Estrogens in aquatic environments pose significant ecological and health risks due to their cumulative effects rather than individual impacts. This study investigates the voltammetric behavior of estrone (E1), 17β-estradiol (E2), estriol (E3), and 17α-ethinylestradiol (EE2), presenting a cost-effective and straightforward method for their simultaneous determination. Using differential pulse voltammetry (DPV) with a boron-doped diamond electrode, the method demonstrates high precision (deviations under 4%) and a linear dynamic range of 15.35-134.55 µmol·L-1. Integration of a vacuum evaporation step reduced detection limits to 10-8 mol·L-1, enabling effective analysis of real water samples. This optimized approach ensures practical applicability for monitoring total estrogen content in aquatic systems, providing an accessible and reliable alternative to conventional methods.

• Klíčová slova
• boron-doped diamond electrode, differential pulse voltammetry, estradiol, estriol, estrone, ethinylestradiol,
• MeSH
• chemické látky znečišťující vodu * analýza   MeSH
• elektrochemické techniky * metody   MeSH
• elektrody MeSH
• estradiol analýza   MeSH
• estriol analýza   MeSH
• estrogeny * analýza   MeSH
• estron analýza   MeSH
• ethinylestradiol analýza   MeSH
• limita detekce MeSH
• voda chemie   analýza   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• chemické látky znečišťující vodu * MeSH
• estradiol MeSH
• estriol MeSH
• estrogeny * MeSH
• estron MeSH
• ethinylestradiol MeSH
• voda MeSH

Some freshwater phytoplankton species have been suggested to produce estrogenic compounds in concentrations which could cause adverse effects to aquatic biota, while other studies showed no estrogenic effects after exposure to phytoplankton extracts or pointed out possible sources of the overestimation of the estrogenic activity. This study aimed to clarify these research inconsistencies by investigating estrogenicity of biomass extracts from both environmental freshwater blooms and laboratory cyanobacterial and algae cultures by in vitro reporter bioassay. Biomasses of 8 cyanobacterial and 3 algal species from 7 taxonomic orders were extracted and tested. Next to this, samples of environmental water blooms collected from 8 independent water bodies dominated by phytoplankton species previously assessed as laboratory cultures were tested. The results showed undetectable or low estrogenicity of both freshwater blooms and laboratory cultures with E2 equivalent concentration (EEQ) in a range from LOQ up to 4.5 ng EEQ/g of dry mass. Moreover, the co-exposure of biomass extracts with environmentally relevant concentration of model estrogen (steroid hormone 17β-estradiol; E2), commonly occurring in surface waters, showed simple additive interaction. However, some of the biomass extracts elicited partially anti-estrogenic effects in co-exposure with higher E2 concentration. In conclusion, our study documents undetectable or relatively low estrogenic potential of biomass extracts from both environmental freshwater blooms and studied laboratory cultured cyanobacterial and algae species. Nevertheless, in case of very high-density water blooms, even this low estrogenicity (detected for two cyanobacterial species) could lead to EEQ content in biomass reaching effect-based trigger values indicating potential risk, if recalculated per water volume at field sites. However, these levels would not occur in water under realistic environmental scenarios and the potential estrogenic effects would be most probably minor compared to other toxic effects caused by massive freshwater blooms of such high densities.

• Klíčová slova
• Anti-Estrogenicity, Aquatic ecosystem, Cyanobacteria, Estrogen, Freshwater bloom, Phytoplankton,
• MeSH
• estradiol toxicita   MeSH
• estrogeny * toxicita   analýza   MeSH
• estron MeSH
• fytoplankton MeSH
• sinice * MeSH
• voda MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• estradiol MeSH
• estrogeny * MeSH
• estron MeSH
• voda MeSH

