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Biological monitoring of metals with special references to the early stages of the life cycle.

In summary, the biological monitoring of toxic metals in the early stages of the life cycle has not yet received the attention it deserves. Most metals have simply not been studied at all. Some successful applications with mercury, lead, and cadmium have been reviewed here. But even for these widely studied metals, there is a great need to study physiological models for metal disposition in developing animals.

Animals↗

Monitoring biological effects of contamination in marine fish along French coasts by measurement of ethoxyresorufin-O-deethylase activity.

The use of bioindicators to evaluate exposure to the biological effects of chemical pollutants in marine organisms constitutes a new tool in the monitoring field. The establishment of a North Sea monitoring network in 1991, involving such international organizations as the North Sea Task Force, the International Council for the Exploration of the Sea, and the Intergovernmental Oceanography Commission, led French researchers to develop an enzymatic biomarker to monitor biological effects within the National Observation Network. The biomarker, ethoxyresorufin-O-deethylase (EROD), dependent on the CP450 system, has been monitored biannually since 1992 in several species of fish (Callionymus lyra, Limanda limanda, Serranus sp., Mullus barbatus) in two coastal sites particularly exposed to industrial and domestic pollution. A rapid method is used to assay EROD enzymatic activity determined along a pollution gradient, and results are interpreted on a microplate reader. The strategy of this approach is to assess the effects on the marine ecosystem during prolonged exposure to specific pollutants such as polyaromatic hydrocarbons, polychlorinated biphenyls, and dioxins.

Animals↗

Variability in biological monitoring of solvent exposure. I. Development of a population physiological model.

Biological indicators of exposure to solvents are often characterised by a high variability that may be due either to fluctuations in exposure or individual differences in the workers. To describe and understand this variability better a physiological model for differing workers under variable industrial environments has been developed. Standard statistical distributions are used to simulate variability in exposure concentration, physical workload, body build, liver function, and renal clearance. For groups of workers exposed daily, the model calculates air monitoring indicators and biological monitoring results (expired air, blood, and urine). The results obtained are discussed and compared with measured data, both physiological (body build, cardiac output, alveolar ventilation) and toxicokinetic for six solvents: 1,1,1-trichloroethane, trichloroethylene, tetrachloroethylene, benzene, toluene, styrene, and their main metabolites. Possible applications of this population physiological model are presented.

Adipose Tissue↗

Environmental and biological monitoring of benzene exposure in a cohort of Italian taxi drivers.

An integrated approach based on ambient and biological monitoring, the latter including both biomarkers of exposure and susceptibility, was applied to characterize benzene exposure in a group of 37 taxi drivers of the city of Parma (Italy). Airborne benzene concentrations were assessed by 24 h personal sampling and work-shift sampling inside the taxicab using passive samplers (Radiello). Benzene metabolites, trans,trans-muconic acid (t,t-MA) and S-phenylmercapturic acid (S-PMA), and urinary cotinine as biomarker of smoking habits were measured by isotopic dilution liquid chromatography tandem mass spectrometry in both pre-shift (PS) and end-of-shift (EOS) samples. Urinary benzene (U-B) levels were determined by solid-phase microextraction gas chromatography-mass spectrometry in EOS samples. Relevant polymorphisms of microsomal epoxide hydrolase, NAD(P)H:quinone oxidoreductase, glutathione S-transferases M1-1 (GSTM1), T1-1, and A1 were characterized by PCR-based methods. Mean airborne benzene concentration was 5.85 +/- 1.65 microg/m3, as assessed by 24 h personal sampling integrating for work-shift, indoor or general environment activities. Significantly, higher benzene concentrations were detected in the taxicab during the work-shift (7.71 +/- 1.95 microg/m3, p < 0.005). Smokers eliminated significantly higher concentrations of U-B and S-PMA than non-smokers in EOS samples [geometric mean (geometric S.D.): 2.58 (4.23) versus 0.44 (1.79) microg/l for U-B; 3.79 (1.50) versus 2.14 (1.87) microg/gcreat. for S-PMA, p < 0.002]. Within smokers, S-PMA concentrations significantly increased at the end of the work-shift compared to pre-shift values (p < 0.05). t,t-MA showed a similar behaviour, although differences were not significant. In the narrow range examined, no correlation was observed between air benzene concentration and urinary biomarkers. All benzene biomarkers but EOS t,t-MA were correlated with U-cotinine (p < 0.05). GSTM1 polymorphism significantly modulated S-PMA excretion, as subjects bearing the GSTM1pos genotype [3.61 (1.15) microg/gcreat.] excreted significantly higher S-PMA concentrations than GSTM1null subjects [2.19 (1.18) microg/gcreat., p < 0.05].

