Grantee Research Project Results
1998 Progress Report: Solid-Matrix Luminescence Analysis of B[a]P-DNA Adducts
EPA Grant Number: R824100Title: Solid-Matrix Luminescence Analysis of B[a]P-DNA Adducts
Investigators: Hurtubise, Robert J.
Institution: University of Wyoming
EPA Project Officer: Packard, Benjamin H
Project Period: October 1, 1995 through September 30, 1998
Project Period Covered by this Report: October 1, 1997 through September 30, 1998
Project Amount: $431,400
RFA: Exploratory Research - Chemistry and Physics of Water (1995) RFA Text | Recipients Lists
Research Category: Water , Safer Chemicals , Land and Waste Management
Objective:
To develop the principles and methods for the solid-matrix luminescence characterization and determination of benzo[a]pyrene-DNA adducts, for the capillary electrophoresis separation of benzo[a]pyrene metabolites, and for the laser-induced detection of benzo[a]pyrene metabolites and benzo[a]pyrene-DNA adducts in capillary electrophoresis.Progress Summary:
A capillary-zone electrophoresis (CZE) method was developed for the determination of the pKa values for four hydroxy-benzo[a]pyrenes, 1-hydroxy-pyrene, and 3-hydroxy-benzanthracene. These results are important because of the difficulty in determining the pKa values of large molecular weight metabolites and in predicting the migration properties in CZE. A major study was undertaken to develop a rapid and simple CZE method for the separation of metabolites associated with benzo[a]pyrene, pyrene, and benzanthracene. A good separation was achieved with y-cyclodextrin-modified micellar electrokinetic chromatography. However, by employing nonaqueous CZE, a much better separation of the metabolites was achieved. Two nonaqueous CZE methods were developed. One method involved a seven minute compound-class separation of the metabolites, and the other CZE approach separated the mixture of nine of the metabolites which were structurally quite different. The previous method would easily lend itself to the laser-induced fluorescence detection of the metabolites separated by CZE. In other work, solid-matrix phosphorescence (SMP) data were obtained for the direct determination of B[a]P-DNA adducts. It was shown that a linear relationship could be acquired between the SMP intensity and percentage of the diolepoxide in DNA for DNA samples modified at 0.0001, 0.0005, and 0.001%. In addition, SMP was easily detected from DNA samples modified with diolepoxides at levels of 0.00001% and 0.00005%. The previous two samples are at levels of adduct content that are found in humans. The results of these experiments are very significant because they provide a simple and direct means for the determination of B[a]P-DNA adducts.Future Activities:
Methods will be developed for the characterization and determination of the metabolites of B[a]P by laser-induced fluorescence after separation by capillary electrophoresis. Solid-matrix phosphorescence methods will be developed for the determination of B[a]P-DNA adducts in purified samples from humans.Journal Articles on this Report : 3 Displayed | Download in RIS Format
| Other project views: | All 25 publications | 15 publications in selected types | All 15 journal articles |
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Li M, Hurtubise RJ. Characterization and quantitation of benzo[a]pyrene-DNA adducts by solid-matrix luminescence. Analytical Letters 1998;31(3):491-510. |
R824100 (1998) R824100 (Final) |
not available |
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Tjioe SW, Hurtubise RJ. Solid-matrix fluorescence and phosphorescence detection and characterization of benzo[a]pyrene-DNA adducts with Whatman 1 and Whatman 1PS filter paper. Applied Spectroscopy 1998;52(3):414-419. |
R824100 (1998) R824100 (Final) |
not available |
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Xu X, Hurtubise RJ. Determination of the pK(a) values of polycyclic aromatic hydrocarbon metabolites by capillary zone electrophoresis. Journal of Liquid Chromatography & Related Technologies 1999;22(5):669-679. |
R824100 (1998) R824100 (Final) |
not available |
Supplemental Keywords:
Health, Scientific Discipline, Water, Engineering, Risk Assessments, Chemistry, Environmental Chemistry, Engineering, Chemistry, & Physics, Physics, soil contaminants, heavy atom effects, solid matrix luminescence, human exposure, electrophoretic studies, sediment toxicity, capillary zone, DNA adducts, humidity, physicochemicalProgress 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.