Styrene In Blood Research Articles

Analysis of biomonitoring data to assess employer compliance with OSHA's permissible exposure limits for air contaminants.

The Occupational Safety and Health Administration (OSHA) regulates exposures to hazardous chemicals in workplace air. When contemporaneous exposure measurements are unavailable, retrospective analysis of biomarkers could provide valuable information about workers' exposures. Single-compartment pharmacokinetic (PK) models were created to relate the concentration of a chemical in the air to the concentration of the chemical or its metabolite in workers' blood or urine. OSHA utilized the PK models in investigations of three fatal incidents in which workers were exposed to nickel carbonyl, methyl bromide, or styrene. To obtain the minimum plausible estimate of each exposure, OSHA used conservative assumptions about parameters such as workers' inhalation rates, baseline levels of biomarker, and chemicals' volumes of distribution. OSHA analyzed a worker's urinary nickel concentration and concluded that his 8-htime-weighted average exposure to nickel carbonyl was at least 0.06 mg/m3 . Analysis of a worker's postexposure, premortem blood bromide level revealed that his exposure to methyl bromide was at least 181 mg/m3 . Post-mortem blood styrene measurements suggested that a third worker's exposure to styrene was at least 625 mg/m3 . These exposures exceeded OSHA's permissible exposure limits of 0.007 mg/m3 for nickel carbonyl, 80 mg/m3 for methyl bromide, and 426 mg/m3 for styrene. OSHA successfully cited the three employers for violations of chemical exposure limits. Analysis of biomarkers via PK modeling enables retrospective evaluations of workers' acute exposures to hazardous chemicals. These techniques are useful to occupational regulators who assess employer compliance with mandatory exposure limits.

Environmental Styrene Exposure and Sensory and Motor Function in Gulf Coast Residents.

Background:Although styrene is an established neurotoxicant at occupational exposure levels, its neurotoxicity has not been characterized in relation to general population exposures. Further, occupational research to date has focused on central nervous system impairment.Objective:We assessed styrene-associated differences in sensory and motor function among Gulf coast residents.Methods:We used 2011 National Air Toxics Assessment estimates of ambient styrene to determine exposure levels for 2,956 nondiabetic Gulf state residents enrolled in the Gulf Long-term Follow-up Study, and additionally measured blood styrene concentration in a subset of participants 1 to 2 y after enrollment (). Participants completed an enrollment telephone interview and a comprehensive test battery to assess sensory and motor function during a clinical follow-up exam 2 to 4 y later. Detailed covariate information was ascertained at enrollment via telephone interview. We used multivariate linear regression to estimate continuous differences in sensory and motor function, and log-binomial regression to estimate prevalence ratios for dichotomous outcomes. We estimated associations of both ambient and blood styrene exposures with sensory and motor function, independently for five unique tests.Results:Those participants in the highest 25% vs. lowest 75% of ambient exposure and those in the highest 10% vs. lowest 90% of blood styrene had slightly diminished visual contrast sensitivity. Mean vibrotactile thresholds were lower among those in the highest vs. lowest quartile of ambient styrene and the highest 10% vs. lowest 90% of blood styrene ( log microns; 95% CI: , and log microns; 95% CI: , , respectively). The highest vs. lowest quartile of ambient styrene was associated with significantly poorer postural stability, and (unexpectedly) with significantly greater grip strength.Discussion:We observed associations between higher styrene exposure and poorer visual, sensory, and vestibular function, though we did not detect associations with reduced voluntary motor system performance. Associations were more consistent for ambient exposures, but we also found notable associations with measured blood styrene. https://doi.org/10.1289/EHP3954

Determinants of environmental styrene exposure in Gulf coast residents.