Acetylcysteine↗

Design and evaluation of a breath-analysis system for biological monitoring of volatile compound.

To ensure the health and safety of workers, integrated industrial hygiene methodologies often include biological monitoring of the workers to help understand their exposure to chemicals. To this end, a field-portable breath-analysis system was developed and tested to measure selected solvents in exhaled air. The exhaled breath data were evaluated using a physiologically based pharmacokinetic (PBPK) model to relate exposure to tissue dose. The system was designed to monitor workers every time they entered or left a work environment--a vast improvement over current 8-hour integrated monitoring strategies. The system combines (1) chemical dosimeters to measure airborne contaminant levels (analyzed in the field/ workplace); (2) real-time breath analysis to quantitate exposure; and 3) PBPK models to estimate internal target tissue dose. To evaluate the system, field tests were conducted at two locations: (1) at an incinerator in Tennessee monitoring benzene and toluene exposures; and (2) a waste repackaging facility in Washington State where hexane, trimethylbenzene, and methylene chloride was monitored. Exhaled breath was sampled and analyzed before and after each specific job task, which ranged from 15 min to 8 hours in duration. In both field studies several volunteers had posttask breath levels higher than pretask levels. The greatest increase corresponded to 573 ppb for methylene chloride and 60 ppb for toluene. Compared with breath analysis, the chemical dosimeters underpredicted the dosimetry, particularly for longer sampling intervals when the volume of air sampled may have diluted exposures. The results of the field studies illustrate the utility of monitoring workers for exposures throughout the day, particularly when job-specific tasks may indicate a potential for exposure.

Air Pollutants, Occupational↗

Biological monitoring and medical screening at the workplace in the EC countries.

Health surveillance in the work environment involves continuous biological monitoring and medical screening, with the purpose of primary and secondary prophylaxis of work-related diseases. Is this screening activity governed by a rationale based on knowledge of dangerous exposure and the availability of valid tests? In the USA, where health surveillance programmes are used extensively, a study has found screening activity to be associated more with plant size than which relevant exposure. This study was done to elucidate the character and extent of use of health surveillance in the work environment in the EC countries with the aid of a questionnaire survey. The chief medical officers of the National Labour Inspectorates supplied information on substances covered by health surveillance programme in the EC member states, together with the legislative status and numbers of exposed workers. Belgium, France, Italy and the former Federal Republic of Germany made extensive use of health surveillance programmes in cases of known exposure to metals, organic solvents, carcinogenic and genotoxic substances, mineral dust, ionizing radiation, and biological agents. Denmark and Holland ran national programmes only for substances covered by EC directives, while England, Ireland, Spain, and Portugal comprised an intermediate user group. The result suggest that the use of health surveillance is related more to the national choice of standard regulatory instruments than to relevant exposure.

Environmental Monitoring↗

Biological monitoring of aromatic diisocyanates in workers exposed to thermal degradation products of polyurethanes.

