Ecotoxicological characterization of the antiepileptic drug carbamazepine using eight aquatic species: baseline study for future higher tier tests.

Ecotoxicological effects of the antiepileptic drug carbamazepine (CBZ) were investigated in one primary producer (Desmodesmus subspicatus) and seven invertebrate species (Daphnia magna, Daphnia pulex, Ceriodaphnia dubia, Gammarus fossarum, Potamopyrgus antipodarum, Lumbriculus variegatus and Chironomus riparius) using OECD and US EPA guidelines for chronic toxicity testing. The present data set was used to conduct a hazard assessment for CBZ including confirmatory data. While most of our results were in accordance with previous studies, published effect data for C. dubia and D. pulex could not be confirmed, even though they have previously been considered to be the most sensitive invertebrate species to CBZ exposure. The non-biting midge, C. riparius, was the most sensitive test organism in the present study. From an EC10 of 406 µg/L and a no observed effect concentration (NOEC) of 400 µg/L, a predicted no effect concentration (PNEC) of 8 µg/L was calculated. With regard to realistic predicted and measured environmental concentrations, the environmental risk can be considered as low for CBZ when the assessment is based on laboratory-based effect data. To conduct a refined and more realistic assessment, this study provides foundational data for two future, higher tier studies: one multiple-stressor experiment and one mesocosm study.

Nontarget Analysis via LC-QTOF-MS to Assess the Release of Organic Substances from Polyurethane Coating.

A novel approach was investigated for the assessment of leaching from a one-component polyurethane (1C-PU) coating used for hydraulic structures using nontarget analysis via LC-QTOF-MS. Leaching behavior of the 1C-PU coating was studied using experiments in which the coating was exposed to water for defined periods (6 h, 24 h, 3 d, 14 d). Three hardening durations for the 1C-PU coating were tested (0 h, 24 h, 14 d) as well as two water matrices (ultrapure water and river water), including a successive water renewal. Dissolved organic carbon, total nitrogen bound and nontarget measurements via LC-QTOF-MS showed that under all tested conditions organic substances were leached out of the 1C-PU coating. The shorter the hardening duration and the longer the leaching duration, the higher were the number and quantities of the eluted substances, while the influence of the water matrix was minor with respect to substance elution. Based on the MS2 spectra from the LC-QTOF-MS measurements, 30 substances released from the 1C-PU coating were tentatively identified. These substances belong to five chemical groups: derivatives of (i) N-(tosyl)carbamate, (ii) p-toluenesulfonamide (PTSA), (iii) 4,4'-methylenediphenyl diisocyanate (4,4'-MDI), (iv) toluenediisocyanate (TDI), and (v) oligo(ethylene) ([C2H4O]n) as well as p-toluenesulfonic acid. The identity of seven substances was confirmed by authentic reference standards, all of which exhibited an elevated bacterial toxicity to Aliivibrio fischeri. p-Toluenesulfonic acid was present in a German canal (Teltowkanal) with concentrations of up to 11 µg L-1.

Micropollutant degradation via extracted native enzymes from activated sludge.

