Objective: The purpose of this study was to determine the degradation of 10% carbamide peroxide within the first hour of use and the effect of pellicle on the degradation of carbamide peroxide in vivo. Method and materials: Fifteen subjects were assigned to wear nightguard trays filled with bleaching gel for 0.5, 2.5, 5.0, 10.0, 20.0, 40.0, and 60.0 minutes under 2 conditions: (1) subjects did not receive a prophylaxis before each bleaching treatment (pellicle was assumed to be present on the tooth surface); and (2) subjects received prophylaxis to remove pellicle from the tooth surface. At each evaluation time, 3 types of sample were collected: (1) a 'grab sample,' a sample of gel taken with a spatula from the reservoir of the tray on the maxillary right central incisor; (2) the remaining gel from the tray; and (3) the remaining gel scraped from the teeth. The collected gel samples were analyzed for the amount of peroxide using the United States Pharmacopeia-accepted method. Subjects were asked not to swallow the saliva during treatment but to expectorate into a beaker. The collected saliva was analyzed to determine the amount of peroxide. This sample represented the approximate amount of peroxide that subject would ingest during treatment. Results: Pellicle did not affect the degradation of carbamide peroxide for the teeth or grab samples. The degradation of carbamide peroxide was exponential, except during the first 5 minutes, when the degradation rate was much higher. The saliva collected from subjects during use of the bleaching tray revealed an average of 2.1 mg of carbamide peroxide for 1 hour of bleaching treatment. Conclusion: The degradation rate is not affected by the removal of the pellicle.