Urinary levels of proteins and metabolites in workers exposed to toluene

Summary We measured urinary excretion of albumin and retinol-binding proteins to investigate the occurrence of early renal dysfunction in 45 paint workers exposed principally to toluene, and in the same number of unexposed control subjects matched individually for sex and age. Two biological indicators of personal toluene absorption, namely urine hippuric acid and o-cresol, were also measured in the exposed subjects. A significantly higher level and increased prevalence of elevated retinol-binding protein in the urine of exposed workers was found, whereas no significant difference in urinary albumin concentration was seen between the two groups. Urinary concentrations of retinol-binding protein was correlated (r = 0.399, P < 0.006) with that of o-cresol, but not with hippuric acid or employment duration. The results suggest a dose-dependent early tubular effect due to toluene exposure that might be useful for monitoring individuals exposed to toluene at work.     

甲苯接触者生物监测指标研究——尿中马尿酸和邻甲酚浓度

对32名职业接触甲苯、18名志愿受试者和77名非职业接触甲苯者的尿中马尿酸及邻甲酚的测定,发现在非接触者中,尿中马尿酸存在日间波动,邻甲酚排出量极少。工人和志愿者接触甲苯后,尿中马尿酸即开始上升,到脱离时达高峰,以后迅速下降,4小时左右降到正常本底水平,班末尿中马尿酸浓度与空气浓度相关(工人:r=0.64,志愿者:r=0.78)。尿中邻甲酚在低浓度接触者中,难以检出,但在高浓度接触时,班末尿中邻甲酚与空气浓度相关(工人;r=0.63,志愿者;r=0.65)。马尿酸和邻甲酚作为甲苯生物监测指标可结合使用。     

Effect of ethanol,cimetidine and propranolol on toluene metabolism in man

Summary In a climatic exposure chamber four healthy volunteers were exposed to 100ppm toluene, 100ppm toluene + ethanol, 100ppm toluene + cimetidine, and 100ppm toluene + propranolol for 7h each at random over four consecutive days. A control experiment and 3.5 h of exposure to 200 ppm toluene were also performed. Ethanol inhibited toluene metabolism by 0.5 as expressed by the urinary excretion of two of the metabolites of toluene, namely o-cresol and hippuric acid. In agreement with this, the mean alveolar concentration of toluene was greater by 1.7 during ethanol exposure; 45 min after discontinuation of exposure the increase was by 3.3. Neither cimetidine nor propranolol changed toluene metabolism significantly. The results indicate that ethanol may prolong the time interval in which toluene is retained in the human body in persons simultaneously exposed to ethanol and toluene. When using o-cresol or hippuric acid in biological monitoring of persons occupationally exposed to toluene, the consumption of ethanol should be considered.Supported by grants from the Working Environment Fund, Denmark     

Method for simultaneous determination of six metabolites of toluene, xylene and ethylbenzene, and its application to exposure monitoring of workers in a printing factory with gravure machines

For the biological monitoring of exposure to solvent composed of toluene, xylene, and ethylbenzene used in a printing factory with gravure machines, we developed a HPLC method for the simultaneous determination of urinary metabolites of this solvent, i.e. hippuric acid, o-, m-, and p-methylhippuric acid, mandelic acid and phenylglyoxylic acid. Except for phenylglyoxylic acid, urinary concentrations of the metabolites determined by the present method correlated well with the air concentrations of the respective solvent components. Hence the present method is useful in monitoring solvent exposure. In 91 workers of the printing factory and 53 control subjects, we also determined the concentrations of some phenolic metabolites and confirmed that o-cresol is a useful indicator for monitoring toluene exposure.     

Exposure to toluene increases the urinary excretion of D-glucaric acid.

