Occurrence, variation and behaviour of nonylphenol (NP) and octylphenol (OP) were studied in surface water and groundwater in Guiyang, Guizhou Province, southwestern China. Discharge of wastewater from Guiyang City was the main source of alkylphenols (APs) entering the aquatic environment. The concentrations of NP and OP in river water ranged from 40 to 1582 ng L(-1) and from below the lowest limit of detection (LOD) to 67 ng L(-1), respectively. NP and OP were also detected in groundwater. Both NP and OP exhibited spatial and temporal variations in river water and groundwater. It was found that concentrations of NP and OP in river water was low upstream and dramatically increased downstream, and higher concentration of NP was found in winter compared to that in summer. Proportions of NP and OP were trapped by suspended particulate matter (SPM), which accounted for 7.6-50.0% and 3.4-25.6% of their total concentration in the river water system, respectively. Seasonal changes in water flow were responsible for the temporal variations of APs. To determine the behaviour of APs along the river, a mass balance equation based on chloride was used. The results showed that a mixing process was the predominant factor to determine upstream APs concentrations; while the discharge of wastewater controlled the concentrations of APs downstream. Considering the adverse effect of APs on organisms, combined effect modeling was used to assess the toxicity to fish. It was found that the predicted mixture effect for APs in river water on fish vitellogenin induction was low upstream and medium downstream, respectively.