Bisphenols, parabens, alkylphenols and triclosan are anthropogenic substances with a phenolic group that have been introduced to the environment in recent decades. As they possess hormone-like effects, they have been termed endocrine disruptors (EDs), and can interfere with steroid pathways in organisms. To evaluate the potential impact of EDs on steroid biosynthesis and metabolism, sensitive and robust methods enabling the concurrent measurement of EDs and steroids in plasma are needed. Of crucial importance is the analysis of unconjugated EDs, which possess biological activity. The aim of the study was to develop and validate LC-MS/MS methods with and without a derivatization step for the analysis of unconjugated steroids (estrone-E1, estradiol-E2, estriol-E3, aldosterone-ALDO) and different groups of EDs (bisphenols, parabens, nonylphenol-NP and triclosan-TCS), and compare these methods on a set of 24 human plasma samples using Passing-Bablok regression analysis. Both methods were validated according to FDA and EMA guidelines. The method with dansyl chloride derivatization allowed 17 compounds to be measured: estrogens (E1, E2, E3), bisphenols (bisphenol A-BPA, BPS, BPF, BPAF, BPAP, BPZ, BPP), parabens (methylparaben-MP, ethylparaben-EP, propylparaben-PP, butylparaben-BP, benzylparaben-BenzylP), TCS and NP, with lower limits of quantification (LLOQs) between 4 and 125 pg/mL. The method without derivatization enabled 15 compounds to be analyzed: estrogens (E1, E2, E3), ALDO, bisphenols (BPA, BPS, BPF, BPAF, BPAP, BPZ), parabens (MP, EP, PP, BP, BenzylP) with LLOQs between 2 and 63 pg/mL, and NP and BPP in semiquantitative mode. Adding 6 mM ammonium fluoride post column into mobile phases in the method without derivatization achieved similar or even better LLOQs than the method with the derivatization step. The uniqueness of the methods lies in the simultaneous determination of different classes of unconjugated (bioactive) fraction of EDs together with selected steroids (estrogens + ALDO in the method without derivatization), which provides a useful tool for evaluating the relationships between EDs and steroid metabolism.

• Klíčová slova
• Aldosterone, Bisphenol, Estrogen, Nonylphenol, Paraben, Triclosan,
• MeSH
• benzhydrylové sloučeniny analýza   MeSH
• chromatografie kapalinová MeSH
• endokrinní disruptory * analýza   MeSH
• estrogeny analýza   MeSH
• estron analýza   MeSH
• lidé MeSH
• parabeny analýza   MeSH
• tandemová hmotnostní spektrometrie metody   MeSH
• triclosan * analýza   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• benzhydrylové sloučeniny MeSH
• endokrinní disruptory * MeSH
• estrogeny MeSH
• estron MeSH
• parabeny MeSH
• triclosan * MeSH

Stagnant freshwaters can be affected by anthropogenic pollution and eutrophication that leads to massive growth of cyanobacteria and microalgae forming complex water blooms. These can produce various types of bioactive compounds, some of which may cause embryotoxicity, teratogenicity, endocrine disruption and impair animal or human health. This study focused on potential co-occurrence of estrogenic and retinoid-like activities in diverse stagnant freshwaters affected by phytoplankton blooms with varying taxonomic composition. Samples of phytoplankton bloom biomass and its surrounding water were collected from 17 independent stagnant water bodies in the Czech Republic and Hungary. Total estrogenic equivalents (EEQ) of the most potent samples reached up to 4.9 ng·g-1 dry mass (dm) of biomass extract and 2.99 ng·L-1 in surrounding water. Retinoic acid equivalent (REQ) measured by in vitro assay reached up to 3043 ng·g-1 dm in phytoplankton biomass and 1202 ng·L-1in surrounding water. Retinoid-like and estrogenic activities at some sites exceeded their PNEC and effect-based trigger values, respectively. The observed effects were not associated with any particular species of cyanobacteria or algae dominating the water blooms nor related to phytoplankton density. We found that taxonomically diverse phytoplankton communities can produce and release retinoid-like compounds to surrounding water, while estrogenic potency is likely related to estrogens of anthropogenic origin adsorbed to phytoplankton biomass. Retinoids occurring in water blooms are ubiquitous signalling molecules, which can affect development and neurogenesis. Selected water bloom samples (both water and biomass extracts) with retinoid-like activity caused effects on neurodifferentiation in vitro corresponding to those of equivalent all-trans-retinoic acid concentrations. Co-occurrence of estrogenic and retinoid-like activities in stagnant water bodies as well as the potential of compounds produced by water blooms to interfere with neural differentiation should be considered in the assessment of risks associated with water blooms, which can comprise complex mixtures of natural and anthropogenic bioactive compounds.

