Analytical and Bioanalytical Chemistry

, Volume 408, Issue 10, pp 2515–2525 | Cite as

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

  • Darina Lankova
  • Katerina Urbancova
  • Radim J. Sram
  • Jana Hajslova
  • Jana Pulkrabova
Research Paper


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.

Graphical abstract

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)


Monohydroxylated metabolites of polycyclic aromatic hydrocarbons SRM 3673 Tandem mass spectrometry Ultra-high-performance liquid chromatography Urine 



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.

Compliance with ethical standards

The study has been approved by the appropriate ethics committee and has been performed in accordance with the ethical standards.

Conflict of interest

The authors declare that they have no competing interests.

Informed consent

Informed consent was obtained from all participants for being included in the study.

Supplementary material

216_2016_9350_MOESM1_ESM.pdf (1.3 mb)
ESM 1 (PDF 1353 kb)


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Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Darina Lankova
    • 1
  • Katerina Urbancova
    • 1
  • Radim J. Sram
    • 2
  • Jana Hajslova
    • 1
  • Jana Pulkrabova
    • 1
  1. 1.Faculty of Food and Biochemical Technology, Department of Food Analysis and NutritionUniversity of Chemistry and Technology, PraguePrague 6Czech Republic
  2. 2.Institute of Experimental Medicine AS CRPrague 4Czech Republic

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