The acquired dental pellicle helps prevent erosion, but the protection level is unknown. This in situ study tested whether a two-hour pellicle protects against different erosive challenges by orange juice. Subjects wore palatal appliances loaded with either enamel or dentin specimens. Pellicle was allowed to form, or not (control), on the surfaces of the specimens intra-orally for 2 hrs before the erosive challenges of 0 (control), 10, 20, and 30 minutes' duration. Specimens were randomly removed from the appliances after each challenge. Percentage of surface microhardness change (%SMC) was determined for the enamel specimens, and that of mineral loss and lesion depth for the dentin specimens. Enamel specimens with the pellicle showed a significantly lower %SMC, only after the 10-minute challenge. No protection was found for dentin. It was concluded that the acquired pellicle reduced dental erosion, but that this effect was limited to the less severe erosive challenge on enamel surfaces.