• Klíčová slova
• Endocrine disruption, Estrogenic activity, Neural differentiation, Phytoplankton water bloom, Retinoid-like activity,
• MeSH
• estrogeny analýza   MeSH
• estron MeSH
• eutrofizace MeSH
• fytoplankton MeSH
• komplexní směsi MeSH
• lidé MeSH
• retinoidy * MeSH
• rostlinné extrakty MeSH
• sinice * MeSH
• tretinoin MeSH
• voda MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• estrogeny MeSH
• estron MeSH
• komplexní směsi MeSH
• retinoidy * MeSH
• rostlinné extrakty MeSH
• tretinoin MeSH
• voda MeSH

Aquatic biotests are important tools targeting various effects in ecotoxicology, including endocrine disruption. Unintentional exposure of bioassay organisms to endocrine disruptors during cultivation or testing may interfere with assessed endpoints. We illustrate this issue on the example of laboratory phytoplankton cultivation, where possible sources of estrogenic compounds have been revealed. Fifty-four blank samples (water and fresh or cultivated growth media) were assessed by in vitro biotests for their estrogenicity, and major known estrogens originating from plastic materials, bisphenol A and alkylphenols, were analyzed in selected samples. The samples of freshly prepared growth medium elicited weak estrogenic response in bioassays and some samples of the aerated media caused responses even above the 50% of maximum of the reference compound (17β-estradiol, E2), while the samples from diverse laboratory water sources did not show significant estrogenic activity. The results identified substances contained in the growth medium as minor but reproducible contributors to estrogenicity in the cultivations. Sporadic but significant effects (up to 4.9 ng E2 equivalent/L) can be ascribed to compounds released from the used plastic materials during aeration of the cultivations. The potential sources of unintentional exposure to estrogenic compounds need to be considered in aquatic cultivations and biotests, since they could impact their outcomes, especially in arrangements assessing reproduction or whole life cycle biotests, or production of bioactive compounds by phytoplankton. The findings emphasize the necessity to assess all relevant blanks, ideally by sensitive high throughput in vitro assays that reflect also unknown pollutants and minimize all potential sources of background contamination. In vitro assays show very good applicability for this purpose since they enable to screen for any background estrogenicity of the used media and materials without the need of analyzing individual compounds, which often might not be known.

• Klíčová slova
• Aeration, Alkylphenols, Aquatic medium, Bisphenol A, Estrogenicity,
• MeSH
• chemické látky znečišťující vodu * toxicita   MeSH
• endokrinní disruptory * analýza   toxicita   MeSH
• estrogeny analýza   toxicita   MeSH
• fytoplankton MeSH
• monitorování životního prostředí metody   MeSH
• plastické hmoty MeSH
• voda MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• chemické látky znečišťující vodu * MeSH
• endokrinní disruptory * MeSH
• estrogeny MeSH
• plastické hmoty MeSH
• voda MeSH

A novel coating based on hybrid monolith with metal-organic framework (MOF) onto conventional Teflon-coated magnetic stir bars was developed. For this purpose, the external surface of the Teflon stir bar was firstly vinylized in order to immobilize a glycidyl methacrylate (GMA)-based polymer onto the magnet. Then, an amino-modified MOF of type MIL-101 (NH2-MIL-101(Al)) was covalently attached to the GMA-based monolith. After the synthesis process, several parameters affecting extraction of target estrogens by stir bar sorptive extraction (SBSE) including pH, ionic strength, extraction time, stirring rate, desorption solvent, and desorption time were also investigated. The resulting hybrid monolith was evaluated as SBSE sorbent for extraction of three estrogens (estrone, 17β-estradiol, estriol) and synthetic 17β-ethinylestradiol from water and human urine samples followed by HPLC with fluorescence detection (excitation and emission wavelengths, 280 and 310 nm, respectively). Under the optimal experimental conditions, the analytical figures of the method were established, achieving satisfactory limits of detection in the range of 0.015-0.58 µg L-1, recovery results ranging from 70 to 95% with RSD less than 6%, and precision values (intra- and inter-extraction units) below 6%.

• Klíčová slova
• Estrogens, HPLC-fluorescence detection, Hybrid monolith, Metal–organic framework, PTFE magnet, Stir bar; Extraction,
• MeSH
• chemické látky znečišťující vodu analýza   MeSH
• chromatografie kapalinová MeSH
• endokrinní disruptory analýza   MeSH
• epoxidové sloučeniny chemie   MeSH
• estrogeny analýza   MeSH
• lidé MeSH
• methakryláty chemie   MeSH
• porézní koordinační polymery chemie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• chemické látky znečišťující vodu MeSH
• endokrinní disruptory MeSH
• epoxidové sloučeniny MeSH
• estrogeny MeSH
• glycidyl methacrylate MeSH Prohlížeč
• methakryláty MeSH
• porézní koordinační polymery MeSH

