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

Polyethoxylated nonylphenol (NPEOx) surfactants and nonylphenol (NP) as their major degradation product as well as some synthetic and natural estrogens and progestogens have been reported to be present in freshwater systems, mainly at the vicinity of urban discharges and sewage treatment plants (STPs), at levels high enough to exhort estrogenicity to wildlife. To determine both presence and effects of such compounds in two tributaries of the Llobregat river (NE Spain), water samples and carp, Cyprinus carpio, were collected from selected sites along a transect, for chemical and biological determinations, respectively. Also influent and effluent water from several STPs, discharging into these rivers, was collected for its chemical characterization. NP and NPEO were determined by solid-phase extraction (SPE)followed by liquid chromatography-mass spectrometry (CC-MS). Representative estrogens, both natural (estradiol, estriol, estrone) and synthetic (ethynyl estradiol, mestranol, diethylstilbestrol), progestogens (norethindrone, levonorgestrel), and the natural hormone progesterone were determined by off line SPE followed by LC-diode array detection (DAD)-MS. High levels of NP were encountered in all water samples (up to 600 mug/L), whereas only a few samples gave quantifiable levels of estrogens and progestogens (ng/L range). Western blot analysis of male carp plasma vitellogenin (VTG), using a polyclonal antibody raised in the cyprinid Koi carp, detected this protein in all samples, 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 induction in fish (r=0.75).