Workers at a printing plant exposed to low concentrations of toluene (43-401 mg/m3, median 155 mg/m3) had increased urinary D-glucaric acid (3.55-5.12 mmol/mol creatinine) excretion at the end of the shift compared with controls (2.45-3.35 mmol/mol creatinine). No increase was found after the summer holiday (1.92-2.89 mmol/mol creatinine) but excretion had increased two weeks later (4.05-5.55 mmol/mol creatinine). These changes in the excretion of D-glucaric acid were not correlated to levels of exposure, to changes of urinary hippuric acid and o-cresol half lives (three to eight hours), nor to o-cresol/hippuric acid concentration ratios when measured at the end of daily exposure. Since a significant intra and interindividual variability of urinary D-glucaric acid was found in all groups, urinary D-glucaric acid excretion is suitable to monitor group but not individual exposure.     

Exposure to toluene increases the urinary excretion of D-glucaric acid

Workers at a printing plant exposed to low concentrations of toluene (43-401 mg/m3, median 155 mg/m3) had increased urinary D-glucaric acid (3.55-5.12 mmol/mol creatinine) excretion at the end of the shift compared with controls (2.45-3.35 mmol/mol creatinine). No increase was found after the summer holiday (1.92-2.89 mmol/mol creatinine) but excretion had increased two weeks later (4.05-5.55 mmol/mol creatinine). These changes in the excretion of D-glucaric acid were not correlated to levels of exposure, to changes of urinary hippuric acid and o-cresol half lives (three to eight hours), nor to o-cresol/hippuric acid concentration ratios when measured at the end of daily exposure. Since a significant intra and interindividual variability of urinary D-glucaric acid was found in all groups, urinary D-glucaric acid excretion is suitable to monitor group but not individual exposure.     

Effects of smoking and drinking habits on urinary o-cresol excretion after occupational exposure to toluene vapor among chinese workers

The relationship between the time-weighted average intensity of exposure to toluene and o-cresol concentration in shift-end urine was investigated in nearly 500 factory workers of both sexes in China, together with a similar number of nonexposed control subjects. Toluene concentration (25 ppm as geometric mean and 550 ppm as the maximum) was monitored by diffusive sampling using carbon cloth as adsorbent followed by gas chromatographic (GC) analysis. o-Cresol (up to 7 mg/1) was measured by GC after acid hydrolysis of samples. Urinary o-cresol levels correlated significantly (r = 0.69–0.77; p < 0.01) with toluene exposure in men, women and the two sexes in combination, regardless of correction for urine density. When compared with hippuric acid, however, o-cresol was less sensitive as an indicator of exposure to toluene and is not a suitable biological marker for detecting low level toluene exposure. Since urinary o-cresol level was significantly reduced by smoking, drinking, and the two habits combined, it cannot be considered reliable as an indicator of exposure to toluene. © 1994 Wiley-Liss, Inc.     

Exposure of workers to a mixture of toluene and xylenes. I. Metabolism.

The urinary excretion of hippuric acid and methylhippuric acid was studied in workers (233 subjects; 122 men and 111 women) exposed to toluene and xylenes in combination and in non-exposed controls (281 subjects; 141 men and 140 women) recruited from the same factories or factories of the same regions. Smoking and drinking habits of the subjects were obtained by medical interviews. From each worker, one urine sample was collected at the end of a shift and analysed for hippuric and methylhippuric acids by high performance liquid chromatography. Air samples for the estimation of toluene and xylenes were collected with diffusive personal samplers. There was a linear correlation between the time weighted average exposure either to toluene or xylene isomers and the concentrations of hippuric acid or methylhippuric acid isomers in urine. Essentially no difference was found in the correlation between quantitative exposure and excretion in the three xylene isomers. Comparison of the slopes of regression lines indicated the absence of metabolic interaction between toluene and xylenes at the measured concentrations. The metabolism of toluene and xylenes was significantly reduced among smokers or drinkers compared with non-smokers and non-drinkers.     

