Urinary 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 as a test of exposure

Ogata, M., Tomokuni, K., and Takatsuka, Y. (1970).Brit. J. industr. Med.,27, 43-50. Urinary 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 as a test of exposure. Twenty-three male volunteers were exposed in groups of four or five to toluene and m- and p-xylene vapour for periods of 3 hours or of 7 hours with one break of an hour. Urine was collected at hourly intervals for several hours, and thereafter all urine was collected until 18 hours after the end of the exposure period, and was analysed for hippuric and methylhippuric acids. It was shown that hippuric acid was excreted equivalent to 68% of the toluene absorbed, and m-methylhippuric acid equivalent to 72% of the m-xylene absorbed. Up to hydrocarbon concentrations of 200 ppm the total quantity of hippuric acids excerted was proportional to the total exposure (ppm × hours). In descending order of precision the following were also related to exposure: rate of excretion during the exposure period; concentrations of hippuric acid in urine corrected to constant urine density; and concentrations in urine uncorrected for density. The last could not be used to calculate exposure, but the others could be to give screening tests to show whether workmen could have been exposed to concentrations greater than the maximum allowable.

The effects of exposure on blood pressure, pulse rate, flicker value, and reaction time were measured. There were some variations which suggested that the MAC of toluene should be set higher than 200 ppm.

     

Quantitative determination in urine of hippuric acid and m- or p-methylhippuric acid, metabolites of toluene and m- or p-xylene

Ogata, M., Tomokuni, K., and Takatsuka, Y. (1969).Brit. J. industr. Med.,26, 330-334. Quantitative determination in urine of hippuric acid and m- or p-methylhippuric acid, metabolites of toluene and m- or p-xylene. Improved and more specific methods for the quantitative determination of hippuric and m- and p-methylhippuric acids in urine are described. The acids were extracted from urine with ether/ethanol, which was dried with silica gel, or with ethyl acetate. After removing the solvent by evaporation coloured azlactones were formed by reaction with p-dimethylaminobenzaldehyde in acetic anhydride (DAB, only usable after ether/ethanol extraction) or benzenesulphonyl chloride in pyridine, and the absorbances were measured. The sensitivities were about 4 μg./ml. of urine using DAB, and 20 μg./ml. of urine using benzenesulphonyl chloride reagent. Separation of hippuric and methylhippuric acids was achieved by paper and thin-layer chromatography before estimation. A spot test is also described.     

Significance of urinary hippuric acid determination as an index of toluene exposure

Ikeda, Masayuki, and Ohtsuji, Hatsue (1969).Brit. J. industr. Med.,26, 244-246. Significance of urinary hippuric acid determination as an index of toluene exposure. Urine samples from 118 male workers in photogravure printing factories were analysed for hippuric acid. The urinary levels of hippuric acid were proportional to the environmental concentrations of toluene, although within wide variations. The urinary concentration of hippuric acid corresponding to 200 p.p.m. of toluene was 3·5 g./litre (specific gravity 1·016) or 4·3 g./g. creatinine.     

Hippuric acid, phenol, and trichloroacetic acid levels in the urine of Japanese subjects with no known exposure to organic solvents

Ikeda, M., and Ohtsuji, H. (1969).Brit. J. industr. Med.,26, 162-164. Hippuric acid, phenol, and trichloroacetic acid levels in the urine of Japanese subjects with no known exposure to organic solvents. Urine samples from 36 male and 30 female university students and 31 male factory workers with no known exposure to industrial organic solvents were analysed for hippuric acid, phenol, and trichloroacetic acid, which are the major metabolites of toluene, benzene, and trichloroethylene respectively. The normal levels were less than 1·4 g./l. for hippuric acid, less than 80 mg./l. for phenol, and less than 1 mg./l. for trichloroacetic acid. No evidence was obtained to suggest that correction for urine concentration with either specific gravity or creatinine concentration minimizes individual variation of metabolite levels.     

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.     