Exposure to diisocyanates was assessed by biological monitoring among workers exposed to the thermal degradation products of polyurethanes (PURs) in five PUR-processing environments. The processes included grinding and welding in car repair shops, milling and turning of PUR-coated metal cylinders, injection moulding of thermoplastic PUR, welding and cutting of PUR-insulated district heating pipes during installation and joint welding, and heat-flexing of PUR floor covering. Isocyanate-derived amines in acid-hydrolysed urine samples were analysed as perfluoroacylated derivatives by gas chromatography mass spectrometry in negative chemical ionisation mode. The limits of quantification (LOQs) for the aromatic diamines 2,4- and 2,6-toluenediamine (2,4- and 2,6-TDA) and 4,4'-methylenedianiline (4,4'-MDA) were 0.25 nmol l(-1), 0.25 nmol l(-1) and 0.15 nmol l(-1), respectively. The LOQ for the aliphatic diamines hexamethylenediamine (HDA), isophoronediamine (IpDA) and 4,4'-diaminodicyclohexyl methane (4,4'-DDHM) was 5 nmol l(-1). TDA and MDA were detected in urine samples from workers in car repair shops and MDA in samples from workers welding district heating pipes. The 2,4-TDA isomer accounted for about 80% of the total TDA detected. No 2.6-TDA was found in the urine of non-exposed workers. The highest measured urinary TDA and MDA concentrations were 0.79 nmol mmol(-1) creatinine and 3.1 nmol mmol(-1) creatinine, respectively. The concentrations found among non-exposed workers were 0.08 nmol mmol(-1) creatinine for TDA and 0.05 nmol mmol(-1) creatinine for MDA (arithmetic means). Exposure to diisocyanates originating from the thermal degradation of PURs are often intermittent and of short duration. Nevertheless, exposure to aromatic diisocyanates can be identified by monitoring diisocyanate-derived amines in acid-hydrolysed urine samples.

Adult↗

Immunochemical techniques in biological monitoring.

Immunoassays are analytical methods that detect interactions between antibodies and antigens. Immunoassays were used originally to detect large biological molecules. The new generation of these antibody-based assays can detect small synthetic compounds. As a result, immunoassays are being developed specifically for biomarkers of exposure and effect to environmentally prevalent chemicals. Immunochemical detection of parent compounds in blood and tissues, metabolites in excreta, and adducts with DNA and protein have been successfully performed by several investigators. Although there is great potential for use of immunoassays in biological monitoring studies, the limitations of these analyses must be fully understood to prevent improper evaluation of the acquired data. This review will cover some of the background material necessary to understand how an antibody-based assay is developed. The differences between polyclonal and monoclonal antibody-based assays and the importance of antibody class, affinity, specificity, and cross-reactivity must be considered in both study design and data analysis.

Antibodies↗

Biological monitoring of occupational exposure to cyclohexane by urinary 1,2- and 1,4-cyclohexanediol determination.

OBJECTIVES: This article reports the results obtained with the biological and environmental monitoring of occupational exposure to cyclohexane using 1,2-cyclohexanediol (1,2-DIOL) and 1,4-DIOL in urine. The kinetic profile of 1,2-DIOL in urine suggested by a physiologically based pharmacokinetic (PBPK) model was compared with the results obtained in workers. METHODS: Individual exposure to cyclohexane was measured in 156 workers employed in shoe and leather factories. The biological monitoring of cyclohexane exposure was done by measurement of 1,2-DIOL and 1,4-DIOL in urine collected on different days of the working week. In all, 29 workers provided urine samples on Monday (before and after the work shift) and 47 workers provided biological samples on Thursday at the end of the shift and on Friday morning. Another 86 workers provided biological samples at the end of the work shift only on Monday or Thursday. RESULTS: Individual exposure to cyclohexane ranged from 7 to 617 mg/ m3 (geometric mean value 60 mg/m3). Urinary concentrations of 1,2-DIOL (geometric mean) were 3.1, 7.6, 13.2, and 6.3 mg/g creatinine on Monday (pre- and postshift), Thursday (postshift) and Friday (pre-shift), respectively. The corresponding values recorded for 1,4-DIOL were 2.8, 5.1, 7.8, and 3.7 mg/g creatinine. A fairly close, statistically significant correlation was found between environmental exposure to cyclohexane and postshift urinary 1,2-DIOL and 1,4-DIOL on Monday. Data collected on Thursday and Friday showed only a poor correlation to exposure with a wide scatter. Both metabolites have a urinary half-life of close to 18 h and accumulate during the working week. CONCLUSIONS: Comparison between data obtained from a PBPK model and those found in workers suggests that 1,2-DIOL and 1,4-DIOL are urinary metabolites suitable for the biological monitoring of industrial exposure to cyclohexane.

Cyclohexanes↗

Chromium bond detection in isolated erythrocytes: a new principle of biological monitoring of exposure to hexavalent chromium.

