The aim of this study was to determine the leakage of SuperEBA and intermediate restorative material (IRM) in root canal samples, with or without orthograde filling, by evaluating bovine serum albumin (BSA) microleakage using spectrophotometry. Thirty-five single-rooted teeth were divided into five groups, instrumented, and had apices resected. Root-end cavities in groups I and II were filled with SuperEBA and IRM. The samples from the groups III, IV, and V were filled with gutta-percha and sealer. In groups IV and V, root-end cavities were filled with SuperEBA and IRM. After 60 days, the greatest microleakage of BSA was observed in group II (4.1 +/- 0.0011 ng), followed by group III (3.4 +/- 0.0064 ng), and then group I (2.6 +/- 0.0019 ng). Samples from groups IV and V leaked the least (0.7 +/- 0.0014 ng). Significantly less leakage (p < 0.05) occurred in samples filled with orthograde and root-end fillings than did in samples filled only with an orthograde approach and the samples with IRM root-end fillings.