Biological monitoring of low benzene exposure in Italian traffic policemen

A comparative evaluation of urinary biomarkers was carried out to characterize benzene exposure in a group of 100 traffic policemen of the city of Parma (Italy). All subjects were monitored once, in two consecutive days characterized by similar climatic conditions but preceded by two windy days. Benzene ambient concentration measured by municipal air monitoring stations was 1 microg/m(3) (Day 1) and 2 microg/m3 (Day 2). Personal exposure to ambient concentrations of benzene, toluene, ethylbenzene and xylene (BTEX) was assessed by using Radiello((R)) passive-diffusive samplers in a subgroup of 24 workers. Benzene metabolites, t,t-muconic acid (t,t-MA) and S-phenylmercapturic acid (S-PMA) were determined by isotopic dilution liquid chromatography-tandem mass spectrometry on spot urine samples collected at the end of the shift. Urinary benzene (U-B) was determined by solid-phase microextraction gas chromatography-mass spectrometry. Airborne benzene concentration expressed as median [and interquartile range] was 6.07 [0.28-9.53] microg/m(3), as assessed by personal sampling. Urinary concentrations of biomarkers in the whole group were 41.8 [34.1-89.8] microg/g creatinine for t,t-MA, 0.67 [0.23-1.32] microg/g creatinine for S-PMA, and 0.16 [0.13-0.26] microg/l for U-B. Smokers eliminated significantly higher concentrations of unchanged BTEX and benzene metabolites than non-smokers (p < 0.05). When traffic policemen were distinguished into indoor (n=31) and outdoor workers, no significant differences were observed for either airborne benzene or urinary biomarkers. Significantly lower concentrations of S-PMA and U-B were determined in samples collected at Day 1 as compared to Day 2 (p < 0.0001 and p = 0.003, respectively) suggesting that these biomarkers are enough sensitive and specific to detect changes in airborne benzene concentration even at few microg/m(3).     

Correlation between environmental and biological monitoring of exposure to benzene in petrochemical industry operators

The present work was aimed to study in petrochemical industry operators the correlation, if any, between environmental exposure to low levels of benzene and two biological exposure indexes in end-shift urine, i.e. trans, trans-muconic acid (t,t-MA) and S-phenylmercapturic acid (SPMA). Exposure to benzene was assessed in 133 male subjects employed in outdoor operations in a petrochemical plant, using personal passive-diffusive air samplers worn at the breathing zone; adsorbed benzene was determined by GC-FID analysis. S-PMA was determined by a new HPLCMS/MS method, after (quantitative) acidic hydrolysis of the cysteine conjugate precursor. t,t-MA was measured by an HPLC-UV method. Smoking habits were assessed by means of a self-administered questionnaire.     

Construction of a database of benzene biological monitoring

Biological monitoring of occupational exposure to benzene has been conducted in the petroleum, steel and chemical industries. The urinary benzene-specific biomarker, S-phenylmercapturic acid (PMA), was quantified in post-shift samples using a sensitive enzyme-linked immunosorbent assay (ELISA) and expressed as a function of urinary creatinine concentration. The assay, based on a PMA-specific antiserum, is sufficiently sensitive to measure PMA levels in non-occupationally exposed control subjects. The assay delivers batch results in a timely manner which may be as short as 3 h. Samples were analysed from groups of workers engaged in coke oven combustion processes, petroleum refining and decontamination of a benzene land spill. The construction of a database of results provides an index of benzene uptake as a consequence of the respective work processes and tasks and readily enables benchmarking exercises aimed at comparing degrees of exposure across segments of industry.     