Mutual metabolic suppression between benzene and toluene in man

Summary The exposure intensity during a shift and the metabolite levels in the shift-end urine were examined in male workers exposed to either benzene (65 subjects; the benzene group), toluene (35 subjects; the toluene group), or a mixture of both (55 subjects; the mixture group). In addition, 35 non-exposed male workers (the control group) were similarly examined for urinary metabolites to define background levels. A linear relationship was established between the intensity of solvent exposure and the corresponding urinary metabolite levels (i.e. phenol, catechol and quinol from benzene, and hippuric acid and o-cresol from toluene) in each case when one of the three exposed groups was combined with the control group for calculation. Comparison of regression lines in combination with regression analysis disclosed that urinary levels of phenol and quinol (but not catechol) were lower in the mixture group than in the benzene group when the intensities of exposure to benzene were comparable, indicating that the biotransformation of benzene to phenolic compounds (excluding catechol) in man is suppressed by co-exposure to toluene. Conversely, metabolism of toluene to hippuric acid was suppressed by benzene co-exposure. Conversion of toluene to o-cresol was also reduced by benzene, but to a lesser extent. The significance of the present findings on the mutual suppression of metabolism between benzene and toluene is discussed in relation to solvent toxicology and biological monitoring of exposure to the solvents.     

Biological monitoring for occupational exposure to toluene

A study was undertaken to examine the relationship between exposure of workers to toluene in the work environment and biological indicators of toluene exposure. The biological indicators studied were toluene in expired air, toluene in blood obtained by the finger prick method, and urinary hippuric acid. The study was undertaken in a factory in Singapore that manufactures speakers for audio systems. A total of 86 female workers exposed to toluene at the workplace and a control group of workers not exposed to toluene were examined. All of them were teetotalers, were nonsmokers, and gave no history of chronic drug usage. The 8-hr time-weighted average exposure level of toluene ranged from 1.6 ppm to 263 ppm. The study showed the expected toluene levels in finger prick blood was 1.4 micrograms/mL after an 8-hr exposure to 100 ppm of toluene. Toluene concentration in expired air of 16 ppm after an 8-hr exposure to 100 ppm compared favorably with other studies. The toluene in blood/expired air ratio was observed to be lower than in other studies. In this study, the expected urinary hippuric acid level for a 100-ppm exposure to toluene was 2.7 g/g creatinine. This level is higher than that recorded in other studies. The results showed that at low levels of toluene, urinary hippuric acid is not a valuable indicator of exposure. Toluene in expired air is the most reliable biological indicator of exposure to toluene.     

Measurement of toluene and xylene metabolites by gas chromatography

Summary A new method for analyzing urinary hippuric and methyl hippuric acids by gas chromatography was developed. As well the direct analysis of hippuric acids as the determination of their alkaline hydrolytic products, benzoic and toluic acids, are described. Results of the two different methods are compared and discussed. The method was applied to workers occupationally exposed to paints containing mainly toluene and xylene. The urinary hippuric acid concentrations were related to the level of toluene and xylene in blood.     

甲苯的慢性危害和尿中的马尿酸作为职业性接触的生物学监测指标探讨

本文对１４０名接触以甲苯为主的有机溶剂的包漆工（平均接触工龄８．３９年）进行了横断面调查，发现头昏、头痛、失眠、乏力、腹隐痛等症状的出现率，神衰综合征和慢性咽炎的患病率明显高放对照组（Ｐ＜０．０５或Ｐ＜０．０１）。常规肝功能和血清碱性磷酸酶（Ｓ－ＡＫＰ）无明显改变（Ｐ＞０．０５）。包漆工班末尿马尿酸平均浓度显著高於班前和对照组班末的平均浓度（分别Ｐ＜０．０５，Ｐ＜０．０１）。男、女包漆工班末尿马尿酸平均浓度均高于班前（分别Ｐ＜０．０５，Ｐ＜０．０１）。空气中苯系物的浓度均在最高允许浓度范围内。结果提示，长期接触低浓度的以甲苯为主的有机溶剂，对中枢神经系统有不利影响。尿中马尿酸水平可作为反映工人接触低浓度甲苯的有用的、生物学监测指标。     

Somatosensory evoked potentials in workers exposed to toluene and styrene.