Background:In a previous study of exposure to oil-related chemicals in Gulf coast residents, we measured blood levels of volatile organic compounds. Levels of styrene were substantially elevated compared to a nationally representative sample. We sought to identify factors contributing to these levels, given the opportunities for styrene exposure in this community.Methods:We measured blood styrene levels in 667 Gulf coast residents and compared participants’ levels of blood styrene to a nationally representative sample. We assessed personal and environmental predictors of blood styrene levels using linear regression and predicted the risk of elevated blood styrene (defined as above the National Health and Nutrition Examination Survey 95th percentile) using modified Poisson regression. We assessed exposure to styrene using questionnaire data on recent exposure opportunities and leveraged existing databases to assign ambient styrene exposure based on geocoded residential location.Results:These Gulf coast residents were 4–6 times as likely as the nationally representative sample to have elevated blood styrene levels. The change in styrene (log ng/mL) was 0.42 (95% CI: 0.34, 0.51) for smoking, 0.34 (0.09, 0.59) for time spent in vehicles and 1.10 (0.31, 1.89) for boats, and −0.41 (−0.73, −0.10) for fall/winter blood draws. Residential proximity to industrial styrene emissions did not predict blood styrene levels. Ambient styrene predicted elevated blood styrene in subgroups.Conclusions:Personal predictors of increasing blood styrene levels included smoking, vehicle emissions, and housing characteristics. There was a suggestive association between ambient and blood styrene. Our measures of increased regional exposure opportunity do not fully explain the observed elevated blood styrene levels in this population.

Environmental styrene exposure and neurologic symptoms in U.S. Gulf coast residents

BackgroundStyrene is an established neurotoxicant at occupational levels, but effects at levels relevant to the general population have not been studied. We examined the neurologic effects of environmental styrene exposure among U.S. Gulf coast residents. MethodsWe used National Air Toxics Assessment (NATA) 2011 estimates of ambient styrene concentrations to assign exposure levels for 21,962 non-diabetic Gulf state residents, and additionally measured blood styrene concentration in a subset of participants (n = 874). Neurologic symptoms, as well as detailed covariate information, were ascertained via telephone interview. We used log-binomial regression to estimate prevalence ratios (PR) and 95% confidence intervals (95% CI) for cross-sectional associations between both ambient and blood styrene levels and self-reported neurologic symptoms. We estimated associations independently for ten unique symptoms, as well as for the presence of any neurologic, central nervous system (CNS), or peripheral nervous system (PNS) symptoms. We also examined heterogeneity of associations with estimated ambient styrene levels by race and sex. ResultsOne-third of participants reported at least one neurologic symptom. The highest quartile of estimated ambient styrene was associated with one or more neurologic (PR, 1.12; 95% CI: 1.07,1.18), CNS (PR, 1.17; 95% CI: 1.11,1.25), and PNS (PR, 1.16; 95% CI: 1.09,1.25) symptom. Results were less consistent for biomarker analyses, but blood styrene level was suggestively associated with nausea (PR, 1.78; 95% CI: 1.04, 3.03). In stratified analyses, we observed the strongest effects among non-White participants. ConclusionsIncreasing estimated ambient styrene concentration was consistently associated with increased prevalence of neurologic symptoms. Associations between blood styrene levels and some neurologic symptoms were suggestive. Environmental styrene exposure levels may be sufficient to elicit symptomatic neurotoxic effects.

Effects of Styrene-metabolizing Enzyme Polymorphisms and Lifestyle Behaviors on Blood Styrene and Urinary Metabolite Levels in Workers Chronically Exposed to Styrene.

The aim of this study was to investigate whether genetic polymorphisms of CYP2E1, GSTM1, and GSTT1 and lifestyle habits (smoking, drinking, and exercise) modulate the levels of urinary styrene metabolites such as mandelic acid (MA) and phenylglyoxylic acid (PGA) after occupational exposure to styrene. We recruited 79 male workers who had received chronic exposure in styrene fiberglass-reinforced plastic manufacturing factories. We found that serum albumin was significantly correlated with blood styrene/ambient styrene (BS/AS), urinary styrene (US)/AS, and US/BS ratios as well as urinary metabolites, that total protein correlated with US/MA and US/PGA ratios, and that low density lipoprotein (LDL)-cholesterol significantly correlated with US/BS, US/MA, and US/PGA ratios. Multiple logistic regression analyses using styrene-metabolizing enzyme genotypes and lifestyle habits as dependent variables and blood and urine styrene concentrations and urine styrene metabolite levels as independent variables revealed that CYP2E1*5 was associated with the MA/US ratio and GSTM1 with US/BS, that a smoking habit was associated with US/AS and MA/US ratios and MA and PGA levels, and that regular exercise was correlated with PGA/US. In conclusion, the results suggested that genetic polymorphisms of styrene-metabolizing enzymes, lifestyle behaviors, and albumin and LDL-cholesterol serving as homeostasis factors together are involved in styrene metabolism.