Environmental and biological monitoring of benzene,toluene, ethylbenzene and xylene (BTEX) exposure in residents living near gas stations

ABSTRACT

The volatile organic compounds benzene, toluene, ethylbenzene, and xylene (BTEX) are emitted into the atmosphere at gas stations (GS) leading to chronic exposure of nearby residents, which raises public health concerns. This study aimes at determining the contribution of GS emissions to BTEX exposure in nearby residents. Three Control and Exposed areas to BTEX emissions from GS were defined in a medium-sized European city (Porto, Portugal). BTEX atmospheric levels were determined in Control and Exposed areas using passive samplers deployed outdoors (n = 48) and indoors (n = 36), and human exposure was estimated for 119 non-smoking residents using the first urine of the day. Results showed that median BTEX outdoor and indoor concentrations were significantly higher for Exposed than Control areas, with exception of ethylbenzene and xylene indoor concentrations, where no marked differences were found. Comparison of urinary concentrations between Exposed and Control residents demonstrated no significant differences for benzene and ethylbenzene, whereas levels of toluene and xylene were significantly higher in Exposed residents. No marked correlation was obtained between atmospheric BTEX concentrations and urinary concentrations. Data indicate the potential impact on air quality of BTEX emissions from GS, which confirms the importance of these findings in urban planning in order to minimize the impact on health and well-being of surrounding populations.     

S-Phenylcysteine formation in hemoglobin as a biological exposure index to benzene

Benzene is metabolized to intermediates that bind to hemoglobin, forming adducts. These hemoglobin adducts may be usable as biomarkers of exposure. In this paper, we describe the development of a gas chromatography/mass spectroscopy assay for quantitating the binding of the benzene metabolite, benzene oxide, to cysteine groups in hemoglobin. We used this assay to study the hemoglobin adduct, S-phenylcysteine (SPC), in the blood of rats and mice exposed to benzene either by inhalation or by gavage. We were able to detect SPC in the hemoglobin of exposed rats and mice, to show the linearity of the exposure dose-response relationship, and to establish the sensitivity limits of this assay. For the same exposure regime, rats showed considerably higher levels of SPC than did mice. As yet, we have not been able to detect SPC in the globin of humans occupationally exposed to benzene. We attempted to determine whether the SPC found in hemoglobin originated from the metabolism of benzene within or outside of the red blood cell. We hypothesized that the greatest red blood cell metabolism would be associated with peripheral reticulocytes, which retain high metabolic capacity. After exposing rats to benzene, we isolated the red blood cells and used discontinuous Percoll gradients to fractionate them into age groups. No differences in SPC levels were found among any of the fractions, suggesting that the SPC found in globin originates from the metabolism of benzene to benzene oxide in a location external to the red blood cell. To our knowledge, this is the first demonstration of the nonenzymatic binding of the benzene metabolite, benzene oxide, to protein. The products of the interaction of benzene oxide with protein are particularly appealing as potential biomarkers. These adducts are stable both in vivo and during isolation, and because high background levels in unexposed individuals are unlikely, the presence of such adducts should be a specific indicator of exposure to benzene. If SPC can be detected in humans, the present assay, with some modification, may be amenable to routine monitoring of worker exposures to benzene.     

Aspects of biological monitoring of exposure to glycol ethers

Glycol ethers are frequently used as solvents, detergents, and emulsifiers alone or as components in industrial and consumer products. The monomethyl and monoethyl ethers of ethylene glycol, and their acetate esters, are teratogenic and embryotoxic and cause testicular damage in laboratory animals, while the monobutyl ether causes hemolysis of the red blood cells. The adverse effects are attributed to the acid metabolites methoxy-, ethoxy- and butoxyacetic acid, respectively. The glycol ethers may readily enter the body by inhalation as well as dermal uptake. Biological monitoring of exposure to glycol ethers has therefore been suggested. This paper reviews physical properties, occurrence, analysis, toxicity, and toxicokinetics of the most common glycol ethers and then discusses toxicokinetic aspects of biological monitoring. The effect of physical exercise and the relative importance of respiratory and percutaneous absorption on the internal exposure to glycol ethers are illustrated. Monitoring the acid metabolite in urine is suggested as the best index of exposure. Intra- and inter-individual variability, dose-dependent toxicokinetics, and metabolic induction and inhibition are examples of possible sources of error in the estimation of internal exposure from the urinary excretion of acid metabolite.     

Reference values and action levels of biological monitoring in occupational exposure.

