Analytical and Bioanalytical Chemistry

, Volume 383, Issue 4, pp 638–644 | Cite as

Quantification of urinary conjugates of bisphenol A, 2,5-dichlorophenol, and 2-hydroxy-4-methoxybenzophenone in humans by online solid phase extraction–high performance liquid chromatography–tandem mass spectrometry

  • Xiaoyun Ye
  • Zsuzsanna Kuklenyik
  • Larry L. Needham
  • Antonia M. Calafat
Original Paper


Urinary concentrations of phenols or their metabolites have been used as biomarkers to assess the prevalence of exposure to these compounds in the general population. Total urinary concentrations, which include both free and conjugated (glucuronide and sulfated) forms of the compounds, are usually reported. From a toxicologic standpoint, the relative concentrations of the free species compared with their conjugated analogs can be important because conjugation may reduce the potential biologic activity of the phenols. In this study, we determined the percentage of glucuronide and sulfate conjugates of three phenolic compounds, bisphenol A (BPA), 2,5-dichlorophenol (2,5-DCP), and 2-hydroxy-4-methoxybenzophenone (benzophenone-3, BP-3) in 30 urine samples collected between 2000 and 2004 from a demographically diverse group of anonymous adult volunteers. We used a sensitive on-line solid phase extraction–isotope dilution–high performance liquid chromatography–tandem mass spectrometry method. These three phenols were detected frequently in the urine samples tested. Only small percentages of the compounds (9.5% for BPA, and 3% for 2,5-DCP and BP-3) were excreted in their free form. The percentage of the sulfate conjugate was about twice that of the free compound. The glucuronide conjugate was the major metabolite, representing 69.5% (BPA), 89% (2,5-DCP), and 84.6% (BP-3) of the total amount excreted in urine. These results are in agreement with those reported before which suggested that BPA-glucuronide was an important BPA urinary metabolite in humans. To our knowledge, this is the first study describing the distribution of urinary conjugates of BP-3 and 2,5-DCP in humans.


