ABSTRACTThis study evaluated the effect of feed supplementation with chia and canola oils as a substitute for soybean oil on the composition of Nile tilapia muscle tissue using chemometrics. Diets were supplemented with 2.1% of each oil and were provided to fish for 15 and 30 days. Compared to soybean oil, supplementation with canola and chia oils significantly increased (P < 0.05) the contents of eicosapentaenoic acid (EPA, 20:5n-3), docosapentaenoic acid (DPA, 22:5 n-3), and docosahexaenoic acid (DHA, 22:6 n-3) in Nile tilapia fillet. At 30 days, DHA content increased 97% in Nile tilapia fed chia oil and 91% in treatment with canola oil. The highest EPA content correlated to treatment with chia oil (7.33 mg 100 g−1). The long-chain polyunsaturated fatty acids (LC-PUFAs) precursors, linoleic acid and α-linolenic acid, were observed to increase according to treatment type and feed supplementation duration. The principal component analysis resulted in a two-principal-component model that described 92.07% of the total data variance. Also, it highlighted that the replacement of soybean oil with canola and chia oils in Nile tilapia diets contributed to increasing the n-3 LC-PUFA concentration in Nile tilapia fillets, improving its nutritional value.