To study the interfacial activities of zwitterionic surfactant against crude oil, four alkyl hydroxyl sulfobetaines were synthesized. The effects of surfactant structure, concentration, salinity of formation water, and crude oil composition on interfacial activities were discussed. It shows that the octadecyl hydroxyl sulfobetaine can achieve ultralow interfacial tension at the concentrations no less than 0.001%, exhibiting a good interfacial activity and a very strong antidilution ability. The salinity of formation water has no obvious effect on lowering the interfacial tension and the polar component in crude oil is very crucial for the production of ultralow interfacial tension. This work is important for enhanced oil recovery.