Tramadol, a central analgesic acting on serotonin neurotransmission, is often co-used with ondansetron, a 5-HT3 antagonist, for the management of postoperative pain to decrease nausea and vomiting. The aim of the study is to test the hypothesis that this drug combination raises tramadol requirement by patient-controlled analgesia (PCA). Forty patients undergoing hernioplasty or thyroidectomy were enrolled in a randomized, controlled study and allocated to receive ondansetron 4 mg i.v. (n = 20) or saline (n = 20). At 0, 1, 2, 4 and 24 h of PCA, tramadol consumption was evaluated. Tramadol consumption (mg × kg–1 × h–1) was higher in the ondansetron group at the 2-hour time point compared to the control group (0.24 ± 0.1 vs. 0.17 ± 0.16; p = 0.01). Our study suggests that ondansetron acutely reduces the analgesic efficacy of tramadol.