Grantee Research Project Results
2003 Progress Report: Biomarkers for Air Pollutants: Development of Hemoglobin Adduct Methodology for Assessment of Exposure to Butadienes and Polycyclic Aromatic Hydrocarbons, SEER project of SIP: Experimental Program To Stimulate Competitive Research (EPSCoR) From The Commonwealth Of Kentucky
EPA Grant Number: R829419E02Title: Biomarkers for Air Pollutants: Development of Hemoglobin Adduct Methodology for Assessment of Exposure to Butadienes and Polycyclic Aromatic Hydrocarbons, SEER project of SIP: Experimental Program To Stimulate Competitive Research (EPSCoR) From The Commonwealth Of Kentucky
Investigators: Hurst, Harrell E. , Myers, Steven R.
Institution: University of Louisville
EPA Project Officer: Chung, Serena
Project Period: October 1, 2002 through September 30, 2004 (Extended to September 30, 2005)
Project Period Covered by this Report: October 1, 2002 through September 30, 2003
Project Amount: $219,287
RFA: EPSCoR (Experimental Program to Stimulate Competitive Research) (2000) RFA Text | Recipients Lists
Research Category: EPSCoR (The Experimental Program to Stimulate Competitive Research)
Objective:
The objective of this research project is to develop a methodology that can measure biomarkers of systemic exposures to chloroprene (2-chloro-1,3-butadiene; CAS# 126-99-8) and selected polycyclic aromatic hydrocarbons ([PAH]: fluoranthene, CAS# 205-44-0; benzo(a)pyrene, CAS# 50-32-8).
Progress Summary:
During Year 2 of the project, research focused on the development of working assays for the measurement of hemoglobin (Hb) adducts formed from reactive metabolites of the toxics. As this protein is abundant in blood, persists for up to 4 months in humans, and exhibits nucleophilic sites that serve as surrogates for molecular sites of toxic actions, Hb adducts may serve as biomarkers of significant exposure to these compounds. Standards were synthesized by reaction of valine, 13C-valine, or the tripeptide valine-tyrosine-valine, with a synthetic epoxide metabolite of chloroprene, (1-chloroethenyl)oxirane (CEO). Model Hb adducts were produced using mouse erythrocytes exposed to CEO in vitro. An assay for chloroprene epoxide-Hb adducts was developed with selected ion monitoring gas chromatographic/mass spectrometry. Following in vitro CEO exposure, hemolysis, and precipitation of mouse globin, this method relies on sequential reactions of Edman cleavage of adducted N-terminal valine and trimethylsilylation of hydroxyl groups produced by epoxide opening during adduct formation. Adducts from PAH exposure have been analyzed after acid hydrolysis of labile PAH-Hb carboxylate adducts. Kinetics of in vitro reactions of CEO with N-terminal valine, and of PAH epoxides with various Hb sites, have been characterized in mouse red cell Hb. If sufficient assay sensitivity can be realized, these assays may enable the assessment of exposure and the formation of toxic metabolites of these air pollutants.
Future Activities:
We will attempt to determine the utility of the assays for assessment of exposure, which will require treatment of mice with the parent compounds to enable in vivo formation of electrophilic metabolites and assessment of the extent of epoxide formation. These studies are required, as usefulness of the potential biomarkers depends on the sensitivity of the analytical methods, extent of adducts formed in vivo, and alternative detoxification reactions.
Journal Articles:
No journal articles submitted with this report: View all 14 publications for this projectSupplemental Keywords:
carcinogen, chemicals, water, watersheds, sediments, dioxin, metals, aquatic, lifecycle analyses, biology, analytical, surveys, ecosystem protection, environmental exposure and risk, health, sustainable industry, sustainable business, biochemistry, health risk assessment, monitoring/modeling, risk assessments, state, cleaner production, pollution prevention, Kentucky, KY, polychlorinated biphenyls, PCBs, polycyclic aromatic hydrocarbons, PAHs, air emissions, air pollution, alternative materials, amphibians, amphibian bioindicator, amphibian decline, amphibian population, animal model, biomarker based exposure inference, biomarker measurements, biomarkers, butadiene, causal mechanisms, clean technology, developmental stability, ecosystem health, exposure, exposure assessment, hazardous emissions, human exposure, human health risk, pollution prevention, pollution prevention research., Sustainable Industry/Business, RFA, Health, Ecosystem Protection/Environmental Exposure & Risk, Scientific Discipline, Geographic Area, Health Risk Assessment, EPA Region, Risk Assessments, State, amphibians, Environmental Chemistry, Biochemistry, cleaner production/pollution prevention, clean technology, hazardous emissions, pollution prevention, Kentucky (KY), butadiene, air emissions, human exposure, amphibian bioindicator, population decline, animal model, exposure, causal mechanisms, amphibian decline, alternative materials, Region 3, biomarker measurements, air pollution, biomarker, biomarker based exposure inference, human health risk, PAH, pollution prevention research, biomarkersProgress and Final Reports:
Original AbstractThe perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Conclusions drawn by the principal investigators have not been reviewed by the Agency.