Determination of Phenanthrene and Hydroxyphenanthrenes in Various Biological Matrices at Trace Levels using Gas Chromatography-Mass Spectrometry

Abstract

A fast and efficient multiresidue extraction-purification procedure was developed for low levels (ppb range) of phenanthrene and hydroxyphenanthrene in biological matrices, in order to quantify phenanthrene and metabolites in blood, milk, urine, and biological tissues of lactating goats. Detection and identification of the analytes (phenanthrene and 1-, 2-, 3-, 4-, and 9-hydroxyphenanthrene) were achieved using gas chromatography coupled to mass spectrometry. Deuterium-labeled phenanthrene was used as internal standard for phenanthrene and 2-OHfluorene for metabolites. The developed method includes enzymatic hydrolysis, liquid-liquid extraction, and Envi-Chrom P SPE column purification. Analyses were performed in the selected ion monitoring mode to achieve ad hoc sensitivity in accordance with analyte concentrations in food samples. Detection limits were between 2.3 and 5.1 ng/mL (ppb) for milk samples, 0.5 and 2.5 ng/mL for urine and blood samples, and 1.9 and 8.0 ng/g for tissue samples. This original multiresidue and multimatrix analytical methodology was applied to metabolism studies and polycyclic aromatic hydrocarbon (PAH) risk assessment. We demonstrated, for the first time, that metabolites were present in milk. These results suggested that usual PAHs analysis methods based on the detection of native molecule are not representative of the real contamination of biological matrices.

Development of a gas chromatography/mass spectrometry method to quantify several urinary monohydroxy metabolites of polycyclic aromatic hydrocarbons in occupationally exposed subjects

“…Limits of quantification (LOQ) were in the 0.1-1.4 g/l range ( Table 2). These levels are lower than levels previously reported for gas chromatography coupled with low-resolution mass spectrometry assay [7,32], while they are of course higher than those obtained with high-resolution mass spectrometry [25].…”

Effect of Exposure to Soil-Bound Polycyclic Aromatic Hydrocarbons on Milk Contaminations of Parent Compounds and Their Monohydroxylated Metabolites

“…The apparent decrease in metabolite concentration in milk after 7-14 days of exposure suggests that biotransformation of PAHs into metabolites other than the monohydroxylated forms may have occurred. Indeed, Pyr is mainly monohydroxylated into its 1-OH Pyr form, whereas Phe can be monohydroxylated to different forms, which may spread output concentrations over several metabolites (30). Fully understanding the decreased concentrations of metabolites needs further studies as other metabolic pathways cannot be excluded.…”

Metabolism of [D10]Phenanthrene to Tetraols in Smokers for Potential Lung Cancer Susceptibility Assessment: Comparison of Oral and Inhalation Routes of Administration

“…The EI-MS/MS conditions were as follows: source temperature, 200°C; emission current, 110 A. Selected reaction monitoring with a collision energy of 14 eV, electron energy of Ϫ55 eV, and Ar collision gas pressure of 1.0 mTorr was used to detect HOPhe-trimethylsilyl ethers (HOPhe-TMS), [D 10 ]Phe analysis was performed essentially as described, with modifications (Grova et al, 2005). A 5-ml aliquot of urine or 1-ml aliquot of plasma was placed in a 10-ml centrifuge tube containing 50:50 (v/v) of cyclohexane/ethyl acetate and [ 13 C 6 ]Phe (1 ng) as internal standard.…”