James D. Prah

Body Burden Measurements and Models to Assess Inhalation Exposure to Methyl Tertiary Butyl Ether (MTBE)

Biomarkers of methyl tertiary butyl either (MTBE) exposure and the partitioning of inhaled MTBE into the body were investigated in a human chamber study. Two subjects were exposed to an environmentally relevant nominal 5,011 micrograms/m3 (1.39 ppm) MTBE for 1 hour, followed by clean-air exposure for 7 hours. Breath and blood were simultaneously sampled, while total urine was collected at prescribed times before, during, and after the exposure. Mass-balance and toxicokinetic analyses were conducted based upon the time series measurement of multiple body-burden endpoints, including MTBE in alveolar breath, and MTBE and tertiary butyl alcohol (TBA) in venous blood and urine. The decay of MTBE in the blood was assessed by fitting the post-exposure data to a 2- or 3-exponential model that yielded residence times(tau) of 2-3 min, 15-50 min, and 3-13 h as measured by alveolar breath, and 5 min, 60 min, and 32 h as evaluated from venous blood measurements. Based on observations of lower than expected blood and breath MTBE during uptake and a decreasing blood-to-breath ratio during the post-exposure decay period, we hypothesize that the respiratory mucous membranes were serving as a reservoir for the retention of MTBE. The decay data suggest that 6-9% of the MTBE intake may be retained by this non-blood reservoir. The compartmental modeling was further used to estimate important parameters that define the uptake of inhaled MTBE. The first of these parameters is f, the fraction of C(air) exhaled at equilibrium, estimated as 0.60 and 0.46 for the female and male subject, respectively. The second parameter is the blood-to-breath partition coefficient (P) estimated as approximately 18. The product of these parameters provides an estimate of the blood concentration at equilibrium as 8-11 times the air concentration. Blood TBA lagged MTBE levels and decayed more slowly (tau = 1.5-3 h), providing a more stable indication of longer term integrated exposure. The concentration ranges of MTBE and TBA in urine were similar to that of the blood, ranging from 0.37 to 15 micrograms/L and 2 to 15 micrograms/L, respectively. In urine, MTBE and TBA by themselves bore little relationship to the exposure. However, the MTBE:TBA ratio followed the pattern of exposure, with peak values occurring at the end of the exposure (20- and 60-fold greater than pre-exposure values) before decaying back to pre-exposure levels by the end of the 7-h decay period. Urinary elimination accounted for a very small fraction of total MTBE elimination (< 1%).

Exposure reconstruction for reducing uncertainty in risk assessment: example using MTBE biomarkers and a simple pharmacokinetic model

Abstract Adverse health risks from environmental agents are generally related to average (long-term) exposures. Because a given individuals contact with a pollutant is highly variable and dependent on activity patterns, local sources and exposure pathways, simple ‘snapshot’ measurements of surrounding environmental media may not accurately assign the exposure level. Furthermore, susceptibility to adverse effects from contaminants is considered highly variable in the population so that even similar environmental exposure levels may result in differential health outcomes in different individuals. The use of biomarker measurements coupled to knowledge of rates of uptake, metabolism and elimination has been suggested as a remedy for reducing this type of uncertainty. To demonstrate the utility of such an approach, we invoke results from a series of controlled human exposure tests and classical first-order rate kinetic calculations to estimate how well spot measurements of methyl tertiary butyl ether and the primary metabolite, tertiary butyl alcohol, can be expected to predict different hypothetical scenarios of previous exposures. We found that blood and breath biomarker measurements give similar results and that the biological damping effect of the metabolite production gives more stable estimates of previous exposure. We also explore the value of a potential urinary biomarker, 2-hydroxyisobutyrate suggested in the literature. We find that individual biomarker measurements are a valuable tool in reconstruction of previous exposures and that a simple pharmacokinetic model can identify the time frames over which an exogenous chemical and the related chemical biomarker are useful. These techniques could be applied to broader ranges of environmental contaminants to assess cumulative exposure risks if ADME (Absorption, Distribution, Metabolization and Excretion) is understood and systemic biomarkers can be measured.

