Oxygen (δ18O) and hydrogen (δD) stable isotopes in the surface waters of the Huai River basin were analyzed in this study. Results indicated the northern waters had higher δ18O and δD than the southern waters, the water δ18O and δD increased along the water flow directions. These variations mostly resulted from the spatial differences of precipitation and evaporation. Comparing with published different continents' river water δ18O data, this study suggests that evaporation effect is a more plausible interpretation than altitude effect as the cause of δ18O increasing from upriver to downriver waters. This region's local surface water line (LSWL, δD = 5.36δ18O − 18.39; r2 = 0.84) represents one of the first presented LSWLs in eastern China. The correlation between d-excess and δ18O demonstrates this region is dominated by the Pacific oceanic moisture masses in summer. Comparing the various LSWLs from eastern China and eastern United States river waters, this study proposes a hypothesis that the water LSWLs slopes of lower latitude regions may be less than those of higher latitude regions within similar topographic areas. This hypothesis may be tested in other geographically comparable coupled areas in the world if corresponding large-scale data can be found.