ENDOCRINE DISRUPTORS FROM COMBUSTION AND VEHICULAR EMISSIONS: IDENTIFICATION AND SOURCE NOMINATION
Brian Gullett1*, Jeff Ryan1, Paul Lemieux1, Carolyn Acheson2, Michael DeVito3, James Rabinowitz3, Sukh Sidhu4, Richard Striebich4, Joy Klosterman4
1National
U.S. Environmental Protection Agency Risk Management Research Laboratory, Air Pollution Prevention and Control Division (MD-65), Research Triangle Park, NC 27711 2National Risk Management Research Laboratory, Cincinnati, OH 45268 3National Health and Environmental Effects Research Laboratory, Toxicology Division, Research Triangle Park, NC 27711 4University 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”
EDCs are 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?
Benzene
CH2(S)+C2H2
-H
C2H4+C2H3
-H
+H2CCCH H2CCCH+CH3
-H2 +H +H
-H2
CH2CHCHCH2
+H
-H +H (C=C-C=C and C-C=C=C) -H +H
CH2CHCCH2
H2CCCH3 +H2CCCH or +H2CCCH3
CH3 CH3
Simple, low-Cnumber HC’s
C3H4 (allene)
-H2 +H
H2C=C=CR H2C-C=CR R=H (propargyl) R=CH3 (1-methylallenyl) CH3
C3H5 (allyl)
-H
C2H3+CH3
Branched Aromatics
After Marinov, Pitz, Westbrook, Castaldi and Senkan, 1996
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 Bioassays
– domestic waste burning, diesel trucks (HDDVs), forest fires, fireplaces, and woodstove….others. – Yeast estrogen assay – CALUX – Vitellogenin mRNA Assay
•
Sample fractionation to isolate target compounds
– HPLC technique (L. Brooks) – Capillary Electrophoresis – TIE method (G. Ankley) – diagram/description – Structure clues
• •
Multi Dimensional Gas Chromatography QSAR, Statistical analyses
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
Exhaust T
On-Road Diesel Dioxin Sampling Train
Heated Filter Box Glass Tube
Plastic Nut with Teflon Washer
Filter
T P
High Volume Vacuum Pump Pumps Orifice
P T T
½ Inch Teflon Tubing
SiO 2
SS Water Ice Jacket Water XAD-2 Module Water Knockout
V Pump Dry Gas Meter
T P Thermocouple Differential Pressure Transmitter Vacuum Gauge
V
Cooler
Ice Bath
Coiled SS Tubing
“Uncontrolled” Burning Sources
Burning of Domestic Waste
Waste Types
Wheat Straw Testing
CA PCDD/F, PCB Sources?
FIREPLACE
WOODSTOVE
SAMPLE CHARACTERIZATION Yeast Estrogen Screen
Environmental Estrogens Responding to Yeast Screen
• Row A = Bisphenol Row C = Genistein Row E = Nonyphenol Row F = Octylphenol Row H = 17β-Estradiol Rows B,D,and G (blank) • Deep Red Color indicates Estrogenic Activity • Yellow indicates Background (β-gal)
Gas Chromatography/Mass Spectroscopy
Abundance
Time (min)
Combustion Pollutant Analysis via Multidimensional Gas Chromatography (courtesy of UDRI)
F I D Cryotrap and release
Conventional GC-MS
*
4m 30 m Non-polar Primary column MSD
34.00
36.00
38.00
40.00
42.00
44.00
46.00
48.00
50.00
52.00
54.00
Polar secondary column Ion-trap Mass Spectromet er
MDGC heartcut of region
39.80 39.90 40.00 40.10 40.20 40.30 40.40 40.50
*
40.60 40.70 40.80 40.90 41.00
Multidimensio nal GC Assembly
RESULTS
Examples of oxygenates in diesel extracts using MDGC-MS (scanning mode)
HC O HC O
•
Benzaldehyde
HC O
•Hydroxybenzaldehyde
OH O
•
Methylbenzaldehyde
CH3
•Naphthalene dione
O
O
•
Dibenzofuran
O
•Hydroxybiphenyl
OH
•
Xanthene
•Naphthalenecarboxaldehyde
H C O
Examples of Oxygenates from Barrel Burn Extract
( ) denotes Match Quality out of 100
O
RESULTS
•hydroxy cyclopentanone (7) •benzaldehyde (87) •substituted cyclohexanone (72)
HO
O
OH
OH
O
•benzene ethanol 3methoxy (52)
R
O
O
O
•dihydroxy dimethyl benzaldehyde (5)
OH
•benzoic acid (64)
HO
OH O
O
•4-ethyl-2-methoxy phenol (90)
HO
•long chain carboxylic acids •Hydroxy methoxy phenyl ethanone (72)
O R
HO
O O
OH
HO
•phenol (80) •methyl quinol (90) •alkyl phenols (72 - 91)
R OH
•benzendiol (90)
HO
O
•dimethyl anisole (27)
O
O
•hydroxy methoxy phenyl propanone
HO
O
OH
O
•eugenol (96)
O OH
•methyl benzenediol (78)
O
OH
OH
•methoxy methyl phenol (93) •hydroxy phenyl butadiene (64)
O
•ethyl vanillin (5)
HO
•methoxy propyl phenol (64)
O
O HO
OH
•methoxy propenyl phenol (90)
HO
•vanillin (78)
O
O OH
RESULTS
Examples of PAH from Barrel Burn Extract
( ) denotes Match Quality out of 100
F
•fluoro-biphenyl (72)
•biphenylene (72) •acenaphthalene (80)
•fluorene (90)
•biphenyl (83)
R
•bibenzyl (74) •phenanthrene or anthracene (72) •fluoranthene or
•methyl fluorene (74)
•me-biphenyl (80) •dimethyl phenanthrene (64) •tetramethyl phenanthrene (38)
•phenyl naphthalene (69) •methyl phenanthrene (43)
pyrene (72) •vinyl naphthalene (48)
RESULTS
Examples of Oxy-PAH from Barrel Burn Extract
( ) denotes Match Quality out of 100
O
•benzofuran (83)
•benzophenone (87)
O
•methyl benzofuran (72)
O
O
O
•fluoren-one (72)
•phthalic anhydride (86)
O
O
ENDOCRINE DISRUPTORS FROM COMBUSTION AND VEHICULAR EMISSIONS: IDENTIFICATION AND SOURCE NOMINATION
Brian Gullett1*, Clyde Owens1, Jeff Ryan1, Paul Lemieux1, Carolyn Acheson2, Michael DeVito3, James Rabinowitz3, Sukh Sidhu4, Richard Striebich4, Joy Klosterman4
U.S. Environmental Protection Agency 1National Risk Management Research Laboratory, Air Pollution Prevention and Control Division (MD-65), Research Triangle Park, NC 27711 2National Risk Management Research Laboratory, Cincinnati, OH 45268 3National Health and Environmental Effects Research Laboratory, Toxicology Division, Research Triangle Park, NC 27711 4University of Dayton Research Institute, 300 College Park, Dayton, OH 45469 *919-541-1534, 919-541-0554 (fax), gullett.brian@epa.gov