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Recently, quantitative analyses of food web structure based on carbon and nitrogen stable isotopes are widely applied to environmental assessments as well as ecological researches of various ecosystems, particularly rivers and streams. In the present study, we analyzed carbon and nitrogen stable isotope ratios of POM (both planktonic and attached forms), zooplankton, benthic macroinvertebrates and fish collected from 6 sites located at Nakdong River. Samples were collected from upstream areas of 5 weirs (Sangju, Gangjeong-Goryeong, Dalseong, Hapcheon-Changnyeong, and Changnyeong-Haman Weirs) and one downstream area of Hapcheon-Changnyeong Weir in dry season (June) and after rainy season (September). We suggested ranges of their carbon and nitrogen stable isotope ratios and calculated their trophic levels in the food web to compare their temporal and spatial variations. Trophic levels of organisms were relatively higher in Sangju Weir located at upper part of Nakdong River, and decreased thereafter. However, the trophic levels were recovered at the Changnyeong-Haman Weir, the lowest weir in the river. The trophic level calculated by nitrogen stable isotope ratios showed more reliable ranges when they were calculated based on zooplankton than POM used as baseline. The suggested quantitative ecological information of the majority of biological communities in Nakdong River would be helpful to understand the response of river food web to environmental disturbances and can be applied to various further researches regarding the quantitative approaches for the understanding food web structure and function of river ecosystems as well as restoration.