A new derivatization procedure for the analysis of hippuric acid and m-methyl-hippuric acid by gas chromatography

Summary The industrial solvents, toluene and xylene, have physicochemical properties that can be hazardous to the workers exposed. Since hippuric acid and m-methyl-hippuric acid represent the products of toluene and xylene biotransformation in urine, they are used as biological markers in studies on occupational exposure to these solvents. Several methods have been used to determine hippuric acid and m-methyl-hippuric acid —either based on gas chromatography or on high-performance liquid chromatography. In this study we propose the derivatization of hippuric acid and methyl-hippuric acid using methanol in acid medium (HCl), a low-cost reagent with a low level of toxicity. The method has been routinely used in our laboratory for 1 year and has proven to be a reliable procedure for the biological control of occupational exposure to toluene and/or xylene.     

A rapid colorimetric method for the determination of phenylglyoxylic and mandelic acids: Its application to the urinalysis of workers exposed to styrene vapour

Ohtsuji, H., and Ikeda, M.(1970).Brit. J. industr. Med.,27, 150-154. A rapid colorimetric method for the determination of phenylglyoxylic and mandelic acids. Its application to the urinalysis of workers exposed to styrene vapour. A rapid colorimetric method has been developed for the determination of the styrene metabolites, phenylglyoxylic and mandelic acids, in urine. Ether extracts of acidified urine containing the two acids were evaporated to dryness in a test tube, and a mixture of sulphuric acid and formalin (100:1, v/v) was added for colour development. Factors necessary to convert optical extinction to gravimetric units were determined. When urinesamples from workers exposed to up to 30 ppm of styrene were analysed for phenylglyoxylic and mandelic acids together with hippuric acid, it was found that the phenylglyoxylic acid level provided the most sensitive index of styrene exposure and that the optical extinction at 350 nm was practically proportional to phenylglyoxylic acid concentration. No significant increase in hippuric acid levels was observed.     

Urinary levels of proteins and metabolites in workers exposed to toluene

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 less than 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.     

Hippuric acid and ortho-cresol as biological indicators of occupational exposure to toluene

Industrial exposure to toluene was studied in a group of 18 subjects working in a printing plant, exposed only to this solvent. Environmental monitoring was carried out using personal samplers for the whole work-shift. Urine samples were collected for the determination of hippuric acid and ortho(o)-cresol before toluene exposure, at the end of the work-shift, and 5, 9, and 17 h after the end of the work-shift. The values of two metabolites in all the urinary samples were corrected for g creatinine and specific gravity (1.024). Toluene time weighted average (TWA) concentrations ranged from 51 to 221 mg/m³ (7-h samples; two samplings lasting 3.5 h each). Urinary hippuric acid and o-cresol values at the end of the work-shift were significantly higher than the prework-shift values. Both hippuricuria and o-cresoluria end-of-work-shift values, corrected for creatinine and specific gravity, were significantly related to the mean daily environmental concentration of toluene, the correlation being weaker for o-cresol. Correlation coefficients were 0.88 and 0.84 for hippuric acid and 0.63 and 0.62 for o-cresol after correction for creatinine and specific gravity, respectively. No significant relationship was observed between environmental exposure and the values of the two urinary metabolites 5, 9, and 17 h after the end of the work-shift. Extrapolated values from the linear regression analysis at 375 mg/m³ were in good agreement with the biological exposure index (BEI) suggested by ACGIH for hippuric acid. We conclude that determination of hippuric acid in urine samples collected at the end of the work-shift can be used for routine biological monitoring of exposure to toluene, even at low levels. O-cresol seems to be a less reliable indicator of toluene exposure.     

Investigation of exposure of ships'' painters to organic solvents

Mikulski, P. I., Wiglusz, Renata, Bublewska, Anna, and Uselis, J. (1972).Brit. J. industr. Med.,29, 450-453. Investigation of exposure of ships' painters to organic solvents. An analysis was carried out of the hazard to ships' painters working at different concentrations of toluene and xylene vapours in air. The authors point out that it is possible to use a joint determination of hippuric and methylhippuric acids for the assessment of mixed exposure. The problem of benzene contamination of solvents is considered. During the investigations exposure to toluene and xylene was found to impair the excretion of uric acid.     

