Molecular insights into the formation and remobilization potential of nonextractable anthropogenic organohalogens in heterogeneous environmental matrices.

Anthropogenic organohalogens (AOHs) are toxic and persistent pollutants that occur ubiquitously in the environment. An unneglectable portion of them can convert into nonextractable residues (NER) in the natural solid substances. NER-AOHs are not detectable by conventional solvent-extraction, and will get remobilized through changes of surrounding environment. Consequently, the formation and fate of NER-AOHs should be investigated comprehensively. In this study, solvent extraction, sequential chemical degradation and thermochemolysis were applied on different sample matrices (sediments, soils and groundwater sludge, collected from industrial areas) to release extractable and nonextractable AOHs. Covalent linkages were observed most favorable for the hydrophilic-group-containing monocyclic aromatic AOHs (HiMcAr-AOHs) (e.g. halogenated phenols, benzoic acids and anilines) incorporating into the natural organic matter (NOM) as NER. Physical entrapment mainly contributed to the NER formation of hydrophobic monocyclic aromatic AOHs (HoMcAr-AOHs) and polycyclic aromatic AOHs (PcAr-AOHs). The hypothesized remobilization potential of these NER-AOHs follow the order HiMcAr-AOHs > HoMcAr-AOHs/ aliphatic AOHs > PcAr-AOHs. In addition, the NOM macromolecular structures of the studied samples were analyzed. Based on the derived results, a conceptual model of the formation mechanisms of NER-AOHs is proposed. This model provides basic molecular insights that are of high value for risk assessment and remediation of AOHs.

Formation and Fate of Point-Source Nonextractable DDT-Related Compounds on Their Environmental Aquatic-Terrestrial Pathway.

Nonextractable residues (NER) are pollutants incorporated into the matrix of natural solid matter via different binding mechanisms. They can become bioavailable or remobilize during physical-chemical changes of the surrounding conditions and should thus not be neglected in environmental risk assessment. Sediments, soils, and groundwater sludge contaminated with DDXs (DDT, dichlorodiphenyltrichloroethane; and its metabolites) were treated with solvent extraction, sequential chemical degradation, and thermochemolysis to study the fate of NER-DDX along different environmental aquatic-terrestrial pathways. The results showed that DDT and its first degradation products, DDD (dichlorodiphenyldichloroethane) and DDE (dichlorodiphenyldichloroethylene), were dominant in the free extractable fraction, whereas DDM (dichlorodiphenylmethane), DBP (dichlorobenzophenone), and DDA (dichlorodiphenylacetic acid) were observed primarily after chemical degradation. The detection of DDA, DDMUBr (bis( p-chlorophenyl)-bromoethylene), DDPU (bis( p-chlorophenyl)-propene) and DDPS (bis( p-chlorophenyl)-propane) after chemical treatments evidenced the covalent bindings between these DDXs and the organic matrix. The identified NER-DDXs were categorized into three groups according to the three-step degradation process of DDT. Their distribution along the different pathways demonstrated significant specificity. Based on the obtained results, a conceptual model of the fate of NER-DDXs on their different environmental aquatic-terrestrial pathways is proposed. This model provides basic knowledge for risk assessment and remediation of both extractable and nonextractable DDT-related contaminations.

Development and testing of a fractionated filtration for sampling of microplastics in water.

A harmonization of sampling, sample preparation and detection is pivotal in order to obtain comparable data on microplastics (MP) in the environment. This paper develops and proposes a suitable sampling concept for waterbodies that considers different plastic specific properties and influencing factors in the environment. Both artificial water including defined MP fractions and the discharge of a wastewater treatment plant were used to verify the derived sampling procedure, sample preparation and the subsequent analysis of MP using thermal extraction-desorption gas chromatography - mass spectrometry (TED-GC-MS). A major finding of this paper is that an application of various particle size classes greatly improves the practical handling of the sampling equipment. Size classes also enable the TED-GC-MS to provide any data on the MP size distribution, a substantial sampling property affecting both the necessary sampling volume and the optimal sampling depth. In the artificial water with defined MP fractions, the recovery rates ranged from 80 to 110%, depending on the different MP types and MP size classes. In the treated wastewater, we found both polyethylene and polystyrene in different size classes and quantities.

Degree of phenyl chlorination of DDT-related compounds as potential molecular indicator for industrial DDT emissions.

