Aims: To describe levels of sexual desire across the menopausal transition (MT) and early postmenopause (PM), including effects of age, MT-related factors, health, stress, symptoms (hot flash, sleep, mood), and social opportunity factors. Methods: A subset of Seattle Midlife Women's Health Study (SMWHS) participants who provided data during the early reproductive, early and late menopausal transition stages, or postmenopause (n ¼ 286), including menstrual calendars for staging the MT, annual health reports between 1990 and 2005, and morning urine samples assayed for estrone glucuronide (E 1 G), testosterone (T), and follicle-stimulating hormone (FSH) was included. Multilevel modeling using the R program was used to test factors related to sexual desire. Results: Women experienced a significant decrease in sexual desire during the late MT stage ( p < 0.01) and early PM ( p < 0.0001). Those with higher urinary E 1 G and T reported significantly higher levels of sexual desire, whereas those with higher FSH levels reported significantly lower sexual desire ( p < 0.0001, 0.06, and 0.0002, respectively). Women using hormone therapy also reported higher sexual desire ( p ¼ 0.02). Those reporting higher perceived stress reported lower sexual desire ( p < 0.0001), but history of sexual abuse did not have a significant effect. Those most troubled by symptoms of hot flashes, fatigue, depressed mood, anxiety, difficulty getting to sleep, early morning awakening, and awakening during the night also reported significantly lower sexual desire ( p range from <0.03 to 0.0001), but there was no effect of vaginal dryness. Women with better perceived health reported higher sexual desire ( p < 0.0001), and those reporting more exercise and more alcohol intake also reported greater sexual desire ( p < 0.0001). Having a partner was associated with lower sexual desire. Conclusions: Clinicians working with women traversing the MT should be aware that promoting healthy sexual functioning among midlife women requires consideration of their changing biology as well as ongoing life challenges.