Correlation of xylene exposure and methyl hippuric acid excretion in urine among paint industry workers

Different calculations of methyl hippuric acid excretion in urine were correlated to the time-weighted average (TWA) of the xylene exposure of a complete workday for 40 paint industry workers exposed to 12 different solvents. The 8-h TWA xylene exposure varied between 0 and 865 (median 69) mg/m3. The amount of methyl hippuric acid excreted in about 24 h showed only a slightly higher linear correlation to the xylene exposure than the amount of methyl hippuric acid excreted per hour during the latter part of the workshift among the 37 subjects exposed to TWA xylene air concentrations of 0-200 mg/m3. It was concluded that the methyl hippuric acid excretion rate during the latter part of the workshift can be used for crude xylene exposure categorizations.     

Clinical and histopathological changes of the nasal mucosa induced by occupational exposure to sulphuric acid mists

Aims: To assess potential alterations of the nasal mucosa by clinical and histopathological evaluation of workers exposed to sulphuric acid mists at anodising plants, correlating the findings with duration of exposure and sulphuric acid concentrations in the air, and comparing them with a control group.

Methods: Fifty two workers from five plants underwent a clinical evaluation (standard questionnaire, clinical, and ear, nose, and throat examination including nasal endoscopy). For the histopathological study, 20 of the 52 subjects (study group) were randomly selected, as well as 11 unexposed subjects (control group), matched by sex, age, and smoking habits. Nasal biopsy specimens were obtained from the anterior septum mucosa and the anterior curvature of the middle turbinate in each individual. A total of 56 nasal mucosa specimens (37 in the study group and 19 in the control group) were evaluated with regard to normal respiratory epithelium or metaplastic epithelium, atypia or dysplasia, and alterations of the lamina propria.

Results: The histopathological study revealed squamous metaplasia in 29 (79%) and atypia in 13 (35%) of the 37 study group samples. No association was found between exposure duration and the clinical and histopathological variables, but a significant association was found between sulphuric acid concentrations higher than 200 µg/m³ and pale mucosal patches and ulcerations in the exposed subjects. Logistic regression analysis showed that the exposed subjects had a fivefold risk of developing atypia compared with the unexposed subjects.

Conclusions: Workers exposed to sulphuric acid mists presented with a high incidence of nasal symptoms, and macroscopic and microscopic changes of the nasal mucosa, including squamous atypia and dysplasia. The risk for these histopthological lesions increased with higher sulphuric acid concentrations in the air, revealing an exposure-response relation.

     

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     

Urinary excretion of total trichloro-compounds, trichloroethanol, and trichloroacetic acid as a measure of exposure to trichloroethylene and tetrachloroethylene

Ikeda, M., Ohtsuji, H., Imamura, T., and Komoike, Y. (1972).Brit. J. industr. Med.,29, 328-333. Urinary excretion of total trichloro-compounds, trichloroethanol, and trichloroacetic acid as a measure of exposure to trichloroethylene and tetrachloroethylene. To investigate the relation between trichloroethylene and tetrachloroethylene concentrations in working environments and metabolite concentrations in urine, a series of surveys was conducted at 17 workshops where the vapour concentration in the air of each workshop was relatively constant. Urine samples collected from 85 male workers were analysed for total trichloro-compounds (TTC), and trichloroacetic acid (TCA). Trichloroethanol (TCE) was estimated by difference. Statistical analyses of the data revealed that the urinary concentrations of both TTC and TCE were proportional to the atmospheric concentration of trichloroethylene. The concentration of TCA was also related to the vapour concentration up to 50 p.p.m. but not at higher concentrations. Further calculations suggested that only one-third of the trichloroethylene absorbed through the lungs was excreted in the urine during working time.

In tetrachloroethylene exposure, urinary metabolite levels increased until the atmospheric concentration of the solvent reached 50 to 100 p.p.m., but little increase occurred at higher concentration. This observation was further confirmed by experimental exposure of rats. The toxicological significance of changes in the metabolism of the two solvents is discussed in relation to the possible necessity of reducing the threshold limit value from the current value of 100 p.p.m.