Zayandeh Rood river is the most important river in central Iran supplying water for a variety of uses including drinking water for approximately three million inhabitants. The study aimed to investigate the quality of water concerning the presence of pharmaceutical active compounds (PhACs) and hormonelike compounds, which have been only poorly studied in this region. Sampling was performed at seven sites along the river (from headwater sites to downstream drinking water source, corresponding drinking water, and treated wastewater) affected by wastewater effluents, specific drought conditions, and high river-water demand. The targeted and nontargeted chemical analyses and in vitro bioassays were used to evaluate the presence of PhACs and hormonelike compounds in river water. In the samples, 57 PhACs and estrogens were detected with LC-MS/MS with the most common and abundant compounds valsartan, carbamazepine, and caffeine present in the highest concentrations in the treated wastewater in the concentrations of 8.4, 19, and 140 μg/L, respectively. A battery of in vitro bioassays detected high estrogenicity, androgenicity, and AhR-mediated activity (viz., in treated wastewater) in the concentrations 24.2 ng/L, 62.2 ng/L, and 0.98 ng/L of 17β-estradiol, dihydrotestosterone and 2,3,7,8-TCDD equivalents, respectively. In surface water samples, estrogenicity was detected in the range of <0.42 (LOD) to 1.92 ng/L of 17β-estradiol equivalents, and the drinking water source contained 0.74 ng/L of 17β-estradiol equivalents. About 19% of the estrogenicity could be explained by target chemical analyses, and the remaining estrogenicity can be at least partially attributed to the potentiation effect of detected surfactant residues. Drinking water contained several PhACs and estrogens, but the overall assessment suggested minor human health risk according to the relevant effect-based trigger values. To our knowledge, this study provides some of the first comprehensive information on the levels of PhACs and hormones in Iranian waters.

• Klíčová slova
• Drugs, Endocrine disruptors, In vitro biotest, LC-MS/MS, Nontargeted screening,
• MeSH
• chemické látky znečišťující vodu * analýza   MeSH
• chromatografie kapalinová MeSH
• estrogeny analýza   MeSH
• léčivé přípravky * MeSH
• lidé MeSH
• odpadní voda MeSH
• pitná voda * MeSH
• tandemová hmotnostní spektrometrie MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Írán MeSH
• Názvy látek
• chemické látky znečišťující vodu * MeSH
• estrogeny MeSH
• léčivé přípravky * MeSH
• odpadní voda MeSH
• pitná voda * MeSH

Estrogenic compounds are widely released to surface waters and may cause adverse effects to sensitive aquatic species. Three hormones, estrone, 17β-estradiol and 17α-ethinylestradiol, are of particular concern as they are bioactive at very low concentrations. Current analytical methods are not all sensitive enough for monitoring these substances in water and do not cover mixture effects. Bioassays could complement chemical analysis since they detect the overall effect of complex mixtures. Here, four chemical mixtures and two hormone mixtures were prepared and tested as reference materials together with two environmental water samples by eight laboratories employing nine in vitro and in vivo bioassays covering different steps involved in the estrogenic response. The reference materials included priority substances under the European Water Framework Directive, hormones and other emerging pollutants. Each substance in the mixture was present at its proposed safety limit concentration (EQS) in the European legislation. The in vitro bioassays detected the estrogenic effect of chemical mixtures even when 17β-estradiol was not present but differences in responsiveness were observed. LiBERA was the most responsive, followed by LYES. The additive effect of the hormones was captured by ERα-CALUX, MELN, LYES and LiBERA. Particularly, all in vitro bioassays detected the estrogenic effects in environmental water samples (EEQ values in the range of 0.75-304 × EQS), although the concentrations of hormones were below the limit of quantification in analytical measurements. The present study confirms the applicability of reference materials for estrogenic effects' detection through bioassays and indicates possible methodological drawbacks of some of them that may lead to false negative/positive outcomes. The observed difference in responsiveness among bioassays - based on mixture composition - is probably due to biological differences between them, suggesting that panels of bioassays with different characteristics should be applied according to specific environmental pollution conditions.

• Klíčová slova
• Bioassay, Chemical mixture, Endocrine disrupting compound (EDC), Environmental quality standard (EQS), Estrogenicity, Hormone mixture,
• MeSH
• biotest MeSH
• chemické látky znečišťující vodu * analýza   toxicita   MeSH
• endokrinní disruptory * analýza   MeSH
• estrogeny analýza   toxicita   MeSH
• estron MeSH
• monitorování životního prostředí MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• chemické látky znečišťující vodu * MeSH
• endokrinní disruptory * MeSH
• estrogeny MeSH
• estron MeSH

