<p>Aim was to develop and evaluate omega-3 fatty acid rich nanoemulsions using flaxseed oil (FO) as the core lipid source, stabilized with alpha-lipoic acid and SDG-lignan. Two formulations, containing 20% and 25% FO, were prepared using optimized emulsifier blends. Characterization included measurement of droplet size, zeta potential, polydispersity index, and nutritional profiling. Morphological validation was performed through transmission electron microscopy. Stability was assessed under accelerated, real-time, and refrigerated conditions along with oxidative stability and Rancimat analysis. Acute oral toxicity study (Organization for Economic Co-operation and Development guideline-425) was carried out in female Wistar rats. Erythrocyte fatty acid composition was analyzed after 30 days of supplementation. Consumer acceptability was assessed using a nine-point hedonic scale. The formulations exhibited mean droplet sizes of 99.95 ± 38.34nm (FO-A) and 134.40 ± 6.76nm (FO-C). Total omega-3 fatty acid content recorded in FO-A was 10.83 ± 1.52% and 18.53 ± 0.01% in FO-C with the omega-6 to omega-3 ratio remaining below one. Rancimat analysis demonstrated enhanced oxidative stability compared to raw flaxseed oil. Stability studies confirmed a shelf life of 60, 150, and 240 days under accelerated, real-time, and refrigerated storage conditions, respectively. Toxicological evaluation showed no mortality, adverse clinical symptoms, or organ damage based on histopathological evaluation of vital organs. Supplementation improved erythrocyte bioconversion of polyunsaturated fatty acids compared to the control group. Sensory evaluation indicated superior acceptability in milk-based matrix compared to water. The developed flaxseed oil nanoemulsions are safe, stable, and effective carriers of omega-3 fatty acids with potential applications as oral supplements or food fortification ingredients.</p> Graphical abstract <p></p>