The purpose of this study was to evaluate and compare the sealing ability of ZOE, FR and Sealapex, as a root canal sealer. Ninety six extracted anterior or premolar human teeth with single root were instrumented by using conventional method. After instrumentation, the teeth were divided into three groups and thirty two teeth in each group were filled as follows: ZOE group: ZOE cement as a root canal sealer in combination with guttapercha cone FR group: FR root canal sealer in combination with guttapercha cone Sealapex group: Sealapex root canal sealer in combination with guttapercha cone $^{45}Ca$ in the form of calcium chloride, was employed as the tracer in this study and produce the auto-radiograph. The depth of isotope penetration into the root canal was evaluated by method by Yates and Hembree at the intervals of 1 day, 7 days, 14 days and 30 days. The following conclusions were derived from the results obtained; 1. After 1 day, ZOE group exhibited the greatest penetration degree of radioisotope (p < 0.05). 2. After 14 days, there was some difference of the radioisotope penetration between each group, but its difference was not significant statistically. 3. After 30 days Sealapex group exhibited the least penetration degree of radioisotope (p < 0.025). 4. In ZOE and Sealapex groups, there was no change of the degree of radioisotope penetration with time.