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In this study, a cellulose acetate (CA) membrane is formed as an interference rejection membrane on a glucose sensor to measure glucose in saliva. Glucose in saliva is successfully measured in vivo without any pretreatment of human saliva. A mouthguard (MG) glucose sensor is developed to monitor salivary glucose, which is reported to be correlated with the blood glucose level. Salivary components of ascorbic acid (AA) and uric acid (UA) hinder the accurate measurement of the glucose concentration of human saliva. CA-coated electrodes are prepared to investigate the interference rejection membrane. To measure hydrogen peroxide, which is a reaction product of glucose oxidase, effects of AA and UA are examined. Characteristics of the fabricated biosensor are examined on the basis of artificial saliva. The as-developed MG sensor can quantify the glucose concentration in the range of 1.75-10 000 μmol/L, which includes a salivary sugar concentration of 20-200 μmol/L. For the measurement of saliva samples collected from healthy subjects, the output corresponding to the concentration is confirmed; this suggests the possibility of glucose measurement. This MG glucose sensor can provide a useful method for the unrestricted and noninvasive monitoring of saliva glucose for the management of diabetes patients.