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RECORD NUMBER: 12 OF 123

Main Title Bioassay-Directed Fractionation of 1-Nitropyrene Metabolites: Generation of Mutagrams by Coupling Reverse-Phase HPLC with Microsuspension Mutagenicity Assays.
Author Lewtas, J. ; King, L. C. ; Williams, K. ; Ball, L. M. ; DeMarini, D. ;
CORP Author Health Effects Research Lab., Research Triangle Park, NC. ;North Carolina Univ. at Chapel Hill. Dept. of Environmental Sciences and Engineering.
Publisher 1990
Year Published 1990
Report Number EPA/600/J-90/543;
Stock Number PB91-242537
Additional Subjects Mutagens ; Mutagenicity tests ; Bioassay ; Liquid chromatography ; Salmonella typhimurium ; Metabolic activation ; Lung ; Rabbits ; Reprints ; Nitropyrenes
Holdings
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Status
NTIS  PB91-242537 Some EPA libraries have a fiche copy filed under the call number shown. 07/26/2022
Collation 11p
Abstract
The authors performed bioassay-directed fractionation of a model complex mixture (rabbit lung S9-generated metabolites of (14)C-radiolabeled 1-nitropyrene) by assaying reverse-phase HPLC fractions using two microsuspension mutagenicity assays. A forward-mutation assay measuring mutation at the gpt locus (8-azaguanine resistance) in Salmonella typhimurium TM677 was performed in a total volume of 100 micro1, and a reverse-mutation assay measuring mutation at the hisD3052 allele in S. typhimurium TA98 was performed in a total volume of 200 micro1. HPLC fractions were collected every 30 s for 45 min, resulting in 90 fractions per run. The HPLC chromatogram (absorbance at 280 nm) and the (14)C profile were compared to the mutagenicity profiles (mutagrams) and to the mutagenic potencies of pure metabolites studied separately. The results indicate that a fine dissection of the mutagenic fractions can be obtained by coupling HPLC to microsuspension mutagenicity assays. Differences observed between the mutagrams generated by the two bacterial strains were most likely due to metabolic (nitroreductase) differences between the two strains. Themethod should be generally applicable to the bioassay-directed chemical analysis of complex mixtures. (Copyright (c) Oxford University Press.)