Skip to main content

Advertisement

SpringerLink
Log in

Quantitative determination of free and total bisphenol A in human urine using labeled BPA glucuronide and isotope dilution mass spectrometry

  • Research Paper
  • Published:
Analytical and Bioanalytical Chemistry Aims and scope Submit manuscript

Abstract

Bisphenol A (BPA) is a widely used industrial chemical in the manufacturing of polycarbonate plastic bottles, food and beverage can linings, thermal receipts, and dental sealants. Animal and human studies suggest that BPA may disrupt normal hormonal function and hence, potentially, have negative effects on the human health. While total BPA is frequently reported, it is recognized that free BPA is the biologically active form and is rarely reported in the literature. The objective of this study was to develop a sensitive and improved method for the measurement of free and total BPA in human urine. Use of a labeled conjugated BPA (bisphenol A-d6 β-d-glucuronide) allowed for the optimization of the enzymatic reaction and permitted an accurate determination of the conjugated BPA concentration in urine samples. In addition, a 13C12-BPA internal standard was used to account for the analytical recoveries and performance of the isotope dilution method. Solid-phase extraction (SPE) combined with derivatization and analysis using a triple quadrupole GC-EI/MS/MS system achieved very low method detection limit of 0.027 ng/mL. BPA concentrations were measured in urine samples collected during the second and third trimesters of pregnancy in 36 Canadian women. Total maternal BPA concentrations in urine samples ranged from not detected to 9.40 ng/mL (median, 1.21 ng/mL), and free BPA concentrations ranged from not detected to 0.950 ng/mL (median, 0.185 ng/mL). Eighty-six percent of the women had detectable levels of conjugated BPA, whereas only 22 % had detectable levels of free BPA in their urine. BPA levels measured in this study agreed well with data reported internationally.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. United States Environmental Protection Agency (2013) Bisphenol A (BPA) action plan summary. http://www.epa.gov/opptintr/existingchemicals/pubs/actionplans/bpa.html.

  2. Calafat AM, Ye X, Wong LY, Reidy JA, Needham LL (2008) Exposure of the U.S. population to bisphenol A and 4-tertiary-octylphenol: 2003–2004. Environ Health Perspect 116(1):39–44. doi:10.1289/ehp.10753

    Article  CAS  Google Scholar 

  3. Melzer D, Rice NE, Lewis C, Henley WE, Galloway TS (2010) Association of urinary bisphenol a concentration with heart disease: evidence from NHANES 2003/06. PLoS One 5(1):e8673

    Article  Google Scholar 

  4. Lang IA, Galloway TS, Scarlett A, Henley WE, Depledge M, Wallace RB, Melzer D (2008) Association of urinary bisphenol A concentration with medical disorders and laboratory abnormalities in adults. JAMA 300(11):1303–1310

    Article  CAS  Google Scholar 

  5. Hengstler JG, Foth H, Gebel T, Kramer PJ, Lilienblum W, Schweinfurth H, Völkel W, Wollin KM, Gundert-Remy U (2011) Critical evaluation of key evidence on the human health hazards of exposure to bisphenol A. Crit Rev Toxicol 41(4):263–291

    Article  CAS  Google Scholar 

  6. Völkel W, Colnot T, Csanády GA, Filser JG, Dekant W (2002) Metabolism and kinetics of bisphenol a in humans at low doses following oral administration. Chem Res Toxicol 15(10):1281–1287

    Article  Google Scholar 

  7. Tominaga T, Negishi T, Hirooka H, Miyachi A, Inoue A, Hayasaka I, Yoshikawa Y (2006) Toxicokinetics of bisphenol A in rats, monkeys and chimpanzees by the LC-MS/MS method. Toxicology 226(2–3):208–217

    Article  CAS  Google Scholar 

  8. Fernandez MF, Arrebola JP, Taoufiki J, Navalon A, Ballesteros O, Pulgar R, Vilchez JL, Olea N (2007) Bisphenol-A and chlorinated derivatives in adipose tissue of women. Reprod Toxicol 24(2):259–264

    Article  CAS  Google Scholar 

  9. Stahlhut RW, Welshons WV, Swan SH (2009) Bisphenol A data in NHANES suggest longer than expected half-life, substantial nonfood exposure, or both. Environ Health Perspect 117(5):784–789

