Tooth bleaching is a popular aesthetic treatment to modify the color of teeth. Despite the extensive literature concerning the subject, there is still no consensus regarding the application mode of the different bleaching agents and their effect on enamel. Therefore, this study evaluated the influence of different bleaching protocols on whitening efficiency and enamel superficial hardness. Bovine enamel fragments were embedded in acrylic resin and wet-sanded to obtain a flat buccal surface. The specimens were then randomly divided into 6 groups (n=10), based on the bleaching material [HP Maxx 35% (35% hydrogen peroxide), HP Blue 35% (35% hydrogen peroxide + Ca) and Whiteness Perfect 10% (10% carbamide peroxide)] and application mode (3 applications of 15 min, 1 application of 45 min, 1 application of 1h30 or 1 application of 3h30). The color and superficial hardness were assessed before and after bleaching. The color was assessed by means of a digital spectrophotometer, using CIELab parameters. Vickers hardness was determined using a load of 200g for 10s. Data were statistically analyzed by one-way ANOVA with repeated measures and Tukey?s test (? = 0.05). Concerning the color alteration, the groups were similar to each other, except for at-home bleaching protocols, which were statistically different from each other. Similarly, there was a reduction in hardness values comparing the initial and final periods, with lowest final hardness for the at-home bleaching protocol group applied for 3h30. It can be concluded that the bleaching protocols influenced the final color and enamel surface hardness, evidencing that lower gel concentrations applied for longer periods promoted greater variation in color and lower final hardness.