Although uncoupling metabolism is a possible measure to reduce sludge generation during the activated sludge process for treating organic wastewater, the fate of the chemical uncoupler in the activated sludge system and its residual toxicity have not been widely studied. In this work, 2,4,6-trichlorophenol (TCP), a typical metabolic uncoupler, was used as an example to investigate the distribution and persistence of chemical uncouplers in an activated sludge system treating municipal wastewater. In addition, the cytotoxicity of residual TCP in the effluent was also evaluated using Vero cells. Results showed that ranges of residual TCP in the effluent were 0.5-1.0, 0.9-1.4, and 1.3-2.4 mg L -1 when the TCP in the feed was 2, 4, and 6 mg L -1 , respectively. TCP concentrations in the sludge phase significantly exceeded those in the water phase, due to the accumulation effect of sludge on TCP. TCP was biodegraded to some extent during the operating process, and the degradation efficiency increased with the number of operating cycles. About 0.07 mg g -1 sludge (dry weight) in the cell interior is a critical concentration for efficient inhibition of excess sludge generation. Although the acute cytotoxicity of TCP at concentrations less than 2.0 mg L -1 in the effluent was relatively low, concentrations greater than 2.5 mg L -1 had a significant effect on the morphology and proliferation of Vero cells. Accumulative and chronic effects of trace TCP in effluents on the ecosystem should be further studied. Dosages of more than 4 mg L -1 TCP (corresponding to 2.5 mg L -1 residual concentration) for sludge reduction in sequencing batch reactors treating municipal wastewater is not recommended due to acute toxicity.