Analytical and Bioanalytical Chemistry

, Volume 401, Issue 3, pp 995–1002

Measurement of bisphenol A, bisphenol A ß-d-glucuronide, genistein, and genistein 4′-ß-d-glucuronide via SPE and HPLC-MS/MS

Original Paper

DOI: 10.1007/s00216-011-5151-8

Cite this article as:
Coughlin, J.L., Winnik, B. & Buckley, B. Anal Bioanal Chem (2011) 401: 995. doi:10.1007/s00216-011-5151-8


Bisphenol A (BPA) is a synthetic industrial reactant used in the production of polycarbonate plastics, and genistein is a natural phytoestrogen abundant in the soybean. Current studies investigating the endocrine-disrupting effects of concomitant exposures to BPA and genistein have warranted the development of an analytical method for the simultaneous measurement of BPA and genistein, as well as their primary metabolites, bisphenol A ß-d-glucuronide (BPA gluc) and genistein 4′-ß-d-glucuronide (genistein gluc), respectively. All four analytes were extracted from rat plasma via solid phase extraction (SPE). Three SPE cartridges and four elution schemes were tested. Plasma extraction using Bond Elut Plexa cartridges with sequential addition of ethyl acetate, methanol, and acetonitrile yielded optimal average recoveries of 98.1 ± 1.8% BPA, 94.9 ± 8.0% genistein, 91.4 ± 6.1% BPA gluc, and 103 ± 6.1% genistein gluc. Identification and quantification of the four analytes were performed by a validated HPLC-MS/MS method using electrospray ionization and selective reaction monitoring. This novel analytical method should be applicable to the measurement of BPA, genistein, BPA gluc, and genistein gluc in urine, cultures, and tissue following in vivo exposures. While reports of the determination of BPA and genistein independently exist, the simultaneous optimized extraction and detection of BPA, genistein, BPA gluc, and genistein gluc have not previously been reported.


BPA and genistein co-exposure scenario. BPA-laden polycarbonate plastic baby bottle filled with soymilk, a rich source of genistein, provides a classic exposure scenario to young children—a population that is particularly vulnerable to the effects of endocrine-disrupting compounds


Bisphenol A (BPA) Genistein Bisphenol A ß-d-glucuronide Genistein 4′-ß-d-glucuronide HPLC Mass spectrometry Electrospray ionization Chromatographic techniques 



Electrospray ionization


High-performance liquid chromatography


Mass spectrometry


Tandem MS


Relative standard deviation

Supplementary material

216_2011_5151_MOESM1_ESM.pdf (101 kb)
ESM 1(PDF 101 kb)

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Janis L. Coughlin
    • 1
    • 2
    • 3
  • Bozena Winnik
    • 1
  • Brian Buckley
    • 1
    • 2
    • 3
  1. 1.Environmental and Occupational Health Sciences InstituteA Joint Institute of Rutgers University and the University of Medicine and Dentistry of New Jersey (UMDNJ)PiscatawayUSA
  2. 2.Joint Graduate Program of ToxicologyRutgers UniversityPiscatawayUSA
  3. 3.University of Medicine and Dentistry of New JerseyPiscatawayUSA