[Evolution of industrial toxicology toward vanishing doses and the human genome]. / Evoluzione della tossicologia industriale tra dosi effimere e genoma umano.

BACKGROUND: This article aims to discuss the influence that the application of the recent discoveries in genomics will have on the theory and practice of industrial toxicology in developed post-industrial countries. It is stressed that the recent advances in toxicogenomics can be integrated into the existing wealth of knowledge on the toxic properties of industrial chemicals to improve the efficacy of prevention of toxicological risk. METHODS AND RESULTS: The understanding of the biochemical and physiological mechanisms underlying susceptibility or resistance to the toxic effects of industrial xenobiotics, and in particular to carcinogens, allows us to split the epidemiologically derived relationship linking the frequency of disease in the exposed population to the level of workplace contamination into a set of sequential sub-relationships linking: a) the exposure level to that of workplace contamination; b) the internal dose to the exposure level; c) the biological effect (e.g., measured through biochemical markers of early effect) to the internal dose; d) the frequency of disease to that of observation of early biochemical effects. Each of the cited relationships is affected by a degree of uncertainty due to the variability of biological response among the examined individuals, which in turn requires a definition of the statistical limits for the association functions between the variables. As a consequence, the possibility of investigating the individual biochemical and physiological steps in the causal mechanism that links toxic exposure to disease does not necessarily lead to an increase in the information potential of biological monitoring, since the uncertainty due to inter-individual variability is amplified through the sequence of causal relationships to the point that the data from biological monitoring become valueless with regard to the prediction of the frequency or probability of disease. This is particularly true when exposure to 'low doses' is investigated, as is now increasingly frequent in post-industrial developed countries, where workplace contamination is now greatly reduced to levels which may be borderline with those in the general environment. Thus at the low-dose end of the range of contamination and exposure values there is an area where, for statistical reasons consequent to the heterogeneity of examined populations, a quantitative prediction of internal exposure due to environmental contamination, of biological adverse effects due to exposure levels and of frequency of disease due to the extent or frequency of biological effects is no longer reliably possible. This in turn impairs the preventive efficacy of biological monitoring. CONCLUSIONS: A closer integration between industrial toxicology and state-of-the-art molecular genetics derived from the recent sequencing of the human genome is the way to overcome the limitations described. In particular, the individual subjects in the examined populations can be classified with regard to some genetically controlled characters relevant to the biotransformation of xenobiotics and to DNA repair and the statistical analysis of data can be performed on more homogeneous subpopulations, in order to decrease inter-individual variability of biochemical and physiological response. This in turn increases the predictive power of the biological markers, both of dose and effect, and improves the efficacy of prevention, e.g., by highlighting oversensitive subpopulations or lifestyles which can increase the risk of occupational and environmental disease.

Comparison between blood and urinary toluene as biomarkers of exposure to toluene.

OBJECTIVES: To compare blood toluene (TOL-B) and urinary toluene (TOL-U) as biomarkers of occupational exposure to toluene, and to set a suitable procedure for collection and handling of specimens. METHOD: An assay based on headspace solid-phase microextraction (SPME) was used both for the determination of toluene urine/air partition coefficient (lambdaurine/air) and for the biological monitoring of exposure to toluene in 31 workers (group A) and in 116 non-occupationally exposed subjects (group B). Environmental toluene (TOL-A) was sampled during the work shift (group A) or during the 24 h before specimen collection (group B). Blood and urine specimens were collected at the end of the shift (group A) or in the morning (group B) and toluene was measured. RESULTS: Toluene lambdaurine/air was 3.3 +/- 0.9. Based on the specimen/air partition coefficient, it was calculated that the vial in which the sample is collected had to be filled up to 85% of its volume with urine and 50% with blood in order to limit the loss of toluene in the air above the specimen to less than 5%. Environmental and biological monitoring of workers showed that the median personal exposure to toluene (TOL-A) during the work-shift was 80 mg/m3, the corresponding TOL-B was 82 microg/l and TOL-U was 13 microg/l. Personal exposure to toluene in environmentally exposed subjects was 0.05 mg/m3, TOL-B was 0.36 microg/l and TOL-U was 0.20 microg/l. A significant correlation (P < 0.05) was observed between TOL-B or TOL-U and TOL-A (Pearson's r = 0.782 and 0.754) in workers, but not in controls. A significant correlation was found between TOL-U and TOL-B both in workers and in controls (r = 0.845 and 0.681). CONCLUSION: The comparative evaluation of TOL-B and TOL-U showed that they can be considered to be equivalent biomarkers as regards their capacity to distinguish workers and controls and to correlate with exposure. However, considering that TOL-U does not require an invasive specimen collection, it appears to be a more convenient tool for the biological monitoring of exposure to toluene.

