Practical applications: The development of multilayer emulsions can be extremely useful to the encapsulation of lipophilic bioactive molecules for food applications, such as polyunsaturated fatty acids (PUFAs), flavors, vitamins, etc. Moreover, this technique provides a system for the creation of O/W emulsions with improved stability to environmental stresses, protection of oil phase, and controlled and triggered release of microencapsulated compounds. In this work, the design of stable O/W emulsions containing PUFAs rich oil droplets surrounded by multiple layer interfacial membranes from food grade ingredients, such as lupin proteins (AluProt-CGNA), chitosan, and xanthan gum membranes; has been performed. The designed emulsion proved to be stable under different environmental stresses found in many food products. Moreover, thermal stability exhibited by these emulsions could also be an important characteristic for its use in food products.