We conducted a randomized controlled trial to assess the feasibility and safety of performing gynecological single-port transumbilical laparoscopic-assisted adnexal surgery without urethral catheterization in a day surgery setting. A total of 153 patients with adnexal disease were enrolled in this prospective randomized controlled trial (RCT). All subjects performed single-port transumbilical laparoscopic-assisted adnexal surgery between March 2021 and July 2022 in a day surgery center. After completion of the baseline survey, participants were randomized into one of three groups. Participants were randomized into one of three groups: uncatheterized (n = 51), intermittent catheterized (n = 51), or indwelling catheterized (n = 51). The primary outcomes were the incidence of lower urinary tract symptoms (LUTS) and microscopic hematuria, and the secondary outcomes included the incidence of urinary tract infection (UTI), the incidence of urinary retention, the incidence of bladder injury, the time till first urination, the time till first ambulation, the time till first exhaust, the time till first feeding and Kolcaba comfort score. The incidence of postoperative LUTS in the uncatheterized group (17.65%) was lower than that in the intermittent catheterized group (52.94%) and the indwelling catheterized group (84.31%), and there was significant difference between the two catheterized groups (P < 0.001). In the patients without vaginal manipulation, the incidence of microscopic hematuria in the uncatheterized group (0%) was lower than that in the intermittent catheterized group (37.50%) and the indwelling catheterized group (38.89%) (P < 0.05). There were no significant differences in the first urination time, first ambulation time, first exhaust time, first feeding time, and comfort score among the three groups (P > 0.05). Moreover, no urinary retention, UTI and bladder injury were recorded in the three groups. Gynecological single-port laparoscopic adnexal surgery without urinary catheter is safe and feasible in a day surgery ward, which can reduce the incidence of postoperative LUTS and microscopic hematuria.