High-performance thin-layer chromatography-umu combined with nontarget analysis-a sensitive genotoxicity screening...

Monitoring of genotoxic chemicals released into the water cycle or formed through transformation processes is critical to prevent harm to human health. The development of the high-performance thin-layer chromatography (HPTLC)-umu bioassay combines sample separation and detection of genotoxic substances in the low ng/L concentration range. In this study, raw, process, and drinking water samples from 11 different waterworks in Germany were analyzed using the HPTLC-umu. A genotoxic response was detected in three out of 42 samples (one raw water sample and two drinking water samples). Genotoxicity testing using the micronucleus and p53-CALUX (chemical activated luciferase gene expression) assays revealed genotoxic effects for the raw water sample and one of the drinking water samples. Fortunately, only minor genotoxic effects could be detected in the two drinking water samples, with estimated 4-nitroquinoline-N-oxide equivalency concentrations of 0.12 ng/L and 0.17 ng/L, respectively. After extraction of the genotoxic zones from HPTLC plates, identification attempts were conducted with nontarget analysis (NTA). A prioritization approach based on chlorine and bromine isotope pattern revealed several halogenated features for both drinking water samples. These tentatively identified substances were previously reported in the literature as possible carcinogens. Although the detected features could not be conclusively identified, the presence of halogenated features in the genotoxic effective zone of the HPTLC-umu assay strongly suggests that the chlorine disinfection process in both waterworks was responsible for the formation of the genotoxic substances in the drinking water samples. These findings emphasize that the HPTLC-umu in combination with additional genotoxicity assays and NTA can be used to evaluate even relatively clean samples, such as drinking water.