Analytical and Bioanalytical Chemistry

, Volume 410, Issue 16, pp 3871–3883 | Cite as

Rapid and sensitive determination of nine bisphenol analogues, three amphenicol antibiotics, and six phthalate metabolites in human urine samples using UHPLC-MS/MS

  • Yuan Yao
  • Yijun Shao
  • Ming Zhan
  • Xiaoli Zou
  • Weidong Qu
  • Ying Zhou
Research Paper


Bisphenol analogues, amphenicol antibiotics, and phthalate have widely aroused public concerns due to their adverse effects on human health. In this study, a rapid and sensitive method for determination of nine bisphenol analogues, three amphenicol antibiotics, and six phthalate metabolites in the urine based on ultra-high-performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry was developed and validated. The sample pretreatment condition on the base of mixed-mode anion-exchange (Oasis MAX) SPE was optimized to separate bisphenol analogues and amphenicol antibiotics from phthalate metabolites: the former were detected with a mobile phase of 0.1% ammonium water solution/methanol containing 0.1% ammonium water solution in negative mode, whereas the latter were determined with a mobile phase of 0.1% acetic acid solution/acetonitrile containing 0.1% acetic acid in negative mode. The limits of detection were less than 0.26 ng/mL for bisphenol analogues, 0.12 ng/mL for amphenicol antibiotics, and 0.14 ng/mL for phathalate metabolites. The recoveries of all target analytes in three fortification levels ranged from 72.02 to 117.64% with the relative standard deviations of no larger than 14.51%. The matrix effect was adjusted by isotopically labeled internal standards. This proposed method was successfully applied to analyze 40 actual urines and 13 out of 18 studied compounds were detected.

Graphical abstract

Simultaneous determination of nine bisphenol analogues, three amphenicol antibiotics, and six phthalate metabolites in human urine samples


Bisphenol analogues Amphenicol antibiotics Phthalate metabolites Mixed-mode solid-phase extraction Ultra performance liquid chromatography-tandem mass spectrometry Urine sample 



This study was supported by the National Natural Science Foundation of China (No. 81373089), Scientific Research Foundation of Shanghai Municipal Commission of Health and Family Planning (No. 201540053), the National Science Fund for Distinguished Young Scholars of China (No. 81325017), and the Key Program of the National Natural Science Foundation of China (No. 81630088).

Compliance with ethical standards

The study was approved by the Institutional Review Board (IRB) of the School of Public Health, Fudan University (ref: IRB#2013-03-0437). Written informed consent was obtained from all participants and the parents/LAR of the participants.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

216_2018_1062_MOESM1_ESM.pdf (1.1 mb)
ESM 1 (PDF 1077 kb)