Headspace solid-phase microextraction for the determination of benzene, toluene, ethylbenzene and xylenes in urine.

A method for the determination of benzene, toluene, ethylbenzene and xylenes (BTEX) in urine of people exposed to these airborne pollutants present in the living environment, has been described. Solid-phase microextraction has been used for sampling BTEX from the headspace of urine and gas chromatography-mass spectrometry has been applied for the selective analysis of chemicals. The method has the following features: small volume of urine (2 ml) needed, linearity in the range of interest (from the limit of detection up to 5000 ng/l) with coefficient of correlation > or =0.998, limit of detection in the range 12-34 ng/l, good repeatability (coefficient of variation 2-7%), high specificity. The stability of the urine sample during storage (-20 degrees C) was evaluated: BTEX remained stable for up to 2 months. The assay has been successfully applied to the biological monitoring of two subjects environmentally exposed to airborne BTEX in an urban area.

[Biological monitoring of exposure to solvents: a method for the gas-chromatographic determination of aromatic hydrocarbons in the blood and urine]. / Monitoraggio biologico dell'esposizione a solventi: metodo per la determinazione gascromatografica degli idrocarburi aromatici nel sangue e nell'urina.

A gas chromatographic procedure with dynamic head-space purge and trap preconcentration (HSGC) and FID detection for blood and urinary benzene, toluene, ethylbenzene and xylenes (BTEX) determination at low level exposure is described. Critical steps (sample collection, calibration, HSGC conditions, contamination control) are discussed. The calibration curve is linear in the range 50 ng/l-500 micrograms/l; the calculated detection limit is 50 ng/l for all the considered aromatic hydrocarbons (AH) both in blood and urine; the within-day precision, calculated as variation coefficient (CV) at 400 ng/l and 40 micrograms/l (n = 6) was respectively CV = 13% and CV = 6% for all the studied analytes. The recovery rate was in the range 29-70%, depending on the hydrocarbon and matrix (blood or urine) considered. The procedure was applied to the biological monitoring of 151 workers occupationally or environmentally exposed to BTEX. Occupationally exposed subjects showed blood AH levels of 2-4 order of magnitude higher than environmentally exposed subjects. In white-collar workers exposed to BTEX urban pollution a significant difference in blood and urine levels of AH was observed between nonsmokers and smokers. Nonsmokers showed blood AH median values of respectively benzene = 241 ng/l, toluene = 759 ng/l, ethylbenzene = 140 ng/l, xylenes = 604 ng/l. Significatively higher BTEX blood values were observed in smokers after a median consumption of 5 cigarettes in 5 hours; observed median values were respectively: benzene = 365 ng/l toluene = 1327 ng/l, ethylbenzene = 233 ng/l, xylenes = 794 ng/l.

Biological and environmental monitoring of exposure to airborne benzene and other aromatic hydrocarbons in Milan traffic wardens.

Environmental and biological monitoring of airborne aromatic hydrocarbons has been performed in 20 policemen working as traffic wardens exposed to motor vehicle exhausts and in 19 peers employed as clerks. Airborne benzene, toluene, ethylbenzene and xylene concentrations, measured during the workshift, resulted in significantly higher outdoor than indoor concentrations (benzene and related aromatic hydrocarbons mean values, respectively of 53 and 350 micrograms/m3 vs. 29 and 180 micrograms/m3). Blood benzene, toluene, ethylbenzene and xylene concentrations did not differ significantly between indoor and outdoor workers; no differences were found between values obtained at the beginning (07:30 h) and the end of shift (00:30) in either group. Blood hydrocarbon concentrations seem to reflect airborne pollution, whilst the blood benzene concentration determined after the workshift poorly reflects airborne benzene morning peaks. Endshift blood benzene mean concentration in smokers (462 ng/l, n = 9) differs significantly from non-smokers (292 ng/l, n = 39).