In this study, 3 types of zwitterionic phospholipid biosurfactants LDP(S), CDP(S) and CTDP(S) were prepared from 3 different raw materials such as rapeseed oil, coconut oil and cottonseed oil respectively. The structure of the resulting phospholipid biosurfactants was elucidated by FT-IR, 1H NMR and 13C NMR spectroscopies and their interfacial properties have been examined such as CMC, static surface tension, wetting property, solution stability, and foam property. Interfacial property measurement and prescription test in cosmetic formulation prepared with the newly synthesized biosurfactants revealed that CDP(S) biosurfactant possesses excellent mildness and superior interfacial properties, indicating the potential applicability in cosmetic product formulations.