There is evidence that highly unsaturated omega-3 fatty acids present in fish oils can provide a cancer-protective effect; however, when added to laboratory rodent formulations, these fatty acids are subject to rapid and/or extensive oxidation and other chemical changes by exposure to air, light, or heat during processing of pellets or when stored for various lengths of time. An animal diet with 16% refined fish oil and 4% corn oil was commercially prepared with antioxidants (butylated hydroxytoluene and butylated hydroxyquinone in addition to alpha-tocopherol) present, and precautions were taken to prevent oxidation at all stages of production and handling. Fatty acid composition of dried powdered diet as well as freshly processed dried pellets was analyzed from four lots at the beginning and end of a 45-day feeding period. Additionally, fatty acid analyses were carried out on pellets dried by both vacuum and air techniques, and pellets were left exposed to air at room temperature for 96 hours. No statistical difference in overall fatty acid composition was observed among lots due to pelletization or drying procedures. Moreover, 4 days of exposure of pellets to air at ambient temperatures and 45 days of freezer storage had no significant effect on their fatty acid composition. It would appear that rodent diets containing omega-3 fatty acids can be reliably prepared commercially and safely substituted for diets prepared manually and in bulk on a weekly basis.