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Simultaneous analysis of PAH urinary mono- and dihydroxylated metabolites by GC-MS-MS following SPE and two-stage derivatization

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Abstract

A new gas chromatography–tandem mass spectrometry method for the determination of mono- and dihydroxylated polycyclic aromatic hydrocarbon metabolites (OH-PAHs and diol-PAHs) in urine was developed and validated. Various sample preparation procedures were compared, namely liquid–liquid extraction (LLE), dispersive solid-phase extraction (dSPE), and SPE, alone or combined. A novel two-stage derivatization approach using 2 silylation reagents was developed, and an experimental procedure design was used to optimize the programmed temperature vaporization–solvent vent injection (PTV-SV) GC parameters. The method focused on 11 target compounds resulting from four- to five-ring suspected carcinogenic PAHs. SPE was identified as an acceptable and more convenient extraction method for all tested metabolites, with extraction rates ranging from 63 to 86% and relative standard deviations lower than 20%. The two-stage derivatization approach successfully allowed first the derivatization of OH-PAHs by MTBSTFA (N-tert-butyldimethylsilyl-N-methyltrifluoroacetamide) and then diol-PAHs by BSTFA (N,O-bis(trimethylsilyl)trifluoroacetamide) in a single run. The limits of quantification were in the range of 0.01–0.02 μg l−1 for OH-PAHs and 0.02–0.2 μg l−1 for diol-PAHs. The intra- and interday precisions were lower than 10%. The method was applied to determine PAH metabolites in urine collected at the beginning and at the end of the working week from 6 workers involved in aluminum production. The mean diol-PAH levels at the end of the week were 10 to 20 times higher (0.86–2.34 μg g−1 creatinine) than those of OH-PAHs (0.03–0.30 μg g−1). These results confirmed the usefulness of this new analytical technique for detecting and characterizing metabolic patterns of PAHs in urine and assessing carcinogenic occupational exposures.

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Funding

This study was funded by the French sanitary agency ANSES (Agence nationale de sécurité sanitaire de l’alimentation, de l’environnement et du travail), Project n° 2014-1-176.

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Authors and Affiliations

  1. Université Grenoble Alpes, CNRS Grenoble INP, CHU Grenoble Alpes, TIMC, 38000, Grenoble, France

    Marie Marques, Anne Maitre, Christine Demeilliers & Renaud Persoons

  2. Département de Pharmachimie Moléculaire, UMR CNRS 5063, Université Grenoble Alpes, 38000, Grenoble, France

    Luc Choisnard

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  1. Marie Marques
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Marie Marques and Renaud Persoons contributed to the analysis and interpretation of the results. All authors listed have made substantial, direct, and intellectual contributions to the work and approved it for publication.

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Correspondence to Renaud Persoons.

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Marques, M., Maitre, A., Choisnard, L. et al. Simultaneous analysis of PAH urinary mono- and dihydroxylated metabolites by GC-MS-MS following SPE and two-stage derivatization. Anal Bioanal Chem 413, 6823–6835 (2021). https://doi.org/10.1007/s00216-021-03638-4

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