Blubber plays different biological roles in cetaceans, including structural support, physiological demands (thermoregulation, streamlining, buoyancy, etc.), and energy storage. As such, biochemical composition, especially lipid, in the blubber may be vertically stratified in order to effectively assist with these functions. We analysed lipid content, class and fatty acid composition in the blubber of by-catch minke whale (Balaenoptera acutorostrata; n = 11), long-beaked common dolphin (Delphinus capensis; n = 3) and Pacific white-sided dolphin (Lagenorhynchus obiquidens; n = 2) from Korean waters. Lipid content for all three species ranged from 55 to 96% of wet weight, with a dominance of triacylglycerols. Vertical stratification of blubber lipid content was different between species. For minke whale and longbeaked common dolphin, it increased from inner (near the muscle) to outer (near the skin) layer, whereas it decreased for Pacific white-sided dolphin. Thirty-one fatty acids were found in all blubber samples and vertical stratification of fatty acid composition was similar among all three species. Among grouped fatty acids, monounsaturates were the most abundant, gradually increasing from inner to outer blubber layer, whereas polyunsaturates and saturates decreased. Such vertical stratification of lipid content and composition suggests that the modification and accumulation of lipid occur in the blubber itself in order to help cetaceans with their specific physiological needs.