Biomonitoring for Exposure Assessment to Styrene in the Fibreglass Reinforced Plastic Industry: Determinants and Interferents.

Fifty-eight workers exposed to styrene were monitored in four fibreglass reinforced plastic industries of Central Italy. The aim of the study was to explore the factors that can influence the levels of styrene exposure biomarkers of the workers and the aspects that might interfere with the exposure assessment measures, such as the co-exposure to acetone. Personal monitoring of professional exposure to airborne styrene and acetone was carried out by Radiello samplers and GC/MS analysis. Biological monitoring was performed by the determination of urinary metabolites, mandelic (MA), and phenylglyoxylic (PGA) acids with HPLC/MS/MS and unmetabolized styrene in saliva and venous blood by HS/GC/MS. The median values of the four sites ranged between 24.1 to 94.0mg m(-3) and 7.3 to 331.1mg g(-1) creatinine for airborne styrene and MA + PGA, respectively. A good linear correlation was found between styrene in air and its urinary metabolites (r = 0.854). The median value for airborne styrene was found to exceed the (Threshold Limit Value - Time Weighted Average) of 85 mg m(-3) in one site for all the workers and in two if only moulders are considered. The multiple linear regression model showed that the determinants of urinary MA + PGA excretion were the type of process, workers' tasks, level of acetone co-exposure, and the use of respiratory protection devices. Data show that the simultaneous exposure to acetone modify the styrene metabolism with a reduction in the levels of (MA + PGA) excreted. A significant linear log-correlation was found between salivary levels of styrene and blood concentration (r = 0.746) sampled at the same t x time.

Using population physiologically based pharmacokinetic modeling to determine optimal sampling times and to interpret biological exposure markers: The example of occupational exposure to styrene

BackgroundBiomonitoring of chemicals in the workplace provides an integrated characterization of exposure that accounts for uptake through multiple pathways and physiological parameters influencing the toxicokinetics. ObjectivesWe used the case of styrene to (i) determine the best times to sample venous blood and end-exhaled air, (ii) characterize the inter-individual variability in biological levels following occupational exposure and (iii) propose biological limit values using a population physiologically based pharmacokinetic (PBPK) model. MethodsWe performed Monte Carlo simulations with various physiological, exposure and workload scenarios. Optimal sampling times were identified through regression analyses between levels in biological samples and 24-h area under the arterial blood concentration vs. time curve. We characterized the variability in levels of styrene in biological samples for exposures to a time weighted average (TWA) of 20ppm. ResultsSimulations suggest that the best times to sample venous blood are at the end of shift in poorly ventilated workplaces and 15min after the shift in highly ventilated workplaces. Exhaled air samples are most informative 15min after the shift. For a light workload, simulated styrene levels have a median (5th–95th percentiles) of 0.4mg/l (0.2–0.6) in venous blood at the end of shift and 0.5ppm (0.3–0.8) in exhaled air 15min after the end of shift. ConclusionThis study supports the current BEI® of the ACGIH of 0.2mg/l of styrene in venous blood at the end of shift and indicates a biological limit value of 0.3ppm in end-exhaled air 15min after the end of shift.