Internal stress to chromium is only relevant in occupational medicine if it is due to the handling of hexavalent chromium. Cr(VI) ions, after uptake by inhalation or percutaneously are carried in the blood plasma and penetrate--depending on the concentration--into the erythrocytes. Due to the intracellular reduction to Cr(III) and the concurrent intracellular protein binding, the erythrocytes represent an easily accessible target organ for quantitative chromium determination after occupational exposure to Cr(VI) compounds. The results of an earlier experimental study indicate that human plasma too is capable of spontaneous reduction of Cr(VI) ions of up to 2 ppm to Cr(III). This plasma reduction capacity (PRC) can be increased and accelerated considerably by adding ascorbic acid (AA). These findings were supported in this investigation by proving a decreased binding of Cr(VI) inside the erythrocytes under the effect of AA. This leads to the assumption that only those Cr(VI) concentrations can penetrate the membrane of the erythrocytes and enter the cell which either come into contact with the membrane during the reduction process or exceed this limit concentration of 2 ppm. Only in these two instances can corresponding chromium findings be analyzed in isolated and washed erythrocytes. These results are compared with those obtained by conventional methods, such as Cr determination in the blood and/or urine. Our findings indicate that a single determination of chromium concentration in the erythrocytes will permit the monitoring of critical cases of Cr(VI) exposure. This is a new type of biological monitoring in the sense of a condensed longitudinal study, in order to find out whether threshold concentrations have been respected over a given period.

Air Pollutants, Occupational↗

Exposure of coke-oven workers to polycyclic aromatic hydrocarbons based on biological monitoring results.

This study assessed the exposure of coke-oven workers to polycyclic aromatic hydrocarbons (PAH) based on two methods of biological monitoring (urinary mutagenicity by Ames test and urinary 1-hydroxypyrene) and concentrations of benzo(a)pyrene in the air collected by personal sampling. The coke-oven workers were classified into four job categories. There was no significant correlation among three methods applied. According to all methods, the workers most exposed to PAH were those working at the topside area of the coke plant, Coke-oven workers excreted significantly more mutagenic substances and 1-hydroxypyrene (HpU) in urine than the controls. Mutagenic substances in urine of coke-oven workers and the control group were detected only with TA98 after metabolic activation. Median HpU levels of coke-oven workers classified into three job categories (A, B, C) exceeded the biological exposure limit. The effect of smoking on urinary mutagenicity and urinary 1-hydroxypyrene was statistically significant.

Air Pollutants, Occupational↗

Biological monitoring of exposure to organophosphorus insecticides by assay of urinary alkylphosphates: influence of protective measures during manual operations with treated plants.

Biological monitoring was carried out by assaying urinary dimethylated alkylphosphates [dimethyldithiophosphate (DMDTP), dimethylthio-phosphate (DMTP), and dimethylphosphate (DMP)] in 11 workers exposed to chlorpyrifos-methyl and azinphosmethyl during operations in a previously sprayed peach orchard. The subjects were divided into groups on the basis of the protective clothing worn. The results were compared with those of a reference group of 99 subjects not occupationally exposed to organophosphorus insecticides. The hand-wash liquid of the workers was also analyzed to evaluate skin contamination. Significantly higher levels of urinary excretion of alkylphosphates were found in all groups than in unexposed controls (Student's t test). A good correlation was found between quantities of the active ingredients on the hands and urinary excretion of total dimethylated alkylphosphates (r = 0.788) and of DMTP (r = 0.749) and DMP (r = 0.790) alone. The correlation between azinphos-methyl on the hands and urinary excretion of DMDTP was poor (r = 0.069). Under the working conditions investigated, the main route of absorption seems to be via the skin. Respiratory absorption, however, also appears significant in view of the difference in urinary excretion of dimethylated alkylphosphates found between subjects with and without face masks.

Adult↗

Biological monitoring of cadmium exposure: reliability of spot urine samples.