Effect of low level carbon monoxide on compensatory tracking and event monitoring

Experiments by Putz et al. concerning the effect of carbon monoxide (CO) exposure on compensatory tracking and monitoring in healthy young men were replicated. Task and procedural variables were reproduced as closely as practical. Subjects were exposed to either room air or 100 ppm CO. Mean carboxyhemoglobin (COHb) levels in the high CO exposure groups were 5.1% for Putz et al. (70 ppm exposure), and 8.24% for the present study (100 ppm exposure). In both studies elevated COHb produced a statistically significant increase over time in log mean absolute deviation scores (tracking error) with respect to control groups. The magnitude of the effect was smaller in the present study, perhaps because of subtle methodological or training differences between studies. The relationship between task difficulty and magnitude of CO-induced dysfunction remains unresolved. In contrast to Putz et al., no statistically significant effect of COHb in monitoring behavior was found. The failure to replicate this feature of may reflect the large differences in baseline performance, and higher variance in the present study.

Compensatory tracking in humans with elevated carboxyhemoglobin

A total of seventy-four men divided into five groups performed a compensatory tracking task for 4 hr with group mean carboxyhemoglobin (COHb) levels of 0.9, 5.6, 6.1, 11.4 or 16.6%. In all but one group, COHb was formed in approximately 5 min by having the subject breathe a high concentration of carbon monoxide (CO) from a Douglas bag. An appropriate low level of CO in the exposure/test chamber maintained the elevated COHb for the 4-hr duration of the experiment. A slow COHb formation group, not exposed to CO during bag breathing, which was only exposed to low-level CO in the exposure/testing chamber, served as a positive control. Even though mean tracking error scores increased in a nearly monotone manner as a function of COHb, the effects were not statistically significant. Even at the highest dose, the increase in tracking error was small. The increased mean error score due to rapid formation was virtually the same as that due to slow formation.

Trigeminal sensitivity to contact chemical stimulation: A new method and some results

Acquisition of knowledge about the chemical senses might be hastened if chemicals or techniques that permitted stimulation of either the common chemical sense, mediated by the trigeminal nerve, or the sense of smell, mediated by the olfactory nerve without concomitant stimulation of the other sense, were used. Flowing the nonodorized output of an olfactometer into one naris of subjects while velopharyngeal closure was maintained produced effluent flow from the contralateral naris. To prevent odorous molecules from reaching the olfactory epithelium, the contralateral naris was stimulated by applying diluted and undiluted chemical solutions directly to the septal squamous epithelium downstream from the olfactory mucosa. A series of five geometric dilutions of three chemicals and one saturated solution of a fourth were used. The weakest dilution of each chemical was an adequate olfactory stimulus. Two were known trigeminal stimulants (acetic acid and butanol), and two were rated low in trigeminal stimulatory capacity (phenethyl alcohol and vanillin). The results showed that this method was useful in determining the trigeminal threshold for some chemicals. When this method was used, saturated aqueous solutions of vanillin and phenethyl alcohol were clearly not detectable by the trigeminal receptors. The application of the undiluted vanillin and phenethyl alcohol yielded a clear sensation. This result may imply that there are no pure olfactory stimuli.

A computer-controlled vapor-dilution olfactometer

A computer-controlled olfactometer with several innovations leading to closer control over the stimulus parameters and minimization of nonolfactory cues is presented. A microcomputer is used to control the stimulus duration and interstimulus interval. Electronic mass flow con-trollers are employed to maintain close control over the flow in the pure air line and the stimulus-bearing line. Both humidity and temperature are closely regulated. Using a nasal catheter, this stimulus delivery system coupled with velopharyngeal closure and oral breathing permits accurate specification of the quantity of stimulus material actually entering a subject’s nares. The design is such that the computer programming, patch panel interface, and threaded tubing allow versatility in the type of olfactometric study that can be conducted. This system is amenable for threshold, electrophysiological, and sensory adaptation studies of olfaction.

