N-Acetylaspartate (NAA) is an important marker of neuronal function and viability that can be measured using magnetic resonance spectroscopy (MRS). In this paper, we proposed a method to measure NAA synthesis using proton MRS with infusion of uniformly (13)C-labeled glucose, and demonstrated its feasibility in an in vivo study of the rat brain. The rate of (13)C-label incorporation into the acetyl group of NAA was measured using a localized, long echo-time proton MRS method. Signals from the (13)C satellites of the main NAA methyl protons at 2.02 ppm were continuously monitored for 10h. Quantification of the data based on a linear kinetic model showed that NAA synthesis rate in isoflurane-anesthetized rats was 0.19+/-0.02 micromol/g/h (mean+/-standard deviation, n=12).