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Bioassay-directed fractionation and sub-fractionation for mutagenicity and chemical analysis of diesel exhaust particles

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Author: Mutlu, E. · Warren, S.H. · Matthews, P.P. · King, C. · Linak, W.P. · Kooter, I.M. · Schmid, J.E. · Ross, J.A. · Ian Gilmour, M. · DeMarini, D.M.
Source:Environmental and Molecular Mutagenesis, October, 9, 54, 719-736
Identifier: 480073
Keywords: Environment · Combustion emissions · Complex mixtures · Salmonella · aromatic amine · aromatic nitro compound · polycyclic aromatic hydrocarbon · article · bioassay · car · chemical analysis · chemical composition · controlled study · elution · exhaust gas · fractionation · high performance liquid chromatography · mutagenicity · nonhuman · particulate matter · professional knowledge · standard · Salmonella · Urban Development · Built Environment · Earth & Environment · AEC - Applied Environmental Chemistry · EELS - Earth, Environmental and Life Sciences


Several types of diesel exhaust particles (DEPs) have been used for toxicology studies, including a highorganic automobile DEP (A-DEP) from Japan, and a low-organic forklift DEP developed by the National Institute of Standards and Technology (N-DEP). However, these DEPs were not characterized extensively for chemical composition or sub-fractionated and tested extensively for mutagenicity. We collected a compressor-generated DEP (C-DEP) and characterized it by conducting bioassay-directed fractionation of the extractable organics in Salmonella and correlating the results by hierarchical clustering with the concentrations of 32 polycyclic aromatic hydrocarbons (PAHs). Relative to A- and N-DEP, the mutagenic potency of C-DEP was intermediate in TA100 1S9 (PAH mutagenicity) but was lowest in TA98 – S9 (nitroarene mutagenicity). More than 50% of the mass of the extractable organics of C-DEP eluted in the nonpolar fraction 1, and only -20% eluted in the moderately polar Fractions 2 and 3. However, most of the mutagenicity eluted in Fractions 2 and 3, similar to A-DEP but different from N-DEP. HPLC-derived mutagrams of 62 sub-fractions per fraction confirmed that most of the mutagenicity was due to moderately polar compounds. The diagnostic strains identified a strong role for PAHs, nitroarenes, aromatic amines, and oxy-PAHs in the mutagenicity of C-DEP. Hierarchical clustering confirmed the importance of oxy-PAHs but not that of nitroarenes. To our knowledge this is the first use of hierarchical clustering to correlate chemical composition with the mutagenicity of a complex mixture. The chemical analysis and mutagenicity of C-DEP described here makes C-DEP suitable for additional toxicological studies.