Polar compounds dominate in vitro effects of sediment extracts.

Sediment extracts from three polluted sites of the river Elbe basin were fractionated using a novel online fractionation procedure. Resulting fractions were screened for mutagenic, aryl hydrocarbon receptor (AhR)-mediated, transthyretin (TTR)-binding, and estrogenic activities and their potency to inhibit gap junctional intercellular communication (GJIC) to compare toxicity patterns and identify priority fractions. Additionally, more than 200 compounds and compound classes were identified using GC-MS/MS, LC-MS/MS, and HPLC-DAD methods. For all investigated end points, major activities were found in polar fractions, which are defined here as fractions containing dominantly compounds with at least one polar functional group. Nonpolar PAH fractions contributed to mutagenic and AhR-mediated activities while inhibition of GJIC and estrogenic and TTR-binding activities were exclusively observed in the polar fractions. Known mutagens in polar fractions included nitro- and dinitro-PAHs, azaarenes, and keto-PAHs, while parent and monomethylated PAHs such as benzo[a]pyrene and benzofluoranthenes were identified in nonpolar fractions. Additionally, for one sample, high AhR-mediated activities were determined in one fraction characterized by PCDD/Fs, PCBs, and PCNs. Estrone, 17β-estradiol, 9H-benz[de]anthracen-7-one, and 4-nonylphenol were identified as possible estrogenic and TTR-binding compounds. Thus, not only nonpolar compounds such as PAHs, PCBs, and PCDD/Fs but also the less characterized and investigated more polar substances should be considered as potent mutagenic, estrogenic, AhR-inducing, TTR-binding, and GJIC-inhibiting components for future studies.

[1]  Rolf Altenburger,et al.  Effect‐directed analysis of sediment‐associated algal toxicants at selected hot spots in the River Elbe basin with a special focus on bioaccessibility , 2009, Environmental toxicology and chemistry.

[2]  Brian G. Brownlee,et al.  Comparison of the AmesSalmonellaAssay and Mutatox Genotoxicity Assay for Assessing the Mutagenicity of Polycyclic Aromatic Compounds in Porewater from Athabasca Oil Sands Mature Fine Tailings , 1999 .

[3]  P. White.  The genotoxicity of priority polycyclic aromatic hydrocarbons in complex mixtures. , 2002, Mutation research.

[4]  Werner Brack,et al.  Effect‐directed fractionation and identification of cytochrome P4501A‐inducing halogenated aromatic hydrocarbons in a contaminated sediment , 2002, Environmental toxicology and chemistry.

[5]  K. Yamauchi,et al.  Endocrine disrupting chemicals: interference of thyroid hormone binding to transthyretins and to thyroid hormone receptors , 2003, Molecular and Cellular Endocrinology.

[6]  S. Nesnow,et al.  Bacterial mutagenicity of new cyclopenta-fused cata-annelated polycyclic aromatic hydrocarbons, and identification of the major metabolites of benz[j]acephenanthrylene formed by Aroclor-treated rat liver microsomes. , 1989, Mutation research.

[7]  W. Busby,et al.  Mutagenicity of cyclopenta-fused polynuclear aromatic hydrocarbons and a non-polar fraction from a fuel combustion sample in a Salmonella forward mutation assay without exogenous metabolic activation. , 1997, Mutation research.

[8]  Rolf Altenburger,et al.  Effect-directed analysis of contaminated sediments with partition-based dosing using green algae cell multiplication inhibition. , 2009, Environmental science & technology.

[9]  Cytotoxicity and aryl hydrocarbon receptor‐mediated activity of N‐heterocyclic polycyclic aromatic hydrocarbons: Structure‐activity relationships , 2006, Environmental toxicology and chemistry.

[10]  J. McLachlan,et al.  Potential mechanisms of thyroid disruption in humans: interaction of organochlorine compounds with thyroid receptor, transthyretin, and thyroid-binding globulin. , 1999, Environmental health perspectives.

[11]  G. Cass,et al.  Human-cell mutagens in respirable airborne particles in the northeastern United States. 1. Mutagenicity of fractionated samples. , 2004, Environmental science & technology.

[12]  S. Sievers,et al.  Vertical profile of PCDD/Fs, dioxin-like PCBs, other PCBs, PAHs, chlorobenzenes, DDX, HCHs, organotin compounds and chlorinated ethers in dated sediment/soil cores from flood-plains of the river Elbe, Germany. , 2007, Chemosphere.

[13]  J. Trosko,et al.  Concentrations of methylated naphthalenes, anthracenes, and phenanthrenes occurring in Czech river sediments and their effects on toxic events associated with carcinogenesis in rat liver cell lines , 2007, Environmental toxicology and chemistry.

[14]  K. Van den Berg.  Interaction of chlorinated phenols with thyroxine binding sites of human transthyretin, albumin and thyroid binding globulin. , 1990, Chemico-biological interactions.

