We investigated the isotopic composition of the Urumqi River and documented seasonal variability attributable to the mixing of various flow sources. Next, we applied these isotopic signals to partition the sources and studied their temporal variability in summer. The isotope hydrology separation results indicated that groundwater is the dominant streamflow source (approximately 62.7%) in the Urumqi River. Precipitation is an important source for the Urumqi River; approximately 19.1-20.7% of the runoff came from precipitation during summer and early autumn. In summer, approximately 21.1% of the runoff is derived from glacial meltwater. In summer, with the increasing distance to the glacier front, groundwater accounts for a larger and larger percentage of the river water, and the contributions of precipitation and glacial meltwater gradually diminish. Throughout 2012, the proportions of precipitation and glacial meltwater in the streamflow were 17.6% and 14.7%, respectively, and only 5% of the streamflow was derived from snowmelt.