Betaine surfactants with lipophilic groups of different lengths were synthesized in this research and the dynamic interfacial tension (IFT) between solutions of these surfactants and three kinds of crude oil from Shengli Oilfield are measured. The results indicated that, for Gudao and Gudong heavy oil, cetyl dimethyl hydroxyl sulfobetaine (SBET-16) was the most efficient in lowering the IFT in the case of no alkalis, while for Shengtuo heavy oil, cetyl dimethyl carboxymethyl betaine (CBET-16) was best. SBET-16 with the concentration of 0.003–0.1% and 0.005–0.1% can reduce the oil/water IFT to ultra-low for Gudao and Gudong oil respectively, CBET-16 with the concentration of 0.005–0.1% can lower the oil/water IFT to ultra-low for Shengtuo oil. These results showed that for different oils, an oil displacement agent with high capacity to lower the oil/water interfacial tension may be obtained only by changing the molecular structure of betaine surfactant. This study can be used to guide the design of surfactants for alkaline-free combination flooding.