Health hazards among workers in plastic industry

Styrene is a basic building block for manufacturing thousands of products throughout the world. The present study aimed to (1) detect the presence of styrene and/or its metabolites in the workers in one of the Egyptian plastic factories; (2) demonstrate some common health effects of styrene exposure among the same group by some laboratory investigations and compare them with the unexposed healthy individuals; and (3) correlate the duration of styrene exposure and its level in the blood with the severity of the demonstrated health effects. This study was conducted in one of Egyptian plastic factories. The exposed group was 40 male workers, ranging in age from 18 to 33 years (23.20 ± 4.09), working 12 h/day with 1 day off, and working without any protective equipment. A control group of 50 unexposed healthy males matched with the exposed group for age (21-35 yrs (23.40 ± 4.05)), sex, socioeconomic status, and smoking habit is selected. Written individual consent is obtained from all participants followed by (a) a full medical and occupational history and full clinical examination; (b) ventilatory function tests: forced vital capacity (FVC), slow vital capacity, forced expiratory volume in the 1st second (FEV₁)%, FEV₁/FVC%, peak expiratory flow, and mid-expiratory flow 25-75%; (c) analyses of β₂ microglobulin; blood styrene level; and urinary mandelic acid; and (d) cytogenetic study. The study results showed a statistically significant difference between the exposed and the control groups as regard the blood styrene level, urinary mandelic acid level, β₂ microgloblin in urine, and chromosomal study. The study also showed a statistically significant correlation between the duration of styrene exposure and ventilatory function parameters, also between the duration of styrene exposure and some detectable chromosomal aberrations. Our study recommends the implementation of preemployment and periodic medical examinations and health education programs using personal protective equipments and following the recommended allowable concentrations of styrene exposure.

Low level occupational exposure to styrene: Its effects on DNA damage and DNA repair

The present study aimed to evaluate the effects of styrene exposure at levels below the recommended standards of the Threshold Limit Value (TLV-TWA(8)) of 20 ppm (ACGIH, 2004) in reinforced-fiberglass plastics workers. Study subjects comprised 50 exposed workers and 40 control subjects. The exposed workers were stratified by styrene exposure levels, i.e. group I (<10 ppm, <42.20 mg/m(3)), group II (10-20 ppm, 42.20-84.40 mg/m(3)), and group III (>20 ppm, >84.40 mg/m(3)). The mean styrene exposure levels of exposed workers were significantly higher than those of the control workers. Biomarkers of exposure to styrene, including blood styrene and the urinary metabolites, mandelic acid (MA) and phenylglyoxylic acid (PGA), were significantly increased with increasing levels of styrene exposure, but were not detected in the control group. DNA damage, such as DNA strand breaks, 8-hydroxydeoxyguanosine (8-OHdG), and DNA repair capacity, were used as biomarkers of early biological effects. DNA strand breaks and 8-OHdG/10(5)dG levels in peripheral leukocytes of exposed groups were significantly higher compared to the control group (P<0.05). In addition, DNA repair capacity, determined by the cytogenetic challenge assay, was lower in all exposed groups when compared to the control group (P<0.05). The expression of CYP2E1, which is involved in styrene metabolism, in all styrene exposed groups, was higher than that of the control group at a statistically significant level (P<0.05). Levels of expression of the DNA repair genes hOGG1 and XRCC1 were significantly higher in all exposed groups than in the control group (P<0.05). In addition to styrene contamination in ambient air, a trace amount of benzene was also found but, the correlation between benzene exposure and DNA damage or DNA repair capacity was not statistically significant. The results obtained from this study indicate an increase in genotoxic effects and thus health risk from occupational styrene exposure, even at levels below the recommended TLV-TWA(8) of 20 ppm.

Biomarkers of nucleic acid oxidation, polymorphism in, and expression of, hOGG1 gene in styrene-exposed workers

