title: Estrogenicity profile and estrogenic compounds determined in river sediments by chemical analysis, ELISA and yeast assays.
abstract: An effects-directed strategy was applied to bed sediments of a polluted tributary in order to isolate and identify the major estrogenic chemicals it discharges into the River Po, the principal Italian watercourse. Sediment extract was concentrated by solid phase extraction and then fractioned into 10 fractions by reversed phase high performance liquid chromatography (RP-HPLC). Estrogenic activity of whole extract and fractions were determined using a recombinant yeast assay containing the human estrogen receptor (YES). The 10 fractions and whole extract were analysed for target compounds, e.g. estrone (E1), 17beta-estradiol (E2), estriol (E3), 4-nonylphenol (NP), 4-tert-octylphenol (t-OP), bisphenol A (BPA), using both liquid chromatography-tandem mass spectrometry (LC-MS/MS) and non-competitive enzyme-linked immunosorbent assays (ELISA). The YES assay determined high estrogenic activity in whole sediment (15.6 ng/g EE2 equivalents), and positive results for fractions nr 1, 2, 6, 7 and 8. E1, E3 and NP were the main estrogenic chemicals, however, other unidentified compounds contributed to sediment estrogenicity, particularly for polar fractions nr 1 and 2. A GC-MS screening performed in scan mode identified other potential contributors such as phthalates (DBP, BBP), and OP isomers. A next sampling campaign extended to other tributaries and receiving stretches of the River Po confirmed E1, E3 and NP as major estrogenic chemicals potentially threatening other sites of the main river. In general, target compound ELISAs have been shown to be suitable tools for a rapid screening of wide areas or large numbers of environmental samples for estrogenic risk. The potential for interferences suggests however to use cautiously the concentration values obtained from some of the immunoassays.
authors.affiliation: Water Research Institute, National Council of Research, Brugherio (Milan), Italy. vigano@irsa.cnr.it
authors.forename: Luigi
authors.initials: L
authors.lastname: Viganò
authors.forename: Emilio
authors.initials: E
authors.lastname: Benfenati
authors.forename: Anne
authors.initials: A
authors.lastname: van Cauwenberge
authors.forename: Janne K
authors.initials: JK
authors.lastname: Eidem
authors.forename: Claudio
authors.initials: C
authors.lastname: Erratico
authors.forename: Anders
authors.initials: A
authors.lastname: Goksøyr
authors.forename: Werner
authors.initials: W
authors.lastname: Kloas
authors.forename: Silvia
authors.initials: S
authors.lastname: Maggioni
authors.forename: Alberta
authors.initials: A
authors.lastname: Mandich
authors.forename: Ralph
authors.initials: R
authors.lastname: Urbatzka
mesh_terms: D001681:Biological Assay; D002851:Chromatography, High Pressure Liquid; D004784:Environmental Monitoring; D004797:Enzyme-Linked Immunosorbent Assay; D004967:Estrogens / Q000032:analysis* / Q000737:chemistry; D019015:Geologic Sediments / Q000737:chemistry*; D006801:Humans; D011960:Receptors, Estrogen / Q000378:metabolism; D045483:Rivers / Q000737:chemistry*; D052616:Solid Phase Extraction; D014874:Water Pollutants, Chemical / Q000032:analysis* / Q000737:chemistry; D015003:Yeasts / Q000235:genetics / Q000378:metabolism
chemical_list: D004967:Estrogens; D011960:Receptors, Estrogen; D014874:Water Pollutants, Chemical