Individual variability in the effects of opioid analgesics such as fentanyl remains a major challenge for tailored pharmacological treatment including postoperative analgesia. This study aimed to establish a new real-time method for detecting the effects of fentanyl and their individual differences in the preoperative period, using the pressure pain threshold (PPT) and Narcotrend index (NTI) test. Eighty women undergoing elective surgery under general anesthesia were enrolled in this randomized, double-blinded, placebo-controlled study to receive either intravenous fentanyl (Group F) or saline (Group S). Before (T1) and 5 (T2) and 10 min (T3) after intravenous injection, the PPT, NTI, respiratory rate, heart rate, blood pressure, and pulse oxygen saturation were measured. The initial time at which the Narcotrend index showed a decline was also recorded. In total, 40 patients in Group S and 38 patients in Group F were included in the final analysis. At 5 min and 10 min after intravenous fentanyl administration, the analgesic effect was determined by measuring the PPT, which was significantly increased (P < 0.001), and the sedative effect was detected using the NTI, which was significantly decreased (P < 0.001). The distribution of percentage changes of the PPT and NTI showed individual differences. At T2 and T3, the absolute changes in NTI and PPT were positively correlated (r = 0.444 at T2, P = 0.005; r = 0.332 at T3, P = 0.042). Through the PPT and NTI, it was feasible to easily detect the effects of fentanyl and their individual differences in real time before induction of anesthesia in the operation room. This method could potentially be applied to preoperatively determine patients' sensitivity to fentanyl.