Combining passive samplers and biomonitors to evaluate endocrine disrupting compounds in a wastewater treatment plant by LC/MS/MS and bioassay analyses C. Liscio a , E. Magi a, * , M. Di Carro a , M.J.-F. Suter b , E.L.M. Vermeirssen b a Dipartimento di Chimica e Chimica Industriale, Universita ` di Genova, via Dodecaneso, 31,16146 Genova, Italy b Eawag, Swiss Federal Institute of Aquatic Science and Technology, U ¨ berlandstrasse 133, 8600 Du ¨bendorf, Switzerland Passive sampling and biomonitoring were used to determine the amounts of endocrine disrupting compounds in wastewaters. article info Article history: Received 11 February 2009 Received in revised form 24 April 2009 Accepted 27 April 2009 Keywords: Endocrine disrupting compounds (EDCs) Passive sampler Biomonitor Mussels Wastewater abstract Two types of integrative sampling approaches (passive samplers and biomonitors) were tested for their sampling characteristics of selected endocrine disrupting compounds (EDCs). Chemical analyses (LC/MS/ MS) were used to determine the amounts of five EDCs (nonylphenol, bisphenol A, estrone, 17b-estradiol and 17a-ethinylestradiol) in polar organic chemical integrative samplers (POCIS) and freshwater mussels (Unio pictorum); both had been deployed in the influent and effluent of a municipal wastewater treat- ment plant (WWTP) in Genoa, Italy. Estrogenicity of the POCIS samples was assessed using the yeast estrogen screen (YES). Estradiol equivalent values derived from the bioassay showed a positive corre- lation with estradiol equivalents calculated from chemical analyses data. As expected, the amount of estrogens and EEQ values in the effluent were lower than those in the influent. Passive sampling proved to be the preferred method for assessing the presence of these compounds since employing mussels had several disadvantages both in sampling efficiency and sample analyses. Ó 2009 Elsevier Ltd. All rights reserved. 1. Introduction Over the last two decades the presence in the environment of compounds with estrogenic and androgenic properties that can alter the normal functioning of the endocrine system in wildlife and humans has emerged as a major environmental issue. It has generated a considerable amount of attention among the world- wide scientific community, and the potential adverse effects of these endocrine disrupting compounds (EDCs) have been well documented in the literature (Hill et al., 2002; Madsen et al., 2004). Furthermore, scientific results are continuously feeding into regulatory measures, some compounds have been banned, and appropriate tools are needed to study the presence of these compounds in discharges to the aquatic environment. Natural estrogens (such as estrone, E1, and 17b-estradiol, E2) and synthetic estrogens (e.g. 17a-ethinylestradiol, EE2, the active component of oral contraceptives) are very powerful EDCs. They derive mainly from excreta of humans and livestock (Williams et al., 2003). However, man-made chemicals such as nonylphenol (NP) and bisphenol A (BPA) are also known to influence the hormonal system of aquatic organisms (Fromme et al., 2002; Madsen et al., 2004). Alkylphenols and BPA are used to manufacture flame retar- dants, household surfactants, pesticide formulations, industrial products and polycarbonate and epoxy resins. EDCs from domestic, agricultural or industrial sources are released directly or indirectly into the aquatic environment (Birkett and Lester, 2003). Wastewater treatment plants (WWTPs) are important sources of pollution, since EDCs are not fully removed by the treatment processes. The occurrence of these EDCs in WWTPs has been well documented in numerous studies worldwide (Nakada et al., 2006; Vogelsang et al., 2006), including the Medi- terranean area (Gomez et al., 2007; Gonzalez et al., 2007). Monitoring for these compounds can be challenging due to the rapid dilution of the effluents and thus requires sensitive analytical techniques, intensive sampling programs and large sample volumes. Analytical methods for the determination of EDCs in aqueous matrices, such as wastewater and river water, have often been described in the literature and are mostly based on spot sampling followed by laboratory-based extraction and analysis (Ce ´ spedes et al., 2005; Quednow and Pu ¨ ttmann, 2008). This approach, however, provides only an instantaneous measurement of pollutant levels and suffers from the uncertainty of short- and long-term concentration variations. In order to obtain a time-integrated evaluation of the bioavail- ability of different types of chemicals, bivalve molluscs have been used as bioindicators. Bivalves are filter-feeding organisms and are * Corresponding author. Tel.: þ39 010 3536187; fax: þ39 010 3536190. E-mail address: magie@chimica.unige.it (E. Magi). Contents lists available at ScienceDirect Environmental Pollution journal homepage: www.elsevier.com/locate/envpol 0269-7491/$ – see front matter Ó 2009 Elsevier Ltd. All rights reserved. doi:10.1016/j.envpol.2009.04.034 Environmental Pollution 157 (2009) 2716–2721