Effektbasert identifisering av nye miljøgifter i norsk miljø Effect-directed identification of emerging contaminants (Eceda) Kevin V. Thomas Norsk institutt for vannforskning 19. november 2008 1
Rumsfeldism "Reports that say that something hasn't happened are always interesting to me, because as we know, there are known knowns; there are things we know we know. We also know there are known unknowns; that is to say we know there are some things we do not know. But there are also unknown unknowns - the ones we don't know we don't know." 19. november 2008 2
Samarbeids prosjekt NIVA (NO) Martine Muusse*, Katherine Langford, Kevin V Thomas & Knut-Erik Tollefsen UiO (NO) Ketil Hylland NVH (NO) JannecheUtneSkåre& Elisabeth Lie DCU (IRE) Brett Paul UFZ (DE) Werner Brack USEPA (USA) Robert Burgess & Kay Ho UiU (S) Peter Haglund 19. november 2008 3
Nye miljøgifter Eksempler: Narkotika Legemidler Flammehemmer Nanopartikler Biocider Polyfluorerte forbindelser (PFCer) Tilsetningstoffer 19. november 2008 4
Hvordan identifiseres Effects-directed analysis 19. november 2008 5
Eksempel: Narkotika Narkotika forbruk (Norge) Forbruker: Cannabis: 138,000 Kokain: 24,000 Årlig bruk: Cannabis: 35,000 kg Kokain: 1 000 kg PEC: Cannabis: 10 µg/l Kokain: 0.03 µg/l MEC: Kokain: 0.15 µg/l Benzoylecgonin 0.02 µg/l 19. november 2008 6
Effects-directed analysis (EDA) Complex mixture Biological analysis Chemical analysis Fractionation Biological analysis Responsible toxicant 19. november 2008 7
Litt historik. Først rapportert i 1979 (Parkhurst et al.) 1988 adoptert i USA USEPA (som TIE) > 300 publikasjoner 50+ biologisk tester 19. november 2008 8
Matrices! 19. november 2008 9
Eceda: In vitro biotester In vitro cell bioassays: sensitive, specific, and rapid bio-analytical tools that are useful in characterising responses in complex environmental mixtures. Examples: Yeast estrogen screen (YES) Yeast androgen screen (YAS) CALUX: dioxin-like T-screen: Thyroid ABC test: Antibiotika E2 Standard Blank Sample A Sample B Sample C HIGH CONCENTRATION LOW CONCENTRATION 19. november 2008 10
Eceda: Prøver 19. november 2008 11
CALUX: dioksin likende toksisitet Mode of action in native cell PCDDs PCBs PCDFs PAHs Ah-R Ah-R Ah-R Ah-R Ah-R DRE Gene-x DRE Luciferase gene RNA RNA Cytochrome p450 protein Luciferase Detoxification Light Mode of action in DR-CALUX cell Dioxin Responsive- Chemically Activated Luciferase Expression (DR-CALUX). Used for the screening of samples for dioxinlike activity. TEQ CALUX Validated & intercalibrated 19. november 2008 12
Fraksjonering: enkel (RP) 6 5 Alkyl phenol C9 Triclosan Pyrene Log Kow 4 3 2 Methylparaben Benzophenone Diethyltoluamide Atrazine Metolachlor Fluorene Acenaphthene 1 Vanilline Nicotine 0 Caffeine -1 0 5 10 15 20 25 30 Fraction number 19. november 2008 13
Identifikasjon Non-polar GC-HR ToF MS (EI&CI) in full scan. GC(xGC)-TOF MS Polar LC-MS n QTOF Orbitrap 19. november 2008 14
Fungere denne teknikken? 19. november 2008 15
Eksempel fra Norge: Nye endokrine forstyrrelser Largest wastewater stream in offshore oil & gas production (~ 340 x 10 6 m 3 yr -1 ). Complex composition: small amounts of dispersed oil dissolved organic compounds (including h/c) organic acids phenols production chemicals inorganic compounds 19. november 2008 16
Androgenreseptor antagonister 9000 8000 7000 6000 5000 4000 3000 2000 1000 Androgen receptor antagonist activity (ug flutamide/l) 0 Grane A Gullfaks B Heidrun Oseberg C Snorre A Statfjord A Statfjord B Statfjord C Troll B Åsgård B Platform Estimated discharge to Norwegian sector: 10 000 kg Flutamide! Tollefsen, K.E. et al. (2007). Mar. Poll. Bulletin, 54, 277-283. 19. november 2008 17
Accurate mass: useful! 19. november 2008 18
Hva ellers finnes? Morphine Triclosan 19. november 2008 19
Konklusjoner EDA er en måte å identifisere nye miljøgifter Teknikken fungere spesielt bra med in vitro biologisk tester Teknikken allerede har identifisert nye endokrine forstyrrelser i norsk miljø Prosjekt er i gang med prøvekolleksjon og identifisering av dioksin likende stoffer i mose 19. november 2008 20
Takk for meg! 19. november 2008 21