Mutagenicity in Surface Waters: Synergistic Effects of Carboline Alkaloids and Aromatic Amines

For decades, mutagenicity has been observed in many surface waters with a possible link to the presence of aromatic amines. River Rhine is a well-known example of this phenomenon but responsible compound(s) are still unknown. To identify the mutagenic compounds, we applied effect-directed analysis (EDA) utilizing novel analytical and biological approaches to a water sample extract from the lower Rhine. We could identify 21 environmental contaminants including two weakly mutagenic aromatic amines, and the known alkaloid comutagen norharman along with two related β-carboline alkaloids, carboline, and 5-carboline, which were reported the first time in surface waters. Results of mixture tests showed a strong synergism of the identified aromatic amines not only with norharman, but also with carboline and 5-carboline. Additionally, other nitrogen-containing compounds also contributed to the mutagenicity when aromatic amines were present. Thus, comutagenicity of β-carboline alkaloids with aromatic amines is shown to occur in surface waters. These results strongly suggest that surface water mutagenicity is highly complex and driven by synergistic mechanisms of a complex compound mixture (of which many are yet unidentified) rather than by single compounds. Therefore, mixture effects should be considered not only from mutagens alone, but also including possible comutagens and nonmutagenic compounds.