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A novel strategy for the determination of polycyclic aromatic hydrocarbon monohydroxylated metabolites in urine using ultra-high-performance liquid chromatography with tandem mass spectrometry

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Abstract

In this study, a novel analytical approach for the determination of 11 monohydroxylated polycyclic aromatic hydrocarbon metabolites (OH-PAHs) in urine was developed and validated. The rapid, simple and high-throughput sample preparation procedure based on ethyl acetate extraction and subsequent purification by dispersive solid-phase extraction (d-SPE) employing a Z-Sep sorbent is used for the first time. For the identification/quantification of target compounds, ultra-high-performance liquid chromatography (U-HPLC) interfaced with tandem mass spectrometry (MS/MS) was applied. The results of validation experiments performed on the Standard Reference Material (SRM) 3673 (organic contaminants in non-smokers’ urine) were in accordance with the certified values. The method recoveries ranged from 77 to 114 % with the relative standard deviation lower than 20 % and the quantification limits in the range of 0.010–0.025 ng mL−1 (except for benzo[a]pyren-3-ol with 0.9 ng mL−1). Within the pilot study, the new method was used for the analysis of OH-PAHs in 50 urine samples. The concentrations of ΣOH-PAHs were in the range of 0.87–63 ng mL−1 (1600–33,000 ng g−1 creatinine), with naphthalen-2-ol (2-OH-NAP) and phenanthren-1-ol (1-OH-PHEN) being the most abundant exposure biomarkers detected in all samples.

Flow-chart of the analytical method for the determination of major OH-PAHs in urine including the concentrations of detected metabolites in real samples (n = 50)

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Acknowledgments

This study was funded by the Czech Science Foundation Project No. 13-13458S. The authors would like to thank the gynaecological and maternity ward at the Hospital and Polyclinic Karviná for organising the logistic and collection of samples. The ‘Operational Program Prague – Competitiveness’ (CZ.2.16/3.1.00/22197) and ‘National Program of Sustainability’ (NPU I (LO) MSMT - 34870/2013) are also gratefully acknowledged.

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Authors and Affiliations

  1. Faculty of Food and Biochemical Technology, Department of Food Analysis and Nutrition, University of Chemistry and Technology, Prague, Technicka 3, 166 28, Prague 6, Czech Republic

    Darina Lankova, Katerina Urbancova, Jana Hajslova & Jana Pulkrabova

  2. Institute of Experimental Medicine AS CR, Videnska 1083, 142 20, Prague 4, Czech Republic

    Radim J. Sram

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  1. Darina Lankova
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Correspondence to Jana Pulkrabova.

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Lankova, D., Urbancova, K., Sram, R.J. et al. A novel strategy for the determination of polycyclic aromatic hydrocarbon monohydroxylated metabolites in urine using ultra-high-performance liquid chromatography with tandem mass spectrometry. Anal Bioanal Chem 408, 2515–2525 (2016). https://doi.org/10.1007/s00216-016-9350-1

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