Somatosensory evoked potentials (SEPs) were used to evaluate possible subclinical impairment of the nervous system due to occupational exposure to toluene and styrene. A group of 36 rotogravure printers with severe exposure to toluene, 20 workers with severe exposure to styrene in a glass laminate manufacturing plant, and a comparison group of healthy subjects were studied. The severity of exposure was documented by measurements of toluene and styrene concentrations in breathing zone air, by hippuric acid concentration in urine in the group exposed to toluene, and by urinary mandelic acid concentration in the group exposed to styrene. Somatosensory evoked potentials were measured by stimulation of the median nerve at the wrist and the tibial nerve at the ankle. Peripheral conduction velocities (CVs) in both extremities and central conduction time (CCT) after tibial nerve stimulation were significantly decreased in both exposed groups. Significantly prolonged latencies of peripheral and cortical SEPs to median nerve stimulation as well as cortical SEPs to tibial nerve stimulation were found in workers exposed to styrene. Some abnormalities in SEPs at peripheral or spinal and cortical levels were found in eight workers exposed to toluene and six workers exposed to styrene. Of these, in three workers exposed to toluene and two to styrene increased CCT and delayed latencies of cortical responses at normal conduction values in the periphery were found. A trend for increased frequency of abnormal SEPs with duration of exposure to toluene and styrene and alcohol abuse was found. Abnormalities in SEPs in the exposed groups are most probably of multifactorial origin. Central SEP abnormalities in both exposed groups could indicate early signs of subclinical dysfunction at spinal and cortical levels and could be due to toluene or styrene exposure probably potentiated by alcohol consumption in the group exposed to toluene.     

Distribution of urinary hippuric acid concentrations by ALDH2 genotype.

OBJECTIVES--To clarify the relation between the genetic polymorphism of ALDH2 (low Km aldehyde dehydrogenase) and toluene metabolism. METHODS--The study subjects were 253 toluene workers (192 men and 61 women with an age range of 18-66). The genotypes of ALDH2 were classified by artificial restriction fragment length polymorphism into the homozygous genotype of normal ALDH2 (NN), the homozygous genotype of an inactive ALDH2 (DD), and the heterozygous genotype of normal and inactive ALDH2 (ND). The concentrations of hippuric acid (HA), the main metabolite of toluene, was determined in urine specimens of 253 toluene workers. The HA measurements in previous occupational health examinations were also referenced. The HA concentrations corrected for creatinine (HA/C) were compared with the biological exposure index (BEI) for toluene, which is 2.5 g/g creatinine. To estimate the toluene exposures, urinary o-cresol concentrations were also determined and compared with another BEI for toluene--that is, 1.0 mg urinary o-cresol/g creatinine. RESULTS--Incidence of each genotype in the toluene workers was almost the same as that in non-exposed controls who lived in the same area as the toluene workers. The incidence of each of the three genotypes also did not differ by smoking habit. Mean urinary HA concentrations were not significantly different in the groups with the different genotypes of ALDH2. The HA concentrations of > 70% of the 890 total samples were < 1.0 g/l. The number of urine samples > 3.0 g/l was 28 (5.4%) in the NN group and 19 (6.4%) in the ND group. No urine samples in the DD group were > 3.0 g/l HA. The distribution of urinary HA in the DD group was significantly different from those in both the NN and ND groups (P < 0.05). Seven (4.9%) of the 136 total specimens in the NN group and four (4.7%) of the 82 total specimens in the ND group exceeded the BEI. There were, however, no urine specimens that exceeded the BEI in the DD group. The maximum HA concentration after correction for creatinine in the DD group was 1.86 g/g creatinine. The percentages of urine specimens in which o-cresol concentrations exceeded this BEI were 14.3% in the NN group, 9.1% in the ND group, and 15.4% in the DD group. Therefore, the exposure rate for all three genotypic groups of workers was almost the same. CONCLUSIONS--The HA concentrations of toluene workers with ALDH2 DD genotype were lower than those of the NN and ND genotypes when they were exposed to relatively high concentrations of toluene. The exposures of the DD group were suspected to be underestimates because they were based on the BEI for the NN genotype.     

