Using model-based screening to help discover unknown environmental contaminants.

Of the tens of thousands of chemicals in use, only a small fraction have been analyzed in environmental samples. To effectively identify environmental contaminants, methods to prioritize chemicals for analytical method development are required. We used a high-throughput model of chemical emissions, fate, and bioaccumulation to identify chemicals likely to have high concentrations in specific environmental media, and we prioritized these for target analysis. This model-based screening was applied to 215 organosilicon chemicals culled from industrial chemical production statistics. The model-based screening prioritized several recognized organosilicon contaminants and generated hypotheses leading to the selection of three chemicals that have not previously been identified as potential environmental contaminants for target analysis. Trace analytical methods were developed, and the chemicals were analyzed in air, sewage sludge, and sediment. All three substances were found to be environmental contaminants. Phenyl-tris(trimethylsiloxy)silane was present in all samples analyzed, with concentrations of ∼50 pg m(-3) in Stockholm air and ∼0.5 ng g(-1) dw in sediment from the Stockholm archipelago. Tris(trifluoropropyl)trimethyl-cyclotrisiloxane and tetrakis(trifluoropropyl)tetramethyl-cyclotetrasiloxane were found in sediments from Lake Mjøsa at ∼1 ng g(-1) dw. The discovery of three novel environmental contaminants shows that models can be useful for prioritizing chemicals for exploratory assessment.

Hydroxylated and methoxylated polybrominated diphenyl ethers and polybrominated dibenzo-p-dioxins in red alga and cyanobacteria living in the Baltic Sea.

Hydroxylated polybrominated diphenyl ethers (OH-PBDEs) and methoxylated polybrominated diphenyl ethers (MeO-PBDEs) are present in the ecosystem of the Baltic Sea. OH-PBDEs are known to be both natural products from marine environments and metabolites of the anthropogenic polybrominated diphenyl ethers (PBDEs), whereas, MeO-PBDEs appear to be solely natural in origin. Polybrominated dibenzo-p-dioxins (PBDDs) are by-products formed in connection with the combustion of brominated flame retardants (BFRs), but are also indicated as natural products in a red alga (Ceramium tenuicorne) and blue mussels living in the Baltic Sea. The aims of the present investigation were to quantify the OH-PBDEs and MeO-PBDEs present in C. tenuicorne; to verify the identities of PBDDs detected previously in this species of red alga and to investigate whether cyanobacteria living in this same region of the Baltic Sea contain OH-PBDEs, MeO-PBDEs and/or PBDDs. The red alga was confirmed to contain tribromodibenzo-p-dioxins (triBDDs), by accurate mass determination and additional PBDD congeners were also detected in this sample. This is the first time that PBDDs have been identified in a red alga. The SigmaOH-PBDEs and SigmaMeO-PBDEs concentrations, present in C. tenuicorne were 150 and 4.6 ng g(-1) dry weight, respectively. In the cyanobacteria 6 OH-PBDEs, 6 MeO-PBDEs and 4 PBDDs were detected by mass spectrometry (electron capture negative ionization (ECNI)). The PBDDs and OH-PBDEs and MeO-PBDEs detected in the red alga and cyanobacteria are most likely of natural origin.

Screening for genotoxicity using the DRAG assay: investigation of halogenated environmental contaminants.

The DRAG test is a rapid high-throughput screening assay for detection of repairable adducts by growth inhibition of Chinese hamster ovary cells (CHO) characterized by different defects in DNA repair. A more pronounced growth inhibition caused by a certain DNA-reactive substance in a repair-deficient cell line (EM9, UV4 and UV5) as compared to wild-type cells (AA8) is interpreted as a consequence of their inability to repair induced DNA lesions. Thus, the use of such cell lines in the DRAG test may provide information of the type of DNA lesions induced by a certain genotoxic substance. To select optimal assay conditions, as well as to provide a mechanistic basis for interpreting the results, the model compounds benzo(a)pyrene-7,8-dihydrodiol-9,10-epoxide (BPDE), ethyl methanesulfonate (EMS), mitomycin C (MMC) and hydrogen peroxide (H2O2) were used. These agents can induce bulky adducts, alkyl adducts, cross-links and oxidative damage, respectively. The specificity of the DRAG test constitutes an important prerequisite for its practical use in a broader context. To assess this aspect, we have investigated the genotoxic and cytotoxic properties of a selection of metabolites of and isomers from polychlorinated biphenyls (PCB) and polybrominated diphenyl ethers (PBDE), along with a few other halogenated compounds. All these compounds have been detected as pollutants in the external environment, and for most of them there is no convincing evidence of mutagenicity from conventional assays. As could be predicted from their mode of action, BPDE, MMC, and EMS were all found to be more toxic in the repair-deficient cell lines compared with wild-type cells. The results with H2O2 were inconclusive, and the PCB metabolite 4,4'-diOH-CB80 only exhibited borderline activity, while all other halogenated compounds, or their metabolites, were found to be inactive. In conclusion, the DRAG assay could provide a robust and useful tool when screening large numbers of potentially genotoxic agents, while in addition providing mechanistic information. However, the usefulness of the selected cell lines to detect oxidative damage may be limited.