Complex mixtures of contaminants from multiple sources, including agriculture, industry or wastewater enter aquatic environments and might pose hazards or risks to humans or wildlife. Targeted analyses of a few priority substances provide limited information about water quality. In this study, a combined chemical and effect screening of water quality in the River Bosna, in Bosnia and Herzegovina was carried out, with focus on occurrence and effects of contaminants of emerging concern. Chemicals in water were sampled at 10 sites along the Bosna River by use of passive sampling. The combination of semipermeable membrane devices (SPMDs) and polar organic chemical integrative samplers (POCIS) enabled sampling of a broad range of contaminants from hydrophobic (PAHs, PCBs, OCPs) to hydrophilic compounds (pesticides, pharmaceuticals and hormones), which were determined by use of GC-MS and LC-MS (MS). In vitro, cell-based bioassays were applied to assess (anti)androgenic, estrogenic and dioxin-like potencies of extracts of the samplers. Of a total of 168 targeted compounds, 107 were detected at least once. Cumulative pollutant concentrations decreased downstream from the city of Sarajevo, which was identified as the major source of organic pollutants in the area. Responses in all bioassays were observed for samples from all sites. In general, estrogenicity could be well explained by analysis of target estrogens, while the drivers of the other observed effects remained largely unknown. Profiling of hazard quotients identified two sites downstream of Sarajevo as hotspots of biological potency. Risk assessment of detected compounds revealed, that 7 compounds (diazinon, diclofenac, 17β-estradiol, estrone, benzo[k]fluoranthene, fluoranthene and benzo[k]fluoranthene) might pose risks to aquatic biota in the Bosna River. The study brings unique results of a complex water quality assessment in a region with an insufficient water treatment infrastructure.

• Klíčová slova
• Contaminants of emerging concern - passive sampling, Hazard profiling - water quality monitoring, In vitro bioassay - endocrine disruption,
• MeSH
• biotest MeSH
• chemické látky znečišťující vodu analýza   MeSH
• dioxiny analýza   MeSH
• endokrinní disruptory analýza   MeSH
• estrogeny analýza   MeSH
• monitorování životního prostředí * MeSH
• odpadní voda MeSH
• organické látky analýza   MeSH
• pesticidy analýza   MeSH
• polychlorované bifenyly analýza   MeSH
• polychlorované dibenzodioxiny analýza   MeSH
• polycyklické aromatické uhlovodíky analýza   MeSH
• řeky chemie   MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Bosna a Hercegovina MeSH
• Názvy látek
• chemické látky znečišťující vodu MeSH
• dioxiny MeSH
• endokrinní disruptory MeSH
• estrogeny MeSH
• odpadní voda MeSH
• organické látky MeSH
• pesticidy MeSH
• polychlorované bifenyly MeSH
• polychlorované dibenzodioxiny MeSH
• polycyklické aromatické uhlovodíky MeSH

This study aimed at demonstrating that effect-based monitoring with passive sampling followed by toxicity profiling is more protective and cost-effective than the current chemical water quality assessment strategy consisting of compound-by-compound chemical analysis of selected substances in grab samples. Passive samplers were deployed in the Dutch river delta and in WWTP effluents. Their extracts were tested in a battery of bioassays and chemically analyzed to obtain toxicity and chemical profiles, respectively. Chemical concentrations in water were retrieved from publicly available databases. Seven different strategies were used to interpret the chemical and toxicity profiles in terms of ecological risk. They all indicated that the river sampling locations were relatively clean. Chemical-based monitoring resulted for many substances in measurements below detection limit and could only explain <20% of the observed in vitro toxicity. Effect-based monitoring yielded more informative conclusions as it allowed for ranking the sampling sites and for estimating a margin-of-exposure towards chronic effect ranges. Effect-based monitoring was also cheaper and more cost-effective (i.e. yielding more information per euro spent). Based on its identified strengths, weaknesses, opportunities, and threats (SWOT), a future strategy for effect-based monitoring has been proposed.

• Klíčová slova
• River Meuse, River Rhine, Speedisk, in vitro bioassay, in vivo bioassay, multiple substance potentially affected fraction of species (msPAF), priority compounds, silicone rubber, species-sensitivity distribution (SSD), toxic pressure, wastewater treatment plant (WWTP),
• MeSH
• androgeny analýza   toxicita   MeSH
• biotest MeSH
• chemické látky znečišťující vodu analýza   toxicita   MeSH
• estrogeny analýza   toxicita   MeSH
• kvalita vody MeSH
• monitorování životního prostředí metody   MeSH
• mutageny analýza   toxicita   MeSH
• řeky chemie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Nizozemsko MeSH
• Názvy látek
• androgeny MeSH
• chemické látky znečišťující vodu MeSH
• estrogeny MeSH
• mutageny MeSH