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Zjišťování potenciálu chemických látek používaných k ochraně proti ultrafialovému záření (UV filtrů) způsobovat anti-progestagenní aktivitu ve vodním prostředí
SMĚŠNÁ, Tereza
The substances used for protection against UV radiation (UV filters) are widely used in industry and also in personal care products. It has been reported from many countries around the world that they enter the aquatic environment. The presence of UV filters in surface waters can pose a risk to aquatic organisms. The aim of this thesis was to test a range of substances from the group of UV filters for anti-progestagenic activity, detect this activity in the aquatic environment, detect UV filters in the aquatic environment, and based on this information, evaluate the potential of organic UV filters to cause anti-progestagenic activity in the aquatic environment. In this thesis, 20 organic UV filters were tested with in vitro bioassay anti-PR-CALUX. In this bioassay cells were exposed to pure substances (UV filters) and reference substance mifepristone. Samples were taken by grab (active) sampling from wastewater treatment plant in Tábor and from surface waters (outdoor swimming pools and Lužnice River). Samples were extracted using solid phase extraction. These extracts from waste and surface waters were tested for progestagenic and anti-progestagenic activity with the (anti-)PR-CALUX bioassay and the concentrations of UV filters were measured using mass spectrometry in the extracts. Samples were also tested for cytotoxicity with the resazurin reduction assay. Nine organic UV filters have shown anti-progestagenic activity, but compared to the reference substance, the activities exhibited by UV filters were several orders of magnitude lower. Ethylhexyl methoxycinnamate, benzophenone-3, and octocrylene were the most potent anti-progestagens. Progestagenic activity was not found at any of the sites. Anti-progestagenic activity was found in 60% of the environmental extracts, and the concentrations ranged from 2.6 to 4.5 ng/L equivalents of mifepristone. Anti-progestagenic activity was detected in the influent of wastewater into the treatment plant, in the recipient of the Lužnice River, and in ponds used for fish farming and also as outdoor swimming pools, namely the Podroužek and Štilec ponds. Thirteen organic UV filters were identified by chemical analysis at the tested sites. The UV filters octocrylene and ethylhexyl methoxycinnamate were found in the highest concentrations, these two UV filters contributed the most to the overall anti-progestagenic activity. At the wastewater treatment plant influent, UV filters contributed 49% to the total anti-progestagenic activity. In the recipient of the Lužnice River and outdoor swimming pools, the contribution of UV filters to this activity was very small, which means that this activity in surface waters will be caused by the occurrence of other substances than in wastewater. In ponds, it can be assumed that wastewater treatment plants will not be the source of these substances. Further research should be aimed at detecting the impact of the occurrence of organic UV filters in the aquatic environment on organisms, but also at identifying substances causing anti-progestagenic activity in the aquatic environment.
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Mapování výskytu (anti-)progestagenních aktivit v povrchových vodách České republiky
KRULOVÁ, Jana
The aim of the bachelor thesis was to map the occurrence of substances with (anti-)progestagenic activity in surface waters of the Czech Republic. Samples were taken at 21 localities using POCIS samplers. Localities included the so-called closure profiles (end of the river basin) and significant sampling points along the profile of the river Elbe. The samples were transported to the laboratory, where they were extracted. (Anti-)PR-CALUX in vitro bioassay was used to detect (anti-)progestagenic activity. First, the cells were plated on a microtiter plate and, after 24 hours of incubation, exposed to a calibration series of the reference substance ORG 2058 for progestagenic activity or the reference substance mifepristone for anti-progestagenic activity and a series of diluted surface water extracts. After 24 hours of exposure, cell luminescence was measured in relative light units. The resulting hormonal activity was expressed in ng/l equivalents of the reference substance (ORG 2058 or mifepristone). Progestagenic activity was below the limit of quantification in the samples from all 21 sampling sites (<6-11 ng/l ORG 2058 equivalents) sampled in the year 2017. Anti-progestagenic activity was detected at 6 localities, ranging from 12 to 33 ng/l mifepristone equivalents. For comparison, 3 samples from the year 2016 were analysed. Anti-progestagenic activity was detected in the range from 14 to 48 ng/l mifepristone equivalents. Progestagenic activity was below the limit of quantification at all 3 localities. The occurrence of anti-progestagenic activity was detected mainly at localities where the chemical industry is in the vicinity. There are companies that can use certain substances with anti-progestagenic activity in the production of their products, such as polycyclic musk compounds, brominated flame retardants or bisphenols. The occurrence of anti-progestagenic activity in these localities could indicate the insufficient ability of industrial wastewater treatment plants to degrade substances exhibiting anti-progestagenic activity. The occurrence of anti-progestagenic activity in surface waters may have a negative impact on aquatic organisms, especially vertebrates. Exposure of these organisms to substances with anti-progestagenic activity could lead to reduced fertility and impaired gonad development.
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