The pesticide DDT (1-chloro-4-[2,2,2-trichloro-1-(4-chlorophenyl)ethyl]benzene) and its degradates are among the most persistent and abundant organochlorine contaminates in the environment, and DDT is still being produced in several Asian countries. In this study, we report for the first time on the detection of DDT-related compounds with one additional or missing chlorine atom at the phenyl group (DDX±Cl) in sediment and soil samples taken in the vicinity of former and current DDT production sites. These congeneric compounds most likely originate from production residues disposed of into the environment. In order to ensure an adequate identification and quantification of this novel organic pollutant group, individual DDX±Cl were synthesized as reference compounds by simulating an impure production of DDT in the laboratory. In contrast to DDX±Cl with (chloro)alkyl moieties, DDX±Cl with (chloro)alkenyl moieties cannot be unambiguous assigned by gas-chromatographic/mass spectrometric (GC/MS) fragmentation and elution orders. The occurrence of DDX±Cl in environmental samples allows to draw conclusions about the purity of the production process in the associated production sites. Moreover, they potentially can serve as molecular indicators to differentiate between industrial DDT emissions and insecticidal applications of DDT. This hypothesis has yet to be confirmed by further research.

Hexabromocyclododecane diastereomers in fish and suspended particulate matter from selected European waters-trend monitoring and environmental quality standard compliance.

The brominated flame retardant hexabromocyclododecane (HBCD) was monitored in fish and sediment (from one lake) or suspended particulate matter (SPM; from five rivers) at European freshwater sites to study the effects of reduction measures implemented by HBCD producers and users in recent years. Bream (Abramis brama) were sampled annually between 2007 and 2013 in the rivers Götaälv/SE, Rhône/FR, Western Scheldt/NL, Mersey/UK, and Tees/UK and in Lake Belau/DE. Sediment/SPM was taken every second year between 2008 and 2014. HBCD was analyzed by LC/MS/MS allowing the determination of the alpha-, beta-, and gamma-diastereomers. For most sites, a decrease in ∑HBCD was observed in fish (e.g., in the Rhône and Western Scheldt by about 80 and 60%, respectively, with significantly decreasing trends, p < 0.01). In the Rhône, HBCD also decreased in SPM. At the sampling site in the Tees which was impacted by a former HBCD point source, fish HBCD levels decreased only after a major flood event in 2013. While fish data indicate a decline in environmental HBCD concentrations at most sites with diffuse emissions, SPM data were less conclusive. The European environmental quality standard for HBCD in fish of 167 µg kg-1 wet weight was met by all fish samples in 2013.

Spatial and temporal trends of metals and arsenic in German freshwater compartments.

Cadmium, lead, mercury, copper, nickel, zinc, and arsenic were analyzed in suspended particulate matter (SPM), zebra mussels, and bream sampled yearly under the program of the German Environmental Specimen Bank (ESB) in the rivers Rhine, Elbe, Danube, Saar, Mulde, and Saale and in Lake Belau. Temporal and spatial trends were analyzed, correlations between metal levels in different specimen types assessed, and sampling sites ranked according to their metal levels by calculating a Multi-Metal Index (MMI) for every specimen type and site. SPM: Highest metal loads were detected in Mulde, Saale, and Elbe right downstream of the Saale confluence. In the Elbe, metal loads in SPM were mostly highest in the upper and middle section of the river while in Rhine and Saar concentrations increased downstream. Temporal trends since 2005 were detected only at three sites. Zebra mussel: MMIs were highest in the tidal section of the Elbe and the lower Rhine and lowest in Lake Belau and the upper Danube. Different temporal trends were detected since the early 1990s depending on site and metal. Bream: As, Pb, Cu, and Hg were analyzed in muscle tissue and Pb, Cd, Cu, and Zn in liver. For both tissues, MMIs were highest in Mulde and Saale and the lower and middle Elbe. Since the early 1990s, Hg, Pb, and Cu decreased in bream muscle at many sites while As increased at 6 of the 17 sites. The findings indicate that Hg, Pb, and Cu have obviously decreased in many freshwater ecosystems in recent years, whereas As and Ni levels have increased at several sites. Metal levels and temporal trends mostly differed between the specimen types under investigation and only few correlations between specimen types were detected. This underlines the importance of including different components of an ecosystem when assessing its environmental quality.

Retrospective study of triclosan and methyl-triclosan residues in fish and suspended particulate matter: results from the German Environmental Specimen Bank.

