The concentrations and magnetization transfer ratios (MTRs) in gray matter (GM) and white matter (WM) of N-acetyl aspartate (NAA), creatine (Cr), choline (Cho), myo-inositol (Ins), and glutamate plus glutamine (Glx) were investigated using magnetic resonance spectroscopic imaging (MRSI). The macromolecule (MM) baseline was studied separately using a metabolite-nulling inversion. Three data sets were collected from a pointresolved spectroscopy (PRESS)-selected volume (TE/TR ؍ 30/ 3000 ms) of human frontal lobe in vivo: one with MT pulses applied, one with an inversion pulse to null small metabolites, and one with no inversion or MT pulses. The MM signal, which was analyzed by integrating the metabolite-nulled spectrum between 0 and 3 ppm, was estimated to be 38% higher in GM than in WM. MM subtraction decreased the signal-to-noise ratio (SNR) and also decreased the reliability of LCModel quantification of most metabolites, but may have improved the accuracy of quantification of Glx. Glx and Cr were both found to correlate strongly with the GM volume fraction of the voxels. Cr showed the highest MTR, but the other metabolites also showed some attenuation of signal when the MT pulses were applied. With the use of magnetic resonance spectroscopic imaging (MRSI), many groups have shown that metabolite concentrations differ between gray matter (GM) and white matter (WM) (1-6). This difference is most marked for glutamate (2,6) or glutamateϩglutamine (Glx) (4). However, the quantification of Glx is often hampered by the contribution of macromolecule (MM) resonances at the same frequencies to the signal detected.The distribution of MM between WM and GM has been studied less extensively, and with conflicting results. One MRSI study conducted at 2T suggested there was little or no difference in MM signal between GM and WM (7), while two more recent studies using single voxels at 1.5T suggested that MM signal may be greater in GM than in WM (8,9).One method of discriminating between MMs and small metabolites is to precede the acquisition with an inversion pulse, with an inversion time chosen to null the signal from small more mobile metabolites (10). The resultant MM spectrum can then be examined to gain insight into the macromolecular makeup of the cells, and can also be subtracted from an otherwise identical spectrum collected without the inversion pulse to yield a spectrum of small metabolites with reduced effects of MM contamination.It would be helpful to be able to characterize not only the concentration of metabolites in different tissues, but also their local cellular environment. As MRI contrasts based on the magnetization transfer (MT) and diffusion of water have proven useful, interest in gaining similar information about the MT and diffusion of metabolites is increasing. Most metabolite signals have been shown to undergo a small attenuation following the application of MT presaturation pulses (11,12). The relative size of this effect in GM and WM has not been investigated previously.In the current study we use...