A novel transdermal formulation of fentanyl‐containing dipropylene glycol droplets dispersed in a silicone matrix with a rate‐controlling membrane was developed. Healthy male subjects (n = 24) received repeated 72‐hour applications of fentanyl (50 μg/h) as the novel matrix and the conventional reservoir formulations in a randomized, 2‐way crossover study. Blood samples were collected, and serum concentrations of fentanyl were assayed using liquid chromatography with mass spectrometry detection. The mean area under the curve (AUCτ) and peak concentrations (Cmax) of the matrix formulation were 84 838 pg·h/mL and 1680 pg/mL, respectively. Ratio and 90% confidence intervals of AUCτ and Cmax between the 2 formulations were within 80% to 125%. Adherence of the matrix formulation was higher than the reservoir formulation (62.5 vs 56.2%, P < .0001), without affecting skin irritation. Vital signs and adverse events of the 2 formulations were similar in nature and frequency. The novel matrix formulation displayed enhanced adherence and resulted in similar pharmacokinetics and tolerability as the reservoir formulation.