PURPOSE: The purpose of this study was to compare the effect of current density on penetration depth, tissue concentration and transdermal transport of methylene blue(MB) by iontophoretic transdermal delivery. METHODS: Twenty-four male Sprague-Dawley rats were randomly divided into 1 mA($0.11mA/cm^2$), 2 mA($0.22mA/cm^2$), 4 mA($0.44mA/cm^2$), and 8 mA($0.89mA/cm^2$) groups. These rats were exposed to anodic iontophoresis of 1% MB using a direct current for 15 minutes. The penetration depth were measured using light microscopy from cryosections of skin tissue. The tissue concentration and transdermal transport were measured using biochemical analysis from target skin tissues. The data were analyzed with one-way analysis of variance. RESULTS: The significant differences in the penetration depth, tissue concentration and transdermal transport were detected among the groups(p<.001). Post hoc comparisons of the penetration depth, tissue concentration and transdermal transport of he 2 mA, 4 mA, and 8 mA iontophoresis groups were greater than in the 1 mA iontophoresis group(p<.05). There was no significant difference, however, among 2 mA, 4 mA, and 8 mA iontophoresis group. CONCLUSION: There was no difference in the efficiency of iontophoresis from 2 mA($0.22mA/cm^2$) to 8 mA($0.89mA/cm^2$). Higher current density can cause skin injury and discomfort sensation. In general, $0.5mA/cm^2$ is proposed to be the maximum iontophoretic current which should be used on human. The appropriate current amplitude should be selected by considering the safety current density and the depth of the target tissue.