Temporal variations of polybrominated dibenzo-p-dioxin and methoxylated diphenyl ether concentrations in fish revealing large differences in exposure and metabolic stability.

The concentrations of polybrominated dibenzo-p-dioxins (PBDDs) and polybrominated methoxylated diphenyl ethers (MeO-PBDEs) were investigated in perch (Perca fluviatilis) collected from a Baltic Sea background contaminated area between 1990 and 2005. No temporal trend was found, but large variations were observed - up to 5-fold and 160-fold differences in MeO-PBDE and PBDD concentrations, respectively - between consecutive years, suggesting that retention of these compounds, particularly the PBDDs, is limited. Examination of the congener profiles using principal component analysis (PCA) and correlation analysis indicated that MeO-PBDEs without adjacent substituents (6-MeO-BDE47) or with two adjacent substituents (2'-MeO-BDE68 and 6-MeO-BDE90) are retained more than MeO-PBDEs with three adjacent substituents (6-MeO-BDE85 and 6-MeO-BDE99) and that 1,3,6,8-tetraBDD and 1,3,7,9-tetraBDD are retained more than the other PBDDs which have vicinal hydrogen. Debromination could explain the limited retention of 6-MeO-PBDE85 and 6-MeO-BDE99 and the absence of 2-MeO-BDE123 and 6-MeO-BDE137, and cytochrome P-450 mediated oxidation could explain the limited retention of PBDDs containing vicinal hydrogen. The levels of organobromines, especially MeO-PBDEs, were found to covary with water conditions related to primary production, for example temperature, depth visibility, and inorganic nutrient concentrations, which also favor fish productivity. The results suggest natural production of MeO-PBDEs and PBDDs and imply that they fluctuate considerably over time, as do common marine toxins in fish. Thus, assessments of human and environmental risk should consider both the average and peak concentrations of these contaminants in marine biota.

Brominated phenols, anisoles, and dioxins present in blue mussels from the Swedish coastline.

INTRODUCTION: Naturally occurring hydroxylated polybrominated diphenyl ethers (OH-PBDEs), their methoxylated counterparts (MeO-PBDEs), and polybrominated dibenzo-p-dioxins (PBDDs), together with their potential precursors polybrominated phenols (PBPs) and polybrominated anisoles (PBAs), were analyzed in blue mussels (Mytilus edulis) gathered along the east coast (bordering the Baltic Sea) and west coast of Sweden (bordering the North Sea). Brown algae (Dictyosiphon foenicolaceus) and cyanobacteria (Nodularia spumigena) from the Baltic Sea, considered to be among the primary producers of these compounds, were also analyzed for comparison. MATERIALS AND METHODS: The samples were liquid-liquid extracted, separated into a phenolic and a neutral fraction, and subsequently analyzed by gas chromatography-mass spectrometry (GS-MS). RESULTS AND DISCUSSION: The levels of OH-PBDEs, MeO-PBDEs and PBDDs were significantly higher in Baltic Sea mussels than in those from the west coast, whereas the levels of PBPs and PBAs displayed the opposite pattern. The blue mussels from the Baltic Sea contained high levels of all analyzed substances, much higher than the levels of, e.g., polybrominated diphenyl ethers. In addition, the GC-MS chromatogram of the phenolic fraction of the west coast samples was dominated by four unknown peak clusters, three of which were tentatively identified as dihydroxy-PBDEs and the other as a hydroxylated-methyl-tetraBDE. CONCLUSIONS: Clearly, all of the compounds analyzed are natural products, both in the Baltic and the North Sea. However, the geographical differences in composition may indicate different origin, e.g., due to differences in the occurrence and/or abundance of various algae species along these two coasts or possibly a more extensive dilution on the west coast.

Polybrominated and mixed brominated/chlorinated dibenzo-p-dioxins in sponge (Ephydatia fluviatilis) from the Baltic Sea.

