BACKGROUND: The age-related decline of dehydroepiandrosterone (DHEA) has prompted research on its experimental replacement in women. Although no relationship to sexual functioning in healthy women has been shown to date, DHEA replacement has potential for affecting sexual response. METHODS: To investigate DHEA effects, 16 sexually functional postmenopausal women participated in a randomized, double-blind, crossover protocol in which oral administration of DHEA (300 mg) or placebo occurred 60 minutes before the presentation of an erotic video segment. Blood DHEA sulfate (DHEAS) changes, subjective and physiological sexual responses, as well as affective responses were measured in response to videotaped neutral and erotic video segments. RESULTS: The concentration of DHEAS increased 2-5-fold following DHEA administration in all 16 women. Subjective ratings across DHEA and placebo conditions showed significantly greater mental (p < 0.016) and physical (p < 0.036) sexual arousal to the erotic video with DHEA vs. placebo. Positive affect also increased during the erotic video across drug conditions. Vaginal pulse amplitude (VPA) and vaginal blood volume (VBV) demonstrated a significant increase (p < 0.001) between neutral and erotic film segments within both conditions (DHEA and placebo) but did not differentiate drug conditions. CONCLUSION: In sum, increases in mental and physical sexual arousal ratings significantly increased in response to an acute dose of DHEA in postmenopausal women.