This study aimed to determine the anti-erosive effects of xylitol, fluoride and a xylitol/fluoride combination used as an additive in an acidic drink or as mouthrinse after enamel was exposed to an acidic drink, in vitro. Human third molars were divided into 7 groups (A-G). Samples from groups A to D were immersed for 5 min in orange juice only (A), orange juice plus either 25% xylitol (B), F(-) 1 ppm (C) or a 25% xylitol/F(-) 1 ppm combination (D), respectively. Samples from groups E to G were immersed in orange juice for 5 min and then in either 40% xylitol (E), F(-) 227 ppm (F) or a 40% xylitol/F(-) 227 ppm combination (G), for 1 min respectively. This process was performed four times daily for 14 days. Mineral loss was determined from the lesion depth and surface hardness. Erosion depth progressively increased in all groups, except E, where erosion depth was significantly lower than group A. Surface microhardness progressively decreased in all groups, except E, where hardness was significantly higher than group A. This study demonstrated that addition of xylitol, fluoride or a xylitol/fluoride combination to an acidic drink or post-treatment with fluoride or a xylitol/fluoride combination could reduce, but not prevent, enamel erosion.