Fish powder consists of a lipid-dense matrix wherein long-chain n-3 polyunsaturated fatty acids (PUFA) — eicosapentaenoic acid (EPA, 20:5 n-3) and docosahexaenoic acid (DHA, 22:6 n-3) — are encapsulated within a collapsed protein–lipid configuration that withstands solvent infiltration. This research evaluated three extraction methods used on the same fish powder: conventional solvent extraction (CSE, 240 min), microwave-assisted extraction (MAE, 20 min), and ultrasonication-assisted extraction (UAE, 30 min). Every treatment was conducted in three replicates and assessed for oil yield, EPA, DHA, total omega-3 content, extraction efficiency, peroxide value (PV), and free fatty acid (FFA) levels. UAE achieved the highest oil yield (19.20 ± 0.20% compared to 17.60 ± 0.20% for MAE and 14.30 ± 0.20% for CSE), the greatest EPA (12.77 ± 0.15%), DHA (21.57 ± 0.25%), and total omega-3 levels (34.33 ± 0.40% of the extracted oil), the best extraction efficiency (91.2%), along with the lowest PV (2.90 ± 0.10 meq O2/kg oil) and FFA (1.21 ± 0.03%). One-way ANOVA revealed significant differences among methods across all six parameters (p < 0.001; F = 273.00–706.92), and Tukey's HSD identified each pairwise contrast as significant (HSD range 0.059–0.533). When measured absolutely, UAE yielded 6.59 g of omega-3 per 100 g of powder compared to 5.22 g (MAE) and 3.41 g (CSE), reflecting a 93.3% enhancement over the traditional method. MAE provided the greatest efficiency per minute of processing (4.27 percentage points/min compared to 3.04 for UAE and 0.30 for CSE). Thus, UAE is advised when recovery and oxidative quality dictate the process selection; MAE is preferred when throughput is the limiting factor.