Concentration-dilution of spot urine samples is a shortcoming of the biological monitoring of industrial xenobiotics. To ascertain whether the adjustment of urinary cadmium measured in spot samples is appropriate, urine samples were taken three times, once a week for 3 successive weeks, from 25 welders employed in the manufacture of jewelry (total 75 samples). Cadmium, creatinine, specific gravity, total urinary solutes, urinary volume and urinary flow rate were measured in 12-h collections and in spot samples taken immediately afterwards. Creatinine and total urinary solutes showed high inverse correlation with urinary flow rate (r = -0.858 and r = -0.768 respectively). Urinary cadmium displayed a similar trend but the correlation was not significant (r = -0.145). Creatinine adjustment of urinary cadmium values in spot samples increased the correlation with the same index in timed samples adjusted for urinary volume (r = -0.808) or urinary flow rate (r = 0.821) compared with non-adjustment (r = 0.732 and r = 0.738, respectively). Creatinine adjustment of spot sample values is also suitable for a wide range of urinary concentrations; discarding excessively diluted or concentrated urines, correlation of urine samples improved for non-adjusted or specific gravity-adjusted values, whereas no changes were observed for creatinine-adjusted values.

Adolescent↗

Biological monitoring of pyrethroids in blood and pyrethroid metabolites in urine: applications and limitations.

The objective of this study was to perform biological monitoring of subjects who are occupationally exposed to pyrethroids. The study group consisted of 30 pest control operators exposed to cyfluthrin, cypermethrin or permethrin. After exposure, 24-h urine samples were collected and 20 ml of blood was drawn. The pyrethroid metabolites cis- and trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropanecarboxylic acid, 3-phenoxybenzoic acid and fluorophenoxybenzoic acid were determined in the urine samples (limit of detection: 0.5 micrograms/l) by GC MS and the pyrethroids in plasma (limit of detection: 5 micrograms) GC-ECD. The concentrations of metabolites in the urine of the pest control operators ranged between < 0.5 micrograms/l and 277 micrograms/l urine. The concentrations of cyfluthrin, cypermethrin and permethrin in the plasma were below the limits of detection (< 5 micrograms/l). To test if the metabolites are specific for pyrethroid exposure, they were determined in the urine of non-exposed subjects (n = 40). In no case could pyrethroid metabolites be detected. A cyfluthrin elimination experiment showed that cyfluthrin metabolites are eliminated following first-order kinetics (t 1/2 = 6.4 h). Storage experiments demonstrate that frozen urine samples (-21 degrees C) show no significant losses of metabolites within a year. In contrast, pyrethroids stored in plasma are susceptible to further biodegeneration.

Adult↗

Environmental and biological monitoring of polycyclic aromatic hydrocarbons (PAHs) in coke plants and other workplaces.

The authors report the results obtained from environmental and biological monitoring of PAHs at different workplaces. Pyrene and benzo(a)pyrene were chosen as indicators of PAHs pollution and their concentrations were determined in the working environments. Two urinary metabolites of these hydrocarbons: 1-hydroxypyrene and 3-hydroxybenzo(a)pyrene, were also studied. PAHs exposure was studied at the following workplaces: a coke plant, a pyrite mine, a railway tunnel and several earthenware factories. The results obtained at the coke plant showed pyrene and benzo(a)pyrene levels (area samplings) at the battery top in the range 1.42-26.57 micrograms/m3 and 0.39-12.85 micrograms/m3 respectively. At the other workplaces lower concentrations of these two compounds were recorded. In the exposed workers, the trend of urinary 1-hydroxypyrene levels was similar to the results of environmental monitoring. The geometric mean of 1-hydroxypyrene concentrations in the urine of coke plant workers was 5.76 micrograms/g creatinine, whereas in the other exposed subjects no geometric mean values higher than 0.60 microgram/g creatinine were obtained. In the case of occupational exposure to low pyrene and benzo(a)pyrene concentrations (pyrite mine, railway tunnel, earthenware factories) the urinary 1-hydroxypyrene levels were similar to those observed in respectively smoking and non-smoking referents.

Coke↗

Formation of N-nitrosoiminodialkanoic acids and their unsuitability as biological monitors for endogenous nitrosation of dipeptides.

