Carla Valla

Comparison between urinary o-cresol and toluene as biomarkers of toluene exposure

The characteristics of urinary o-cresol (o-C) and urinary toluene (TOL-U) as biomarkers of occupational exposure to toluene were comparatively evaluated. One hundred healthy male rotogravure printing workers and 161 male and female control subjects were studied. Personal exposure to airborne toluene (TOL-A) during the shift was determined as a time-weighted average. Simple analytical procedures based on solid phase microextraction followed by gas chromatography/mass spectometry analysis were applied to the determination of end-shift o-C and TOL-U. Median TOL-A was 48 (6.0–162.0) mg/m3 in printers and 0.021 (<0.003–0.137) mg/m3 in controls. o-C was 0.185 (0.032–0.948) mg/g creatinine in printers and 0.027 (<0.006–0.330) mg/g creatinine in the controls. TOL-U was 7.6 (1.8–23.9) μ g/L in printers and 0.140 (0.094–0.593) μ g/L in the controls. According to all indices, exposure to toluene was higher in printers than in the controls. Nevertheless, the distribution of o-C in the two groups partially overlapped, whereas such behavior was not found in TOL-U. Both o-C and TOL-U in printers were correlated with TOL-A (Pearsons on log10-transformed variables r = 0.704 and 0.844, respectively) and with each other (r = 0.683). Smoking habits significantly increased the excretion of o-C but not of TOL-U. From the point of view of sampling conditions and analytical requirements, TOL-U and o-C showed similar properties, but comparison of their intrinsic characteristics showed that TOL-U had higher specificity and sensitivity, lower background values, was better correlated with airborne exposure, and was not influenced by cigarette smoking. Therefore TOL-U may be considered superior to o-C as a biomarker of occupational exposure to toluene.

Determination of hexafluoroisopropanol, a sevoflurane urinary metabolite, by 9-fluorenylmethyl chloroformate derivatization

A reversed-phase HPLC method with fluorescence detection for the quantification of hexafluoroisopropanol (HFIP) in urine is presented. HFIP, a metabolite of the inhalation anesthetic sevoflurane, is excreted mainly in urine as glucuronic acid conjugate. After enzymatic hydrolysis of the glucuronate, primary amino groups of interferent urinary compounds are blocked by reaction with o-phthalic dicarboxaldehyde and 3-mercaptopropionic acid, followed by labeling of HFIP with 9-fluorenylmethyl chloroformate. The derivatization reaction proceeds in a water-acetonitrile (1:1) solution at room temperature with a borate buffer of pH 12.5 as a catalyst. A stable fluorescent derivative of HFIP is formed within 5 min. The HFIP-FMOC derivative is separated by reversed-phase chromatography with isocratic elution on an octadecyl silyl column (33x4.6 mm, 3 microm) and guard column (20x4.0 mm, 40 microm), at 35 degrees C, and detected by fluorescence detection at an excitation wavelength of 265 nm and an emission wavelength of 311 nm. The method detection limit is 40 pg, per 10-microl injection volume, corresponding to 16 microg/l of HFIP in urine. The among-series relative standard deviation is <6% at 200 microg/l (n=6). As a preliminary application, the method was used to detect HFIP concentration in the urine of two volunteers exposed for 3 h to an airborne concentration of sevoflurane in the order of 2 ppm.

Application of Ultraviolet Spectrophotometry to Estimate Occupational Exposure to Airborne Polyaromatic Compounds in Asphalt Pavers

An ultraviolet (UV) spectrophotometric procedure was devised for the determination of polycyclic aromatic compound-oriented organic soluble matter in vapors and particulate collected from emissions of hot asphalt mix. Ultrasonic extraction was carried out with acetonitrile, followed by UV measurements at 254 nm. Polycyclic aromatic compounds (PACs) in volatile and particulate fraction were quantified as phenanthrene or benzo[k]fluoranthene equivalents. A comparison between UV and high-pressure liquid chromatography with fluorescence detection showed that PACs were one to three orders of magnitude higher than the sum of 15 priority polycyclic aromatic hydrocarbons (PAHs); still, significant correlations were found between volatile or particulate PACs and, respectively, total volatile or particulate PAHs. Moreover, in the particulate phase, PACs correlated with total particulate matter quantified by gravimetry. The proposed procedure was employed in a field study for monitoring personal exposure to asphalt emissions of workers engaged in road construction. Observed levels of acetonitrile-soluble PACs in air samples were very low (2–20 μg/m 3 ); however, asphalt pavers were exposed to significantly higher concentrations of volatile PACs than construction workers (geometric mean, 5.9 μg/m 3 vs. 4.1 μg/m 3 ). This method for estimating the global content of volatile or particulate PACs in air samples satisfies our requirements of simplicity and is suitable for conducting an initial screening to assess exposure to airborne polyaromatic organics in asphalt pavers.

Gas chromatography-electron-capture detection of urinary methylhippuric acid isomers as biomarkers of environmental exposure to xylene

Methylhippuric acid isomers (MHAs), urinary metabolites of xylenes, were determined, after clean-up by C18-SPE and esterification with hexafluoroisopropanol and diisopropylcarbodiimide, by GC with ECD detection, on an SPB-35 capillary column (30 m, 0.32 mm I.D., 0.25 microm film thickness, beta = 320). S-benzyl-mercapturic acid was used for internal standardization. Chromatographic conditions were: oven temperature 162 degrees C, for 14.2 min; ramp by 30 degrees C/min to 190 degrees C, for 3.5 min; ramp by 30 degrees C/min to 250 degrees C, for 4 min; helium flow rate: 1.7 ml/min; detector and injector temperature: 300 degrees C. The sample (1 microl) was injected with a split injection technique (split ratio 5:1). MHA recovery was >95% in the 0.5-20 micromol/l range; the limit of detection was <0.25 micromol/l; day-to-day precision, at 2 micromol/l, was Cv<10%. Urinary MHAs were determined in subjects exposed to different low-level sources of xylenes: (a) tobacco smoking habit and (b) BTX urban air pollution (airborne xylene ranging from 0.1 to 3.7 micromol/m3). Study (a) showed a significant difference between urinary MHA median excretion values of nonsmokers and smokers (4.6 micromol/l vs. 8.1 micromol/l, p<0.001). Study (b) revealed a significant difference between indoor workers and outdoor workers (4.3 micromol/l vs. 6.9 micromol/l, p<0.001), and evidenced a relationship between MHAs (y, micromol/mmol creatinine) and airborne xylene (x, micromol/m3) (y = 0.085+0.34x; r = 0.82, p<0.001, n = 56). Proposed biomarkers could represent reliable tools to study very low-level exposure to aromatic hydrocarbons such as those observed in the urban pollution due to vehicular traffic or in indoor air quality evaluation.