Objective To compare and evaluate the efficacy of tooth bleaching with prior application of two different desensitizing agents such as sodium fluoride and 5% potassium nitrate. Materials and methods A total of 108 extracted human maxillary central incisors were stained in black coffee solution and stored in artificial saliva for colour stabilization. The specimens were randomly divided into three groups (n = 36) according to the following protocols: (a) bleaching without desensitizer, (b) bleaching with prior application of sodium fluoride, and (c) bleaching with prior application of 5% potassium nitrate. After fabricating customized trays, desensitizers were applied for 10 minutes followed by 16% carbamide peroxide bleaching gel, which was in contact with the teeth for three hours. The bleaching efficacy was evaluated at baseline (after staining), 3rd day, 7th day, and 14th day using a digital spectrophotometer. Results There was an increase in the overall colour change (∆E) from baseline to 14th day, which was statistically significant at cervical, middle, and incisal thirds of the teeth between the three groups with the sodium fluoride group showing decreased ∆E. Conclusions Carbamide peroxide (16%) showed improved whitening efficacy from baseline to the 14th day with increasing median values at all time periods. The sodium fluoride group showed decreased ∆E value when compared to other groups.