Desmopressin is a synthetic peptide hormone chiefly used for treatment of enuresis in young children. It is available in injectable, intranasal, and oral formulations. While administration by injection is poorly suited for routine use in young children, intranasal and oral administration result in low and variable bioavailability. This study therefore explored the feasibility of administering desmopressin transdermally using Macroflux technology, which uses a microneedle array to overcome the skin barrier. The tips of microneedles in 2-cm2 arrays were covered with a solid coating of various amounts of desmopressin and applied to the skin of hairless guinea pigs for 5 or 15 min. Pharmacologically relevant amounts of desmopressin were delivered after 5 min. Bioavailability was as high as 85% and showed acceptable variability (30%). Immunoreactive serum desmopressin reached peak levels after a Tmax of 60 min. Elimination kinetics for serum desmopressin was similar after transdermal and intravenous (IV) delivery, suggesting the absence of a skin depot. Only 10% of the desmopressin dose loaded onto the microneedle array was found on the skin surface after application. Additionally, the patches were well tolerated. These results suggest that transdermal delivery of desmopressin by Macroflux is a safe and efficient alternative to currently available routes of administration.