Canine skin is a highly dynamic organ that is constantly adapted to changes in its environment. It provides structural, sensory, immunologic, and physiologic functions and contributes an essential barrier function against potential environmental insults. We assessed the changes of canine skin hydration, pH, and protein contents by noninvasive method according to the frequency of application with canine commercial shampoo. Canine commercial shampoo was applied topically every other day, 5 days, and 8 days on different sites, respectively in 8 dogs. Saline-applied site was as a control. Skin hydration was increased and skin surface pH was decreased significantly in canine commercial shampoo-applied site every other day (p < 0.05). Also, skin protein concentration measured by D-squame$^{(R)}$ tape stripping method was increased significantly in canine commercial shampoo-applied site every other day (p < 0.05). There were alterations on every 5 days and 8 days, but not significantly. These results may be recognized that the frequent use of canine commercial shampoo cause abnormalities of the skin barrier function and alteration of stratum corneum integrity/cohesion. Consequently it was suggested that bathing the dog with canine commercial shampoo was appropriate every 5 or 8 days.