Investigations for the detection of genotoxic substances on TLC plates

ABSTRACT

The present study describes modifications to an existing bioautographic method for detecting genotoxic substances on thin-layer chromatography (TLC) plates by using the umu assay. It will be shown that the complex procedure that is described in the literature can be significantly simplified. Using the modified method, it is possible to detect 0.3 ng of the genotoxic substance 4-nitroquinoline-N-oxide. With 5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside (X-gal), a suitable substrate for environmental samples for the detection of the resulting β-galactosidase was found. Experiments showed, in contrast, that indirectly genotoxic substances could not thus far be identified on the TLC plate with overlayer bioautography.

GRAPHICAL ABSTRACT

KEYWORDS:

• Bioautography
• effect-directed analysis
• Genotoxicity
• HPTLC
• Salmonella typhimurium TA1535/pSK 1002
• umuC

Acknowledgments

Both authors contributed equally to this work. The authors would like to thank Dr. Wolfgang Schulz (Zweckverband Landeswasserversorgung) and Prof. Dr. Michael Oehme (Institute for Applied Analytical Chemistry) for their extensive support. Thanks are extended to Dr. Georg Reifferscheid (German Federal Institute of Hydrology, BfG) and Prof. Dr. Klaus Kümmerer (Leuphana University of Lüneburg) for their helpful discussions and Dr. Mandy Schneider and Mr. Steffen Carl (Leuphana University of Lüneburg) for valuable tips on umu testing according ISO 13829:2000.