Bus mass is an important factor that affects fuel consumption and one of the key input parameters associated with automatic shift and hybrid electric vehicle (HEV) energy management strategy. A city bus mass estimation method based on kinetic energy theorem was proposed in this paper. The real-time data including vehicle speed and engine torque were collected by a remote data acquisition system. The samples in the process of being accelerated were selected to conduct vehicle mass estimation at the same bus stop with the same gear. The average estimation error is 2. 92% after the verification by actual data. Compared with the method based on recursive least squares, the algorithm based on kinetic energy theorem requires less sample length and the estimation error is smaller. Therefore, the method is more suitable for the bus mass estimation. The influences of gear, rolling resistance coefficient, wind resistance coefficient and road slope on mass estimation accuracy were analyzed.