Electrochemical enzymatic glucose sensors are intended to measure blood or interstitial fluid glucose concentrations. One class of these glucose sensors are continuous glucose monitors (CGMs), indicated for tracking and trending of glucose concentrations in interstitial fluid and as an adjunct to blood glucose testing. Currently approved CGMs employ a glucose oxidase (GOx) electrochemical detection scheme. Potential interfering agents can impact the accuracy of results obtained by glucose sensors, including CGMs. Seven sugars, seven sugar alcohols, and three artificial sweeteners were in vitro screened for interference with amperometric glucose oxidase (GOx) sensors at concentrations greater than physiologic concentrations. Galactose was investigated further at physiologically relevant concentrations using a custom amperometric system. Furthermore, glucose and galactose calibration experiments were conducted to facilitate multiple enzyme kinetic analysis approaches (Michaelis-Menten and Hill equation) to understand the potential source and mechanism of interference from galactose. Under in vitro testing, except for galactose, xylose and mannose, all screened compounds exhibited interference bias, expressed in mean absolute relative difference (MARD), of ⩽ 20% even at concentrations significantly higher than normal physiologic concentrations. Galactose exhibited, CGM-dependent, MARD of 47-72% and was subjected to further testing. The highest recorded mean relative difference (MRD) was 6.9 ± 1.3% when testing physiologically relevant galactose concentrations (0.1-10 mg/dL). Enzyme kinetic analysis provided calculations of maximum reaction rates ( ), apparent Michaelis constants ( ), and Hill equation h parameters for glucose and galactose substrates for the enzymes in the CGMs. Under the conditions of in vitro screening, 14 of the 17 compounds did not exhibit measuarable interference. Galactose exhibited the highest interference during screening, but did not substantially interfere with CGMs under the conditions of in vitro testing at physiologically relevant concentrations. Enzyme kinetic analysis conducted with galactose supported the notion that (1) the reactivity of GOx enzyme toward nonglucose sugars and (2) the presence of enzymatic impurities (such as galactose oxidase) are two potential sources for sugar interference with GOx glucose sensors, and thus, should be considered during device development.