GC/MS determination of N-phenylvaline, a possible biomarker for benzene exposure in human hemoglobin by the "N-alkyl Edman method".

We report the application of a modified Edman degradation procedure to the analysis of benzene oxide adducts at the N-terminal valine of human hemoglobin (Hb). Benzene oxide is thought to be formed in the liver from benzene and adduct formation with macromolecules is therefore likely to occur. We assumed that benzene oxide could covalently bind to hemoglobin after leaving the hepatic tissue. The "N-alkyl Edman method" was adapted for the approach to investigate this hypothesis. Using capillary gas chromatography/mass spectrometry (GC/MS) with negative chemical ionization, we could not detect N-phenylvaline in blood samples from persons occupationally exposed to benzene. We conclude that adducts of benzene oxide to the N-terminal valine of Hb are not formed in detectable amounts in vivo and consequently are not suitable for biomonitoring purposes. This result clearly indicates that other reactive benzene metabolites have to be taken into account not only in the search for a biomarker but also as the ultimate carcinogenic species.