This study investigated nucleic acid oxidation associated with styrene exposure, mRNA expression levels of hOGG1 gene and the role of the genetic polymorphism Ser326Cys of human 8-oxoguanine DNA N-glycosylase 1 ( hOGG1) in 60 styrene-exposed workers and 50 unexposed clerks. Biomarkers of exposure (styrene in blood, mandelic and phenylglyoxylic acids and 4-vinylphenol in urine) and urinary biomarkers of nucleic acid oxidation, namely 8-oxo-7,8-dihydro-2′-deoxyguanosine (U-8-oxodGuo), 8-oxo-7,8-dihydroguanosine (U-8-oxoGuo) and 8-oxo-7,8-dihydroguanine (U-8-oxoGua) were determined by liquid chromatography–tandem mass spectrometry. The levels of 8-oxodGuo adduct and 2′-deoxyguanosine (dGuo) were measured by HPLC in DNA from white blood cells (WBC). Genomic DNA and RNA from blood samples were used to characterize the Ser326Cys polymorphism and the mRNA expression levels of the hOGG1 gene, respectively, by PCR-based methods. Exposed workers showed lower values of 8-oxodGuo/10 5 dGuo ratio in WBC-DNA but higher concentrations of U-8-oxoGuo compared to controls ( p = 0.002 and p = 0.008, respectively, t-test for independent samples). In the whole group, all urinary biomarkers of nucleic acid oxidation correlated with both the sum of mandelic and phenylglyoxylic acids ( rho > 0.33, p < 0.0001) and 4-vinylphenol ( rho > 0.29, p < 0.001), whereas 8-oxodGuo/10 5 dGuo in WBC showed a negative correlation with exposure parameters ( rho < −0.24, p < 0.02). Subjects bearing the hOGG1 Ser/Ser genotype showed lower values of 8-oxodGuo/10 5 dGuo in WBC than those with at least one variant Cys allele (0.34 ± 0.16 vs 0.45 ± 0.21, p = 0.008). In the subgroup of hOGG1 Ser/Ser subjects, laminators showed lower levels of WBC 8-oxodGuo/10 5 dGuo ratio and significantly higher concentrations of U-8-oxoGua than controls ( p = 0.07 and p = 0.01, respectively, t-test for independent samples). Interestingly, workers showed higher levels of hOGG1 expression compared to controls ( p < 0.0005). Styrene exposure seems to be associated with oxidation damage to nucleic acids, particularly to RNA and with an induction of the BER system.

Trends in Occupational Exposure to Styrene in the European Glass Fibre-Reinforced Plastics Industry

Aim: This study presents temporal trends of styrene exposure for workers in the European glass fibre-reinforced plastics (GRP) industry during the period 1966–2002.Methods: Data of personal styrene exposure measurements were retrieved from reports, databases and peer-reviewed papers. Only sources with descriptive statistics of personal measurements were accepted. The styrene exposure data cover personal air samples and biological monitoring data, that is, urinary styrene metabolites (mandelic acid and/or phenylglyoxylic acid) and styrene in blood. Means of series of measurements were categorized by year, country, production process, job and sampling strategy. Linear mixed models were used to identify temporal trends and factors affecting exposure levels.Results: Personal exposure measurements were available from 60 reports providing data on 24145 1–8-h time-weighted average shift personal air samples. Available data of biological exposure indicators included measurements of mandelic acid in post-shift urine (6361 urine samples being analysed). Trend analyses of the available styrene exposure data showed that the average styrene concentration in the breathing zone of open-mould workers in the European GRP industry has decreased on average by 5.3% per year during the period 1966–1990 and by only 0.4% annually in the period after 1990. The highest exposures were measured in Southern Europe and the lowest exposures in Northern Europe with Central Europe in between. Biological indicators of styrene (mandelic acid in post-shift urine) showed a somewhat steeper decline (8.9%), most likely because urine samples were collected in companies that showed a stronger decrease of styrene exposure in air than GRP companies where no biological measurements were carried out.

