Development of a chemiluminescent enzyme immunoassay for urinary 1-hydroxypyrene

Abstract

A chemiluminescent enzyme-immunoassay for urinary 1-hydroxypyrene has been developed and optimized. The enzymatic activity of horseradish peroxidase-labeled tracer was measured with an enhanced chemiluminescent system and the results were compared with those from conventional colorimetric detection. The method fulfilled all the requirements of accuracy and precision and the detection limit was 0.001 pmol/well, which enabled analysis in less than 1 microL urine. Subjects working in the center of Bologna who were exposed daily to vehicular exhaust gas were studied. Their urinary 1-hydroxypyrene concentrations were compared with the levels of benzo( a)pyrene in air particulate matter. Urinary 1-hydroxypyrene, which ranged from 0.5 to 10 nmol L(-1), correlated poorly with the concentration of benzo( a)pyrene in air particulate matter, which ranged from 5 to 140 ng m(-3). No significant effect of vehicle exhaust gas exposure was observed among the different groups of subjects working in different areas of the town. Thus, at a relatively low level of exposure 1-hydroxypyrene does not seem to be a sensitive biomarker of exposure to polycyclic aromatic hydrocarbons.

Detection of 1-hydroxypyrene in urine by direct fluorometric analysis on a solid sorbing phase. Validation and application of the method to biological monitoring of PAH-exposed persons

“…The mean concentration reported for nonsmoking street policemen (0.07 µmol/mol creatinine) in this city is lower than that found for asphaltexposed paving workers and also within the mean usual concentration range (<0.10 µg/L). These values are very close to our results for Bordeaux street exposures, thus confirming previous findings [2,31,32,33] suggesting that 1-OHPy in urine cannot be used with confidence as a biomarker for nonoccupational PAH exposure. Moreover, these 1-OHPy contents are significantly lower than those 820 Fig.…”

“…The other samples (Urine 1 to 4) have been collected from anonymous volunteers at Bordeaux University Fig. 8 Urinary 1-OHPy concentrations (µmol/mol creatinine) determined at the end of the shift of a volunteer exposed in various urban sites with respect to morning concentrations, and comparison with data (marked with a star) reporting mean 1-OHPy concentrations among street policemen, industrial and road-exposed workers in the city of Budapest [15] reported for smokers (>0.13 µmol/mol creatinine) and are in accordance with other studies [2,34], which leads to the conclusion that urinary concentrations of PAH metabolites are influenced more by smoking habit than by exposure to urban atmospheric pollution.…”

“…However these values are within the mean concentration range (<0.10 µmol/mol creatinine) usually found among people not at all or weakly exposed to atmospheric PAH [9]. It follows that, in agreement with previous work [2,31], the small increase of 1-OHPy concentration reported here with respect to morning mean value does not allow us to conclude whether it results from atmospheric PAH exposure or from other sources such as food intake. Also, for comparison, this figure shows data from the study of Strickland et al [15] concerning persons from the city of Budapest.…”

“…Although urinary 1-OHPy may not exactly reflect the air content of potentially toxic PAH such as benzo[a]pyrene [2], significantly good correlation between air PAH content and urinary 1-OHPy has been reported [2,10]. This justifies the fact that, during the last 15 years, a large number of studies on occupational and environmental PAH exposure have been carried out using 1-OHPy as an appropriate indicator of PAH exposure [3,4,5,6,7,8,9,10,11,12,13,14,15,16,17,18].…”

Investigating the Quantitative Structure-Activity Relationships for Antibody Recognition of Two Immunoassays for Polycyclic Aromatic Hydrocarbons by Multiple Regression Methods

“…Some immunoassay techniques, such as enzyme-linked immunosorbent assay (ELISA) [2,3], fluorescence polarization immunoassay [4], chemiluminescent immunoassay [5], real-time immuno-polymerase chain reaction assay [6] and immunosensor [7], were developed for PAHs analysis. Several ELISA test kits for PAHs have been produced and are commercially available, among which RaPID and Ris C are the most widely studied and used [8].…”