Bio-diesel has high potential as a substitute fuel and renewable energy source in the future and can be directly used in existing diesel enginse. In the present study, a new method has been employed to investigate bio-diesel synthesis in a column reactor packed with cation exchange resin from four types of waste oil. The tested resins included cation exchange resins D002, 002CR, and 732. Cation exchange resin D002 presented more effective catalytic activity than 002CR and 732 in this work. The bio-diesel yield can achieve 85.6-97.1% under the following operating conditions: oil to methanol molar ratio 1: 9, reaction temperature 60 C, and reaction time 4 h. The tested resin D002 showed high operation stability after repeated uses. Compaed with bio-diesel production catalyzed by traditional sulfuric acid, bio-diesel synthesis from waste oil in a packed column reactor has application potentials.