Identification and quantification of urinary benzo[a]pyrene and its metabolites from asphalt fume exposed mice by microflow LC coupled to hybrid quadrupole time-of-flight mass spectrometry

Abstract

Prolonged, extensive exposure to asphalt fume has been associated with several adverse health effects. Inhaled polycyclic aromatic hydrocarbons (PAHs) from asphalt fume exposure have been suspected of inducing such effects. In this study, a bioanalytical method was proposed and evaluated to identify and quantify benzo[a]pyrene and its hydroxy-metabolites. This method is based on coupling a microflow liquid chromatography (LC) to a hybrid quadrupole orthogonal acceleration time-of-flight mass spectrometry (Q-TOFMS). In the experiment, thirty-two B6C3F1 mice were exposed to asphalt fume in a whole body inhalation chamber for 10 days (4 h day⁻¹) and twelve other mice were used as controls. The asphalt fume was generated at 180 °C and the concentrations in the animal exposure chamber ranged 175–182 mg m⁻³. Benzo[a]pyrene and its metabolites of 3-hydroxybenzo[a]pyrene, benzo[a]pyrene-7,8-dihydrodiol(±), benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide(±), and benzo[a]pyrene-7,8,9,10-tetrahydrotetrol(±) in the urine of asphalt fume exposed mice were identified and found at 3.18 ng 100 mL⁻¹, 31.36 ng 100 mL⁻¹, 11.56 ng 100 mL⁻¹, 54.92 ng 100 mL⁻¹, and 45.23 ng 100 mL⁻¹ respectively. The results revealed that the urinary benzo[a]pyrene and its hydroxy-metabolites from exposed mice were at significantly higher levels (p < 0.001) than those from the control groups. Compared with several other technologies such as HPLC-UV and HPLC-fluorescence, the new method is more sensitive and selective, and it can also provide additional useful information on the structures of the metabolites. Hence, this method can be used to perform the assessment and to study the mechanisms of the adverse health effects. The fragmentation patterns established in this study can also be used to identify and quantify PAH metabolites in other biological fluids.