Office of Research and Development U.S. Environmental Protection Agency National Exposure Research Laboratory
Monitoring the Air for Toxic and Genotoxic Compounds
Kim Rogers Why Are We Interested in Monitoring Toxic Vapors ?
• Industrial Contamination Deliberate Accident
• Homeland Security Why Toxicity Monitoring ?
• Simple • Rapid • Sensitive • Adaptable from Commercial Sector • Biochemical / Physiological Relevance What Compounds Are of Interest ?
• Toxic Industrial Chemicals (TICs) High Risk Toxic Gasses Medium Risk Toxic Volatiles
• Chemical Warfare Agents (CWAs) • Non-Traditional Agents (NTAs) Adapt Waste Water Screening Assays for Air Monitoring
• Acute Toxicity Microtox IQ-Tox Daphnia magna
• Genotoxicity DNA Melting/Annealing Analysis Vapor Accumulation Assay Direct Microtox Assay
SPMD DMSO
Exposure Neat Stock Dilution Neat Stock Dilution TIC in in Microtox TIC in Chamber in Microtox Assay DMSO Methanol Assay Reagent Reagent
EC50 AppEC50 SPMD How Does It Work
• Low Density Polyethylene – 10A pores
• Biological Mimic – Triolein, DMSO, Carbon
• After Deployment – Direct assay – Extract with solvent SPMD in 40 mL and 4L Exposure Chambers Microtox Reagent (bacterium) Photobacterium vibrio 300
250
200
150
EC-50 (ppmv) 100 Methylhydrazine
50 fluorophosphate Diisopropyl Methylchlorosilane Chloroacetone Acetone cyanohydrin Acetone Methyl chloroformate 1,2-Dibromoethane Allylamine Formaldehyde 1-Octanethiol Stilbene chloride Sulfuryl Acrolein Tricholoracetyl chloride Tricholoracetyl chloride Methanesulfonyl Phosphorus oxychloride 0 Diketene 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 800
700
600
500 1,2-Dibromoethane
400 cyanohydrin Acetone 1-Octanethiol Methyl chloroformateMethyl Chloroacetone Sulfuryl chloride Sulfuryl Methylchlorosilane Diisopropyl fluorophosphate Diisopropyl
300 chloride Trichloroacetyl Acrolein Phosphorus oxychloride -- 5,400 Phosphorus oxychloride
200 Stilbene Allylamine Concentration Factors
100 Methanesulfonyl chloride Methanesulfonyl Methylhydrazine Formaldehyde Diketene 0 12345678910111213141516 1000
800
600
400 Formaldehyde
Apparent EC-50 (ppbv) 200 Diisopropyl fluorophosphate Diisopropyl Allylamine Methylchlorosilane Methyl chloroformate Chloroacetone Methanesulfonyl chloride Methanesulfonyl Sulfuryl chloride Sulfuryl Stilbene Methylhydrazine 1,2-Dibromoethane Trichloroacetyl chloride Trichloroacetyl 1-Octanethiol Phosphorus oxychloride Phosphorus Acrolein Diketene 0 cyanohydrin Acetone 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 19 Trichlororacetylchloride 18 Methanesulfonylchloride 17 Sulfurylchloride 16 Hypochlorite (bleach) 15 More Effective Phenol 14 with 5 min. Assay Methylchlorosilane 13 Methylchloroformate 12 1,2-Dibromoethane 11 1-Octanethiol 10 Stilbene 9 Listerine More Effective 8 Allylamine with 15 min. Assay X Axis Title 7 Formaldehyde 6 Methylhydrazine 5 Chloroacetone 4 Acetone cyanohydrin 3 Acrolein 2 Diketene 1 Zinc Sulfate -60 -40 -20 0 20 40 60 80 % Relative Change in EC50 Between 5 and 15 min. 100
80
60 % Toxicity 40 % Mass Acrolein
20
Relative Mass and Toxicity (%) and Toxicity Mass Relative 0
0 5 10 15 20 25 Accumulation Time (hr) Daphnia magna Used in the IQ-Tox Test 100
% Daphnia IQ-tox 1 hr % Daphnia 18 hr 80 % Daphnia 24 hr
60
40 EC-50 (ppmv)
20
l e e ne d ne o lein tene ro e thio hloride stilbene ormat c n ac dike l roacet allylamine moetha of oxychlorideocta onyl chlori ury lf chlo u dibro roacetyl chloride sulf o l 1,2- methylchlorosilane hanes methylchlor t trich phosphorus me % Daphnia IQ-tox 1 hr 100 % Microtox 5 min
10
1 EC-50 (ppmv) EC-50
0.1
e e ide n in ne ine r to te ride ride e role e lo m o lorid stilbene la ethane h ychlo ac dik l ch lly o c roac y a yl ox octane thiol hlo on rom ur s c lf ib thylchlorosilanesulf horu esu ,2-d p n 1 me os ha methylchloroformate h t p trichloroacetyl chl me
120 100 80 60 40 20
Fluorescence (fl.u.) 0 0 0.02 0.04 0.06 0.08 Time (a.u)
120 C) o 100 80 60 40 20
Temperature ( 0 0 0.02 0.04 0.06 0.08 Time (a.u) 100 Relative Fluorescence(%)
80
60
40
20
0 10 50 100 150 200 250
Concentration of DNA (ng/mL) 100
80
60
40
Relative Fluorescence (%) 20
0 Control Cyclohexane Toluene Phenol Glutaraldehyde Mitomycin C Genotoxic and Non-Genotoxic Controls
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80 60 40 20 100 fluorescence in decrease Percent 100 Relative Fluorescence(%)
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0 Control 10 20 40 60 80 100 Concentration of Formaldehyde (mM) 100
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Relative Fluorescence (%) Fluorescence Relative 20
0 0 20406080100 Concentration of Formaldehyde (mM) Summary
• Air Sampling • Toxicity Microtox IQ-Tox • Genotoxicity DNA Melting Annealing Analysis Future Directions
•Sampling – Carbon Fabric • Sensitivity
• Bioassay – Genetically Engineered Yeast • Cytotoxicity & Genotoxicity
• Environmental Pollutants – Nanaomaterials
• Environmental Applications – Vapor, Water, Sediment Acknowledgements
• Kumar Ramanathan NRC-Postdoc • Srividia Kailasam NRC-Postdoc • Stacey Harper EPA-Postdoc
• Jim Petty USGS, CERC, MO • Jim Huckins • Robert Gale