Association Between Genetic Polymorphisms in Styrene-Metabolizing Enzymes and Biomarkers in Styrene-Exposed Workers*

Single-nucleotide polymorphisms (SNP) in genes of styrene-metabolizing enzymes could modulate biomarker concentrations in blood or urine after exposure to styrene. Ten SNP were analyzed to study their influence on styrene-specific biomarkers in 89 workers of a fiber-reinforced plastic boat building factory. The internal styrene body burden was analyzed in post-shift blood and urine samples. External styrene exposure was measured by passive samplers. Spearman rank correlations between styrene exposure and biomarkers were calculated and distributions of biomarkers were checked for lognormality. Mixed linear models were applied to analyze the influence of genotypes and styrene exposure, on styrene in blood (Monday and Thursday post-shift) and on phenyglyoxylic acid (PGA; adjusted for day of measurement, Monday to Thursday) due to a lognormal distribution, smoking (current, not current), and use of respirators. Stratified analyzes for workers without and with different types of respirators were also performed. The models of both the subgroups revealed a significant influence dependent on the respirator type that workers used for inhalation protection. An influence of the external styrene concentration on the urinary PGA concentration was not observed. After implementation of the SNP into the model significant lower adjusted means of urinary PGA concentrations were found for GSTP1 105IleVal and CYP2E1 –71TT. For styrene levels in blood no significant effect was observed. A significant influence on styrene levels in blood was correlated with external styrene concentration only in workers without use of respirators. The effects of two SNP on urinary PGA decrease indicated a limited modulating SNP effect. The most effective prevention for styrene exposure was obtained with the wearing of respirators.

Comparison of genotoxic potency of styrene 7,8-oxide with radiation and human cancer risk estimation of styrene using the rad-equivalence approach

Styrene is suspected to cause lympho-hematopoietic malignancies through the formation of styrene 7,8-oxide. However, we are still unable to calculate the cancer risk for workers exposed to styrene using epidemiological data. The aims of this study were to determine the blood dose after styrene exposure and to compare the genotoxic potency of styrene 7,8-oxide and gamma radiation in order to calculate the cancer risk by means of the rad-equivalence approach. Leucocytes of 20 individuals were exposed to 0, 0.1, 0.2 or 0.3 mM styrene 7,8-oxide (1 h) or 1, 2 or 3 gray (=100, 200, 300 rad) gamma radiation. Genotoxicity was evaluated with the cytokinesis-block micronucleus assay. Comparison of the two slopes of the regression lines between micronuclei and dose revealed a genotoxic potency for styrene 7,8-oxide of 37 rad/mMh, corresponding with a median value derived from mutagenicity studies (1, 37, 208 rad/mMh). At exposure levels of 1 ppm styrene, a blood styrene 7,8-oxide concentration between 0.03 x 10(-)(6) and 0.42 x 10(-)(6) mM is to be expected using data of toxicokinetic models and human exposure studies. With the cancer risk per unit dose of gamma radiation as benchmark, we calculated a lifetime risk of acquiring a fatal lympho-hematopoietic cancer of 0.17 in 10(3) workers (between 0.037 x 10(-)(3) and 5.0 x 10(-)(3)) exposed to 20 ppm styrene during 40 years.

An Improved Approach for Accurate Quantitation of Benzene, Toluene, Ethylbenzene, Xylene, and Styrene in Blood

Widespread exposure to benzene, toluene, ethylbenzene, xylene, and styrene (BTEXS) and the potential for this exposure to cause health effects drives the need to develop improved methods for measuring exposure. In this work, we demonstrate our latest assay for quantifying BTEXS in blood and characterize sources of both positive and negative biases. This method involves blood sample collection using common techniques followed by static headspace sampling using solid-phase microextraction and gas chromatography/mass spectrometry analysis. We found that the greatest and unexpected source of positive bias was from contamination of butyl rubber materials used in sample preparation consumables such as Vacutainer stoppers, syringe plungers, and sample vial septa. Conversely, the primary cause of negative bias observed was from the diffusion loss of BTEXS from blood during transfer into sample vials. By minimizing or eliminating these and other sources of bias, we improved method accuracy and precision to within 10% while maintaining low-picogram per milliliter detection. Furthermore, upon comparison of these results with those from other laboratories, we observe substantially lower blood BTEXS levels reported to date for nonoccupationally exposed nonsmokers. A relatively unbiased method, as such, will help elucidate any potential associations between adverse health effects and human exposure to low levels of BTEXS.