     

Excretion of organic chlorine compounds in the urine of persons exposed to vapours of trichloroethylene and tetrachloroethylene

Ogata, M., Yoshiko, T., and Tomokuni, K. (1971).Brit. J. industr. Med.,28, 386-391. Excretion of organic chlorine compounds in the urine of persons exposed to vapours of trichloroethylene and tetrachloroethylene. Male volunteers were exposed to 170 p.p.m. of trichloroethylene vapour either for 3 hours or for 7 hours with one break of 1 hour; or to 87 p.p.m. of tetrachloroethylene vapour for 3 hours. Urine was collected frequently up to 100 hours after the start of exposure, and was analysed for trichloroethanol and trichloroacetic acid. After trichloroethylene exposure, trichloroethanol was excreted most rapidly shortly after exposure ceased, and trichloroacetic acid most rapidly 42 to 69 hours after exposure ceased. Total recoveries of trichloroethylene inhaled, up to 100 hours, were: trichloroethanol, after 3 hours' exposure, 53·1%; after 7 hours' exposure, 44%; trichloroacetic acid, similarly: 21·9% and 18·1%. The effects of exposure on blood pressure, pulse rate, flicker value, and reaction time were measured. The diastolic blood pressure was decreased significantly after 3 hours' exposure to 170 p.p.m. trichloroethylene. After tetrachloroethylene exposure, in 67 hours trichloroacetic acid was excreted to 1·8% tetrachloroethylene retained and an unknown chloride equivalent to 1·0%.

Urine samples from 10 workers in an automobile parts factory were analysed for trichloroethanol and trichloroacetic acid. Trichloroethanol concentrations in the urine taken after work were higher than in the urine taken before work while for trichloroacetic acid the concentrations were reversed, due to the difference in the time course of excretion. The urinary levels of trichloroethanol, trichloroacetic acid, and total trichloro compounds were almost proportional to the environmental concentration of trichloroethylene.

     

Occupational chronic exposure to organic solvents XVI. Ambient and biological monitoring of workers exposed to toluene

Object Ambient air and biological monitoring of an occupational toluene exposure was carried out on a group of 33 workers. Method The biological monitoring of the workers was based on determination of the concentration of toluene in blood and on quantification of the urinary metabolites o-cresol and hippuric acid. All blood and urine samples were collected post-shift. Results The average toluene concentration in the workplace air was 65?ppm, ranging from 13 to 151?ppm. An average concentration of toluene in blood of 911?μg/l was found, corresponding to an average urinary concentration of 2.9?mg/l (2.3?mg/g creatinine) o-cresol and 2.4?g/l (1.9?g/g creatinine) hippuric acid. Both urinary metabolites can be correlated with the concentration of toluene in ambient air and blood, respectively. Conclusions The results of our study indicate that the determination of the urinary o-cresol excretion represents a diagnostically specific and sensitive parameter for the estimation of an individual toluene uptake. In contrast, monitoring of the concentration of hippuric acid in urine cannot be recommended for assessment of individual exposure. To set up a biological tolerance value (BAT) for o-cresol, a urinary concentration of 3?mg/l o-cresol should be in accordance with the current MAK value of 50?ppm toluene.     

Urinary excretion of O-cresol and hippuric acid after toluene exposure in rotogravure printing

Summary In 62 male rotogravure printers, the time-weighted average (TWA) toluene exposure during one workweek ranged from 8 to 496 mg/m³ (median 96). Post-shift urinary excretion of hippuric acid showed a poor correlation with the air toluene concentration. Level of o-cresol excretion ranged from 0.08 to 2.37 mmol/mol creatinine and was associated with the exposure (r _s = 0.57, P<0.0001), although the variation was considerable. However, this metabolite was significantly influenced by smoking habits, both in the workers (0.34 vs 0.10 mmol/mol creatinine after adjustment to zero exposure for the smokers and non-smokers, respectively; P = 0.03) and in 21 unexposed controls (0.18 vs 0.06 mmol/mot creatinine; P = 0.002). The excretion of these metabolites was followed during vacation, when the workers were unexposed. The shared one-compartment half-time was 44h (± SE 30, 82). After 2–4 weeks of vacation, the concentration of o-cresol was significantly higher for the smokers than the non-smokers (0.14 vs 0.06 mmol/mol creatinine; P = 0.02).No smoking-associated difference was found for the urinary hippuric acid concentration. However, there was an association between alcohol consumption and hippuric acid excretion (P = 0.03); no such difference was shown for o-cresol. These results demonstrate that hippuric acid excretion is unsuitable for biological monitoring of toluene exposure when the exposure level is below 200 mg/m³. Also, in spite of the favourable excretion kinetics, the impact of smoking and the large interindividual variation warrant the same conclusion for o-cresol as a means of monitoring low level exposure in an individual worker.     

