The present study was aimed to investigate the impact of Chromium trioxide on biochemical parameters of freshwater fish, Channa punctatus exposed to sub lethal concentration for 7, 15, 30 and 60 days. Aspartate transaminase (AST), Alanine transaminase (ALT), Orthophosphoric Monoester Phospho-hydrolase (OMP) and Orthophosphoric acid Monoester Phospho-hydrolase (OAMP) activities have been analysed for different exposure. Results illustrates that AST, ALT, OMP and OAMP were found to be increases in comparison to control. However, the data’s were collected are statistically significance at p<0.05, P<0.01 and P< 0.001 level.