    Article  CAS  Google Scholar 

  10. Ouchi K, Watanabe S (2002) Measurement of bisphenol A in human urine using liquid chromatography with multi-channel coulometric electrochemical detection. J Chromatogr B Anal Technol Biomed Life Sci 780(2):365–370

    Article  CAS  Google Scholar 

  11. Kim YH, Kim CS, Park S, Han SY, Pyo MY, Yang M (2003) Gender differences in the levels of bisphenol A metabolites in urine. Biochem Biophys Res Commun 312(2):441–448

    Article  CAS  Google Scholar 

  12. He Y, Miao M, Herrinton LJ, Wu C, Yuan W, Zhou Z, Li DK (2009) Bisphenol A levels in blood and urine in a Chinese population and the personal factors affecting the levels. Environ Res 109(5):629–633

    Article  CAS  Google Scholar 

  13. Li X, Ying G-G, Zhao J-L, Chen Z-F, Lai H-J, Su H-C (2013) 4-Nonylphenol, bisphenol-A and triclosan levels in human urine of children and students in China, and the effects of drinking these bottled materials on the levels. Environ Int 52(0):81–86. doi:10.1016/j.envint.2011.03.026

    Article  CAS  Google Scholar 

  14. Brock JW, Yoshimura Y, Barr JR, Maggio VL, Graiser SR, Nakazawa H, Needham LL (2001) Measurement of bisphenol A levels in human urine. J Expo Anal Environ Epidemiol 11(4):323–328. doi:10.1038/sj.jea.7500174

    Article  CAS  Google Scholar 

  15. Markham DA, Waechter JM Jr, Wimber M, Rao N, Connolly P, Chuang JC, Hentges S, Shiotsuka RN, Dimond S, Chappelle AH (2010) Development of a method for the determination of bisphenol A at trace concentrations in human blood and urine and elucidation of factors influencing method accuracy and sensitivity. J Anal Toxicol 34(6):293–303

    Article  CAS  Google Scholar 

  16. Ye X, Pierik FH, Hauser R, Duty S, Angerer J, Park MM, Burdorf A, Hofman A, Jaddoe VW, Mackenbach JP, Steegers EA, Tiemeier H, Longnecker MP (2008) Urinary metabolite concentrations of organophosphorous pesticides, bisphenol A, and phthalates among pregnant women in Rotterdam, the Netherlands: the Generation R study. Environ Res 108(2):260–267

    Article  CAS  Google Scholar 

  17. Ye X, Pierik FH, Angerer J, Meltzer HM, Jaddoe VW, Tiemeier H, Hoppin JA, Longnecker MP (2009) Levels of metabolites of organophosphate pesticides, phthalates, and bisphenol A in pooled urine specimens from pregnant women participating in the Norwegian Mother and Child Cohort Study (MoBa). Int J Hyg Environ Health 212(5):481–491

    Article  CAS  Google Scholar 

  18. Kawaguchi M, Inoue K, Yoshimura M, Ito R, Sakui N, Okanouchi N, Nakazawa H (2004) Determination of bisphenol A in river water and body fluid samples by stir bar sorptive extraction with in situ derivatization and thermal desorption-gas chromatography-mass spectrometry. J Chromatogr B Anal Technol Biomed Life Sci 805(1):41–48. doi:10.1016/j.jchromb.2004.02.005

    Article  CAS  Google Scholar 

  19. Kawaguchi M, Sakui N, Okanouchi N, Ito R, Saito K, Izumi S, Makino T, Nakazawa H (2005) Stir bar sorptive extraction with in situ derivatization and thermal desorption-gas chromatography–mass spectrometry for measurement of phenolic xenoestrogens in human urine samples. J Chromatogr B Anal Technol Biomed Life Sci 820(1):49–57

    Article  CAS  Google Scholar 

  20. Kawaguchi M, Ito R, Okanouchi N, Saito K, Nakazawa H (2008) Miniaturized hollow fiber assisted liquid-phase microextraction with in situ derivatization and gas chromatography–mass spectrometry for analysis of bisphenol A in human urine sample. J Chromatogr B Anal Technol Biomed Life Sci 870(1):98–102

    Article  CAS  Google Scholar 

  21. Fukata H, Miyagawa H, Yamazaki N, Mori C (2006) Comparison of ELISA- and LC-MS-based methodologies for the exposure assessment of bisphenol A. Toxicol Mech Methods 16(8):427–430. doi:10.1080/15376520600697404

