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There is a need for an objective technique to assess the degree of hypoactive sexual desire disorder (HSDD). Recently, we described such a methodology (event-related potential technique [ERP]) based on recording of p300 electroencephalography (EEG) waves elicited by auditory stimuli during synchronous exposure to erotic films.To compare sexual interest of sexually healthy women to females with sexual dysfunction (FSD) using ERP, and to explore whether FSD women with and without HSDD would respond differently to two different types of erotic stimuli—films containing (I) or not containing (NI) sexual intercourse scenes.Twenty-two women with FSD, of which nine had HSDD only, and 30 sexually healthy women were assessed by the Female Sexual Functioning Index. ERP methodology was performed applying erotic NI or I films.Significant differences in percent of auditory p300 amplitude reduction (PR) in response to erotic stimuli within and between all three groups for each film type.PRs to each film type were similar in sexually healthy women (60.6% ± 40.3 (NI) and 51.7% ± 32.3 [I]), while in women with FSD, reduction was greater when viewing the NI vs. I erotic films (71.4% ± 41.0 vs. 37.7% ± 45.7; P = 0.0099). This difference was mainly due to the greater PR of the subgroup with HSDD in response to NI vs. I films (77.7% ± 46.7 vs. 17.0% ± 50.3) than in the FSD women without HSDD group or the sexually healthy women (67.5% ± 38.7 vs. 50.4% ± 39.4 respectively), P = 0.0084. For comparisons, we used the mixed-model one-way analysis of variance.Differences in neurophysiological response patterns between sexually healthy vs. sexually dysfunctional females may point to a specific inverse discrimination ability for sexually relevant information in the subgroup of women with HSDD. These findings suggest that the p300 ERP technique could be used as an objective quantitative tool for libido assessment in sexually dysfunctional women. Vardi Y, Sprecher E, Gruenwald I, Yarnitsky D, Gartman I, and Granovsky Y. The P300 event-related potential technique for libido assessment in women with hypoactive sexual desire disorder. J Sex Med 2009;6:1688-1695.