The primary objectives of biological monitoring are (1) to prevent health impairment, (2) to assist in the assessment of risk, and (3) to evaluate the effectiveness of environmental controls. An efficient way to achieve these objectives would be to enforce the compliance of biological exposure standards at the workplace. However, biological monitoring should be viewed in the total context of control and prevention of work-related diseases, and not merely to comply with permissible standards. Biological monitoring depends very much on the conditions that the chemical is absorbed and how it is metabolised. Genetic diversity could therefore contribute to significant differences in this aspect. Furthermore, many of the reference values established have so far not been fully validated and therefore their usefulness is rather limited. This paper reviews and illustrates using some recent findings to show that biological reference values are influenced not just by the above mentioned issues, but also factors such as (1) health and nutritional status of the exposed population, (2) social and cultural factors, and (3) climatic conditions. Caution has to be taken when considering having an action level for some of the biological reference value. Biological reference values set without considering people, technology, and working conditions would be fraught with difficulties in implementation.     

Environmental and biological monitoring in the estimation of absorbed doses of pesticides

Exposure to pesticides affects most of the population, not only persons occupationally exposed. In a context of high variability of exposure, biological monitoring is important because of the various routes by which exposure can occur and because it assesses both occupational and non-occupational exposure. The main aim of this paper was to critically compare estimates of absorbed dose measured by environmental and biological monitoring in situations in which they could both be applied. The combination of exposure measurements and biological monitoring was found to provide extremely important information on the behaviour of employees, and on the proper use and effectiveness of personal protection equipment.     

Importance of biotransformation pathways for interpreting biological monitoring of exposure.

A detailed characterisation of the biotransformation pathways followed by the chemical of interest is, evidently, of primordial importance to design reliable biomonitoring programs. This short overview discusses some recent studies that are of potential interest for the development of new avenues of research. Four topics are addressed: tissue localisation of the metabolic process; importance of speciation; metabolic interactions and polymorphism and interindividual variability. While some of these studies offer new data that is directly useful for practical application, others raise mainly questions and indicate that there is still a large effort of research to be performed to improve the design and the interpretation of biomonitoring programs.     

Validation protocol and analytical quality in biological monitoring of occupational exposure to antineoplastic drugs

Occupational exposure to antineoplastic cytostatic drugs has been recognized as a potential health hazard since the seventies. Safety guidelines and recommendations have been published in several countries in order to improve operating procedures and keep exposure levels as low as possible. Nevertheless, contamination still occurs. With a view to preventing exposure, standardized authorized methods and a mandatory monitoring system appear to be an essential prerequisite. First of all, reliable analytical methods are necessary, and a wide number of drugs should be monitored, on the basis of the continual change in the treatment protocols. In fact, most of the methods available in the literature were not properly validated, and a lack of details concerning quality assurance is clearly observed. When assessing analytical methods applied for routine testing, validation studies are of utmost importance. Not only performance parameters, such as sensitivity, specificity, LOD and LLQ, should therefore be determined, but also the uncertainty of measurement, which gives a measure of the confidence that can be placed on the results.     

Immune parameters in biological monitoring of pesticide exposure: current knowledge and perspectives.

Exposure to pesticides can cause a number of effects on the immune system, varying from a slight modulation of immune functions to the development of clinical immune diseases. The aim of this study has been reviewing published data on immune effects of pesticides in humans, with particular attention for effects observed in absence of any other change, and to the possibility of identifying a dose effect relationship. Some evidence of immunotoxic effects in man involve organophosphorus compounds, some organochlorine insecticides (OC), some carbamates, some phenoxy herbicides, dithiocarbamates, and pentachlorophenol (PCP). The alterations are usually observed in absence of any other change; in some cases, data suggest the presence of a dose effect relationship. The prognostic significance of the observed changes is still unclear. The Authors propose a tier approach to assess immune effects in humans.     

Reevaluation of benzene exposure for the Pliofilm (rubberworker) cohort (1936-1976).

