Introduction: Supplementation of the diet with fish oil omega-3 fatty acids, especially eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) has been associated with multiple health benefits. This study aimed to determine total lipid and omega-3 content in two fishes from Indonesia, which were Pangasius pangasius (P. pangasius) and Chanos chanos (C. chanos). Methods: Total lipid was extracted from P. pangasius and C. chanos and the lipid content was then analysed using gas chromatography–mass spectrometry (GC-MS). Results: Lipid content of C. chanos (4.63±3.84%) was higher than P. pangasius (3.94±1.43%) but less than that found in Salmo salar (S. salar) which was found to contain 6.98±2.56% lipid. Furthermore, polyunsaturated fatty acid omega-3 (EPA and DHA) analysis showed that C. chanos oil contained 0.36% EPA and 1.17% DHA. These levels are lower than that found in S. salar, often referred to as the “gold standard” for omega-3 fatty acids. Conclusion: C. chanos contains considerable amounts of EPA and DHA. As it is widely available in Indonesia, it may be used as source of omega-3 fatty acids instead of salmon.