MR spectroscopy (MRS) can provide the noninvasive detection of a number of biologically important cellular metabolites. Recently, this technique has been applied to provide unique assessments of maturational changes in brain biochemistry. Significant variations in the concentration of intracellular metabolite levels have been observed in the neonatal brain with anatomic location and with development. These changes are critical to define in normal neonatal populations so as to provide improved understanding of brain maturation as well as to establish normative values for the differentiation of abnormal metabolism in cases of brain injury. This article includes the description of MRS techniques for neonatal studies and results in the study of the premature and term newborn brain.