The effects of gamma irradiation at different doses (0, 3 and 5 kGy) on lipid quality and fatty acid composition in vacuum-packed hot smoked rainbow trout (Oncorhynchus mykiss) fillets during cold storage (2 °C) were investigated. The major fatty acids were identified as palmitic, oleic, linoleic and docosahexaenoic acids (DHA). The fatty acid compositions were not affected by the irradiation process initially. However, the increase on the total saturated fatty acids (SFA) of irradiated fillets was higher than the control group at the end of the storage. While a significant decrease was observed in the control group of total polyunsaturated fatty acids (PUFA), no change was observed in the groups irradiated with 3 and 5 kGy doses at the end of the storage. The TBA values of 0, 3 and 5 kGy irradiated groups were 1.27, 1.46 and 1.58 mg MA / kg, respectively, the PV values were 6.12, 9.18 and 9.97 meq / kg and the FFA values were 5.36%, 5.67% and 6.10%, respectively, at the end of the storage. Using a combination of techniques to various processed or fresh seafood products will likely play a significant role in enhancing the manufacture of safe meals with extended shelf lives.