Please cite this article in press as: G. Provencher, et al., J. Chromatogr. A (2014), http://dx.doi.org/10.1016/j.chroma.2014.04.072 ARTICLE IN PRESS G Model CHROMA-355364; No. of Pages 8 Journal of Chromatography A, xxx (2014) xxx–xxx Contents lists available at ScienceDirect Journal of Chromatography A j o ur na l ho me page: www.elsevier.com/locate/chroma Determination of bisphenol A, triclosan and their metabolites in human urine using isotope-dilution liquid chromatography–tandem mass spectrometry Gilles Provencher a , René Bérubé a , Pierre Dumas a , Jean-Franc ¸ ois Bienvenu a , Éric Gaudreau a , Patrick Bélanger a , Pierre Ayotte a,b,c,∗ a Centre de toxicologie du Québec, Institut national de santé publique du Québec (INSPQ), 945 Wolfe, Québec, QC, Canada G1V 5B3 b Axe Santé des populations et pratiques optimales en santé, Centre de recherche du CHU de Québec, 2875 boul. Laurier, Édifice Delta 2, Bureau 600, Québec, QC, Canada G1V 2M2 c Département de médecine sociale et préventive, Université Laval, Pavillon Ferdinand-Vandry, Québec, QC, Canada G1V 0A6 a r t i c l e i n f o Article history: Received 30 December 2013 Received in revised form 15 April 2014 Accepted 20 April 2014 Available online xxx Keywords: Bisphenol A Triclosan Glucuronide metabolites Sulfate metabolites Isotope dilution LC–MS/MS Human urine a b s t r a c t Bisphenol A (BPA) and triclosan (TCS) are ubiquitous environmental phenols exhibiting endocrine dis- rupting activities that may be involved in various health disorders in humans. There is a need to measure separately free forms and conjugated metabolites because only the former are biologically active. We have developed sensitive methods using isotope-dilution liquid chromatography–tandem mass spec- trometry for individual measurements of free BPA and TCS as well as their metabolites, BPA glucuronide (BPAG), BPA monosulfate (BPAS), BPA disulfate (BPADS), TCS glucuronide (TCSG) and TCS sulfate (TCSS) in urine. Comparative analyses of urine samples from 46 volunteers living in the Quebec City area using the new methods and a GC–MS/MS method previously used in our laboratory revealed very strong correla- tions for total BPA (Spearman’s r s = 0.862, p < 0.0001) and total TCS concentrations (r s = 0.942, p < 0.0001). Glucuronide metabolites were the most abundant BPA and TCS species in urine samples (>94% of total urinary concentrations). Unconjugated TCS concentrations represented a small proportion of total TCS species (median = 1.6%) but its concentration was likely underestimated due to losses by adsorption to the surface of polypropylene tubes used for sample storage. To our knowledge, we are the first to report levels of free, sulfated and glucuronidated TCS levels in human urine. © 2014 Elsevier B.V. All rights reserved. 1. Introduction Bisphenol A (BPA) and triclosan (TCS) are two widely used phenolic compounds with numerous industrial and commercial applications. BPA is the monomer in the production of polycarbo- nate plastics and is also a component of epoxy resins that are used to produce food and beverage packaging materials and dental sealants among others [1]. Trace amounts of BPA have been shown to leach from polycarbonate containers [2–4], which led to the ban of poly- carbonate baby bottles in Canada in 2010. BPA has been found in foodstuffs [5,6], and diet is most likely the major source of exposure to this compound [1,7,8]. TCS is a bactericidal agent added to soap and toothpaste [9,10]. Dermal absorption and ingestion of TCS in ∗ Corresponding author at: Centre de toxicologie du Québec, Institut national de santé publique du Québec (INSPQ), 945 Wolfe, Québec, QC, Canada G1V 5B3. Tel.: +1 418 650 5115x4654; fax: +1 418 654 2148. E-mail address: pierre.ayotte@inspq.qc.ca (P. Ayotte). these personal care products are thought to contribute the most to human exposure [9]. Both BPA and TCS are endocrine disrupting chemicals: BPA is an estrogen receptor agonist and an androgen receptor antag- onist [11,12]. TCS interacts with constitutive androstane and pregnane-X receptors [13] and exhibits antagonistic activity in both estrogen receptor- and androgen receptor-responsive bioassays [14]. Human exposure to these compounds have been associated with various adverse health effects including reproductive and endocrine disorders, cardiovascular disease, obesity and metabolic syndrome, cancer, immune system dysfunction and neurobehav- ioral defects [1,15–17]. However, whether or not these associations indicate an important public health risk is highly controversial. An argument raised by those supporting a trivial health risk from exposure to these phenolic compounds is their rapid bio- transformation to biologically-inactive metabolites [18]. Indeed, following their absorption by the gastro-intestinal tract, free BPA and TCS are rapidly and efficiently conjugated by hepatic uridine diphosphate-glucuronyltransferases to their glucuronidated forms http://dx.doi.org/10.1016/j.chroma.2014.04.072 0021-9673/© 2014 Elsevier B.V. All rights reserved.