The effects of duck oil and lard oil on lipotoxicity induced by saturated long-chain fatty acids were evaluated in HepG2 cells. Lipotoxicity triggered by palmitate, a saturated fatty acid, was inhibited more by duck oil-loaded nanoemulsion (DO-NE) than by lard oil-loaded nanoemulsion (LO-NE) and control nanoemulsion (NE) in HepG2 cells. Accumulation of reactive oxygen species and lipid vacuoles in HepG2 cells induced by palmitate treatment was inhibited by DO-NE but not by LO-NE. Consistently, treatment of HepG2 cells with DO-NE, but not with NE or LO-NE, significantly reduced the expression levels of peroxisome proliferator-activated receptor-γ2 and sterol regulatory element-binding protein-1, which are key regulatory proteins in hepatic lipid accumulation. In addition, the cleavage of poly (ADP-ribose) polymerase and caspase-3 were reduced more by DO-NE than by LO-NE, indicating that DO-NE directly attenuates cellular damage induced by palmitate. Collectively, these results imply that the biological activity of duck oil against palmitate-induced cellular damage is more potent than that of lard oil. PRACTICAL APPLICATIONS: Accumulated lipids in nonadipose tissues, especially the liver, cause lipotoxicity, a pathologic feature of hepatic disorders, by inducing oxidative stress. A nanoemulsion loaded with duck oil, which is a functional food widely consumed by Korean people, inhibited lipotoxicity by suppressing lipid accumulation in HepG2 cells exposed to palmitate, which mimic nonalcoholic fatty liver disease. Thus, we propose that duck oil can be used as a functional food to improve lipid-induced hepatic disorders.