  1. 1.
    Muhamad MS, Salim MR, Lau WJ, Yusop Z. A review on bisphenol A occurrences, health effects and treatment process via membrane technology for drinking water. Environ Sci Pollut Res Int. 2016;23(12):11549–67.CrossRefPubMedGoogle Scholar
  2. 2.
    Guo Y, Alomirah H, Cho HS, Minh TB, Mohd MA, Nakata H, et al. Occurrence of phthalate metabolites in human urine from several Asian countries. Environ Sci Technol. 2011;45(7):3138–44.CrossRefPubMedGoogle Scholar
  3. 3.
    Rubin BS. Bisphenol A: an endocrine disruptor with widespread exposure and multiple effects. J Steroid Biochem Mol Biol. 2011;127(1–2):27–34.CrossRefPubMedGoogle Scholar
  4. 4.
    Swan SH. Environmental phthalate exposure in relation to reproductive outcomes and other health endpoints in humans. Environ Res. 2008;108(2):177–84.CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Diamanti-Kandarakis E, Bourguignon JP, Giudice LC, Hauser R, Prins GS, Soto AM, et al. Endocrine-disrupting chemicals: an Endocrine Society scientific statement. Endocr Rev. 2009;30(4):293–342.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Casals-Casas C, Desvergne B. Endocrine disruptors: from endocrine to metabolic disruption. Annu Rev Physiol. 2011;73:135–62.CrossRefPubMedGoogle Scholar
  7. 7.
    Chen D, Kannan K, Tan H, Zheng Z, Feng YL, Wu Y, et al. Bisphenol analogues other than BPA: environmental occurrence, human exposure, and toxicity-a review. Environ Sci Technol. 2016;50(11):5438–53.CrossRefPubMedGoogle Scholar
  8. 8.
    Caballero-Casero N, Lunar L, Rubio S. Analytical methods for the determination of mixtures of bisphenols and derivatives in human and environmental exposure sources and biological fluids. A review. Anal Chim Acta. 2016;908:22–53.CrossRefPubMedGoogle Scholar
  9. 9.
    Liu HY, Lin SL, Fuh MR. Determination of chloramphenicol, thiamphenicol and florfenicol in milk and honey using modified QuEChERS extraction coupled with polymeric monolith-based capillary liquid chromatography tandem mass spectrometry. Talanta. 2016;150:233–9.CrossRefPubMedGoogle Scholar
  10. 10.
    Hanekamp JC, Bast A. Antibiotics exposure and health risks: chloramphenicol. Environ Toxicol Pharmacol. 2015;39(1):213–20.CrossRefPubMedGoogle Scholar
  11. 11.
    Chen M, Tao L, Collins EM, Austin C, Lu C. Simultaneous determination of multiple phthalate metabolites and bisphenol-A in human urine by liquid chromatography-tandem mass spectrometry. J Chromatogr B Anal Technol Biomed Life Sci. 2012;904:73–80.CrossRefGoogle Scholar
  12. 12.
    Zhang S, Liu Z, Guo X, Cheng L, Wang Z, Shen J. Simultaneous determination and confirmation of chloramphenicol, thiamphenicol, florfenicol and florfenicol amine in chicken muscle by liquid chromatography-tandem mass spectrometry. J Chromatogr B Anal Technol Biomed Life Sci. 2008;875(2):399–404.CrossRefGoogle Scholar
  13. 13.
    Rocha BA, da Costa BR, de Albuquerque NC, de Oliveira AR, Souza JM, Al-Tameemi M, et al. A fast method for bisphenol A and six analogues (S, F, Z, P, AF, AP) determination in urine samples based on dispersive liquid-liquid microextraction and liquid chromatography-tandem mass spectrometry. Talanta. 2016;154:511–9.CrossRefPubMedGoogle Scholar
  14. 14.
    Zhou X, Kramer JP, Calafat AM, Ye X. Automated on-line column-switching high performance liquid chromatography isotope dilution tandem mass spectrometry method for the quantification of bisphenol A, bisphenol F, bisphenol S, and 11 other phenols in urine. J Chromatogr B Anal Technol Biomed Life Sci. 2014;944:152–6.CrossRefGoogle Scholar
  15. 15.
    Wang HX, Wang B, Zhou Y, Jiang QW. Rapid and sensitive analysis of phthalate metabolites, bisphenol A, and endogenous steroid hormones in human urine by mixed-mode solid-phase extraction, dansylation, and ultra-performance liquid chromatography coupled with triple quadrupole mass spectrometry. Anal Bioanal Chem. 2013;405(12):4313–9.CrossRefPubMedGoogle Scholar
  16. 16.
    Heffernan AL, Thompson K, Eaglesham G, Vijayasarathy S, Mueller JF, Sly PD, et al. Rapid, automated online SPE-LC-QTRAP-MS/MS method for the simultaneous analysis of 14 phthalate metabolites and 5 bisphenol analogues in human urine. Talanta. 2016;151:224–33.CrossRefPubMedGoogle Scholar
  17. 17.
    Wang HX, Wang B, Zhou Y, Jiang QW. Rapid and sensitive screening and selective quantification of antibiotics in human urine by two-dimensional ultraperformance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. Anal Bioanal Chem. 2014;406(30):8049–58.CrossRefPubMedGoogle Scholar
  18. 18.
    Kim M, Song NR, Choi JH, Lee J, Pyo H. Simultaneous analysis of urinary phthalate metabolites of residents in Korea using isotope dilution gas chromatography-mass spectrometry. Sci Total Environ. 2014;470-471:1408–13.CrossRefPubMedGoogle Scholar
  19. 19.