Subclinical abnormalities in workers with continuous low-level toluene exposure

Short-term exposure to a high concentration (TWA > 100 ppm) of toluene can cause hepatotocixity and neurotoxicity in humans. Data on the effects of exposure to low levels of toluene, however, are controversial. In addition, few studies on the effects of toluene exposure on the autonomic nervous system have been conducted. Urine samples from 34 male factory workers in Taiwan who were exposed to low levels of toluene either intermittently (n = 13) or continuously (n = 21) were taken on a Monday morning after a 2-day hiatus and at the end of the workweek on Friday evening. Urinary hippuric acid levels were measured using high-performance liquid chromatography (HPLC). A complete blood work-up was also performed for each subject. The prevalence and severity of neurotoxic symptoms were investigated by a self-reported questionnaire, a neuropsychiatric battery, and sympathetic and peripheral nerve function tests. The mean value of urinary hippuric acid corrected for creatinine (Cr) was 0.34 ± 0.18 g/g Cr on Monday morning and 0.43 ± 0.26 g/g Cr on Friday evening. The difference in the mean value of urinary hippuric acid between the two periods (p < 0.01) and the odds ratio of impairment of sympathetic (OR = 4.13, p = 0.11) and peripheral nerves (OR = 6.94, p = 0.074) were higher in workers continuously exposed to toluene. In addition, workers who were continuously exposed to toluene had a lower mean platelet count (216 ± 41 × 10(6) /μL) than workers who were intermittently exposed (252 ± 40 × 10(6)/μL), (p = 0.018). Furthermore, there was a positive relationship between neurological abnormalities and a self-reported neuropsychiatric measurement (r = 0.35-0.66, p < 0.05) in all workers. These data suggest that continuous exposure to low levels of toluene may be associated with sympathetic and peripheral nerve dysfunction and sub-clinical hematological damage. Further research needs to be carried out regarding how chronic exposure to low-levels of toluene affects workers.     

Occupational toluene exposure induces cytochrome P450 2E1 mRNA expression in peripheral lymphocytes

Print workers are exposed to organic solvents, of which the systemic toxicant toluene is a main component. Toluene induces expression of cytochrome P450 2E1 (CYP2E1), an enzyme involved in its own metabolism and that of other protoxicants, including some procarcinogens. Therefore, we investigated the association between toluene exposure and the CYP2E1 response, as assessed by mRNA content in peripheral lymphocytes or the 6-hydroxychlorzoxazone (6OH-CHZ)/chlorzoxazone (CHZ) quotient (known as CHZ metabolic ratio) in plasma, and the role of genotype (5 -flanking region RsaI/PstI polymorphic sites) in 97 male print workers. The geometric mean (GM) of toluene concentration in the air was 52.80 ppm (10-760 ppm); 54% of the study participants were exposed to toluene concentrations that exceeded the maximum permissible exposure level (MPEL). The GM of urinary hippuric acid at the end of a work shift (0.041 g/g creatinine) was elevated relative to that before the shift (0.027 g/g creatinine; p < 0.05). The GM of the CHZ metabolic ratio was 0.33 (0-9.3), with 40% of the subjects having ratios below the GM. However, the average CYP2E1 mRNA level in peripheral lymphocytes was 1.07 (0.30-3.08), and CYP2E1 mRNA levels within subjects correlated with the toluene exposure ratio (environmental toluene concentration:urinary hippuric acid concentration) (p = 0.014). Genotype did not alter the association between the toluene exposure ratio and mRNA content. In summary, with further validation, CYP2E1 mRNA content in peripheral lymphocytes could be a sensitive and noninvasive biomarker for the continuous monitoring of toluene effects in exposed persons.     

