A new method of removing rotenone from contaminated water using molecularly imprinted polymeric microspheres (MIPMs) is described. The MIPMs were prepared by the emulsifier-free polymerization method. The removal efficiency and selective recognition ability of the MIPMs were studied. The highest removal efficiency was observed at pH = 8. Moreover, the MIPMs can be re-used for at least 10 times without losing any removal efficiency. The MIPMs provided a selective, simple, reliable and practicable solution to remove rotenone from contaminated water.