    Cunha SC, Fernandes JO. 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. 2010;83(1):117–25.CrossRefPubMedGoogle Scholar
  20. 20.
    Gustafsson JBC, Uzqueda BHR. The influence of citrate and phosphate on the mancini single radial immunodiffusion technique and suggested improvements for the determination of urinary albumin. Clin Chim Acta. 1978;90(3):249–57.CrossRefPubMedGoogle Scholar
  21. 21.
    Wang B, Wang H, Zhou W, Chen Y, Zhou Y, Jiang Q. Urinary excretion of phthalate metabolites in school children of China: implication for cumulative risk assessment of phthalate exposure. Environ Sci Technol. 2015;49(2):1120–9.CrossRefPubMedGoogle Scholar
  22. 22.
    Regueiro J, Breidbach A, Wenzl T. Derivatization of bisphenol A and its analogues with pyridine-3-sulfonyl chloride: multivariate optimization and fragmentation patterns by liquid chromatography/Orbitrap mass spectrometry. Rapid Commun Mass Spectrom. 2015;29(16):1473–84.CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Commission Decision (2002/657/EC) of 12 August 2002 implementing Council Directive 96/23/EC concerning the performance of analytical methods and the interpretation of results. Off J Eur Commun L221:8–36.Google Scholar
  24. 24.
    Benijts T, Dams R, Lambert W, De Leenheer A. Countering matrix effects in environmental liquid chromatography–electrospray ionization tandem mass spectrometry water analysis for endocrine disrupting chemicals. J Chromatogr A. 2004;1029(1–2):153–9.CrossRefPubMedGoogle Scholar
  25. 25.
    Li M, Wang Q, Zhu J, Li N, Zou X. A simple analytical method of determining 1-hydroxypyrene glucuronide in human urine by isotope dilution with ultra performance liquid chromatography-tandem mass spectrometry. Anal Bioanal Chem. 2017;409(6):1513–8.CrossRefPubMedGoogle Scholar
  26. 26.
    Yang Y, Lu L, Zhang J, Yang Y, Wu Y, Shao B. Simultaneous determination of seven bisphenols in environmental water and solid samples by liquid chromatography-electrospray tandem mass spectrometry. J Chromatogr A. 2014;1328:26–34. Scholar
  27. 27.
    Vela-Soria F, Ballesteros O, Zafra-Gomez A, Ballesteros L, Navalon A. UHPLC-MS/MS method for the determination of bisphenol A and its chlorinated derivatives, bisphenol S, parabens, and benzophenones in human urine samples. Anal Bioanal Chem. 2014;406(15):3773–85.CrossRefPubMedGoogle Scholar
  28. 28.
    Gonzalez-Marino I, Quintana JB, Rodriguez I, Gonzalez-Diez M, Cela R. Screening and selective quantification of illicit drugs in wastewater by mixed-mode solid-phase extraction and quadrupole-time-of-flight liquid chromatography-mass spectrometry. Anal Chem. 2012;84(3):1708–17.CrossRefPubMedGoogle Scholar
  29. 29.
    Salas D, Borrull F, Marce RM, Fontanals N. Study of the retention of benzotriazoles, benzothiazoles and benzenesulfonamides in mixed-mode solid-phase extraction in environmental samples. J Chromatogr A. 2016;1444:21–31.CrossRefPubMedGoogle Scholar
  30. 30.
    Laven M, Alsberg T, Yu Y, Adolfsson-Erici M, Sun H. Serial mixed-mode cation- and anion-exchange solid-phase extraction for separation of basic, neutral and acidic pharmaceuticals in wastewater and analysis by high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry. J Chromatogr A. 2009;1216(1):49–62.CrossRefPubMedGoogle Scholar
  31. 31.
    Yang Y, Guan J, Yin J, Shao B, Li H. Urinary levels of bisphenol analogues in residents living near a manufacturing plant in South China. Chemosphere. 2014;112:481–6.CrossRefPubMedGoogle Scholar
  32. 32.
    Lewis RC, Meeker JD, Peterson KE, Lee JM, Pace GG, Cantoral A, et al. Predictors of urinary bisphenol A and phthalate metabolite concentrations in Mexican children. Chemosphere. 2013;93(10):2390–8.CrossRefPubMedGoogle Scholar
  33. 33.
    Myridakis A, Balaska E, Gkaitatzi C, Kouvarakis A, Stephanou EG. Determination and separation of bisphenol A, phthalate metabolites and structural isomers of parabens in human urine with conventional high-pressure liquid chromatography combined with electrospray ionisation tandem mass spectrometry. Anal Bioanal Chem. 2015;407(9):2509–18.CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Yuan Yao
    • 1
    • 2
  • Yijun Shao
    • 1
    • 2
  • Ming Zhan
    • 3
  • Xiaoli Zou
    • 4
  • Weidong Qu
    • 1
    • 5
  • Ying Zhou
    • 1
    • 2
    • 3
  1. 1.Key Laboratory of Public Health Security, School of Public Health, Ministry of EducationFudan UniversityShanghaiChina
  2. 2.Department of Chemistry in Public Health, School of Public HealthFudan UniversityShanghaiChina
  3. 3.Pudong New Area for Disease Control and PreventionFudan University Pudong Institute of Preventive MedicineShanghaiChina
  4. 4.Department of Sanitary Technology, West China School of Public HealthUniversity of SichuanChengduChina
  5. 5.Department of Environmental Health, School of Public HealthFudan UniversityShanghaiChina

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