The aim of this study was to determine pain during endometrial biopsy by comparing the use of intrauterine instillation of levobupivacaine or paracervical block with a placebo in a randomized, double-masked trial in premenopausal and postmenopausal women. A total of 90 women were enrolled in the study. Patients were allocated to either the control or case groups. Group 1 consisted of 30 cases with intrauterine anesthesia with 5 ml 0.5% levobupivacaine. Group 2 consisted of 30 patients who underwent paracervical block with lidocaine. No analgesic agent was given to the remaining 30 patients; these cases comprised the control group. The primary outcome measures were pain or discomfort experienced during the procedure. When the pain scores of the different groups were compared, the scores in the intrauterine levobupivacaine and paracervical block groups were found to be significantly lower compared to those in the control group. There was no difference between the levobupivacaine and paracervical block groups in terms of pain scores. There was a marked positive correlation between biopsy indications and pain scores. Pain scores were lower in cases with the indication of polymenorrhea, hypermenorrhea and metrorraghia compared to those in the cases with other indications. In conclusion, the transcervical intrauterine instillation of levobupivacaine or paracervical block with lidocaine brings about pain relief during and after endometrial biopsy.