[15]  Werner Brack,et al.  Effect-directed analysis of contaminated sediment from the wastewater canal in Pancevo industrial area, Serbia. , 2009, Chemosphere.

[16]  J. Giesy,et al.  Polychlorinated naphthalenes and other dioxin‐like compounds in Elbe River sediments , 2008, Environmental toxicology and chemistry.

[17]  A D Vethaak,et al.  Development of a stably transfected estrogen receptor-mediated luciferase reporter gene assay in the human T47D breast cancer cell line. , 1999, Toxicological sciences : an official journal of the Society of Toxicology.

[18]  Safe,et al.  Toxic equivalency factors (TEFs) for PCBs, PCDDs, PCDFs for humans and wildlife. , 1998, Environmental health perspectives.

[19]  W. Brack,et al.  Polychlorinated naphthalenes in sediments from the industrial region of Bitterfeld. , 2003, Environmental pollution.

[20]  Werner Brack,et al.  Automated fractionation procedure for polycyclic aromatic compounds in sediment extracts on three coupled normal-phase high-performance liquid chromatography columns. , 2008, Journal of chromatography. A.

[21]  Miroslav Machala,et al.  Inhibition of gap-junctional intercellular communication by environmentally occurring polycyclic aromatic hydrocarbons. , 2002, Toxicological sciences : an official journal of the Society of Toxicology.

[22]  Timo Hamers,et al.  Toxicological profiling of sediments using in vitro bioassays, with emphasis on endocrine disruption , 2004, Environmental toxicology and chemistry.

[23]  R. Santella,et al.  Mutagenicity of some methylated benzo[a]pyrene derivatives. , 1982, Mutation research.

[24]  L. Johnson,et al.  Structure of human transthyretin complexed with bromophenols: a new mode of binding. , 2000, Acta crystallographica. Section D, Biological crystallography.

[25]  G. Gabrielsen,et al.  Recombinant transthyretin purification and competitive binding with organohalogen compounds in two gull species (Larus argentatus and Larus hyperboreus). , 2009, Toxicological sciences : an official journal of the Society of Toxicology.

[26]  W. Brack.  Effect-directed analysis: a promising tool for the identification of organic toxicants in complex mixtures? , 2003, Analytical and bioanalytical chemistry.

[27]  K. Yamauchi,et al.  Effect of diethylstilbestrol on thyroid hormone binding to amphibian transthyretins. , 2000, General and comparative endocrinology.

[28]  J. Sacchettini,et al.  Hydroxylated polychlorinated biphenyls selectively bind transthyretin in blood and inhibit amyloidogenesis: rationalizing rodent PCB toxicity. , 2004, Chemistry & biology.

[29]  W. Thilly,et al.  Bioassay-directed chemical analysis of Los Angeles airborne particulate matter using a human cell mutagenicity assay , 1998 .

[30]  J. Trosko,et al.  Inhibition of gap junctional intercellular communication by noncoplanar polychlorinated biphenyls: inhibitory potencies and screening for potential mode(s) of action. , 2003, Toxicological sciences : an official journal of the Society of Toxicology.

[31]  K. Wakabayashi,et al.  Mutagenic activity of surface soil and quantification of 1,3-, 1,6-, and 1,8-dinitropyrene isomers in soil in Japan. , 2000, Chemical research in toxicology.

[32]  W. Brack,et al.  Sequential fractionation procedure for the identification of potentially cytochrome P4501A-inducing compounds. , 2003, Journal of chromatography. A.

[33]  J. Giesy,et al.  Relative Potencies of Individual Polychlorinated Naphthalenes and Halowax Mixtures To Induce Ah Receptor-Mediated Responses , 2000 .

[34]  M. Mizukami,et al.  Molecular macrocluster formation on silica surfaces in phenol-cyclohexane mixtures. , 2007, Langmuir : the ACS journal of surfaces and colloids.

[35]  J. Giesy,et al.  Comparison of Ah receptor-mediated luciferase and ethoxyresorufin-O-deethylase induction in H4IIE cells: implications for their use as bioanalytical tools for the detection of polyhalogenated aromatic hydrocarbons. , 1996, Toxicology and applied pharmacology.

[36]  E. Zeiger,et al.  Suppressive effects of chemicals in mixture on the Salmonella plate test response in the absence of apparent toxicity. , 1984, Environmental mutagenesis.

[37]  T. Visser,et al.  Biotransformation of brominated flame retardants into potentially endocrine-disrupting metabolites, with special attention to 2,2',4,4'-tetrabromodiphenyl ether (BDE-47). , 2008, Molecular nutrition & food research.

[38]  G. Löfroth,et al.  Nitroarenes: mutagenicity in the Ames Salmonella/microsome assay and affinity to the TCDD-receptor protein , 1982 .

[39]  D. Power,et al.  Disruption of thyroid hormone binding to sea bream recombinant transthyretin by ioxinyl and polybrominated diphenyl ethers. , 2007, Chemosphere.