Evaluation of time-dependent toxicity and combined effects for a series of mono-halogenated acetonitrile-containing binary mixtures.

Mixture and time-dependent toxicity (TDT) was assessed for a series of mono-halogenated acetonitrile-containing combinations. Inhibition of bioluminescence in was measured after 15, 30 and 45-min of exposure. Concentration-response (x/y) curves were determined for each chemical alone at each timepoint, and used to develop predicted x/y curves for the dose-addition and independence models of combined effect. The x/y data for each binary mixture was then evaluated against the predicted mixture curves. Two metrics of mixture toxicity were calculated per combined effect model: (1) an EC-based dose-addition (AQ) or independence (IQ) quotient and (2) the mixture/dose-addition (MX/DA) and mixture/independence (MX/I) metrics. For each single chemical and mixture tested, TDT was also calculated. After 45-min of exposure, 25 of 67 mixtures produced curves that were consistent with dose-addition using the MX/DA metric, with the other 42 being less toxic than predicted by MX/DA. Some mixtures had toxicity that was consistent with both dose-addition and independence. In general, those that were less toxic than predicted for dose-addition were also less toxic than predicted for independence. Of the 25 combinations that were consistent with dose-addition, 22 (88%) mixtures contained chemicals for which the individual TDT values were both >80%. In contrast, of the 42 non-dose-additive combinations, only 2 (4.8%) of the mixtures had both chemicals with individual TDT values >80%. The results support previous findings that TDT determinations can be useful for predicting chemical mixture toxicity.