Estrogenicity determination in sewage treatment plants and surface waters from the Catalonian area (NE Spain)

Polyethoxylated nonylphenol (NPEOx) surfactants and nonylphenol (NP) as the ir major degradation product as well as some synthetic and natural estrogen s and progestogens have been reported to be present in freshwater systems, mainly at the vicinity of urban discharges and sewage treatment plants (STP s), at levels high enough to exhort estrogenicity to wildlife. To determine both presence and effects of such compounds in two tributaries of the Llob regat river (NE Spain), water samples and carp, Cyprinus carpio, were colle cted from selected sites along a transect, for chemical and biological dete rminations, respectively. Also influent and effluent water from several STP s, discharging into these rivers, was collected for its chemical characteri zation. NP and NPEO were determined by solid-phase extraction (SPE)followed by liquid chromatography-mass spectrometry (CC-MS). Representative estroge ns, both natural (estradiol, estriol, estrone) and synthetic (ethynyl estra diol, mestranol, diethylstilbestrol), progestogens (norethindrone, levonorg estrel), and the natural hormone progesterone were determined by off line S PE followed by LC-diode array detection (DAD)-MS. High levels of NP were en countered in all water samples (up to 600 mug/L), whereas only a few sample s gave quantifiable levels of estrogens and progestogens (ng/L range). West ern blot analysis of male carp plasma vitellogenin (VTG), using a polyclona l antibody raised in the cyprinid Koi carp, detected this protein in all sa mples, the VTG increase being more evident at the vicinity of the treatment plants. A certain correlation was also found between NP in water and VTG i nduction in fish (r=0.75).