The aim of the present study was to test the impact of different toothpastes on the prevention of erosion. Enamel demineralization and remineralization were monitored using surface microhardness (SMH) measurements. Human enamel specimens were treated following two different procedures: (1) incubation in toothpaste slurry followed by acid softening and artificial saliva exposure; (2) acid softening followed by incubation in toothpaste slurry and artificial saliva exposure. For the control procedure, toothpaste treatment was excluded. The following toothpastes were tested: Zendium, Sensodyne Proschmelz (Pronamel), Prodent Rocket Power, Meridol and Signal active. Normalized SMH values compared to the baseline (= 1.00) after 1-hour artificial saliva exposure for procedure 1 (respectively for procedure 2) were as follows (mean: 95% CI): Sensodyne Proschmelz 0.97: 0.93, 1.00 (0.92: 0.90, 0.94), Zendium 0.97: 0.94, 1.00 (0.89: 0.83, 0.95), Meridol 0.97: 0.94, 1.00 (0.94: 0.92, 0.96), Signal active 0.94: 0.91, 0.97 (0.95: 0.91, 0.99), Prodent Rocket Power 0.92: 0.90, 0.94 (0.93: 0.89, 0.97) and control 0.91: 0.88, 0.94. Further exposure to artificial saliva for up to 4 h showed no significant improvement of SMH. Regression analyses revealed a significant impact of the applied procedure. Incubation in toothpaste slurries before the acid challenge seems to be favorable to prevent erosion. None of the tested toothpastes showed statistically significant better protection than another against an erosive attack.