Background/aim: In this prospective observational study, we aimed to evaluate the relationship between serum levels of vitamin D and acute postoperative pain scores, as well as opioid analgesic consumption in patients undergoing laparoscopic cholecystectomy. Materials and methods: The study was performed in the Medical Faculty Hospital, from April 2020 to April 2021. Postoperative visual analog scale (VAS) pain scores, total tramadol consumption, number of requests on patient-controlled analgesia (PCA) were compared between the vitamin D deficient (≤20 ng/mL; n = 25) and vitamin D nondeficient (>20 ng/mL; n = 55) groups at five time points (T0: in the recovery room, T1: 1st hour in the ward, T2: 6th hour, T3: 12th hour, and T4: 24th hour). Results: Postoperative VAS pain scores were similar in the vitamin D deficient group at all time points (T0-4), but differed significantly only at the T-0 time point (p = 0.020). The mean cumulative tramadol consumption was significantly higher in the vitamin D deficiency group than in the nondeficiency group (p = 0.005). Vitamin D levels were lower in patients with VAS ≥ 4 at the postoperative T-0 time point (p = 0.009). In the multivariate linear regression analysis, 15.7% of cumulative tramadol consumption was due to vitamin D deficiency (β = –0.188). Conclusion: Our study shows that preoperative low vitamin D level was associated with an increase in acute postoperative pain scores and consumption of opioid analgesics in patients undergoing laparoscopic cholecystectomy. Our findings may be useful for postoperative pain management in patients with vitamin D deficiency.