A procedure was developed to assess the biodegradation of micropollutants in cell-free lysates produced from activated sludge of a municipal wastewater treatment plant (WWTP). This proof-of-principle provides the basis for further investigations of micropollutant biodegradation via native enzymes in a solution of reduced complexity, facilitating downstream protein analysis. Differently produced lysates, containing a variety of native enzymes, showed significant enzymatic activities of acid phosphatase, ß-galactosidase and ß-glucuronidase in conventional colorimetric enzyme assays, whereas heat-deactivated controls did not. To determine the enzymatic activity towards micropollutants, 20 compounds were spiked to the cell-free lysates under aerobic conditions and were monitored via LC-ESI-MS/MS. The micropollutants were selected to span a wide range of different biodegradabilities in conventional activated sludge treatment via distinct primary degradation reactions. Of the 20 spiked micropollutants, 18 could be degraded by intact sludge under assay conditions, while six showed reproducible degradation in the lysates compared to the heat-deactivated negative controls: acetaminophen, N-acetyl-sulfamethoxazole (acetyl-SMX), atenolol, bezafibrate, erythromycin and 10,11-dihydro-10-hydroxycarbamazepine (10-OH-CBZ). The primary biotransformation of the first four compounds can be attributed to amide hydrolysis. However, the observed biotransformations in the lysates were differently influenced by experimental parameters such as sludge pre-treatment and the addition of ammonium sulfate or peptidase inhibitors, suggesting that different hydrolase enzymes were involved in the primary degradation, among them possibly peptidases. Furthermore, the transformation of 10-OH-CBZ to 9-CA-ADIN was caused by a biologically-mediated oxidation, which indicates that in addition to hydrolases further enzyme classes (probably oxidoreductases) are present in the native lysates. Although the full variety of indigenous enzymatic activity of the activated sludge source material could not be restored, experimental modifications, e.g. different lysate filtration, significantly enhanced specific enzyme activities (e.g. >96% removal of the antibiotic erythromycin). Therefore, the approach presented in this study provides the experimental basis for a further elucidation of the enzymatic processes underlying wastewater treatment on the level of native proteins.

Transformation of oxcarbazepine and human metabolites of carbamazepine and oxcarbazepine in wastewater treatment and sand filters.

Carbamazepine (CBZ) and oxcarbazepine (OXC) are widely used anticonvulsants that are extensively metabolized in the human body. The pharmaceuticals and their human metabolites are present in influents and effluents of wastewater treatment plants (WWTPs), in rivers and streams, and in drinking water. In this study, the biodegradation of OXC and its main human metabolite, 10-hydroxy-CBZ (10OHCBZ), and the main human metabolite of CBZ, 10,11-dihydro-10,11-dihydroxy-CBZ (DiOHCBZ), was investigated in contact with activated sludge from a wastewater treatment plant (WWTP) and sand filter material from a waterworks. The transformation of DiOHCBZ, 10OHCBZ, and OXC led to the formation of the following main TPs: 1-(2-benzoic acid)-(1H,3H)-quinazoline-2,4-dione (BaQD), 1-(2-benzoic acid)-(1H,3H)-quinazoline-2-one (BaQM), 9-aldehyde-acridine, 9-carboxylic acid-acridine (9-CA-ADIN), hydroxyl 9-CA-ADIN, acridone (ADON), 11-keto-OXC, and 2,2'-(carbamoylazanediyl)dibenzoic acid. TP formation could be explained by three proposed transformation pathways, including reactions such as oxidation, α-ketol rearrangement, or benzylic acid rearrangement. The results highlight the fact that the TP abundances strongly depend on the concentrations spiked in the lab-scale experiments. BaQD, 9-CA-ADIN, and ADON were detected in WWTP effluents, rivers, and streams. 9-CA-ADIN was found at maximum concentrations in WWTP effluent and rivers up to 920 ± 50 ng L(-1) and 304 ± 26 ng L(-1), respectively. Even in drinking water, BaQD and 9-CA-ADIN were present at concentrations of 26 ± 2 ng L(-1) and 189 ± 3 ng L(-1), respectively.

Direct coupling of thin-layer chromatography with a bioassay for the detection of estrogenic compounds: applications for effect-directed analysis.

The present study investigated the hypothesis that the coupling of high-performance thin-layer chromatography with the yeast estrogen screen (planar-YES, p-YES) can be used as a screening tool for effect-directed analysis. Therefore, the proposed method was challenged for the first time with several real samples from various origins such as sediment pore water, wastewater, and sunscreens. It was possible to detect and quantify estrogenic effects in all investigated sample types, even in the presence of demanding matrixes. Furthermore, the specific agonistic effect of the estrogen receptor activation could be detected in samples exhibiting cytotoxic effects and at cytotoxic levels of analyzed estrogenic compounds, which is not possible with the classic YES. The analysis of samples by the p-YES results in profiles of estrogenic activity. By means of this profiles samples can be compared qualitatively and quantitatively with respect to different compositions of bioactive compounds in mixtures. In conclusion, the p-YES approach seems to have a high potential to be used as a valuable screening tool for various applications in effect-directed analysis.