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Background The chemical quality of drinking water is widely unknown in low-income countries. Objective We conducted an exploratory study in Manhica district (Mozambique) to evaluate drinking water quality using chemical analyses and cell-based assays. Methods We measured nitrate, fluoride, metals, pesticides, disinfection by-products, and industrial organochlorinated chemicals, and conducted the bioassays Ames test for mutagenicity, micronuclei assay (MN-FACS), ER-CALUX, and antiAR-CALUX in 20 water samples from protected and unprotected sources. Results Nitrate was present in all samples (median 7.5 mg/L). Manganese, cobalt, chromium, aluminium, and barium were present in 90-100% of the samples, with median values of 32, 0.6, 2.0, 61, 250 μg/l, respectively. Manganese was above 50 μg/l (EU guideline) in eight samples. Arsenic, lead, nickel, iron, and selenium median values were below the quantification limit. Antimony, cadmium, copper, mercury, zinc and silver were not present. Trihalomethanes, haloacetic acids, haloacetonitriles and haloketones were present in 5-28% samples at levels ≤4.6 μg/l. DDT, dieldrin, diuron, and pirimiphos-methyl were quantified in 2, 3, 3, and 1 sample, respectively (range 12-60 ng/L). Fluoride was present in one sample (0.11 mg/l). Trichloroethene and tetrachloroethene were not present. Samples were negative in the in vitro assays. Significance Results suggest low exposure to chemicals, mutagenicity, genotoxicity and endocrine disruption through drinking water in Manhica population. High concentration of manganese in some samples warrants confirmatory studies, given the potential link to impaired neurodevelopment.

Persistent and mobile (PM) chemicals spread quickly in the water cycle and can reach drinking water. If these chemicals are also toxic (PMT) they may pose a threat to the aquatic environment and drinking water alike, and thus measures to prevent their spread are necessary. In this study, nontarget screening and cell-based toxicity tests after a polarity-based fractionation into polar and non-polar chemicals are utilized to assess and compare the effectiveness of ozonation and filtration through activated carbon in a wastewater treatment and drinking water production plant. Especially during wastewater treatment, differences in removal efficiency were evident. While median areas of non-polar features were reduced by a factor of 270, median areas for polar chemicals were only reduced by a factor of 4. Polar features showed significantly higher areas than their non-polar counterparts in wastewater treatment plant effluent and finished drinking water, implying a protection gap for these chemicals. Toxicity tests revealed higher initial toxicities (especially oxidative stress and estrogenic activity) for the non-polar fraction, but also showed a more pronounced decrease during treatment. Generally, the toxicity of the effluent was low for both fractions. Combined, these results imply a less effective removal but also a lower toxicity of polar chemicals. The behaviour of features during advanced waste and drinking water treatment was used to classify them as either PM chemicals or mobile transformation products (M-TPs). A suspect screening of the 476 highest intensity PM chemicals and M-TPs in 57 environmental and tap water samples showed high frequencies of detection (median80%), which indicates the wide distribution of these chemicals in the aquatic environment and thus supports the chosen classification approach and the more generally applicability of obtained insights.

Persistent and mobile chemicals (PM chemicals) were searched for in surface waters by hydrophilic interaction liquid chromatography (HILIC) and supercritical fluid chromatography (SFC), both coupled to high resolution mass spectrometry (HRMS). A suspect screening was performed using a newly compiled list of 1310 potential PM chemicals to the data of 11 surface water samples from two river systems. In total, 64 compounds were identified by this approach. The overlap between HILIC- and SFC-HRMS was limited (31 compounds), confirming the complementarity of the two methods used. The identified PM candidates are characterized by a high polarity (median logD -0.4 at pH 7.5), a low molecular weight (median 187 g/mol), are mostly ionic (54 compounds) and contain a large number of heteroatoms (one per four carbons on average). Among the most frequently detected novel or yet scarcely investigated water contaminants were cyanoguanidine (11/11 samples), adamantan-1-amine (10/11), trifluoromethanesulfonate (9/11), 2-acrylamido-2-methylpropanesulfonate (10/11), and the inorganic anions hexafluorophosphate (11/11) and tetrafluoroborate (10/11). 31% of the identified suspects are mainly used in ionic liquids, a chemically diverse group of industrial chemicals with numerous applications that is so far rarely studied for their occurrence in the environment. Prioritization of the findings of PM candidates is hampered by the apparent lack of toxicity data. Hence, precautionary principles and minimization approaches should be applied for the risk assessment and risk management of these substances. The large share of novel water contaminants among these findings of the suspect screening indicates that the universe of PM chemicals present in the environment has so far only scarcely been explored. Dedicated analytical methods and screening lists appear essential to close the analytical gap for PM compounds.

