 11% HBA and 1% MPD, and the other individual had 3% HBA and 1% MPD in the urine, expressed as a percentage of MTBE uptake. This indicates that HBA and MPD occur at significantly higher levels in the urine (detected by NMR) than MTBE and TBA (detected by GC). To our knowledge, this is the first characterization of MTBE metabolites, other than TBA, in humans. Further urine, blood, and expired air were collected up to 22 h after exposure, and the toxicokinetics of MTBE, TBA, and acetone were determined by GC. Low relative uptake (39%), a low level of postexposure exhalation of MTBE (17%), and low recovery of TBA in the urine (<1%) were observed. The same subjects had previously been exposed to unlabeled MTBE in a whole-body exposure study [Nihlén, A., et al. (1998) Toxicol. Appl. Pharmacol. 148, 274-280], and the toxicokinetics of MTBE and TBA in this facemask exposure did not differ from the previous whole-body chamber exposure.

Methyltertiary-butyl ether (MTBE) is widely used in gasoline as an oxygenate and octane enhancer. The aim of this study was to evaluate the uptake, distribution, metabolism, and elimination of MTBE in humans. Ten healthy male volunteers were exposed to MTBE vapor (5, 25, and 50 ppm) on three different occasions during 2 h of light physical exercise (50 W). MTBE and the metabolitetertiary-butyl alcohol (TBA) were monitored in exhaled air, blood, and urine. Blood and urine were collected at selected time intervals, during and up to 3 days after the exposure, and analyzed by head space gas chromatography. MTBE in exhaled air was collected with sorbent sample tubes and subsequently analyzed by gas chromatography. The respiratory uptake of MTBE was rather low (42–49%), and the respiratory exhalation was high (32–47%). A relatively low metabolic blood clearance (0.34–0.52 L/h/kg) was seen compared to many other solvents. The kinetic profile of MTBE in blood could be described by four phases, and the average half-lives were 1 min, 10 min, 1.5 h, and 19 h. The post-exposure decay curve of MTBE in urine was separated into two linear phases, with average half-lives of 20 min and 3 h. The average post-exposure half-lives of TBA in blood and urine were 10 and 8.2 h, respectively. The urinary excretion of MTBE and TBA was less than 1% of the absorbed dose, indicating further metabolism of TBA, other routes of metabolism, or excretion. The kinetics of MTBE and TBA were linear up to the highest exposure level of 50 ppm. We suggest that TBA in blood or urine is a more appropriate biological exposure marker for MTBE than the parent ether itself.

The aim of this study was to evaluate acute effects of ethyl tert-butyl ether (ETBE) in man after short-term exposure. ETBE may in the future replace methyl tert-butyl ether, a widely used oxygenate in unleaded gasoline. Eight healthy male volunteers were exposed to ETBE vapor for 2 h at four levels (0, 5, 25, and 50 ppm) during light physical exercise. The subjects rated irritative symptoms, discomfort, and central nervous system effects in a questionnaire. Ocular (eye redness, tear film break-up time, conjunctival epithelial damage, and blinking frequency), nasal (acoustic rhinometry and analysis of inflammatory markers and cells in nasal lavage fluid), and pulmonary (peak expiratory flow, forced expiratory volume in 1 s, forced vital capacity, vital capacity, and transfer factor) measurements were performed. Significantly increased ratings of solvent smell (p = 0.001, repeated-measures ANOVA) were seen during exposures and correlated to exposure levels. Furthermore, significantly elevated ratings of discomfort in throat and airways were seen during and after 50 ppm compared to the control exposure (p = 0.02). Increased nasal swelling (p = 0.001) and blinking frequency (p = 0.01) were noted at all exposure levels, but their magnitudes were not related to exposure levels. A slightly impaired pulmonary function was seen at 25 and 50 ppm, since forced vital capacity (p = 0.02) and vital capacity (p = 0.04) differed significantly from the clean air exposure. Although the impairments seemed to fall within normal inter- and intraindividual variation and have no clinical relevance as such, it cannot be excluded that other individuals may react more severely than eight healthy male volunteers in this study.

