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Determination of 1-hydroxypyrene in human urine by HPLC with electrochemical detection at a boron-doped diamond film electrode

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Abstract

A high-performance liquid chromatographic method with electrochemical detection (HPLC-ED) at a boron-doped diamond film electrode with preliminary separation and preconcentration by solid-phase extraction (SPE) has been developed for the determination of 1-hydroxypyrene (1-HP) in human urine. 1-HP is among the most widely used biomarkers of exposure to polycyclic aromatic hydrocarbons. Optimal HPLC-ED conditions have been found: mobile phase methanol–0.05 mol L−1 phosphate buffer pH 5.0 (80:20, v/v), detection potential +1,000 mV versus Ag/AgCl (3 mol L−1 KCl), and flow rate 0.8 mL min−1. For SPE, LiChrolut® RP-18 E cartridges were used. The extraction yield was (87.0 ± 5.8) % (n = 5). The concentration dependence of 1-HP was measured in the concentration range from 0.01 to 10 μmol L−1 (2.18–2,180 μg L−1) using methanolic solutions resulting from the SPE pretreatment of spiked human urine samples. The limit of detection (signal-to-noise ratio 3) and the limit of quantification (signal-to-noise ratio 10) of the biomarker were 0.013 μmol L−1 (2.84 μg L−1) and 0.043 μmol L−1 (9.39 μg L−1), respectively, which is sufficient for its determination in the urine of persons exposed to polycyclic aromatic hydrocarbons.

Chromatograms of 1-hydroxypyrene (injected 20 μL of methanolic solution resulting from the solid phase extraction pretreatment of spiked human urine sample); concentrations of the analyte in urine: 0 (1), 0.2 (2), 0.4 (3), 0.6 (4), 0.8 (5) and 1.0 μmol L−1 (6). Measured by HPLC with electrochemical detection at the boron-doped diamond film electrode; detection potential +1,000 mV vs. Ag/AgCl (3 mol L−1 KCl); mobile phase: MeOH:0.05 mol L−1 phosphate buffer pH 5.0 (80:20, v/v); flow rate 0.8 mL min−1

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Acknowledgments

Financial support of this work provided by the Ministry of Education, Youth and Sports of the Czech Republic [projects MSM 0021620857 and KONTAKT (AMVIS) project ME 10004 (NEMVAD)], by Charles University in Prague (projects UNCE 2012/44 and SVV 2012-265201), and by the Grant Agency of the Czech Republic (project P206/12/G151) is gratefully acknowledged.

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Authors and Affiliations

  1. Faculty of Science, Department of Analytical Chemistry, UNESCO Laboratory of Environmental Electrochemistry, Charles University in Prague, Hlavova 2030/8, 12843, Prague 2, Czech Republic

    Oksana Yosypchuk, Jiří Barek & Vlastimil Vyskočil

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  1. Oksana Yosypchuk
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  2. Jiří Barek
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  3. Vlastimil Vyskočil
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Correspondence to Vlastimil Vyskočil.

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Published in the special paper collection Instrumental Methods of Analysis (IMA 2011) with guest editors Maria Ochsenkuehn-Petropoulou, Nikos Kallithrakas and Panagiotis Kefalas.

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Yosypchuk, O., Barek, J. & Vyskočil, V. Determination of 1-hydroxypyrene in human urine by HPLC with electrochemical detection at a boron-doped diamond film electrode. Anal Bioanal Chem 404, 693–699 (2012). https://doi.org/10.1007/s00216-012-6140-2

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  • DOI: https://doi.org/10.1007/s00216-012-6140-2

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