Highly sensitive routine method for urinary 3-hydroxybenzo[a]pyrene quantitation using liquid chromatography-fluorescence detection and automated off-line solid phase extraction.
Many workers and also the general population are exposed to polycyclic aromatic hydrocarbons (PAHs), and benzo[a]pyrene (BaP) was recently classified as carcinogenic for humans (group 1) by the International Agency for Research on Cancer. Biomonitoring of PAHs exposure is usually performed by urinary 1-hydroxypyrene (1-OHP) analysis. 1-OHP is a metabolite of pyrene, a non-carcinogenic PAH. In this work, we developed a very simple but highly sensitive analytical method of quantifying one urinary metabolite of BaP, 3-hydroxybenzo[a]pyrene (3-OHBaP), to evaluate carcinogenic PAHs exposure. After hydrolysis of 10 mL urine for two hours and concentration by automated off-line solid phase extraction, the sample was injected in a column-switching high-performance liquid chromatography fluorescence detection system. The limit of quantification was 0.2 pmol L(-1) (0.05 ng L(-1)) and the limit of detection was estimated at 0.07 pmol L(-1) (0.02 ng L(-1)). Linearity was established for 3-OHBaP concentrations ranging from 0.4 to 74.5 pmol L(-1) (0.1 to 20 ng L(-1)). Relative within-day standard deviation was less than 3% and relative between-day standard deviation was less than 4%. In non-occupationally exposed subjects, median concentrations for smokers compared with non-smokers were 3.5 times higher for 1-OHP (p<0.001) and 2 times higher for 3-OHBaP (p<0.05). The two urinary biomarkers were correlated in smokers (ρ=0.636; p<0.05; n=10) but not in non-smokers (ρ=0.09; p>0.05; n=21).
Equipe EPSP, Environnement et Prédiction de la Santé des Populations-Laboratoire TIMC (UMR CNRS 5525), Université Joseph Fourier Grenoble, Faculté de Médecine, Domaine de la Merci, 38700 La Tronche, France.
Damien
D
Barbeau
Anne
A
Maître
Marie
M
Marques
D000293:Adolescent; D000328:Adult; D000368:Aged; D001331:Automation / Q000379:methods; D001580:Benzopyrenes / Q000032:analysis* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005453:Fluorescence; D006801:Humans; D008297:Male; D008875:Middle Aged; D016273:Occupational Exposure / Q000032:analysis*; D011084:Polycyclic Aromatic Hydrocarbons / Q000378:metabolism / Q000652:urine; D011721:Pyrenes / Q000032:analysis / Q000378:metabolism; D012680:Sensitivity and Specificity; D052616:Solid Phase Extraction / Q000379:methods*; D013050:Spectrometry, Fluorescence / Q000379:methods; D016482:Urinalysis / Q000379:methods*; D055815:Young Adult
D001580:Benzopyrenes; D011084:Polycyclic Aromatic Hydrocarbons; D011721:Pyrenes; C021338:3-hydroxybenzo(a)pyrene; C033146:1-hydroxypyrene