The important Iranian Karun River has never been investigated for the presence of potentially endocrine-disrupting chemicals, nonylphenol (NP). In this study, concentrations of NP were measured in water from Karun River and five wastewater discharge points into this river, collected during April to July 2010. The analytes were extracted by solid-phase extraction, and quantitative analyses were performed by HPLC–FLD. NP was detected in water and wastewater samples with 0.17–1.83 and 15.27–21.79 μg/L, respectively. The results showed that the NP content of aqueous phase of all wastewater samples higher than particulate phase, which were detected in the aqueous and particulate phases with mean concentrations of 12.8 ± 2.4 and 5.2 ± 1.2 μg/L, respectively. These data suggest that the NP levels in Karun river water are likely attributable to untreated municipal wastewaters discharged directly into the river. To our knowledge; this is the first study to evaluate NP concentrations in water and wastewater in Iran.