Choline is an essential nutrient participating as the initial substrate in major metabolic pathways. The differential metabolic routing of choline was investigated in MCF7 human breast cancer implanted in nude mice and in the kidney, liver, and brain of these mice. The distribution of metabolites following infusion of [1,2-(13)C]-choline was monitored by (13)C magnetic resonance spectroscopy. This infusion led to an 18-fold increase in plasma choline and to concomitant changes in the content and distribution of choline metabolites. In vivo kinetic studies of the tumor during the infusion demonstrated accumulation of choline in the interstitium and intracellular synthesis of phosphocholine. The amount of unlabeled choline metabolites was 7.1, 4.1, 3.5, and 1.4 micromol/g in the kidney, liver, tumor, and brain, respectively. The variations in the labeled metabolites were more pronounced with high amounts in the kidney and liver (8.0 and 4.3 micromol/g, respectively) and very low amounts in the tumor and brain (0.33 and 0.12 micromol/g, respectively). In the kidney and liver, betaine (unlabeled and labeled) was the predominant choline metabolite. The dominant unlabeled metabolite in breast cancer was phosphocholine and in the brain glycerophosphocholine. Magn Reson Med 46:31-38, 2001.