Predictors of occupational exposure to styrene and styrene-7,8-oxide in the reinforced plastics industry

Objective: To identify demographic and work related factors that predict blood levels of styrene and styrene-7,8-oxide (SO) in the fibreglass reinforced plastics (FRP) industry. Methods: Personal breathing-zone air samples and...

Temporal association between serum prolactin concentration and exposure to styrene

Background: Previous studies have suggested that occupational exposure to styrene is associated with increased serum levels of the anterior pituitary hormone prolactin (PRL). Aims: To test the hypotheses that: (1)...

Analysis of Styrene and its Metabolites in Blood and Urine of Workers Exposed to both Styrene and Acetone

A purge-and-trap gas chromatographic (PT-GC) method for determining styrene concentrations in urine and blood samples has been used in the biological monitoring of workers exposed to styrene and acetone. Blood and urine samples were collected from 34 individuals exposed to both solvents at the end of a 4-h shift and measured for styrene in urine (Su), blood (Sb), and the two major urinary metabolites, mandelic acid (MA) and phenylglyoxylic acid (PGA). A second urine sample was taken at the beginning of the next shift. Environmental exposure was measured using passive personal monitoring and GC. Urinary excretion of MA and PGA was measured by high-performance liquid chromatography. The average exposures to styrene and acetone were 70.5 mg/m3 and 370.5 mg/m3, respectively. In end-of-shift samples there was a significant correlation between concentrations of Su and Sb and the metabolites PGA, MA (r = 0.714 and 0.788, p < 0.001 for Su and r = 0.644 and 0.566, p < 0.005 for Sb). A high correlation between Sb and Su (r = 0.732, p < 0.001) also existed. Poor correlations were found between Su and metabolites in samples collected at the beginning of the next shift (r = 0.491 and 0.474 for PGA and MA, respectively, p < 0.05). There was a better correlation between the biological parameters at the end of the shift and the environmental styrene (r = 0.841 for PGA, r = 0.834 for MA, r = 0.788 for Su, and r = 0.698 for Sb; p < 0.001) compared with those at the start of the shift (r = 0.81 for PGA, 0.675 for MA, and 0.650 for Su; p < 0.001). We found that the concentration of excreted metabolites decreased significantly when environmental concentrations of acetone increased (p < 0.05), particularly at the end of the shift. Although the best correlation with environmental styrene was obtained with the sum of PGA and MA at the end of the shift (r = 0.862, p < 0.001), urine and blood styrene were shown to be more useful biological monitoring indicators because their concentrations were not affected by acetone co-exposure.

DNA adducts, strand breaks and micronuclei in mice exposed to styrene by inhalation

Genotoxic and clastogenic effects of styrene were studied in mice. Male NMRI mice were exposed by inhalation to styrene in concentrations of 750 and 1500 mg/m 3 for 21, 7, 3 and 1 days (6 h/day, 7 days/week). Followed parameters included styrene in blood, specific styrene oxide (SO) induced DNA adducts, DNA strand breaks and micronuclei. The formation of SO induced 7-SO-guanines and 1-SO-adenines in DNA was analysed from lung tissues by two versions of the 32P-postlabelling technique. In lungs after 21 days of exposure to 1500 mg/m 3 the level of 7-SO-guanine was 23.0±11.9 adducts/10 8 normal nucleotides, while 1-SO-adenine was detected at the levels of 0.6±0.2 adducts/10 8 normal nucleotides. Both 7-SO-guanines and 1-SO-adenines strongly correlated with exposure parameters, particularly with styrene concentration in blood ( r=0.875, P=0.0002 and r=0.793, P=0.002, respectively). DNA breaks were measured in peripheral lymphocytes, bone marrow cells and liver cells using comet assay. To discern oxidative damage and abasic sites, endonuclease III was used. In bone marrow of exposed mice slight increase of strand breaks can be detected after 7 days of inhalation. A significant increase was revealed in the endonuclease III-sensitive sites after 21 days of inhalation in bone marrow. In the liver cells inhalation exposure to both concentrations of styrene did not virtually affect either levels of DNA single-strand breaks or endonuclease III-sensitive sites. The inhalation of 1500 mg/m 3 of styrene induced significant increase of micronuclei after 7 days of exposure (10.4±2.5/1000 cells, i.e. twice higher micronuclei frequency than in controls). After 21 days of inhalation no significant difference between the control group and the two exposed groups was observed. Whether the decrease of micronuclei after 21 days of inhalation was due to the inhibition of cell proliferation caused by styrene or due to the natural eliminaton of chromatide fragments, remains to be clarified. An interesting link has been found between DNA single-strand breaks in bone marrow and frequencies of micronuclei ( r=0.721, P=0.028).