    Article  CAS  Google Scholar 

  22. Dekant W, Volkel W (2008) Human exposure to bisphenol A by biomonitoring: methods, results and assessment of environmental exposures. Toxicol Appl Pharmacol 228(1):114–134

    Article  CAS  Google Scholar 

  23. Vandenberg LN, Chahoud I, Heindel JJ, Padmanabhan V, Paumgartten FJ, Schoenfelder G (2010) Urinary, circulating, and tissue biomonitoring studies indicate widespread exposure to bisphenol A. Environ Health Perspect 118(8):1055–1070. doi:10.1289/ehp.0901716

    Article  CAS  Google Scholar 

  24. Matthews JB, Twomey K, Zacharewski TR (2001) In vitro and in vivo interactions of bisphenol A and its metabolite, bisphenol A glucuronide, with estrogen receptors α and β. Chem Res Toxicol 14(2):149–157

    Article  CAS  Google Scholar 

  25. Shimizu M, Ohta K, Matsumoto Y, Fukuoka M, Ohno Y, Ozawa S (2002) Sulfation of bisphenol A abolished its estrogenicity based on proliferation and gene expression in human breast cancer MCF-7 cells. Toxicol in Vitro 16(5):549–556

    Article  CAS  Google Scholar 

  26. Fox SD, Falk RT, Veenstra TD, Issaq HJ (2011) Quantitation of free and total bisphenol A in human urine using liquid chromatography–tandem mass spectrometry. J Sep Sci 34(11):1268–1274

    Article  CAS  Google Scholar 

  27. Ye X, Kuklenyik Z, Needham LL, Calafat AM (2005) 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. Anal Bioanal Chem 383(4):638–644. doi:10.1007/s00216-005-0019-4

    Article  CAS  Google Scholar 

  28. Volkel W, Bittner N, Dekant W (2005) Quantitation of bisphenol A and bisphenol A glucuronide in biological samples by high performance liquid chromatography-tandem mass spectrometry. Drug Metab Dispos 33(11):1748–1757

    Article  Google Scholar 

  29. Volkel W, Kiranoglu M, Fromme H (2008) Determination of free and total bisphenol A in human urine to assess daily uptake as a basis for a valid risk assessment. Toxicol Lett 179(3):155–162

    Article  Google Scholar 

  30. Cunha SC, Fernandes JO (2010) Quantification of free and total bisphenol A and bisphenol B in human urine by dispersive liquid–liquid microextraction (DLLME) and heart-cutting multidimensional gas chromatography-mass spectrometry (MD-GC/MS). Talanta 83(1):117–125

    Article  CAS  Google Scholar 

  31. Volkel W, Kiranoglu M, Fromme H (2011) Determination of free and total bisphenol A in urine of infants. Environ Res 111(1):143–148

    Article  Google Scholar 

  32. Lacroix MZ, Puel S, Collet SH, Corbel T, Picard-Hagen N, Toutain P, Viguie C, Gayrard V (2011) Simultaneous quantification of bisphenol A and its glucuronide metabolite (BPA-G) in plasma and urine: applicability to toxicokinetic investigations. Talanta 85(4):2053–2059

    Article  CAS  Google Scholar 

  33. Kubwabo C, Kosarac I, Stewart B, Gauthier BR, Lalonde K, Lalonde PJ (2009) Migration of bisphenol A from plastic baby bottles, baby bottle liners and reusable polycarbonate drinking bottles. Food Addit Contam Part A Chem Anal Control Expo Risk Assess 26(6):928–937

    Article  CAS  Google Scholar 

  34. Stottmeister E, Heemken OP, Hendel P, Donnevert G, Frey S, Allmendinger H, Sawal G, Jandel B, Geiss S, Donau R, Koch A, Heinz I, Ottaviani M, Veschetti E, Hartl W, Kubwabo C, Benthe C, Tobinski V, Woldmann H, Spilker R (2009) Interlaboratory trial on the analysis of alkylphenols, alkylphenol ethoxylates, and bisphenol A in water samples according to ISO/CD 18857-2. Anal Chem 81(16):6765–6773. doi:10.1021/ac900813m

    Article  CAS  Google Scholar 

  35. United States Environmental Protection Agency (1986) Part 136—Guidelines establishing test procedures for the analysis of pollutants. http://dnr.wi.gov/regulations/labcert/documents/guidance/-LODguide.pdf)

