Aim: This cross-sectional study aims to evaluate the patient's perception of discomfort and pain arising from brass wire and elastomeric separators used in orthodontic appliances among Pakistani adolescents. Materials and methods: A total of 80 patients (40 females and 40 males) aged 18–28 years who had undergone fixed orthodontic therapy were included in this study. Details were gathered through a self-administered questionnaire provided to patients via a single investigator. The discomfort and the pain initiated by the immediate placement of separators until the seventh day were evaluated. Results: Results showed that after immediate placement, brass wire separators caused the highest pain levels (88.6%) compared to elastomeric separators (2.9%). The pain perception for elastomeric separators was higher (71.4%) after 2 days of insertion than that for brass wire separators. A gradual decline in pain from both separators over 7 days was observed. Patients reported discomfort while eating and chewing. The perception of discomfort and pain between genders was not significant (p value > 0.05). Conclusion: After immediate insertion, the brass wire separators caused more discomfort and pain, which lasted until 24 hours than elastomeric separators. The discomfort and the pain associated with elastomeric separators lasted until the fourth day of insertion. Clinical significance: The study indicated the clinical significance of brass wire and elastomeric separators to separate the placement of orthodontic bands.