Effects of the exposure to polycyclic aromatic hydrocarbons or toluene on thiobarbituric acid reactive substance level in elementary school children and the elderly in a rural area]

OBJECTIVES: Polycyclic aromatic hydrocarbons (PAH) and toluene have been reported to induce reactive oxygen species and oxidative stress. This study was performed to investigate the effects of low level exposure to PAHs or toluene on the lipid peroxidation level in elementary school children and the elderly in a rural area. METHODS: Forty seven elementary school children and 40 elderly people who were living in a rural area and not occupationally exposed to PAH or toluene were the subjects of this study. Information about active or passive smoking and diet was obtained using a self-administered questionnaire. The urinary 1-hydroxypyrene (1-OHP), 2-naphthol, hippuric acid and thiobarbituric acid reactive substance (TBARS) concentrations were measured, and these values were corrected with the urinary creatinine concentration. RESULTS: In school children, the geometric means of the urinary 1-OHP, 2-naphthol, hippuric acid and TBARS levels were 0.02 micromol/mol creatinine, 0.47 micromol/mol creatinine, 0.14 g/g creatinine and 0.95 micromol/g creatinine, respectively. Those values for the elderly were 0.07 micromol/mol creatinine, 1.87 micromol/mol creatinine, 0.11 g/g creatinine and 1.18 micro mol/g creatinine, respectively. The mean levels of urinary 1-OHP, 2-naphthol and TBARS were significantly higher in the elderly subjects than in the children. The urinary TBARS level was not correlated with the urinary 1-OHP, 2-naphthol and hippuric acid, but they were correlated with the age of the subjects. CONCLUSIONS: These results suggest that low level inhalation exposure to PAH or toluene does not markedly increase lipid peroxidation, and age is a significant determinant of lipid peroxidation.     

Excretion of hippuric acid and m- or p-methylhippuric acid in the urine of persons exposed to vapours of toluene and m- or p-xylene in an exposure chamber and in workshops, with specific reference to repeated exposures

Ogata, M., Takatsuka, Y., and Tomokuni, K. (1971).Brit. J. industr. Med.,28, 382-385. Excretion of hippuric acid and m- or p-methylhippuric acid in the urine of persons exposed to vapours of toluene and m- or p-xylene in an exposure chamber and in workshops, with specific reference to repeated exposures. Four male volunteers were exposed to 200 p.p.m. of toluene for five one-hour periods separated by one-hour intervals. The excretion curve of hippuric acid showed multi-peaks, and almost concided with a theoretical curve previously described. The fraction of the toluene absorbed which was accounted for as hippuric acid was only slightly lower than after a single exposure.

In a paint spraying shop exposure was measured both from the concentrations of toluene in the air by a Kitagawa detector and from the exceretion of urinary hippuric acid. The results were in general agreement, with a correlation coefficient of 0·67.

Urinary hippuric acid and methylhippuric acid were determined on urines from two workers in a shipbuilding yard who used paint thinned with toluene and xylene. The concentrations of the acids varied from day to day depending on the kind and the duration of work. From the concentrations found the mean concentrations to which the workers were exposed were calculated as a fraction of the maximum allowable concentration (M.A.C.). One worker was, on this evidence, exposed to more than the combined M.A.C. on three days out of six.

     

Chronic occupational exposure to toluene

Summary Chronic occupational exposure to toluene was studied in a factory preparing tarpaulins. Seventy-eight workers were studied; 46 were exposed to various concentrations of toluene in air (20–200 ppm), 32 were unexposed workers in the same factory. In many cases the exposure had lasted for 10–20 years. The urinary hippuric acid excretion at the end of work shift showed good correlations to toluene concentrations in air, and it seems to be a good measure of exposure. The hippuric acid in urine samples collected overnight showed that elimination of toluene still occurs several hours after exposure. Most of the biological parameters measured showed no correlation to toluene exposure. The blood leukocyte count did show slight positive correlations to toluene exposure, but even this parameter stayed inside the range of normal values. The occurrence of chronic diseases, drug using habits, and drinking and smoking habits did not show any correlations to toluene exposure.This study has been supported by the grant of Y. Jahnsson Foundation in Finland     

Alveolar air and blood toluene concentration in rotogravure workers

Summary Eleven workers exposed to toluene inhalation in a rotogravure plant were examined for the determination of the concentration of toluene in alveolar air, the excretion of urinary hippuric acid, and the concentration of toluene in the blood both before and after the 7-h work shift: the research has shown that these three paramters were correlated.The partition coefficient of toluene for blood and air was between 7 and 15.