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Drug Metabolism and Disposition Fast Forward
First published on August 15, 2005; DOI: 10.1124/dmd.105.005454

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Received for publication May 12, 2005.
Revised July 14, 2005.
Accepted for publication August 15, 2005.

Quantitation of bisphenol A and bisphenol A glucuronide in biological samples by HLPC-MS/MS

Wolfgang Volkel 1, Nataly Bittner 1, Wolfgang Dekant 1*

1 Department of Toxicology, University of Wuerzburg

* Address correspondence to: E-mail: dekant{at}toxi.uni-wuerzburg.de

Abstract

Bisphenol A (BPA) is a weak estrogen. Pharmacokinetic studies of BPA have demonstrated a rapid and extensive metabolism of BPA to the non-estrogenic BPA-monoglucuronide (BPA-gluc). Some investigators have reported that BPA was found at parts per billion concentrations in the tissues or urine of humans without known exposure to BPA. This work developed a rapid and sensitive method for the determination of BPA and BPA-gluc in plasma and urine based on LC-MS/MS. The LC-ESI-MS/MS method for quantitation of BPA and BPA-gluc uses stable isotope labelled internal standards. A linear ion trap mass spectrometer permits identification and quantitation of BPA-gluc and BPA without sample workup. Development of separation conditions reduced the BPA-background in solvent samples to below 2.5 pmol/mL for BPA. Limit of quantitation (LOQ) for BPA in control urine was 15 pmol/mL, LOQ for BPA-gluc was 65 pmol/mL. Application of the method to urine samples from human subjects (n = 6) after administration of 25 µg BPA/person (estimated maximum human daily intake) permitted the determination of excretion kinetics for BPA-gluc; BPA was below the LOD in all except two of the samples. In urine or blood samples of human subjects (n = 19) without intentional exposure to BPA, BPA-concentrations were always below the limit of detection ({approx} 2.5 pmol/mL) with or without prior glucuronidase treatment. The results show that care is required for analysis of BPA and its major metabolite BPA-gluc. The LOD obtained and the absence of detectable levels of BPA in samples from individuals suggests that general exposure of humans to BPA is much lower than worst-case exposure scenario developed.


Key words: analytical chemistry, analytical pharmacology/toxicology, chemical toxicology, endocrine regulation, human pharmacokinetics, mass spectrometry, phase II drug metabolism, toxicology


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