Methyl tertiary-butyl ether (MTBE) is widely used in gasoline as an oxygenate and octane enhancer. Acute effects, such as headache, nausea, and nasal and ocular irritation, have been associated with the exposure to gasoline containing MTBE. The aim of this study was to assess acute health effects up to the Swedish occupational exposure limit value, both with objective methods and a questionnaire. Ten healthy male volunteers were exposed to MTBE vapor for 2 h at three levels (5, 25, and 50 ppm), during light physical work (50 W). All subjects rated the degree of irritative symptoms, discomfort, and CNS effects before, during, and after all three exposure occasions using a questionnaire. Answers were given on a 100-mm visual analog scale, graded from "not at all" to "almost unbearable." Ocular (redness, tear film break-up time, self-reported tear film break-up time, conjunctival epithelial damage, and blinking frequency) and nasal (mouth and nasal peak expiratory flow, acoustic rhinometry, biochemical inflammatory markers, and cells in nasal lavage) measurements were performed mainly at the highest exposure level. The ratings of solvent smell increased dramatically (ratings up to 50% of the scale) as the volunteers entered the chamber and declined slowly with time (p < 0.05, repeated-measures ANOVA). All other questions were rated from "not at all" to "hardly at all" (0-10% of the scale) with no significant relation to exposure. The eye measurements showed no effects of MTBE exposure. Blockage index, a measure of nasal airway resistance calculated from the peak expiratory flows, increased significantly after exposure; however, the effect was not related to exposure level. In addition, a nonsignificant tendency of decreased nasal volume was seen in the acoustic rhinometry measurements, but with no clear dose-effect relationship. In conclusion, our study suggests no or minimal acute effects of MTBE vapor upon short-term exposure at relatively high levels.

Partition coefficients are essential to a description of the uptake and distribution of volatile substances in humans and in the development of physiologically based pharmacokinetic models. Liquid/air partition coefficients (lambda) of three ethers, methyl t-butyl ether (MTBE), ethyl t-butyl ether (ETBE), and t-amyl methyl ether (TAME) were determined in vitro by head space-gas chromatography. These ethers, and especially MTBE, are used in unladed gasoline to enhance the oxygen and octane content, and to reduce the output of carbon monoxide during combustion. Partition coefficients of t-butyl alcohol (TBA), a metabolite of MTBE, were determined also. The liquids tested were fresh human blood, water (physiological saline), and olive oil. The lambda(blood/air) values were: 17.7 (95% confidence interval 17.0-18.4) for MTBE; 11.7 (11.3-12.1) for ETBE; and 17.9 (17.3-18.6) for TAME. Corresponding lambda(water/air) values were 15.2 (14.9-15.5), 8.39 (8.19-8.59), and 11.9 (11.7-12.1). The ethers have a higher affinity for oil, the values for lambda(oil/air) being 120 (114-125), 190 (183-197), and 337 (320-354), respectively. As expected, the lambda(blood/air) and lambda(water/air) for TBA were much higher than for the ethers, 462 (440-484) and 603 (590-617), respectively. The lambda(oil/air) was 168 (161-174) for TBA. The interindividual variability of the lambda(blood/air) (10 subjects) was calculated as the coefficient of variation, and estimated as: 14% for MTBE, 20% for ETBE, 20% for TAME, and 30% for TBA. No significant difference was seen in the lambda(blood/air) between the sexes.

Methyl tertiary butyl ether (MTBE) is widely used in gasoline as an oxygenator and octane enhancer. There is also an interest in using the ethyl tertiary butyl (ETBE) and methyl tertiary amyl (TAME) ethers. We measured the blood, water, and olive oil/air partition coefficients in vitro of MTBE, ETBE, TAME and tertiary butyl alcohol (TBA), a metabolite of MTBE and ETBE. The results indicate similar uptake and distribution behavior for the three ethers and a slight affinity for fatty tissues. The partition coefficients of TBA indicate that this metabolite is not excreted via the lungs to any great extent and that it is preferentially distributed in body water. Further, we exposed 10 healthy male volunteers to MTBE vapor at 5, 25 and 50 ppm for 2 h during light physical exercise. Uptake and disposition were studied by measuring MTBE and TBA in inhaled and exhaled air, blood and urine. Low uptake, high post-exposure exhalation, and low blood clearance indicate slow metabolism of MTBE relative to many other solvents. A low recovery of TBA in urine (below 1% of uptake) indicates further metabolism of TBA. The concentration of MTBE and TBA in blood was proportional to exposure level suggesting linear kinetics up to 50 ppm. The half life of 7–10 h in blood and urine indicates that TBA would be more suitable than the parent compound as a biomarker for MTBE exposure. Subjective ratings (discomfort, irritative symptoms, CNS effects) and eye (redness, tear film break-up time, conjunctival damage, blinking frequency) and nose (peak expiratory flow, acoustic rhinometry, inflammatory markers in nasal lavage) measurements indicated no or minimal effects of MTBE.