The purpose of this study is to analyze the characteristics of environment-friendliness of Root Barrier materials based on comprehensive experiments on harmfulness of Root Barrier materials and fish toxicity of Oryzias latipes mortality to verify eco-toxicity of each method of construction and Root Barrier material, which are to be applied by taking eco-toxicity into account when building ecological flows in upper areas on natural and artificial grounds. As a result, the following conclusions have been reached in this study: 1. In regard of the harmfulness analyzed, each material showed a different result of analytical value in each lab tank. Compared to lab tank, pH (la), DO (lb), T-N (VI) and T-coli (la) were in the same grade, but COD, SS, T-P and F-coli were less than that of control, respectively increased or decreased by material were analyzed. 2. In the experiment of fish toxicity, Barrier sheet was found to have 66.7% of fish mortality, indicating strong fish toxicity. Synthetic rubber system root barrier sheet (20.0%) was analyzed to have medium fish toxicity, while Synthetic resin system root barrier-waterproof sheet (3.3%), Synthetic rubber system membrane root barrier sheet (3.3%) and Synthetic resin system root barrier sheet (0.0%) showed relatively lower mortality and fish toxicity. To sum up such results as found in the experiment mentioned so far, the values of harmfulness and root penetration resistance analyzed were different in each lab tank, but there was absolutely little correlation with the mortality gained from the experiment on fish toxicity. In the experiment of fish toxicity, environment-friendly root barrier materials were analyzed, and it was found that Synthetic resin system root barrier sheet, Synthetic resin system root barrier waterproof sheet and Synthetic rubber system membrane root barrier sheet are highly environment-friendly. In contrast, Synthetic rubber system root barrier sheet was found to have medium-level environment-friendliness. Also, Barrier sheet was analyzed to have low environment-friendliness.