This study was performed to evaluate the sealing ability of various retrograde filling materials by using bacterial penetration and dye penetration test. One hundred and forty extracted human teeth with single, straight canals and mature apiece were collected and used for this study. All canals were instrumented using an engine driven Ni-Ti file (ProFile). After removing 3mm from the apex of tooth, a standardized 3mm root end cavity was prepared using an ultrasonic instrument. The 70 teeth were randomly divided into 7 groups : 6 groups for retrograde filling using Super-EBA, ZOE, Chelon-Silver, IRM, ZPC and amalgam. The 7th group was used as a negative control. Nail varnish was applied to all external root surfaces to the level of the reseated root ends to prevent lateral microleakages. The specimens were then sterilized in an ethylene oxide sterilizer for 24 hours. 2 mm of the reseated root was immersed in a culture chamber containing a Tripticase Soy Broth with a phenol red indicator. The coronal access of each specimen was inoculated every 72 hours with suspension of Proteus vulgaris. The culture media were observed every 24hours for color change indicating bacterial contamination. The specimens were observed for 4weeks. The remaining 70 teeth were submitted to a dye penetration test. The canals of all teeth were first sealed with AH26 and obturated using an Obtura II system. Root resection, root end preparation and retrograde filling was performed as above. All specimens were suspended in 2% methylene blue dye for 72 hours before being ion gitudinally split. The degree of dye penetration was then measured using a stereomicroscope at 10 magnification and evaluated. The results were as floows : 1. In the bacterial penetration, the degree of leakage was the lowest in the Super-EBA, followed by, in ascending order, ZOE, Chelon-Silver IRM and ZPC. The amalgam showed highest bacterial leakage of all(p<0.01). 2. In the dye penetration, the degree of microleakage was the lowest in the Chelon-Silver and Super-EBA, followed by, in ascending order, IRM, ZPC. The ZOE and amalgam showed the highest microleakage of all (p<0.05). These results suggested that the eugenol based cement, Super-EBA, have excellent sealing ability as a retrograde filling material.