In Germany, micropollutants that (may) occur in drinking water are assessed by means of the health-related indicator value (HRIV concept), developed by the German Federal Environment Agency. This concept offers five threshold values (≤ 0.01 to ≤ 3 μg l−1) depending on availability and completeness of data regarding genotoxicity, neurotoxicity, and germ cell-damaging potential. However, the HRIV concept is yet lacking integration of endocrine disruptors as one of the most prominent toxicological concerns in water bodies, including drinking water. Thresholds and proposed bioassays hence urgently need to be defined. Since endocrine disruption of ubiquitary chemicals as pharmaceuticals, industrial by-products, or pesticides is a big issue in current ecotoxicology, the aim of this study was to explore endocrine effects, i.e., estrogenic and androgenic effects, as an important, additional toxicological mode of action for the HRIV concept using a hierarchical set of well-known but improved bioassays. Results indicate that all of the 13 tested substances, industrial chemicals and combustion products (5), pharmaceuticals and medical agents (4), and pesticides and metabolites (4), have no affinity to the estrogen and androgen receptor in human U2OS cells without metabolic activation, even when dosed at their water solubility limit, while in contrast some of these substances showed estrogenic effects in the RYES assay, as predicted in pre-test QSAR analysis. Using a specifically developed S9-mix with the U2OS cells, those micropollutants, i.e., Benzo[a]pyrene, 2,4-Dichlorophenol, 3,3-Dichlorbenzidin, 3,4-Dichloranilin, and diclofenac, they show estrogenic effects at the same concentration range as for the yeast cells. Three of the drinking water-relevant chemicals, i.e., atrazine, tributyltin oxide, and diclofenac, caused effects on hormone production in the H295R assay, which can be correlated with changes in the expression of steroidogenic genes. One chemical, 17α-Ethinylestradiol, caused an estrogenic or anti-androgenic effect in the reproduction test with Potamopyrgus antipodarum. Considering these results, a proposal for a test strategy for micropollutants in drinking water regarding potential endocrine effects (hormonal effects on reproduction and sexual development) will be presented to enhance the existing HRIV concept.

3,4,3ʹ,4ʹ-tetrachloroazobenzene (TCAB) is not commercially manufactured but formed as an unwanted by-product in the manufacturing of 3,4-dichloroaniline (3,4-DCA) or metabolized from the degradation of chloranilide herbicides, like propanil. While a considerable amount of research has been done concerning the toxicological and ecotoxicological effects of propanil and 3,4-DCA, limited information is available on TCAB. Our study examined the toxicity of TCAB in comparison to its parent compounds propanil and 3,4-DCA, using a battery of bioassays including in vitro with aryl hydrocarbon receptor (AhR) mediated activity by the 7-ethoxyresorufin- O -deethylase (EROD) assay and micro-EROD, endocrine-disrupting activity with chemically activated luciferase gene expression (CALUX) as well as in vivo with fish embryo toxicity (FET) assays with Danio rerio . Moreover, the quantitative structure activity response (QSAR) concepts were applied to simulate the binding affinity of TCAB to certain human receptors. It was shown that TCAB has a strong binding affinity to the AhR in EROD and micro-EROD induction assay, with the toxic equivalency factor (TEF) of 8.7 × 10 − 4 and 1.2 × 10 − 5 , respectively. TCAB presented to be a weak endocrine disrupting compound with a value of estradiol equivalence factor (EEF) of 6.4 × 10 − 9 and dihydrotestosterone equivalency factor (DEF) of 1.1 × 10 − 10 . No acute lethal effects of TCAB were discovered in FET test after 96 h of exposure. Major sub-lethal effects detected were heart oedema, yolk malformation, as well as absence of blood flow and tail deformation. QSAR modelling suggested an elevated risk to environment, particularly with respect to binding to the AhR. An adverse effect potentially triggering ERβ, mineralocorticoid, glucocorticoid and progesterone receptor activities might be expected. Altogether, the results obtained suggest that TCAB exerts a higher toxicity than both propanil and 3,4-DCA. This should be considered when assessing the impact of these compounds for the environment and also for regulatory decisions.

Effluents from wastewater treatment plants (WWTPs) are a major source of estrogenic compounds to the aquatic environment. In the present work, estrogenic activities of effluents from eight municipal WWTPs in Finland were studied. The main objectives of the study were to quantify the concentrations of selected estrogenic compounds, to evaluate their contribution to estrogenic potency and to test the feasibility of the commercial bioassays for wastewater analysis. The effluent samples were analyzed by two in vitro tests, i.e. ERα-CALUX(®) and ELISA-E2, and by liquid chromatography mass spectrometry for six estrogenic compounds: estrone (E1), 17β-estradiol (E2), estriol (E3), 17α-ethinylestradiol (EE2), 17α-estradiol and bisphenol A (BPA). Estrogenic effects were found in all of the effluent samples with both of the bioassays. The concentrations measured with ELISA-E2 (8.6-61.6 ng/L) were clearly higher but exhibited a similar pattern than those with chemical analysis (E2

This article introduces ‘Tox-Box’, a joint research project designed to develop a holistic approach towards a harmonized testing strategy for exposure- and hazard-based risk management of anthropogenic trace substances in drinking water to secure a long-term drinking water supply. The main task of the Tox-Box consortium is to enhance the existing health-related indicator value concept (German: GOW-Konzept - Gesundheitlicher Orientierungswert) through development and prioritization of additional end point-related testing strategies for genotoxicity, neurotoxicity, germ cell damage, and endocrine effects. In this context, substance-specific modes of action will be identified and characterized. Toxicological data collected by the 12 Tox-Box subprojects will be evaluated and weighted to structure a hierarchical testing strategy for an improved risk assessment. A technical guidance document for exposure and hazard-based risk management of anthropogenic trace substances in drinking water will eventually be prepared.