Nitrosation of dipeptides which do not contain imino acids leads to rearrangement and the formation of N-nitrosoiminodialkanoic acids. The optimum conditions for the nitrosation of dipeptides in buffer solutions occur at pH 2.0 (0.8-3.2% yield) and are not significantly catalysed by thiocyanate. In vitro nitrosation in gastric juice resulted in a lower yield. It was demonstrated that under normal gastric conditions, a maximum yield of 0.1 mumol total N-nitrosoiminodialkanoic acids/0.1 mol dipeptide would occur, 0.1 mol representing a typical dietary intake of dipeptide. This corresponds to a total concentration of approximately 20 micrograms/l N-nitrosoiminodialkanoic acids over a 24 h period. However, this figure may be significantly altered due to the fluctuation of nitrosation catalysts and inhibitors in gastric juice. Further studies showed that N-nitrosoiminodialkanoic acids are quantitatively excreted in urine when fed by gavage to rats. However, co-administration of the precursor dipeptide and nitrite resulted in negligible in vivo formation. The presence of N-nitrosoiminodialkanoic acids in normal human urine was not detected. Thus it was concluded that the monitoring of N-nitrosoiminodialkanoic acids in human urine is not a suitable method for biological monitoring of the endogenous nitrosation of dipeptides.

Animals↗

Hydroxyethylvaline adduct formation in haemoglobin as a biological monitor of cigarette smoke intake.

The ethylene oxide adduct formed on the N-terminal valine in haemoglobin was investigated as a biological monitor of tobacco smoke intake. The modified method developed for the determination of the hydroxyethylvaline adduct (HOEtVal) involved reaction of globin with pentafluorophenyl isothiocyanate, extraction of the HOEtVal thiohydantoin product, derivatization of this by trimethylsilylation and quantitation by capillary gas chromatography with selective ion monitoring mass spectrometry using a tetradeuterated internal standard. The method was applied to globin samples from 26 habitual cigarette smokers and 24 non-smokers. There was a significant correlation between cigarette smoke intake as measured by the average number of cigarettes smoked per day and HOEtVal levels (r = 0.537, p less than 0.01). Background levels were found in non-smokers (mean 49.9 pmol/g Hb, range 22-106 pmol/g Hb). Smoking increased these levels by 71 pmol/g Hb/10 cigarettes per day. Cotinine levels in plasma of the smokers were determined by GC-NPD using 2-methyl-4-nitroaniline as internal standard. For non-smokers cotinine was determined by GC-MS selective ion monitoring using d3-methylcotinine as internal standard. There was no correlation between number of cigarettes smoked per day and cotinine levels (r = 0.297, p greater than 0.05) although cotinine was correlated with HOEtVal (r = 0.43, p less than 0.01). The HOEtVal adduct levels thus appear to be a suitable biomonitor for exposure to hydroxyethylating agents in cigarette smoke, reflecting an integrated dose over the erythrocyte lifetime. This is in contrast to plasma cotinine determinations which reflect only the previous day's exposure to nicotine in smoke.

Biomarkers↗

Biological monitoring of occupational exposure to methyl ethyl ketone.

This study was conducted to evaluate the usefulness of three commonly used methods of biological monitoring for worker exposed to methyl ethyl ketone (MEK) under field conditions using blood, breath and urine. Environmental MEK exposures were measured by personal sampling with carbon-felt dosimeters. The correlation coefficient (r) between the time-weighted average (TWA) MEK concentration in air and the MEK concentration in blood collected at the end of the work shift was 0.85. The correlation coefficient between the TWA MEK level in air and the concentration exhaled in the breath of workers at the end of the work shift was 0.71. The end-of-shift urinary MEK excretion correlated best with the environmental concentration (r = 0.89). Correlations became lower after urine samples had been corrected for urinary creatinine (r = 0.83) or specific gravity (r = 0.73). After 8 h exposure to 200 ppm MEK, the corresponding end-of-shift urinary excretion was 5.1 mumol/l or 4.11 mg/g creatinine. This value is higher than that previously found in some studies, the difference probably being due to the physical activities of the present workers and their extensive skin contact with the solvent. The kinetics of inhaled MEK was also studied in eight subjects. Breath and urine samples were collected during the 8-h work shift on 2 consecutive Mondays. The results showed that urinary MEK excretion rose steadily until the end of exposure, whereas the MEK concentration in exhaled air varied markedly throughout the day.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