The Pliofilm cohort is the most intensely studied group of workers chronically exposed to benzene. Information on this cohort has been the basis for regulations and/or guidelines for occupational and environmental exposure to benzene. Rinsky et al. (1986, 1987) and Crump and Allen (1984) developed different approaches for reconstructing the exposure history of each member of the group. The predicted levels of exposure, combined with the data on the incidence of disease, have been used to estimate benzene's carcinogenic potency. In this paper, recent information from worker interviews and historical records from the National Archives and elsewhere were used to evaluate the accuracy of prior exposure estimates and to develop better ones for the cohort. The following factors were accounted for: (1) uptake of benzene due to short-term, high-level exposure to vapors, (2) uptake due to background concentrations in the manufacturing building, (3) uptake due to contact with the skin, (4) morbidity and mortality data on workers in the Pliofilm process, (5) the installation of industrial hygiene engineering controls, (6) extraordinarily long work weeks during the 1940s, (7) data indicating that airborne concentrations of benzene were underestimated due to inaccurate monitoring devices and the lack of adequate field calibration mated due to inaccurate monitoring devices and the lack of adequate field calibration of these devices, and (8) likely effectiveness of respirators and gloves. Our estimates suggest that Crump and Allen (1984) overestimated the exposure of workers in some job classifications and underestimated others, and that Rinsky et al. (1981, 1986) almost certainly underestimated the exposure of nearly all workers. Airborne concentrations of benzene at the St. Marys facility during the years of its operation were found (on average) to be about half those of the two Akron facilities. Our analysis indicates that short-term, high-level exposure to benzene vapors and dermal exposure significantly increased (by about 25-50%) the total absorbed dose of benzene for some workers. One of the key findings was that, unlike prior analyses, the three facilities probably had significantly different airborne concentrations of benzene, especially during the 1940s and 1950s.     

Characterisation of exposure to total and hexavalent chromium of welders using biological monitoring

Inhalation exposure to total and hexavalent chromium (TCr and HCr) was assessed by personal air sampling and biological monitoring in 53 welders and 20 references. Median inhalation exposure levels of TCr were 1.3, 6.0, and 5.4 μg/m³ for welders of mild steel (MS, <5% alloys), high alloy steel (HAS, >5% alloys), and stainless steel (SS, >26% alloys), respectively. The median exposures to HCr compounds were 0.23, 0.20, and 0.08 μg/m³, respectively. Median concentrations of TCr in urine, blood plasma and erythrocytes were elevated in all welders, compared with the corresponding median concentrations in the reference group (p < 0.005). The TCr levels observed in plasma were two-fold higher in welders of SS and HAS than in welders of MS (p < 0.01). Exposure to HCr as indicated by median total content of Cr in erythrocytes was 10 μg/L in welders of SS, MS and HAS. Uptake of TCr during the shift was confirmed for welders of SS by a median increase of urinary TCr from pre- to post-shift of 0.30 μg/g creatinine. For welders of MS and HAS as a group TCr was not increased.     

The United States Environmental Protection Agency's guidelines for applicator exposure monitoring

The Environmental Protection Agency (EPA)'s proposed guidelines for studies which estimate worker exposure to pesticides during application and related work activities are discussed. These guidelines are intended to aid pesticide registrants in conducting occupational exposure monitoring studies for pesticide applicators at indoor and outdoor sites. The Agency's rationale for requiring exposure to be monitored with passive dosimetry techniques or by biological monitoring is described. The practical and theoretical advantages and disadvantages of both approaches are also discussed.     

Analytical methods of biological monitoring for exposure to pesticides: recent update

Extensive use of synthetic pesticides for agricultural and nonagricultural purposes began in the past 50 years. As a result of their wide and extensive application, exposure to hazardous pesticides is a concern to the general population and occupationally exposed persons. Robust methods are therefore needed for measuring markers of pesticide exposure. This article presents a review of the most recently published analytical methodologies and instrumentations developed for and applied to biological monitoring of exposure to pesticides of various classes. Most of the methods reviewed here are based on chromatography combined with mass spectrometry detection. This work clearly demonstrates that although gas chromatography still appears to be the most widely employed technique for pesticide analysis in various biological samples, recently a trend has been observed toward the use of liquid chromatography coupled with tandem mass spectrometry.     

