Reliable quantification of bisphenol A and its chlorinated derivatives in human urine using UPLC-MS/MS method

Talanta. 2014 Jul:125:284-92. doi: 10.1016/j.talanta.2014.02.064. Epub 2014 Mar 12.


Bisphenol A (BPA), a widespread man-made chemical classified as an endocrine disruptor, is increasingly considered as a major cause of concern for human health. Chlorine present in drinking water may react with BPA to form chlorinated derivatives (ClxBPA), which have demonstrated a heightened level of estrogenic activity. If many epidemiological studies report that more than 90% of people have detectable BPA levels in their urine, then no such study has been undertaken regarding ClxBPA. The purpose of this work is to propose a highly sensitive and accurate analytical method adapted to large-scale biomonitoring studies aimed at assessing exposure to BPA and ClxBPA through the use of human urine. To achieve this, we have comprehensively validated a method using salting-out assisted liquid/liquid extraction (SALLE) coupled to UPLC-MS/MS and isotope dilution quantification, to measure unconjugated BPA and ClxBPA in human urine according to the accepted guidelines. Deutered BPA as well as deutered 2,2'-DCBPA was used as internal standards. The matrix calibration curve ranged from 0.05 to 1.60 ng mL(-1) and from 0.5 to 16.0 ng mL(-1) for ClxBPA and BPA respectively, and provided good linearity (r²>0.99). This method was precise (the intra- and inter-day coefficients of variation were <20% at three different concentrations: 0.05 ng mL(-1), 0.2 ng mL(-1), 0.8 ng mL(-1) and 0.5 ng mL(-1), 2 ng mL(-1), 8 ng mL(-1) for ClxBPA and BPA, respectively) and accurate (bias ranged from -13% to +12%). The limit of quantification, validated at 0.05 ng mL(-1) and 0.5 ng mL(-1) for ClxBPA and BPA respectively when using 300 µL of urine, was found to be suitable for the concentration existing in real samples. The matrix effect and the BPA cross-contamination were also investigated in this study. The analytical method developed in this study is in accordance with the requirements applicable to biomonitoring of BPA and ClxBPA in human urine.

Keywords: Bisphenol A; Bisphenol A chlorinated derivatives; LC–MS/MS; Urine.

MeSH terms

  • Benzhydryl Compounds / urine*
  • Calibration
  • Chemistry Techniques, Analytical
  • Chlorine / analysis*
  • Chromatography, High Pressure Liquid*
  • Humans
  • Phenols / urine*
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Tandem Mass Spectrometry*
  • Urinalysis / standards*
  • Urine / chemistry*


  • Benzhydryl Compounds
  • Phenols
  • Chlorine
  • bisphenol A