Dietary carbohydrate restriction presents a challenge to glucose homeostasis. Despite the popularity of carbohydrate restricted diets, little is known regarding the metabolic effects of carbohydrate restriction. The purpose of this study was to examine changes in whole-body carbohydrate oxidation, glucose availability, endogenous glucose production and peripheral glucose uptake after dietary carbohydrate restriction, without the confounding influence of a negative energy balance. Post-absorptive rates of glucose appearance in plasma [Ra] (i.e.; endogenous glucose production) and disappearance from plasma [Rd] (i.e.; glucose uptake) were measured using isotope dilution methods after a conventional diet (60% CHO, 30% FAT, 10% PRO; kcals=1.3 x resting energy expenditure [REE]) and after 2d and 7d of carbohydrate restriction (5% CHO, 60% FAT, 35% PRO; kcals=1.3 x REE) in 8 subjects (mean±SE; 29±4 yr; BMI=24±1 kg/m²) during a 9d hospital visit. Post-absorptive plasma glucose concentration was reduced (P=0.01) after 2d, but returned to pre-diet levels the next day, and remained at euglycemic levels throughout the diet (5.1±0.2, 4.3±0.3, and 4.8±0.4 mmol/L, for pre-diet, 2d, and 7d, respectively). Endogenous glucose production (glucose Ra) and glucose uptake (glucose Rd) were reduced below pre-diet levels (9.8±0.6 µmol/kg^-1/min^-1) after 2d carbohydrate restriction (7.9±0.3 µmol/kg^-1/min^-1) and remained suppressed after 7d (8.3±0.4 µmol/kg^-1/min^-1) (both P<0.001). A greater suppression in carbohydrate oxidation compared with the reduction in glucose uptake led to an increased (all P<0.05) rate of non-oxidative glucose disposal at 7d (5.2±0.5 µmol/kg^-1/min^-1) compared with 2d (2.7±0.5 µmol/kg^-1/min^-1) and pre-diet (1.6±0.8 µmol/kg^-1/min^-1). In response to eucaloric carbohydrate restriction, a marked increase in non-oxidative glucose disposal may help maintain systemic glucose availability.