Reconstruction of benzene exposure for the Pliofilm cohort (1936-1976) using Monte Carlo techniques

The current cancer slope factor and occupational standards for benzene are based primarily on studies of the rubber hydrochloride (Pliofilm) workers. Previous assessments of this cohort by Rinsky et al. (1981, 1987), Crump and Allen (1984), and Paustenbach et al. (1992) relied on different assumptions about the available industrial hygiene data and workplace practices and processes over time, thereby yielding significantly different estimates of annual benzene exposures for many jobs. Given the inherent limitations and uncertainties involved in estimating historical exposures for this cohort, a probabilistic approach was used to better characterize their likely degree of benzene exposure. Ambient air exposures to benzene were based, in part, on the distribution of air sampling data collected at the Pliofilm facilities and assumptions about how workplace concentrations probably decreased over time as the threshold limit value (TLV) was lowered. The likely uptake of benzene from dermal exposures was estimated based on probability distributions for several exposure factors, including surface area, contact rate and duration, and skin absorption. The assessment also quantitatively accounts for improved engineering controls, extended work hours, incomplete Pliofilm production, and the use and effectiveness of respirators over time. All original data and assumptions are presented in this assessment, as is all new information obtained through additional interviews of former workers. Estimated benzene exposures at the 50th and 95th percentiles are reported as equivalent 8-h time-weighted average (TWA) airborne concentrations for 13 job categories from 1936 to 1965 (Akron I and II facilities) and 1939 to 1976 (St. Mary's facility). Data indicate that estimated equivalent airborne benzene concentrations for St. Mary's workers were highest for four job categories (Neutralizer, Quencher, Knifeman, Spreader), typically ranging from about 50 to 90 ppm during 1939-1946 (lower during 1942-1945), and 10 to 40 ppm during 1947-1976 at the 50th percentile. These estimates are 2-3 times greater than for other jobs in the Pliofilm process, and about 1.5 times less than those estimated at the 95th percentile. Estimates of equivalent airborne benzene concentrations for Akron I and II were about 1.5 times higher than for St. Mary's, but there is less confidence in these estimates, given the lack of industrial hygiene monitoring data for these facilities. Study results suggest that Paustenbach et al. (1992) generally over-estimated exposures for those job categories that had the highest exposure by about a factor of two to four. On the other hand, it was concluded that Rinsky et al. (1981, 1987) under-predicted benzene exposures for most jobs, and Crump and Allen (1984) both under- and overpredicted benzene exposures, depending on the specific job category and time period. The new estimates presented in this analysis incorporate what is considered to be the most likely range of plausible exposure values, and, accordingly, provide a better characterization of the potential workplace exposures for this cohort. These data could be combined with current or future mortality information to calculate a new cancer potency factor or occupational health standard for benzene.     

Urinalysis vs. blood analysis, as a tool for biological monitoring of solvent exposure.

Blood and urine samples were collected at the end of an 8-h workshift from 30 male workers exposed to a mixture of n-hexane, ethyl acetate and toluene (each being about 2 ppm as geometric means) and also from 20 nonexposed male workers. Blood samples were analyzed for n-hexane and toluene, and urine samples were analyzed for n-hexane, toluene, 2,5-hexanedione (both with and without hydrolysis) and hippuric acid. Based on the correlation between biological exposure indicators and solvent concentrations in air, sensitivity as an exposure indicator was compared between solvents in blood and solvents or metabolites in urine in terms of the lowest solvent concentration at which the exposed subjects can be statistically separated from the nonexposed. Both n-hexane and toluene in blood were sensitive enough to detect the exposure at 6.1 ppm and 1.4 ppm, respectively. n-Hexane exposure below 2 ppm was detectable also by urinalysis for 2,5-hexadione without hydrolysis. Urinary hippuric acid, however, failed to detect low toluene exposure under the conditions studied. Of additional interest is the fact that toluene in urine correlated significantly with toluene in air, which apparently deserves further study for confirmation.