Abstract
A method is presented for determining monohydroxy polycyclic aromatic hydrocarbons (OHPAHs) having 2-, 3- and 4-rings in human urine by using high-performance liquid chromatography with fluorescence detection. A urine sample containing conjugates of OHPAHs was hydrolysed in the presence of β-glucuronidase/aryl sulfatase and the solution was cleaned up with a solid-phase extraction (C18 and silica). Eight OHPAHs, namely 1- and 2-hydroxynaphthalenes, 2-hydroxyfluorene, 2-, 3- and 4-hydroxyphenanthrenes, 3-hydroxyfluoranthene and 1-hydroxypyrene, were separated and 1- and 9-hydroxyphenanthrenes co-eluted on an alkylamide-type reversed-phase column with fluorimetric detection. The urinary concentrations of OHPAHs were quantified by using deuterated 1-hydoxypyrene as an internal standard. The method showed good repeatability for inter- and intra-day precisions as well as good linearity of calibration curves (r 2 ranged from 0.996 to 0.999). The limits of detection (S/N=3) were in the range from 2.3 fmol to 2.2 pmol per injection. This method was successfully applied to urine samples from non-smoking taxi drivers, traffic policemen and rural villagers of Chiang Mai, Thailand. The results showed higher urinary concentrations of OHPAHs in rural villagers, consistent with higher respiratory exposure to PAHs.
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IARC; International Agency for Research on Cancer (1983) WHO 32(Part 1):155–161, 225–231
Kizu R, Okamura K, Toriba A, Kakishima H, Mizokami A, Burnstein KL, Hayakawa K (2003) Arch Toxicol 77:335–343
Okamura K, Kizu R, Toriba A, Murahashi T, Mizokami A, Bunstein KL, Klinge CM, Hayakawa K (2004) Toxicology 195:243–254
Jongeneelen FJ, Anzion RB, Henderson PT (1987) J Chromatogr A 413:227–232
Strickland P, Kang D (1999) Toxicol Lett 108:191–199
Bouchard M, Viau C (1999) Biomarkers 4:159–187
Jacob J, Seidel A (2002) J Chromatogr B 778:31–47
Toriba A, Chetiyanukornkul T, Kizu R, Hayakawa K (2003) Analyst 128:605–610
Chetiyanukornkul T, Toriba A, Kizu R, Hayakawa K (2004) Polycycl Aromat Compd 24:474–476
Gmeiner G, Stehlik G, Tausch H (1997) J Chromatogr A 767:163–169
Hoffmann D, Hoffmann I (1997) J Toxicol Environ Health 50:307–364
Traynor GW, Apte MG, Sokol HA, Chuang JC, Tucker WG, Mumford JL (1990) Environ Sci Technol 24:1265–1270
Mitra S, Ray B (1995) Atmos Environ 29:3345–3356
Chuang JC, Callahan PJ, Lyu CW, Wilson NK (1999) J Expo Anal Environ Epidemiol 9:85–98
Ohura T, Sugiyama T, Amagai T, Fusaya M, Matsushita H (2002) J AOAC Int 85:188–202
Kuusimäki L, Peltonen Y, Mutanen P, Peltonen K, Savela K (2004) Int Arch Occup Environ Health 77:23–30
Lintelmann J, Hellemann C, Kettrup A (1994) J Chromatogr B 660:67–73
Angerer J, Mannschreck C, Gündel J (1997) Int Arch Occup Environ Health 69:323–331
Elovaara E, Väänänen V, Mikkola J (2003) Arch Toxicol 77:183–193
Serdar B, Waidyanatha S, Zheng Y, Rappaport SM (2003) Biomarkers 8:93–109
Smith CJ, Huang W, Walcott CJ, Turner W, Grainger J, Patterson DG (2002) Anal Bioanal Chem 372:216–220
Chetiyanukornkul T, Toriba A, Kizu R, Makino T, Nakazawa H, Hayakawa K (2002) J Chromatogr A 961:107–112
Bonsnes RW, Taussky HH (1945) J Biol Chem 158:581–591
Kamiya M, Toriba A, Onoda Y, Kizu R, Hayakawa K (2005) Food Chem Toxicol 43:1017–1027
Toriba A, Kuramae Y, Chetiyanukornkul T, Kizu R, Makino T, Nakazawa H, Hayakawa K (2003) Biomed Chromatogr 17:126–132
Grimmer G, Dettbarn G, Naujack KW, Jacob J (1991) Intern J Environ Anal Chem 43:177–186
Jongeneelen FJ (2001) Ann Occup Hyg 45:3–13
Ezzati M, Kammen DM (2002) Environ Health Perspect 110:1057–1068
Smith KR, Samet JM, Romieu L, Bruce N (2000) Thorax 55:518–532
Vatanasapt V, Martin N, Sriplung H, Chindavijak K, Sontipong S, Sriamporn H, Parkin DM, Ferlay J (1995) Cancer Epidemiol Biomarkers Prev 4:475–483
Acknowledgments
This study was supported, in part, by the Industrial Technology Research Grant Program in 2005 from New Energy and Industrial Technology Development Organization (NEDO) of Japan (ID: 05A21705a), and by the Kanazawa University 21-Century COE Program. We thank Assoc. Prof. Dr. Tippawan Prapamontol, Assoc. Prof. Dr. Prasak Thavornyutthikarn and Parinya Phanuwet, Chiang Mai University, for the sample collection in Thailand.
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Chetiyanukornkul, T., Toriba, A., Kameda, T. et al. Simultaneous determination of urinary hydroxylated metabolites of naphthalene, fluorene, phenanthrene, fluoranthene and pyrene as multiple biomarkers of exposure to polycyclic aromatic hydrocarbons. Anal Bioanal Chem 386, 712–718 (2006). https://doi.org/10.1007/s00216-006-0628-6
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DOI: https://doi.org/10.1007/s00216-006-0628-6