Bioaccumulation and ROS generation in liver of freshwater fish, goldfish Carassius auratus under HC Orange No 1 exposure.

HC Orange No. 1 (CAS No. 54381-08-7, 2-nitro-4'-hydroxydiphenylamine) is used as a color additive in hair dyes. In this study, laboratory experiment was carried out to determine HC Orange No. 1 bioaccumulation and oxidative stress in the liver of freshwater fish, goldfish Carassius auratus. Fish were exposed to different concentrations (0.05, 0.1, 0.2, 0.5, and 1.0 mg/L) of HC Orange No. 1 for 10 days, with one group assigned as control. The accumulation of HC Orange No. 1 in liver increased with the exposure concentration (R(2) = 0.94). A secondary spin trapping technique was used followed by electron paramagnetic resonance (EPR) analysis to study the reactive oxygen species (ROS) production. On the basis of the hyperfine splitting constants and shape of the EPR spectrum, the ROS generated in fish liver after exposure was identified as hydroxyl radical (*OH). There is a good correlation between the exposure concentrations and *OH generation (R(2) = 0.92). The *OH signal intensity of the EPR spectrum showed a significant increase (P < 0.05) when the HC Orange No. 1 concentration was 1.0 mg/L, compared with that of the control. A good positive relationship (R(2) = 0.95) was found between the *OH formation and accumulation level of HC Orange No. 1 in liver. The changes of the activities of catalase (CAT), superoxide dismutase (SOD), glutathione S-transferase (GST), and contents of reduced glutathione (GSH) were also detected. These observations indicated a possible mechanism of oxidative stress induced by HC Orange No. 1 on fish.