A retrospective monitoring of triclosan (TCS; period 1994-2003 and 2008) and its potential transformation product methyl-triclosan (MTCS; period 1994-2008) was performed using archived fish samples from German rivers (16 sites, including Elbe and Rhine). At four of these sites suspended particulate matter (SPM) was also investigated covering the period 2005-2007. Samples were analyzed by GC/MS, either directly (MTCS) or after derivatization (TCS). TCS burdens of fish muscle tissue ranged from <0.2-3.4 ng g(-1) ww (wet weight; corresponding to <2-69 ng g(-1) lw, lipid weight) without apparent concentration trends over time. MTCS was detected at considerably higher concentrations in fish ranging from 1.0-33 ng g(-1) ww (47-1010 ng g(-1) lw) and increased until about 2003-2005. Thereafter, concentrations generally were lower, although at some sites single higher values were observed in recent years. In SPM, decreasing MTCS concentrations in the range 1-4 ng g(-1) dry weight were detected while TCS was always below the limit of quantification. Assuming that MTCS concentrations are correlated to TCS consumption, the observed decrease in MTCS levels may be partly a result of the voluntary renunciation of TCS use in detergents for, e.g., laundry or dishwashing declared by a manufacturers' association in 2001. Because of a lack of ecotoxicity studies for MTCS, a QSAR-derived predicted no effect concentration (PNEC) was compared to averaged ambient water concentrations of fish which were calculated from maximum tissue residues by applying an appropriate bioconcentration factor from literature. Since these calculated water concentrations were below the PNEC it is assumed that MTCS alone poses no immediate risk to aquatic organism. The conversion to a PNEC for SPM organisms and comparison with detected SPM levels of MTCS also revealed no risk.

Toxicity, dioxin-like activities, and endocrine effects of DDT metabolites--DDA, DDMU, DDMS, and DDCN.

BACKGROUND, AIM, AND SCOPE: 2,2-bis(chlorophenyl)-1,1,1-trichloroethane (DDT) metabolites, other than those routinely measured [i.e., 2,2-bis(chlorophenyl)-1,1-dichloroethylene (DDE) and 2,2-bis(chlorophenyl)-1,1-dichloroethane (DDD)], have recently been detected in elevated concentrations not only in the surface water of Teltow Canal, Berlin, but also in sediment samples from Elbe tributaries (e.g., Mulde and Havel/Spree). This was paralleled by recent reports that multiple other metabolites could emerge from the degradation of parent DDT by naturally occurring organisms or by interaction with some heavy metals. Nevertheless, only very few data on the biological activities of these metabolites are available to date. The objective of this communication is to evaluate, for the first time, the cytotoxicity, dioxin-like activity, and estrogenicity of the least-studied DDT metabolites. METHODS: Four DDT metabolites, p,p'-2,2-bis(chlorophenyl)-1-chloroethylene (DDMU), p,p'-2,2-bis(chlorophenyl)-1-chloroethane (DDMS), p,p'-2,2-bis(4-ch1oropheny1)acetonitrile (DDCN), and p,p'-2,2-bis(chlorophenyl)acetic acid (DDA), were selected based on their presence in environmental samples in Germany such as in sediments from the Mulde River and Teltow Canal. O,p'-DDT was used as reference in all assays. Cytotoxicity was measured by neutral red retention with the permanent cell line RTG-2 of rainbow trout (Oncorhynchus mykiss). Dioxin-like activity was determined using the 7-ethoxyresorufin-O-deetylase assay. The estrogenic potential was tested in a dot blot/RNAse protection-assay with primary hepatocytes from male rainbow trout (O. mykiss) and in a yeast estrogen screen (YES) assay. RESULTS: All DDT metabolites tested revealed a clear dose-response relationship for cytotoxicity in RTG-2 cells, but no dioxin-like activities with RTL-W1 cells. The dot blot/RNAse protection-assay demonstrated that the highest non-toxic concentrations of these DDT metabolites (50 µM) had vitellogenin-induction potentials comparable to the positive control (1 nM 17ß-estradiol). The estrogenic activities could be ranked as o,p'-DDT > p,p'-DDMS > p,p'-DDMU > p,p'-DDCN. In contrast, p,p'-DDA showed a moderate anti-estrogenic effect. In the YES assay, besides the reference o,p'-DDT, p,p'-DDMS and p,p'-DDMU displayed dose-dependent estrogenic potentials, whereas p,p'-DDCN and p,p'-DDA did not show any estrogenic potential. DISCUSSION: The reference toxicant o,p'-DDT displayed a similar spectrum of estrogenic activities similar to 17ß-estradiol, however, with a lower potency. Both p,p'-DDMS and p,p'-DDMU were also shown to have dose-dependent estrogenic potentials, which were much lower than the reference o,p'-DDT, in both the vitellogenin and YES bioassays. Interestingly, p,p'-DDA did not show estrogenic activity but rather displayed a tendency towards anti-estrogenic activity by inhibiting the estrogenic effect of 17ß-estradiol. The results also showed that the p,p'-metabolites DDMU, DDMS, DDCN, and DDA do not show any dioxin-like activities in RTL-W1 cells, thus resembling the major DDT metabolites DDD and DDE. CONCLUSIONS: All the DDT metabolites tested did not exhibit dioxin-like activities in RTL-W1 cells, but show cytotoxic and estrogenic activities. Based on the results of the in vitro assays used in our study and on the reported concentrations of DDT metabolites in contaminated sediments, such substances could, in the future, pose interference with the normal reproductive and endocrine functions in various organisms exposed to these chemicals. Consequently, there is an urgent need to examine more comprehensively the risk of environmental concentrations of the investigated DDT metabolites using in vivo studies. However, this should be paralleled also by periodic evaluation and monitoring of the current levels of the DDT metabolites in environmental matrices. RECOMMENDATIONS AND PERSPECTIVES: Our results clearly point out the need to integrate the potential ecotoxicological risks associated with the "neglected" p,p'-DDT metabolites. For instance, these DDT metabolites should be integrated into sediment risk assessment initiatives in contaminated areas. One major challenge would be the identification of baseline data for such risk assessment. Further studies are also warranted to determine possible additive, synergistic, or antagonistic effects that may interfere with the fundamental cytotoxicity and endocrine activities of these metabolites. For a more conclusive assessment of the spectrum of DDT metabolites, additional bioassays are needed to identify potential anti-estrogenic, androgenic, and/or anti-androgenic effects.

