Gluconeogenesis, indirect oxidation of fructose‐derived glucose, lactate and glycogen synthesis are major pathways for the disposal of a pure oral fructose load, while de novo lipogenesis (DNL) constitutes a minor pathway. The metabolic fate of fructose co‐ingested with glucose and other nutrients has however not been evaluated. To address this issue, we studied 6 healthy subjects over 6 hours after ingestion of test meals composed of 20±0g lipids and 19±0g protein either without sugars (Ctrl), or with 32±1g 13C‐labelled fructose (F), or with 32±1g 13C‐labelled fructose + 32±1g unlabelled glucose (F+G). Subjects consumed a weight‐maintenance diet containing 20% sugar during 3 days preceding the measurements. Fructose oxidation was assessed from 13CO2 production, gluconeogenesis from 13C‐glucose production, and DNL from post‐prandial 13C‐VLDL‐palmitate concentrations. Ctrl F F+G Net Carbohydrate oxidation (g/6h) 21.0 ± 3.3 29.9 ± 2.8 34.5 ± 3.9 Fructose oxidation (g/6h) ‐ 12.5 ± 0.7 11.0 ± 0.5# Gluconeogenesis (g/6h) ‐ 9.1 ± 0.2 6.4 ± 0.3# Net glycogen synthesis (g/6h) ‐ 4.4 ± 2.1 34.9 ± 5.2# Fructose storage (g/6h) ‐ 21.8 ± 1.2 23.1 ± 1.2 iAUC lactate (mM*6h) ‐51.1 ± 19.6 108.3 ± 11.7* 124.9 ± 26.3* iAUC13C‐VLDL‐palmitate (μmol*6h) ‐ 3.9 ± 1.2 3.7 ± 0.6 All P <0.05, * vs Ctrl, # vs F. iAUC: incremental area under the curve Compared with fructose alone, co‐ingestion of glucose and fructose incorporated in a meal reduced fructose oxidation by 12 ± 5%, and gluconeogenesis by 28 ± 12% by increasing net glycogen storage; but did not enhance 13C‐VLDL palmitate responses. Chronic consumption may lead to increase DNL due to limited glycogen storage.