  36. Kang JH, Katayama Y, Kondo F (2006) Biodegradation or metabolism of bisphenol A: from microorganisms to mammals. Toxicology 217(2–3):81–90. doi:10.1016/j.tox.2005.10.001

    Article  CAS  Google Scholar 

  37. Bai X, Shi H, Ye Z, Sun Q, Wang Q, Wang Z (2013) Degradation of bisphenol A by microorganisms immobilized on polyvinyl alcohol microspheres. Front Environ Sci Eng 1–7

  38. Flint S, Markle T, Thompson S, Wallace E (2012) Bisphenol A exposure, effects, and policy: a wildlife perspective. J Environ Manag 104:19–34. doi:10.1016/j.jenvman.2012.03.021

    Article  CAS  Google Scholar 

  39. Lindblom E, Press-Kristensen K, Vanrolleghem PA, Mikkelsen PS, Henze M (2009) Dynamic experiments with high bisphenol-A concentrations modelled with an ASM model extended to include a separate XOC degrading microorganism. Water Res 43(13):3169–3176. doi:10.1016/j.watres.2009.04.030

    Article  CAS  Google Scholar 

  40. Welshons WV, Nagel SC, vom Saal FS (2006) Large effects from small exposures. III. Endocrine mechanisms mediating effects of bisphenol A at levels of human exposure. Endocrinology 147(6 Suppl):S56–S69. doi:10.1210/en.2005-1159

    Article  CAS  Google Scholar 

  41. Richter CA, Birnbaum LS, Farabollini F, Newbold RR, Rubin BS, Talsness CE, Vandenbergh JG, Walser-Kuntz DR, Vom Saal FS (2007) In vivo effects of bisphenol A in laboratory rodent studies. Reprod Toxicol 24(2):199–224. doi:10.1016/j.reprotox.2007.06.004

    Article  CAS  Google Scholar 

  42. Vandenberg LN, Maffini MV, Sonnenschein C, Rubin BS, Soto AM (2009) Bisphenol-A and the great divide: a review of controversies in the field of endocrine disruption. Endocr Rev 30(1):75–95. doi:10.1210/er.2008-0021

    Article  CAS  Google Scholar 

  43. Woodruff TJ, Zota AR, Schwartz JM (2011) Environmental chemicals in pregnant women in the United States: NHANES 2003-2004. Environ Health Perspect 119(6):878–885. doi:10.1289/ehp.1002727

    Article  Google Scholar 

  44. Quiros-Alcala L, Eskenazi B, Bradman A, Ye X, Calafat AM, Harley K (2013) Determinants of urinary bisphenol A concentrations in Mexican/Mexican-American pregnant women. Environ Int 59C:152–160. doi:10.1016/j.envint.2013.05.016

    Article  Google Scholar 

  45. Barr DB, Wilder LC, Caudill SP, Gonzalez AJ, Needham LL, Pirkle JL (2005) Urinary creatinine concentrations in the U.S. population: implications for urinary biologic monitoring measurements. Environ Health Perspect 113(2):192–200

    Article  CAS  Google Scholar 

  46. Cantonwine D, Meeker JD, Hu H, Sanchez BN, Lamadrid-Figueroa H, Mercado-Garcia A, Fortenberry GZ, Calafat AM, Tellez-Rojo MM (2010) Bisphenol a exposure in Mexico City and risk of prematurity: a pilot nested case control study. Environ Health 9:62. doi:10.1186/1476-069X-9-62

    Article  Google Scholar 

  47. Philippat C, Mortamais M, Chevrier C, Petit C, Calafat AM, Ye X, Silva MJ, Brambilla C, Pin I, Charles MA, Cordier S, Slama R (2012) Exposure to phthalates and phenols during pregnancy and offspring size at birth. Environ Health Perspect 120(3):464–470. doi:10.1289/ehp.1103634

    Article  CAS  Google Scholar 

  48. Casas M, Valvi D, Luque N, Ballesteros-Gomez A, Carsin AE, Fernandez MF, Koch HM, Mendez MA, Sunyer J, Rubio S, Vrijheid M (2013) Dietary and sociodemographic determinants of bisphenol A urine concentrations in pregnant women and children. Environ Int 56:10–18. doi:10.1016/j.envint.2013.02.014

