Analysis of Phenanthrols in Human Urine by Gas Chromatography-Mass Spectrometry: Potential Use in Carcinogen Metabolite Phenotyping

• Published in: Cancer epidemiology, biomarkers & prevention Vol. 13; no. 12; pp. 2167 - 2174
• Main Authors: Carmella, Steven G, Chen, Menglan, Yagi, Haruhiko, Jerina, Donald M, Hecht, Stephen S
• Format: Journal Article
• Language: English
• Published: United States American Association for Cancer Research 01-12-2004
• Subjects: Carcinogens - analysis; Carcinogens - metabolism; Gas Chromatography-Mass Spectrometry; Humans; Phenanthrenes - metabolism; Phenanthrols; Phenols - urine; Phenotype; Polycyclic Aromatic Hydrocarbons - urine; Smoking - adverse effects; Urinalysis - methods
• ISSN: 1055-9965; 1538-7755
• DOI: 10.1158/1055-9965.2167.13.12

Phenanthrene is the simplest polycyclic aromatic hydrocarbon (PAH) containing a bay region, a feature closely associated with
carcinogenicity. We have proposed that measurement of phenanthrene metabolites in human urine could be used to identify interindividual
differences in metabolic activation and detoxification of PAH, and that these differences may be related to cancer susceptibility
in smokers and other exposed individuals. Previously, we reported a method for quantitation of r -1, t -2,3, c -4-tetrahydroxy-1,2,3,4-tetrahydrophenanthrene ( trans , anti -PheT) in human urine. trans , anti -PheT is the ultimate product of the diol epoxide metabolic activation pathway of phenanthrene. In this study, we have extended
our carcinogen metabolite phenotyping approach by developing a method for quantitation of phenanthrols in human urine. PAH
phenols such as phenanthrols are considered as detoxification products. After treatment of the urine by β-glucuronidase and
arylsulfatase, a fraction enriched in phenanthrols was prepared by partitioning and solid phase extraction. The phenanthrols
were silylated and analyzed by gas chromatography-positive ion chemical ionization-mass spectrometry with selected ion monitoring.
[ ring - 13 C 6 ]3-phenanthrol was used as an internal standard. Accurate and reproducible quantitation of four phenanthrols, 1-phenanthrol
(1-HOPhe), 2-HOPhe, 3-HOPhe, and 4-HOPhe, was readily achieved. In smokers, mean levels of 1-HOPhe (0.96 ± 1.2 pmol/mg creatinine)
and 3-HOPhe (0.82 ± 0.62 pmol/mg creatinine) were greater than those of 2-HOPhe (0.47 ± 0.29 pmol/mg creatinine), and 4-HOPhe
(0.11 ± 0.07 pmol/mg creatinine). There were no significant differences between the levels of any of the phenanthrols in smokers
and nonsmokers. Total levels of the quantified phenanthrols were highly correlated with those of 3-HOPhe. Ratios of phenanthrene
metabolites representing activation and detoxification were calculated as trans , anti -PheT divided by 3-HOPhe. There was a 7.5-fold spread of ratios in smokers, and a 12.3-fold spread in nonsmokers, suggesting
that this may be a useful parameter for distinguishing individual metabolic responses to PAH exposure.