Extensive use of neonicotinoid insecticides (NNIs) worldwide calls for further knowledge on their environmental occurrence and risk. The present study highlighted the need for more research on long-term exposure of NNIs in aquatic environment, which is important but remains elusive. Time weighted average concentrations of five commonly used NNIs in urban waterways of Guangzhou, China were measured using newly developed polar organic chemical integrative samplers (POCIS). Acetamiprid (from 18.8 ± 1.9 to 157 ± 31 ng/L; mean ± standard deviation), clothianidin (from 14.8 ± 3.7 to 47.6 ± 10.0 ng/L) and imidacloprid (from 32.9 ± 11.6 to 249 ± 19 ng/L) were detectable in all samples. Thiamethoxam was found at 71.4% of the 21 sampling sites (from not detected to 52.4 ± 9.4 ng/L), while thiacloprid was not detected at any site. Vegetable planting and sewage effluent were the main sources of NNIs in surface water in Guangzhou. Probabilistic environmental exposure distributions were subsequently constructed using the measured concentrations and the exceedances of predicted environmental concentrations of NNI to ecological thresholds were assessed. In Guangzhou, 63.5%, 16.2%, 87.8% and 17.2% of acetamiprid, clothianidin, imidacloprid and thiamethoxam, respectively, exceeded an interim chronic threshold of 35 ng/L for NNIs. Further risk assessment and control measures for the use of NNIs are advocated for protecting the integrity of aquatic ecosystems.