Objective To explore the impact of testosterone levels on female sexual function in menopausal transitional and postmenopausal women. Methods A total of 450 menopausal transitional and postmenopausal women who visited Beijing Obstetrics and Gynecology Hospital for the first time from January to December 2023 were recruited, including 201 menopausal transitional and 249 postmenopausal women. Demographic data and medical history were collected, female sexual function and climacteric symptoms were evaluated by using female sexual function index (FSFI) scale and modified Kupperman index (KMI). The sex hormone levels were tested for all women, including total testosterone(TT), free testosterone(FT), bioactive testosterone(BioT), 4-androstenedione(A4) and estrogen. Multiple linear regression is used to explore the impact of testosterone levels on female sexual function. Results Compared with menopausal transitional women, the FSFI scores in postmenopausal women decreased significantly (
P<0.05). Multiple linear regression analysis showed that BioT level was positively correlated with arousal score (
β=0.190,
P<0.05) and FSFI total score (
β=0.178,
P<0.05), FT level was positively correlated with desire (
β=0.166,
P<0.05), satisfaction (
β=0.176,
P<0.05), and FSFI total score (
β=0.126,
P<0.05), while TT level was not associated with FSFI scores. The KMI score is negatively correlated with the FSFI scores (
P<0.05). Conclusions Female sexual dysfunction (FSD) was associated with reduced FT and BioT levels, while not associated with TT level.