Polybrominated dibenzo-p-dioxins (PBDDs) have recently been found in the Baltic Sea at concentrations 1000 times above that of the chlorinated analogs (PCDDs), yet their sources are undefined. Marine production of organobrominated compounds by sponges is well documented. The objective of the current study was to investigate the potential for an aquatic sponge (Ephydatia fluviatilis), common to the Baltic Sea, to produce PBDDs and other organobromine compounds in the field. Mono- to pentaBDDs as well as several mixed brominated/chlorinated dibenzo-p-dioxins (Br/Cl-DDs), PCDDs and methoxylated polybrominated diphenyl ethers (MeO-PBDEs) were quantified in sponge from the SW Baltic. Concentrations of individual PBDDs in the range 1-80 ng per g extractable organic matter were similar as in blue mussels from the Baltic Sea and about 25 000 times higher than 2,3,7,8-tetraCDD. To the best of our knowledge, this is the first time Br/Cl-DDs are reported in biota from a background environment. While this study does not point out sponges as a dominant source, the concentrations of PBDDs in sponge relative to related anthropogenic compounds such as PBDEs and PCDDs as well as the relative abundance of brominated dioxins and furans strengthens the idea of natural production.

Brominated dibenzo-p-dioxins: a new class of marine toxins?

Levels of polybrominated dibenzo-p-dioxins (PBDDs) were measured in marine fish, mussels, and shellfish. PBDDs were nondetectable in samples from freshwater environments, and their levels were successively higher in samples from the marine environments of the Bothnian Bay and Bothnian Sea, the West Coast of Sweden, and the Baltic Proper. In Baltic Proper littoral fish the levels of PBDDs generally exceeded those of their chlorinated analogues (PCDDs). This is alarming as some Baltic fish species already are contaminated by chlorinated dioxins to such an extent that they cannot be sold on the European market. By comparing spatial trends in PBDD and PCDD distributions, and PBDD patterns in fish, mussels, and algae, we show that the PBDDs are probably produced naturally, and we propose a route for their biosynthesis. We further show that the levels of PBDDs are high (ng/g wet weight) in mussels, and that the levels increase over time. Finally, we discuss the possibility that the PBDDs have adverse biological effects, and that the levels are increasing as a result of global warming and eutrophication.

Identification of polybrominated dibenzo-p-dioxins in blue mussels (Mytilus edulis) from the Baltic Sea.

Polybrominated dibenzo-p-dioxins (PBDDs) are known to be formed as byproducts in connection with the manufacture and combustion of products containing brominated flame retardants. However, to date little is known about the occurrence of PBDDs in biological samples. The aim of the present investigation was to examine the presence of PBDDs in blue mussels (Mytilus edulis) from the Baltic Sea employing a procedure adapted for dioxin analysis. Two triBDDs (1,3,7-triBDD and 1,3,8-triBDD) were identified in biota here for the first time. This identification was based on accurate mass determination and comparison of retention times on three gas chromatographic columns of different polarities (PTE 5, SP-2331, and OV1701/heptakis) with synthesized standards, together with comparisons of electron capture negative ionization (ECNI) and electron ionization (EI) mass spectra. In addition, five PBDDs and one polybrominated dibenzofuran (PBDF) were tentatively identified; altogether, one diBDD, three triBDDs, three tetraBDDs, and one triBDFwere detected in the blue mussels. To our knowledge this is the first time PBDDs have been identified in biota of the Baltic Sea. The sigma triBDD concentration in the blue mussels was estimated to be 170 ng/g lipids. The origin of these PBDDs remains unclear, but a plausible hypothesis could be biogenic formation in the marine environment.

Hydroxylated and methoxylated brominated diphenyl ethers in the red algae Ceramium tenuicorne and blue mussels from the Baltic Sea.

Methoxylated polybrominated diphenyl ethers (MeO-PBDEs) and hydroxylated PBDEs (OH-PBDEs) have recently been identified in fish and wildlife from the Baltic Sea. Both OH-PBDEs and MeO-PBDEs are known natural products, while OH-PBDEs also may be metabolites of PBDEs. The aim of the present study was to determine if the red macroalga Ceramium tenuicorne could be a source for MeO- and OH-PBDEs in the Baltic environment. Blue mussels (Mytilus edulis) from the same area were also investigated for their content of MeO- and OH-PBDEs. Seven OH-PBDEs and four MeO-PBDEs were present both in the red macroalga and the blue mussels. The mussels also contained a monochlorinated OH-tetraBDE. One of the compounds, 6-methoxy-2,2',3,4,4',5-hexabromodiphenyl ether, has never been reported to occur in the environment. The identification was based on comparison of relative retention times with reference standards, on two gas chromatographic columns of different polarities, together with comparisons of full-scan electron capture negative ionization (ECNI) and electron ionization (EI) mass spectra. It is shown that MeO-PBDEs and OH-PBDEs are present in algae, but at this stage it could not be confirmed if the compounds are produced by the alga itself or by its associated microflora and/or microfauna.