Structural diversity of organochlorine compounds in groundwater affected by an industrial point source.

Groundwater samples contaminated by an industrial point source were analysed in order to reveal the structural diversity of halogenated organic contaminants. Particular focus was laid on the metabolites and derivatives related to the pesticides DDT (2,2-bis(chlorophenyl)-1,1,1-trichlorethane) and lindane (γ-hexachlorocyclohexane). Additionally, a wide range of chlorinated and brominated xenobiotics were identified. These results represent a high degree of contamination with organochlorine compounds illustrating a considerable structural diversity in groundwater in the vicinity of the industrial plant. The polar DDT-metabolite DDA (2,2-bis(chlorophenyl)acetic acid), which has been neglected in water studies widely, represents the main DDT metabolite analysed in the water samples. Besides DDA, some unknown substances with structural relation to DDA and DDT were detected and identified, in detail 2,2-bis(4-chlorophenyl)acetic acid N-methyl amide (DDAMA) and 2,2-bis(4-chlorophenyl)acetic acid n-butyl ester (DDABE). As an overall implication of this study it has to be demanded that analysis of industrially affected ground waters have to be based on screening analysis for a comprehensive view on the state of pollution.

Non-target screening analysis of river water as compound-related base for monitoring measures.

BACKGROUND, AIM, AND SCOPE: Building up a comprehensive accurate monitoring program requires the knowledge on the contamination in principal, complemented by detailed information on individual contaminants. The selection of pollutants to be considered in monitoring actions is based dominantly on the information available about their environmental relevance (e.g., persistence, bioaccumulation potential, toxicological and ecotoxicological properties) and their occurrence within the affected environmental system. Therefore, this study focused on the identification of organic contaminants in selected German and European rivers to demonstrate the usefulness of a screening approach as complementary base for the compound selection process within monitoring activities. MATERIALS AND METHODS: Gas chromatography-mass spectrometry-based screening analyses were performed on five and six samples from German and European rivers, respectively. Identification of individual contaminants was based on the investigation of mass spectral and gas chromatographic properties compared with databases and reference materials. RESULTS: This study summarized the results of non-target screening analyses applied to river water samples and focused dominantly on, so far, unnoticed organic contaminants. Numerous compounds have been identified belonging to the groups of pharmaceuticals, technical additives, pesticides, personal care products, and oxygen-, nitrogen-, and sulfur-containing compounds of obviously anthropogenic origin. They are discussed in terms of their structural properties, their possible application or usage, and the environmental information available so far. DISCUSSION: Generally, two different groups of compounds have been differentiated that might contribute to potential monitoring programs. Firstly, more specific contaminants characterizing the individual riverine systems have been depicted (e.g., 4-chloro-2-(trifluoromethyl)aniline, di-iso-propylurea). The consideration of these substances in monitoring analyses to be applied to the corresponding catchment areas is recommended in order to monitor the real state of pollution. Secondly, contaminants have been introduced that appeared with higher multiplicity throughout the different river systems (e.g., TMDD, TXIB). Since these compounds tend to obviously have an elevated environmental stability accompanied by a widespread distribution, it is recommended to consider them in international high-scale monitoring programs. CONCLUSIONS: For monitoring purposes, a fundamental knowledge