Oil-in-water emulsions have shown promise as safe and effective adjuvant formulations for vaccines. In particular, formulations consisting of metabolizable oils such as shark-derived squalene and detergents such as egg phosphatidylcholine have been used to produce stable vaccine emulsion formulations. However, there is an emphasis in pharmaceutical regulatory bodies on using synthetic or plant-derived components from sustainable sources instead of animal-derived components. This study compares the physicochemical properties and biological efficacy of emulsions consisting of oil and detergent components from animal, plant, and synthetic sources. In particular, effects of component structure and source on emulsion stability and biological activity are examined. It is shown that oil-in-water emulsions using animal-derived components can be substituted with synthetic or plant-derived materials while still exhibiting satisfactory physicochemical and biological properties.