Possible role of hydroxyl radicals in the oxidation of dichloroacetonitrile by Fenton-like reaction

Abstract

Dichloroacetonitrile (DCAN), is a member of haloacetonitrile group and detected in drinking water supplies as a by-product of chlorination process. The mechanism of DCAN-induced carcinogenesis is believed to be mediated by oxidative bioactivation of DCAN molecules. The present study was designed to investigate if reactive oxygen species (ROS), similar to that generated in biological systems, are capable of oxidative activation of DCAN. A model ROS generation system (Fenton-like reaction; Fe 2+ and H 2O 2) that predominantly produces hydroxyl radical (OH ·) was used. DCAN oxidation was monitored by the extent of cyanide (CN −) release. The results indicate that DCAN was markedly oxidized by this system, and the rate of oxidation was dependent on DCAN concentration. Four-fold increase in H 2O 2 concentration (50–200 mM) resulted in a 35-fold increase in CN − release. The rates of DACN oxidation in presence of various transition metals were in the following order; iron>copper>titanium. DCAN oxidation was enhanced significantly by the addition of vitamin C and sulfhydryl compounds such as glutathione, N-acetyl- l- cysteine, and dithiothreitol (10 mM) to 140, 130, 145 and 136% of control, respectively. Addition of H 2O 2 scavenger; catalase or iron chelator; desferrioxamine (DFO) resulted in a significant decrease in CN − release 47 and 41% of control, respectively. Addition of various concentrations of the free radical scavengers, DMSO, or mannitol, to the incubation mixtures caused a significant decrease in DCAN oxidation, 32 and 50% of control, respectively. Michaelis–Menten kinetic analysis of the rates of this reaction, with or without inhibitors, indicated that ROS mediated oxidation of DCAN was inhibited by catalase ( K i=0.01 mM)>DFO (0.02 mM)>mannitol (0.09 mM)>DMSO (0.12 mM). In conclusion, our results indicate that DCAN is oxidized by a ROS-mediated mechanism. This mechanism may have an important role in DCAN bioactivation and DCAN-induced genotoxicity at target organs where multiple forms of ROS generating systems are abundant.