on the diversity of pollutants is an important precondition, which can be supported by screening analyses. Obviously, numerous organic contaminants have been neglected so far in environmental studies on river water, comprising also investigation on potential harmful effects and, therefore, their implementation in monitoring activities has been hindered. RECOMMENDATIONS AND PERSPECTIVES: Therefore, based on the results of this study, screening analyses should be established as principle tools to improve and complement the substance spectra for monitoring purposes. Secondly, scientific efforts should be strengthened to expand our knowledge on actually appearing organic contaminants in riverine systems.

Analysis of non-extractable DDT-related compounds in riverine sediments of the Teltow Canal, Berlin, by pyrolysis and thermochemolysis.

Eighteen pre-extracted samples derived from a subaquatic riverine sediment core taken from the Teltow Canal, Berlin (Germany), were treated by off-line TMAH-thermochemolysis and subsequently analyzed by GC-MS to investigate release and thermodegradation of non-extractable anthropogenic organic compounds (bound residues). Furthermore, six selected samples from the lower core section were additionally treated by off-line pyrolysis. Due to former investigations of the extractable fraction of Teltow Canal sediments, high amounts of compounds structurally related to the pesticide DDT (1,1,1-trichloro-2,2-bis(4-chlorophenyl)ethane) were anticipated within the nonextractable fraction. It has been shown that DDT-related bound residues can be gathered by pyrolysis and TMAH-thermochemolysis. Among other compounds, the experiments revealed two DDT-related degradation products (DDPU (3,3-bis(4-chlorphenyl)-1-propene) and DDPS (1,1-bis(4-chlorophenyl)propane)) which were detected for the first time in the environment. The latter compounds may represent formerly unknown metabolites or hints for the existence of carbon-carbon incorporated DDT-metabolites. Both methods tend to produce artifacts which complicate the interpretation of the results. With more knowledge on mechanisms occurring during application of pyrolysis and thermochemolysis, both methods can serve as valuable tools for analyzing bound residues in sediments.

DDT-related compounds bound to the nonextractable particulate matter in sediments of the Teltow Canal, Germany.

Sediment samples of the Teltow Canal (Berlin, Germany) were analyzed with respect to extractable and nonextractable organic compounds. The study focused on the identification and quantitation of bound 2,2-bis(chlorophenyl)-1,1,1-trichlorethane (DDT) residues in order to obtain further information about the fate of DDT-derived compounds within the particulate matter of the aquatic environment. Various chemical degradation techniques and a complementary online pyrolysis-GC/MS method were applied to the pre-extracted sediment residues. Generally, the distribution of the bound DDT-related compounds was found to differ distinctly from the substances distribution within the extractable fraction. The main metabolite of the anaerobic degradation pathway (2,2-bis(chlorophenyl)-1,1-dichlorethane, DDD) is most abundant in the sediment extracts but occurred only in insignificant concentrations in the degradation products of all procedures applied. The most abundant DDT-metabolites released after the degradation procedures were 4,4'-DBP, 4,4'-DDA, and 4,4'-DDM. In addition, 4,4'-DDM was detected at rather high concentrations by pyrolytic analysis. The results imply a weak association to the nonextractable particulate matter based on noncovalent interactions for the observed DDT-related contaminants. The release of these compounds was initiated by the modification and degradation of the organic macromolecular matrix as well as of the inorganic material. Furthermore, numerous methoxychlor-related compounds were detected not only in the extracts but also in parts of the hydrolysis products.