Benzene and naphthalene in air and breath as indicators of exposure to jet fuel

Aims: To estimate exposures to benzene and naphthalene among military personnel working with jet fuel (JP-8) and to determine whether naphthalene might serve as a surrogate for JP-8 in studies of health effects.

Methods: Benzene and naphthalene were measured in air and breath of 326 personnel in the US Air Force, who had been assigned a priori into low, moderate, and high exposure categories for JP-8.

Results: Median air concentrations for persons in the low, moderate, and high exposure categories were 3.1, 7.4, and 252 µg benzene/m³ air, 4.6, 9.0, and 11.4 µg benzene/m³ breath, 1.9, 10.3, and 485 µg naphthalene/m³ air, and 0.73, 0.93, and 1.83 µg naphthalene/m³ breath, respectively. In the moderate and high exposure categories, 5% and 15% of the benzene air concentrations, respectively, were above the 2002 threshold limit value (TLV) of 1.6 mg/m³. Multiple regression analyses of air and breath levels revealed prominent background sources of benzene exposure, including cigarette smoke. However, naphthalene exposure was not unduly influenced by sources other than JP-8. Among heavily exposed workers, dermal contact with JP-8 contributed to air and breath concentrations along with several physical and environmental factors.

Conclusions: Personnel having regular contact with JP-8 are occasionally exposed to benzene at levels above the current TLV. Among heavily exposed workers, uptake of JP-8 components occurs via both inhalation and dermal contact. Naphthalene in air and breath can serve as useful measures of exposure to JP-8 and uptake of fuel components in the body.

     

Changes in zinc and serum proteins due to carbon disulphide exposure

El-Gazzar, R., El-Sadik, Y. M., and Hussein, M. (1973).British Journal of Industrial Medicine,30, 284-288. Changes in zinc and serum proteins due to carbon disulphide exposure. Serum zinc and proteins have been studied in 82 workers in the Egyptian rayon industry, of whom 40 were at the time exposed to carbon disulphide (CS₂) and nine had been exposed in the past for a year or more. Thirty-three workers without exposure were used as controls. Carbon disulphide caused depletion of serum zinc by an increase in the rate of zinc excretion and an increase in all serum protein fractions. All these effects are temporary and improve on cessation of exposure.     

Ventilatory function in workers exposed to low levels of toluene diisocyanate: a six-month follow-up

Peters, John M., Murphy, Raymond L. H., and Ferris, Benjamin, G., Jr. (1969).Brit. J. industr. Med.,26, 115-120. Ventilatory function in workers exposed to low levels of toluene diisocyanate: a six-month follow-up. Thirty-four workers exposed to toluene diisocyanate during production of polyurethane foam were examined with a respiratory questionnaire and tests of ventilatory capacity. The tests of pulmonary function were conducted on Monday morning and afternoon and on Tuesday morning and afternoon. Twenty-eight of these 34 workers had been examined with the same tests six months earlier. On Monday a mean change in the one-second forced expiratory volume (F.E.V._1·0) of –0·16 l. occurred that did not return to the baseline value (Monday a.m.) on Tuesday morning. A statistically significant decrease in all the measurements of ventilatory capacity except the forced vital capacity occurred over the six months in the 28 workers. The F.E.V._1·0 fell an average of 0·14 l. and flow rates at 75%, 50%, 25%, and 10% of vital capacity also decreased significantly. There was a highly significant correlation coefficient (r = 0·72) between one-day changes in F.E.V._1·0 (measured six months earlier) and six-month changes in F.E.V._1·0. Workers with respiratory symptoms (cough and/or phlegm) demonstrated greater falls in F.E.V._1·0 than did asymptomatic workers.

All air concentrations of toluene diisocyanate measured during this study were below the threshold limit value (0·02 p.p.m.).