Carboxyhemoglobin and brain blood flow in humans

It has been shown that with increased carboxyhemoglobin (COHb) and associated decrease in blood oxygen-carrying capacity, a compensatory increase in brain-blood flow (BBF) develops. The BBF response in humans has been shown to be quite variable. Two experiments were conducted in which humans were exposed to sufficient carbon monoxide (CO) to produce COHb levels up to 18.4%. BBF was measured by the method of impedance plethysmography. The first was a pilot study in which BBF in 14 men was studied after transient exposure to various concentrations of CO in air. BBF increased as a function of COHb but not to the same extent (or at all) in some subjects. In a confirmatory experiment with 12 men, BBF was measured once per h during a 4-h experiment. All 12 subjects received CO. The variation of the BBF response among subjects was large and statistically significant whereas the variation over time was not significant. Thus it appears that the magnitude of the BBF response is unique for a given subject and differs across subjects. These results may help predict CO-induced behavioral decrements in future studies if subjects whose BBF response to COHb is small or absent are also more susceptible to impairment by acute CO exposure.

A REAL-TIME METHOD TO EVALUATE THE NASAL DEPOSITION AND CLEARANCE OF ACETONE IN THE HUMAN VOLUNTEER

Nasal dosimetry models have become increasingly quantitative as insights into tissue deposition/clearance and computational fluid dynamics have become available. Validation of these models requires sufficient experimental data. However, investigations into respiratory deposition, particularly in human volunteers, have been historically limited due to methodological limitations. To overcome this, a method for evaluating the nasal wash-in, wash-out phenomena of a highly water-soluble compound in human volunteers was developed and characterized. This methodology was assessed using controlled human inhalation exposures to uniformly labeled [13 C]acetone at approximately 1 ppm concentration for 30 min under different breathing maneuvers (inhale nose/exhale nose; inhale nose/exhale mouth; inhale mouth/exhale nose). A small-diameter air-sampling probe inserted in the nasopharyngeal cavity of the volunteer was connected directly to an ion-trap mass spectrometer capable of sampling every 0.8 s. A second ion-trap mass spectrometer simultaneously sampled from the volunteers exhaled breath stream via a breath-inlet device interface. Together, the two mass spectrometers provided real-time appraisal of the [13 C]acetone concentrations in the nasopharyngeal region and in the exhaled breath stream before, during, and after the different breathing maneuvers. The breathing cycle (depth and frequency) and heart rate were concurrently monitored throughout the exposure using a heart-rate monitor and a human plethysmograph to differentiate inhalation and exhalation. Graphical overlay of the plethysmography results with the mass spectrometer measurements show clear quantifiable differences in [13 C]acetone levels at the nasal probe as a function of breathing maneuvers. Breath-by-breath analyses of [13 C]acetone concentrations indicate that between 40 and 75% of the compound is absorbed upon inhalation and nearly all of that absorbed is released back into the breath stream during exhalation.

Decrements in olfactory sensitivity due to ozone exposure

Approximate olfactory thresholds were determined in an exploratory study in which subjects were exposed to ozone (O3). The odorants were butyl alcohol (BA) and acetic acid (AA). Preliminary data suggest (1) temporary increases in thresholds due to O3 exposure, (2) diminution of O3 effects with repeated exposures, and (3) greater and more reliable effects for BA than for AA.

Flow thresholds of nonodorous air through the human naris as a function of temperature and humidity

The process of olfactory sensation is probably a Gestalt consisting of motor, physical, and chemical components formed during the transport of the odor-bearing airstream past the olfactory epithelium. In studying a sensory process, it is often necessary to examine the components independently. Apart from its contribution to the olfactory Gestalt, in dynamic olfactometry airflow may distract from, compete with, or mask chemical sensation. Four nonsmoking male subjects were employed to obtain flow thresholds of nonodorous air at eight temperature/ humidity combinations. The stimulus-generating apparatus was housed in a temperature-regulated oven. All materials in contact with the airstream were thoroughly cleaned and were constructed of nonreactive materials to avoid olfactory cues. An evaluation of the quality of the output demonstrated purity. A Béésy technique was used to deliver stimuli via a nasal catheter to the subject. Thresholds were found to be nearly identical for the lowest humidity levels regardless of the temperature. In general, the closer the temperature/humidity was to physiological normal, the higher the threshold. Since the thresholds were so strongly functions of temperature and humidity, it is likely that transduction was by trigeminal thermoreceptive afferents. The utilization of subthreshold flows in olfactory research may reduce the trigeminal component in olfaction, thus permitting better assessment of the contribution of the olfactory nerve to olfactory sensation.