Highly sensitive routine method for urinary 3-hydroxybenzo[a]pyrene quantitation using liquid chromatography-fluorescence detection and automated off-line solid phase extraction

Abstract

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).

Early markers of cardiovascular disease are associated with occupational exposure to polycyclic aromatic hydrocarbons

“…However, the measurement of urinary 3-OH-BaP and 3-OH-BaA was hampered due to low concentrations in urine. The 3-OH-BaP concentrations are usually 1000 times lower in urine than 1-OH-PYR 23 . Studies on rats showed that the primary route of excretion for benzo[a]pyrene was via feces and only 0.1–0.3% of benzo[a]pyrene is excreted in urine as 3-OH-BaP upon intravenous administration 24 , 25 .…”

Use of Physiologically-Based Pharmacokinetic Modeling to Simulate the Profiles of 3-Hydroxybenzo(a)pyrene in Workers Exposed to Polycyclic Aromatic Hydrocarbons

“…Polypropylene bottles were used for urine collection and samples were stored at −20°C until analysis. Concentrations of 3-OHBaP in urine were measured according to the method published by Barbeau et al [39] and values were corrected for creatinine concentrations.…”

Recent analytical methods for atmospheric polycyclic aromatic hydrocarbons and their derivatives

“…To avoid the inconvenience, an HPLC method that uses deuterated 1-OHPyr as an internal standard has been developed (Chetiyanukornkul et al, 2002). Recently, many analytical procedures using HPLC-FL (Barbeau, Maître, & Marques, 2011;Chetiyanukornkul, Toriba, Kameda, Tang, & Hayakawa, 2006;Toriba, Chetiyanukornkul, Kizu, & Hayakawa, 2003), GC-(HR)MS(/MS) with silylation (Li et al, 2014;Woudneh et al, 2016) or LC(UPLC)-MS(/MS) with (Jacob, Wilson, & Benowitz, 2007;Luo, Gao, & Hu, 2015) or without derivatization (Lankova, Urbancova, Sram, Hajslova, & Pulkrabova, 2016;Onyemauwa et al, 2009) have been developed to determine urinary 1-OHPyr and other PAH metabolites having four to five rings, which are important for toxic PAH exposure assessments.…”