    Article  CAS  Google Scholar 

  49. Braun JM, Kalkbrenner AE, Calafat AM, Bernert JT, Ye X, Silva MJ, Barr DB, Sathyanarayana S, Lanphear BP (2011) Variability and predictors of urinary bisphenol A concentrations during pregnancy. Environ Health Perspect 119(1):131–137. doi:10.1289/ehp.1002366

    Article  CAS  Google Scholar 

  50. Braun JM, Kalkbrenner AE, Calafat AM, Yolton K, Ye X, Dietrich KN, Lanphear BP (2011) Impact of early-life bisphenol A exposure on behavior and executive function in children. Pediatrics 128(5):873–882. doi:10.1542/peds.2011-1335

    Article  Google Scholar 

  51. Donohue KM, Miller RL, Perzanowski MS, Just AC, Hoepner LA, Arunajadai S, Canfield S, Resnick D, Calafat AM, Perera FP, Whyatt RM (2013) Prenatal and postnatal bisphenol A exposure and asthma development among inner-city children. J Allergy Clin Immunol 131(3):736–742. doi:10.1016/j.jaci.2012.12.1573

    Article  CAS  Google Scholar 

  52. Martina CA, Weiss B, Swan SH (2012) Lifestyle behaviors associated with exposures to endocrine disruptors. Neurotoxicology 33(6):1427–1433. doi:10.1016/j.neuro.2012.05.016

    Article  CAS  Google Scholar 

  53. Meeker JD, Cantonwine DE, Rivera-Gonzalez LO, Ferguson KK, Mukherjee B, Calafat AM, Ye X, Anzalota Del Toro LV, Crespo-Hernandez N, Jimenez-Velez B, Alshawabkeh AN, Cordero JF (2013) Distribution, variability, and predictors of urinary concentrations of phenols and parabens among pregnant women in Puerto Rico. Environ Sci Technol 47(7):3439–3447. doi:10.1021/es400510g

    CAS  Google Scholar 

  54. Kosarac I, Kubwabo C, Lalonde K, Foster W (2012) A novel method for the quantitative determination of free and conjugated bisphenol A in human maternal and umbilical cord blood serum using a two-step solid phase extraction and gas chromatography/tandem mass spectrometry. J Chromatogr B Anal Technol Biomed Life Sci 898:90–94. doi:10.1016/j.jchromb.2012.04.023

    Article  CAS  Google Scholar 

  55. Philippat C, Wolff MS, Calafat AM, Ye X, Bausell R, Meadows M, Stone J, Slama R, Engel SM (2013) Prenatal exposure to environmental phenols: concentrations in amniotic fluid and variability in urinary concentrations during pregnancy. Environ Health Perspect 121(10):1225–1231. doi:10.1289/ehp.1206335

    CAS  Google Scholar 

  56. Chevrier J, Gunier RB, Bradman A, Holland NT, Calafat AM, Eskenazi B, Harley KG (2013) Maternal urinary bisphenol a during pregnancy and maternal and neonatal thyroid function in the CHAMACOS study. Environ Health Perspect 121(1):138–144. doi:10.1289/ehp.1205092

    CAS  Google Scholar 

  57. Harley KG, Aguilar Schall R, Chevrier J, Tyler K, Aguirre H, Bradman A, Holland NT, Lustig RH, Calafat AM, Eskenazi B (2013) Prenatal and postnatal bisphenol A exposure and body mass index in childhood in the CHAMACOS cohort. Environ Health Perspect 121(4):514–520. doi:10.1289/ehp.1205548, 520e511-516

    Google Scholar 

Download references

Acknowledgments

This work was supported by Canada’s Chemicals Management Plan.

Author information

Authors and Affiliations

  1. Environmental and Radiation Health Sciences Directorate, Health Canada, Ottawa, ON, K1A 0K9, Canada

    Cariton Kubwabo, Ivana Kosarac & Kaela Lalonde

  2. Department of Obstetrics and Gynecology, McMaster University, Hamilton, ON, L8N 3Z5, Canada

    Warren G. Foster

Authors
  1. Cariton Kubwabo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ivana Kosarac
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Kaela Lalonde
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Warren G. Foster
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Cariton Kubwabo.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Kubwabo, C., Kosarac, I., Lalonde, K. et al. Quantitative determination of free and total bisphenol A in human urine using labeled BPA glucuronide and isotope dilution mass spectrometry. Anal Bioanal Chem 406, 4381–4392 (2014). https://doi.org/10.1007/s00216-014-7829-1

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00216-014-7829-1

Keywords

Search

Navigation