Urinary hydroxypyrene determination for biomonitoring of firefighters deployed at the Fort McMurray wildfire: an inter-laboratory method comparison

Urinary 1-hydroxypyrene (OH-Pyr) is widely used for biomonitoring human exposures to polycyclic aromatic hydrocarbons (PAHs) from air pollution and tobacco smoke. However, there have been few rigorous validation studies reported to ensure reliable OH-Pyr determination for occupational health and risk assessment. Herein, we report an inter-laboratory method comparison for urinary OH-Pyr when using gas chromatography-high-resolution mass spectrometry (GC-HRMS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) on urine specimens collected from firefighters (n = 42) deployed at the 2016 Fort McMurray wildfire. Overall, there was good mutual agreement in urinary OH-Pyr quantification following enzyme deconjugation with an average bias of 39% with no significant deviation from linearity (slope = 1.36; p > 0.05), whereas technical precision (< 12%) and average recovery (> 85%) were acceptable when using a stable-isotope internal standard. Faster analysis times (4 min) were achieved by LC-MS/MS without chemical derivatization, whereas lower detection limits (0.64 ng/L, S/N = 3) was realized with solid-phase extraction prior to GC-HRMS. A median creatinine normalized OH-Pyr concentration of 128 ng/g was measured for firefighters that were below the recommended biological exposure index due to delays between early stages of emergency firefighting and urine sample collection. Similar outcomes were also measured for 3-hydroxyphenanthrene and 9-hydroxyfluorene that were positively correlated with urinary OH-Pyr (p < 0.05), implying similar uptake, distribution, and liver biotransformation processes. Optimal specimen collection strategies post-deployment together with standardized protocols for OH-PAH analysis are critical to accurately evaluate smoke exposure in firefighters, including experimental conditions to ensure quantitative enzyme hydrolysis of urine samples.