Structurally Selective Ozonolysis of <i>p</i> -Phenylenediamines and Toxicity in Coho Salmon and Rainbow Trout

The tire-rubber-derived ozonation product of <i>N</i>-(1,3-dimethylbutyl)-<i>N</i>'-phenyl-<i>p</i>-phenylenediamine (6PPD), <i>N</i>-(1,3-dimethylbutyl)-<i>N</i>'-phenyl-<i>p</i>-phenylenediamine-quinone (6PPD-Q), was recently discovered to cause acute mortality in coho salmon (<i>Oncorhynchus kisutch</i>). <i>para</i>-Phenylenediamines (PPDs) with variable side chains distinct from 6PPD have been identified as potential replacement antioxidants, but their toxicities remain unclear under environmentally relevant ozone conditions. We herein tested the multiphase gas-surface ozone reactivity of four select PPDs [6PPD, <i>N</i>-isopropyl-<i>N</i>'-phenyl-<i>p</i>-phenylenediamine (IPPD), <i>N</i>,<i>N</i>'-diphenyl-<i>p</i>-phenylenediamine (DPPD), and <i>N</i>-phenyl-<i>N</i>'-cyclohexyl-<i>p</i>-phenylenediamine (CPPD)] and evaluated the toxicity of their reaction mixtures in coho salmon, rainbow trout (<i>Oncorhynchus mykiss</i>), and fathead minnow (<i>Pimephales promelas</i>). 6PPD and IPPD were found to rapidly react with ozone, while no significant multiphase ozone reactivity was observed for DPPD or CPPD. The viability of coho salmon CSE-119 cells was strongly affected by the ozonolysis products of 6PPD but not by those of the other three PPDs. Acute mortality was only observed in juvenile rainbow trout that were exposed to oxidized 6PPD, suggesting a common mechanism of toxic action in the two salmonid fish species. This study reports the structurally selective ozone reactivity of PPDs and the unique toxicity of 6PPD ozonolysis mixtures, which demonstrates that other PPDs are potential alternative antioxidants.