Author's personal copy Journal of Chromatography B, 878 (2010) 3363–3369 Contents lists available at ScienceDirect Journal of Chromatography B journal homepage: www.elsevier.com/locate/chromb Determination of Bisphenol A and its chlorinated derivatives in placental tissue samples by liquid chromatography–tandem mass spectrometry I. Jiménez-Díaz a,b , A. Zafra-Gómez a , O. Ballesteros a,∗ , N. Navea b , A. Navalón a , M.F. Fernández b , N. Olea b , J.L. Vílchez a a Research Group of Analytical Chemistry and Life Sciences, Department of Analytical Chemistry, University of Granada, Campus of Fuentenueva, E-18071 Granada, Spain b Laboratory of Medical Investigations, San Cecilio University Hospital, University of Granada, CIBER Epidemiology and Public Health (CIBERESP), E-18071 Granada, Spain article info Article history: Received 7 July 2010 Accepted 24 October 2010 Available online 30 October 2010 Keywords: Bisphenol A Bisphenol A chlorinated derivatives Liquid chromatography–tandem mass spectrometry (LC–MS/MS) Placental tissue analysis abstract The group of compounds commonly called endocrine disruptors covers a wide range of synthetic and nat- ural substances able to alter the normal hormone function of wildlife and humans, consequently causing adverse health effects. Bisphenol A (BPA) and its chlorinated derivatives are some of these compounds. In this work, we propose a new liquid chromatography–tandem mass spectrometry (LC–MS/MS) method to determine these compounds in human placental tissue samples. The method involves an extraction phase of the extracts from the samples using ethyl acetate, followed by a clean-up phase by centrifuga- tion prior to their quantification by LC–MS/MS using an atmospheric pressure chemical ionization (APCI) interface in the negative mode. Deuterated Bisphenol A (BPA-d 16 ) was used as internal standard. Found detection limits (DL) ranged from 0.2 to 0.6 ng g -1 and quantification limits (QL) from 0.5 to 2.0 ng g -1 for Bisphenol A and its chlorinated derivatives, while inter- and intra-day variability was under 8.1%. The method was validated using standard addition calibration and a spike recovery assay. Recovery rates for spiked samples ranged from 97% to 105%. This method was satisfactorily applied to the determination of BPA and its chlorinated derivatives in 49 placental tissue samples collected from women who live in the province of Granada (Spain). © 2010 Elsevier B.V. All rights reserved. 1. Introduction It is well known that about 100 of industrial chemicals have estrogenic activity in addition to their desired chemical properties. Most of these compounds, commonly called endocrine disrupting chemicals (EDCs), are organic compounds widely used, which are ubiquitous in the environment and in biological samples [1–3]. The main effect of these compounds is associated with their capability to mimic or block the action of natural hormones in living organ- isms, including humans. Estrogen mimics have been perhaps the most studied of all types of EDCs and the ones to capture much of the public’s attention as a public health issue. Many researchers hypothesize that exposure to these endocrine disruptors during critical periods of development – in utero or early postnatal life – could cause morphologic and functional alterations in wildlife and humans affecting growth, reproduction and development [4–6]. It has also been postulated that EDCs accumulate in certain human tissues and their effects might pass to the offspring via the placenta and/or breast milk [7]. ∗ Corresponding author. Tel.: +34 958 243326; fax: +34 958 243328. E-mail address: oballest@ugr.es (O. Ballesteros). One of the representative compounds of the EDCs group is BPA. BPA is a compound with high reactivity and is the raw material used in a large amount of manufactured products, such as polycarbonate plastics, epoxy resins used to line metal cans, and in many plastic consumer products including toys, water pipes, drinking contain- ers, eyeglass lenses, sports safety equipment, dental monomers, medical equipment and tubing and consumer electronics [8]. BPA is one of the highest volume chemicals produced worldwide, with an estimated production of 3.9 million tons in 2006 [10] and over 100 tons released into the atmosphere from its annual production [9]. When BPA is present in treated waters, it may react with resid- ual chlorine originally used as a disinfectant, producing chlorinated BPA derivatives depending on the pH of the medium [11]. Polychlo- rinated BPA has been recently identified and biodegradation test using activated sludge revealed that it is not easily biodegraded [12], in this sense, chlorinated BPA derivatives are more cytotoxic than BPA [13] and all of them show estrogenic activity [14]. Estrogenic activity of BPA was first reported in 1993 [15]. BPA affinity for estrogen receptors is 10,000 to 100,000 fold weaker than that of estradiol, so it has been considered a very weak environmen- tal estrogen. Recent in vitro studies have shown that BPA effects are mediated by both genomic and epigenomic estrogen-response mechanisms, with the disruption of the cell function occurring at 1570-0232/$ – see front matter © 2010 Elsevier B.V. All rights reserved. doi:10.1016/j.jchromb.2010.10.021