Association between genetic polymorphisms and biomarkers in styrene-exposed workers

A comprehensive approach to evaluate genotoxic effects induced by styrene exposure was employed in 44 hand-lamination workers in comparison with 18 unexposed controls. The acquired data on single-strand breaks in DNA (SSBs), frequency of chromosomal aberrations and HPRT mutant frequency in peripheral blood lymphocytes were compared to the results on genotyping of some of the xenobiotic-metabolising enzymes (CYP1A1, CYP2E1, epoxide hydrolase and GSTM1, GSTP1 and GSTT1). Multifactorial regression analysis indicated that SSB in DNA were significantly associated with styrene exposure and with heterozygosity in CYP2E1 (5′-flanking region and intron 6; r 2=0.614). The frequency of chromosomal aberrations (CA), as analysed by linear multiple regression analysis, significantly correlated with years of employment ( P=0.004) and with combinations of epoxide hydrolase ( EPHX) genotypes (exon 3, Tyr/His and exon 4, His/Arg), where individuals with low and medium activity EPHX genotypes exhibited higher frequencies of CA than those with high activity genotypes ( P=0.044, r 2=0.563). Moderately higher HPRT mutant frequencies were detected in styrene-exposed individuals (20.2±25.8×10 −6) as compared to controls (13.3±6.3×10 −6), but this difference was not significant. ANOVA (in the whole set of data) revealed that mutant frequencies at the HPRT gene were significantly associated with years of employment ( F=6.9, P=0.0001), styrene in blood ( F=10.1, P=0.0001), and heterozygosity in CYP2E1 (intron 6; F=13.5, P=0.0008) and GSTP1 (exon 5; F=3.6, P=0.038). In conclusion, our present data suggest that analysed biomarkers of DNA damage may be modulated by polymorphic CYP2E1, EPHX and GSTP1. In our study, styrene-specific DNA and haemoglobin adducts are under investigation. Completing these data with the results of genotyping of metabolising enzymes may provide a useful tool for individual genotoxic risk assessment.

Determination of styrene and styrene-7,8-oxide in human blood by gas chromatography–mass spectrometry

Methods of isotope-dilution gas chromatography–mass spectrometry (GC–MS) are described for the determination of styrene and styrene-7,8-oxide (SO) in blood. Styrene and SO were directly measured in pentane extracts of blood from 35 reinforced plastics workers exposed to 4.7–97 ppm styrene. Using positive ion chemical ionization, styrene could be detected at levels greater than 2.5 μg/l blood and SO at levels greater than 0.05 μg/l blood. An alternative method for measurement of SO employed reaction with valine followed by derivatization with pentafluorophenyl isothiocyanate and analysis via negative ion chemical ionization GC–MS–MS (SO detection limit=0.025 μg/l blood). The detection limits for SO by these two methods were 10–20-fold lower than gas chromatographic assays reported earlier, based upon either electron impact MS or flame ionization detection. Excellent agreement between the two SO methods was observed for standard calibration curves while moderate to good agreement was observed among selected reinforced plastics workers ( n=10). Levels of styrene in blood were found to be proportional to the corresponding air exposures to styrene, in line with other published relationships. Although levels of SO in blood, measured by the direct method, were significantly correlated with air levels of either styrene or SO among the reinforced plastics workers, blood concentrations were much lower than previously reported at a given exposure to styrene. The two assays for SO in blood appear to be unbiased and to have sufficient sensitivity and specificity for applications involving workers exposed to styrene and SO during the manufacture of reinforced plastics.