ENDOCRINE DISRUPTORS FROM COMBUSTION AND VEHICULAR EMISSIONS: IDENTIFICATION AND SOURCE OMINATION Brian Gullett, Jeff Ryan, Paul Lemieux, Carolyn Acheson, Michael DeVito, James abinowitz, SukhSidhu, Richard Striebich, Joy Klosterman U.S. Environmental Protection Agency National Risk Management Research Laboratory, Air Pollution Prevention and Control Division (MD-65), Research Triangle Park, NC 27711 National Risk Management Research Laboratory, Cincinnati, OH 45268 National Health and Environmental Effects Research Laboratory, Toxicology Division, Research Triangle Park, NC 27711 University of Dayton Research Institute, 300 College Park, Dayton, OH 45469 919-541-1534, 919-541-0554 (fax), gullett.brian@epa.gov BACKGROUND •A significant fraction of organic emissions from combustion sources remains uncharacterized •100’s of compounds are emitted –e.g., PAHs, oxygenates, alkyl phenols •We only see what we specifically look for –e.g., PCDD/Fs “dioxins” •EDCsare emitted from combustion sources –e.g., PCDD/F, PCB, Hg •Combustion sources are ubiquitous –constitute a major exposure source GAS PHASE, AROMATIC PRECURSORS – How do we get complex aromatic structures? After Marinov, Pitz, Westbrook, Castaldiand Senkan, 1996 Chart Contact: Brian Gullet at 919-541-1534 OBJECTIVE We will survey combustion sources for potential endocrine disruption activity, try to isolate the compounds responsible for such activity, and attempt to estimate their emission factors. APPROACH •Opportunistic combustion source sampling –domestic waste burning, diesel trucks (HDDVs), forest fires, fireplaces, and woodstove….others. •Bioassays –Yeast estrogen assay –CALUX –Vitellogenin mRNAAssay •Sample fractionation to isolate target compounds –HPLC technique (L. Brooks) –Capillary Electrophoresis –TIE method (G. Ankley) •Multi Dimensional Gas Chromatography –diagram/description •QSAR, Statistical analyses –Structure clues Source Characterization •Done –diesel truck –woodstove –sewage sludge incinerator –domestic waste •Future –pine straw –structural fires –oil spill fire –municipal waste, full scale –DoD diesel truck On-Road Diesel Sampling Chart Contact: Brian Gullet at 919-541-1534 “Uncontrolled” Burning Sources Images Contact: Brian Gullet at 919-541-1534 Burning of Domestic Waste Images Contact: Brian Gullet at 919-541-1534 Waste Types Images Contact: Brian Gullet at 919-541-1534 Wheat Straw Testing Images Contact: Brian Gullet at 919-541-1534 CA PCDD/F, PCB Sources? FIREPLACE WOODSTOVE SAMPLE CHARACTERIZATION Yeast Estrogen Screen Chart Contact: Brian Gullet at 919-541-1534 Environmental Estrogens Responding to Yeast Screen Chart Contact: Brian Gullet at 919-541-1534 •Row A = Bisphenol Row C = Genistein Row E = Nonyphenol Row F = Octylphenol Row H = 17ß-Estradiol Rows B,D,and G (blank) •Deep Red Color indicates Estrogenic Activity •Yellow indicates Background (ß-gal) Gas Chromatography/Mass Spectroscopy Chart Contact: Brian Gullet at 919-541-1534 MDGC heartcut of Chart Contact: Brian Gullet at 919-541-1534 Examples of oxygenates in diesel extracts using MDGC-MS (scanning mode) RESULTS •Benzaldehyde •Methylbenzaldehyde •Dibenzofuran •Xanthene •Hydroxybenzaldehyde •Naphthalene dione •Hydroxybiphenyl •Naphthalene-carboxaldehydeO •hydroxy cyclopentanone (7) •benzaldehyde(87)O •substituted cyclohexanone(72) O •phenol (80) •methyl quinol(90) •alkyl phenols (72 -91) •methoxymethyl phenol (93) •hydroxy phenyl butadiene (64) •benzene ethanol 3-methoxy(52) •benzoic acid (64) •4-ethyl-2-methoxyphenol (90) •benzendiol(90) •dimethylanisole (27) •eugenol(96) •ethyl vanillin (5) •methoxy propenylphenol (90) •dihydroxy dimethyl benzaldehyde(5) •long chain carboxylic acids •Hydroxy methoxy phenylethanone(72) •hydroxymethoxyphenylpropanone •methylbenzenediol (78) •methoxy propyl phenol (64) •vanillin (78) Examples of Oxygenates from Barrel Burn Extract •biphenylene(72) •acenaphthalene(80) •bibenzyl(74) •phenanthreneor anthracene(72) •fluoranthene orpyrene (72) •vinyl naphthalene (48) •fluoro-biphenyl (72) •biphenyl (83) •me-biphenyl (80) •dimethyl phenanthrene(64) •tetramethyl phenanthrene(38) •fluorene(90) •methylfluorene(74) •phenyl naphthalene (69) •methylphenanthrene(43) Examples of PAH from Barrel Burn Extract ( ) denotes Match Quality out of 100 •benzofuran(83) •methylbenzofuran(72) •phthalicanhydride (86) •benzophenone(87) •fluoren-one (72) Examples of Oxy-PAH from Barrel Burn Extract ( ) denotes Match Quality out of 100 ENDOCRINE DISRUPTORS FROM COMBUSTION AND VEHICULAR EMISSIONS: IDENTIFICATION AND SOURCE NOMINATION Brian Gullett, Clyde Owens, Jeff Ryan, Paul Lemieux,Carolyn Acheson, Michael DeVito, James Rabinowitz, Sukh Sidhu, Richard Striebich, Joy Klosterman U.S. Environmental Protection Agency National Risk Management Research Laboratory, Air Pollution Prevention and Control Division (MD-65), Research Triangle Park, NC 27711 National Risk Management Research Laboratory, Cincinnati, OH 45268 National Health and Environmental Effects Research Laboratory, Toxicology Division, Research Triangle Park, NC 27711 University of Dayton Research Institute, 300 College Park, Dayton, OH 45469 919-541-1534, 919-541-0554 (fax), gullett.brian@epa.gov