Purpose: To test the apical leakage prevention performance of three different materials through protein leakage procedures using bovine serum albumin (BSA) and Bradford protein reagent. Materials and Methods: A total of 60 human single-rooted teeth were divided into 4 groups, and conventional root canal filling was done. The root was cut 3 mm from the apex, and a cavity was formed. Proroot MTA (MTA), Fuji II LC (GI), Fuji II LC with XP bond (GIA), and Caviton (CA) were used as experimental materials to fill the cavity in a retrograde filling manner. The extent of BSA leakage was then measured with a ultraviolet visible spectrophotometer 24, 48, and 72 hours after filling. Result: After 24 hours, among the 15 teeth of each group, 2 in MTA, 4 in GI, 3 in GIA, and 7 in CA showed leakage. After 48 hours, 3 in MTA, 5 in GI, 5 in GIA, and 10 in CA had leakage and discoloration. After 72 hours, among the 15 teeth of each group, 3 in MTA, 6 in GI, 5 in GIA, and 10 in CA showed leakage. The leakage in the CA group was greater than that in the MTA group at 48 and 72 hours based on Fisher's exact test (P=0.025), and the difference was statistically significant. Similarly, the leakage in the CA group was greater than that in the MTA group over time based on the Kaplan-Meier survival estimate (P=0.011), and the difference was statistically significant. Conclusion: Glass ionomer, glass ionomer after adhesive application, and MTA all showed leakage. Caviton showed greater leakage compared to MTA 48 and 72 hours after filling, and the difference was statistically significant; thus suggesting that Caviton is not appropriate as retrograde filling material considering its sealing ability.