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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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Monitoring of (anti-)progestagenic activity in wastewater by in vitro bioassay
BERANOVÁ, Petra
The aim of this diploma thesis was to detect (anti-)progestagenic activity in wastewater samples from the influent and effluent of six wastewater treatment plants (WWTPs) located in South Bohemia. Subsequently, the efficiency of the treatment process of this WWTPs was assessed from this point of view. The wastewater from WWTPs was transported to the laboratory and extracted oby solid phase extraction. The eluates were washed, evaporated and dissolved in DMSO. Detection of (anti-)progestagenic activity was performed by using the PR-CALUX in vitro bioassay. Transgenic cells were seeded on well plates and were exposed to ORG2058, reference substance for progestagenic activity, or RU-486, reference substance for antiprogestagenic activity, as well as a number of diluted wastewater extracts. After that, luminiscence of the cells was measured and it was was expressed in relative light units which were a measure of (anti-)progestagenic activity.(Anti-)progestagenic activity was reported in equivalent concentrations of the reference substance. Progestagenic activity in WWTPs influent ranged from below LOQ up to 1.8 ng/l ORG2058 eq. In effluent this activity ranged from below LOQ up to 0.5 ng/l ORG2058 eq. The elimination rate of progestagenic activity ranged from -25% to 100%. Antiprogestagenic activity in WWTPs inffluent was below the LOQ up to 1 ng/l RU-486 eq. In effluent this activity was below LOQ up to 9.7 ng/l RU-486 eq. The elimination rate of antiprogestagenic activity ranged from -50% to 100%.(Anti-)progestagenic activity in waste water has been demonstrated by using the PR-CALUX in vitro bioassay. Negative removal efficiency of some WWTPs was also documented. This is probably caused by biotransformation of some substances which don´t have (anti-)progestagenic activity to substances which have this activity or it is caused by deconjugation of metabolits of compounds with (anti-)progestagenic activity. Antiprogestagenic activity may be more hazardous for organisms living in the aquatic environment than progestagenic activity because of concentrations in WWTPs effluent. However this activity should not be underestimated.
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