The objective of this study was to evaluate the remediation of water containing the insecticide chlorpyrifos by using constructed wetlands (CW) cultivated with Polygonum punctatum, Cynodon spp. and Mentha aquatica, operated under different hydraulic retention times: 24, 48, 96, 144 and 192 h. The system efficiency was based on reduction of the initial concentration of chlorpyrifos and toxicity of the contaminated water. The results showed that constructed wetlands are an excellent alternative for remediation of the insecticide chlorpyrifos in aqueous medium. It was observed that the average overall removal efficiency of the insecticide was 98.6%, and in the first hydraulic retention time, 24 h, chlorpyrifos was removed to levels below the detection limit in all CW. This result is mainly attributed to adsorption and microbial degradation. For the qualitative standard acute toxicity tests with Daphnia similis, for most samples there was a reduction in toxicity greater than 80%. It was reported that the ecotoxicological tests with the effluents of the constructed wetland are a good option as an indicator of the effectiveness of treatments and a promising alternative to complement the physical and chemical analyses.