Glucuronide Sulfate Metabolism Exposure Biomonitoring 


  1. 1.
    Sajiki J, Yonekubo J (2003) Chemosphere 51:55–62CrossRefPubMedGoogle Scholar
  2. 2.
    Howe SR, Borodinsky L, Lyon RS (1998) J Coat Technol 70:69–74CrossRefGoogle Scholar
  3. 3.
    Arenholt-Bindslev D, Breinholt V, Preiss A, Schmalz G (1999) Clin Oral Investig 3:120–125CrossRefPubMedGoogle Scholar
  4. 4.
    Safe SH, Pallaroni L, Yoon K, Gaido K, Ross S, McDonnell D (2002) Environ Health Perspect 110:925–929Google Scholar
  5. 5.
    Safe SH, Pallaroni L, Yoon K, Gaido K, Ross S, Saville B, McDonnell D (2001) Reprod Fertil Dev 13:307–315CrossRefPubMedGoogle Scholar
  6. 6.
    Kuiper GGJM, Lemmen JG, Carlsson B, Corton JC, Safe SH, van der Saag PT, van der Burg P, Gustafsson JA (1998) Endocrinology 139:4252–4263CrossRefPubMedGoogle Scholar
  7. 7.
    Andersen HR, Andersson AM, Arnold SF, Autrup H, Barfoed M, Beresford NA, Bjerregaard P, Christiansen LB, Gissel B, Hummel R, Jorgensen EB, Korsgaard B, Le Guevel R, Leffers H, McLachlan J, Moller A, Nielsen JB, Olea N, Oles-Karasko A, Pakdel F, Pedersen KL, Perez P, Skakkeboek NE, Sonnenschein C, Soto AM, Sumpter JP, Thorpe SM, Grandjean P (1999) Environ Health Perspect 107:89–108CrossRefGoogle Scholar
  8. 8.
    Elsby R, Maggs JL, Ashby J, Park BK (2001) J Pharmacol Exp Ther 297:103–113PubMedGoogle Scholar
  9. 9.
    Snyder RW, Maness SC, Gaido KW, Welsch F, Sumner SCJ, Fennell TR (2000) Toxicol Appl Pharmacol 168:225–234CrossRefPubMedGoogle Scholar
  10. 10.
    Volkel W, Colnot T, Csanady GA, Filser JG, Dekant W (2002) Chem Res Toxicol 15:1281–1287CrossRefPubMedGoogle Scholar
  11. 11.
    Calafat AM, Kuklenyik Z, Reidy JA, Caudill SP, Ekong J, Needham LL (2005) Environ Health Perspect 113:391–395CrossRefGoogle Scholar
  12. 12.
    ATSDR (2004) Toxicological profile for dichlorobenzenes. Draft for Public Comment, Agency for Toxic Substances and Disease Registry, Atlanta, GAGoogle Scholar
  13. 13.
    Tegethoff K, Herbold BA, Bomhard EM (2000) Mutat Res Genet Toxicol Environ Mutagen 470:161–167CrossRefGoogle Scholar
  14. 14.
    Hissink AM, Dunnewijk R, VanOmmen B, VanBladeren PJ (1997) Chem-Biol Interact 103:17–33CrossRefGoogle Scholar
  15. 15.
    Nedelcheva V, Gut I, Soucek P, Frantik E (1998) Chem-Biol Interact 115:53–70CrossRefPubMedGoogle Scholar
  16. 16.
    Hissink AM, Oudshoorn MJ, VanOmmen B, VanBladeren PJ (1997) Toxicol Appl Pharmacol 145:1–9CrossRefPubMedGoogle Scholar
  17. 17.
    CDC (2003) Second National Report on human exposure to environmental chemicals. National Center for Environmental Health, Division of Laboratory Sciences, Centers for Disease Control and Prevention, Atlanta, GAGoogle Scholar
  18. 18.
    Hill RH, Head SL, Baker S, Gregg M, Shealy DB, Bailey SL, Williams CC, Sampson EJ, Needham LL (1995) Environ Res 71:99–108CrossRefGoogle Scholar
  19. 19.
    Schlumpf M, Cotton B, Conscience M, Haller V, Steinmann B, Lichtensteiger W (2001) Environ Health Perspect 109:239–244CrossRefGoogle Scholar
  20. 20.
    Okereke CS, Kadry AM, Abdelrahman MS, Davis RA, Friedman MA (1993) Drug Metab Dispos 21:788–791PubMedGoogle Scholar
  21. 21.
    Felix T, Hall BJ, Brodbelt JS (1998) Anal Chim Acta 371:195–203CrossRefGoogle Scholar
  22. 22.
    Okereke CS, Abdelrahman MS, Friedman MA (1994) Toxicol Lett 73:113–122CrossRefPubMedGoogle Scholar
  23. 23.
    Kim YH, Kim CS, Park S, Han SY, Pyo MY, Yang MH (2003) Biochem Biophys Res Commun 312:441–448CrossRefPubMedGoogle Scholar
  24. 24.
    Hill RH, Shealy DB, Head SL, Williams CC, Bailey SL, Gregg M, Baker SE, Needham LL (1995) J Anal Toxicol 19:323–329PubMedGoogle Scholar
  25. 25.
    Kuklenyik Z, Ekong J, Cutchins CD, Needham LL, Calafat AM (2003) Anal Chem 75:6820–6825CrossRefPubMedGoogle Scholar
  26. 26.
    Lerch O, Zinn P (2003) J Chromatogr A 991:77–97CrossRefPubMedGoogle Scholar
  27. 27.
    Zafra A, del Olmo M, Pulgar R, Navalon A, Vilchez JL (2002) Chromatographia 56:213–218CrossRefGoogle Scholar
  28. 28.
    Rosenfeld JM, Moharir Y, Hill R (1991) Anal Chem 63:1536–1541CrossRefGoogle Scholar
  29. 29.
    Taylor JK (1987) Quality assurance of chemical measurements. Lewis, Chelsea, MIGoogle Scholar
  30. 30.
    Albro PW, Hass JR, Peck CC, Odam DG, Corbett JT, Bailey FJ, Blatt HE, Barrett BB (1981) Drug Metabol Dispos 9:223–225PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Xiaoyun Ye
    • 1
  • Zsuzsanna Kuklenyik
    • 1
  • Larry L. Needham
    • 1
  • Antonia M. Calafat
    • 1
  1. 1.Division of Laboratory Sciences, National Center for Environmental HealthCenters for Disease Control and PreventionAtlantaUSA

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