Metabolomics is a new approach based on the systematic study of the full complement of metabolites in a biological sample. This technology consists of two sequential steps: (1) an experimental technique, based on nuclear magnetic resonance (NMR) spectroscopy or mass spectrometry, designed to profile low-molecular-weight compounds, and (2) multivariate data analysis. The metabolomic analysis of biofluids or tissues has been successfully used in the fields of physiology, diagnostics, functional genomics, pharmacology, toxicology, and nutrition. Recent studies have evaluated how physiological variables or pathological conditions can affect metabolomic profiles of different biofluids in pediatric populations. The overall metabolic status of the neonate is little known. If more information on perinatal/neonatal maturational processes and their metabolic background were available, the management of sick or preterm newborns might be improved. Currently, the use of metabolomics in neonatology is still in the pioneering phase. Meaningful diagnostic information and simple, noninvasive collection techniques make urine a particularly suitable biofluid for metabolomic approach in neonatal medicine, although blood has also been investigated. Different fields of neonatology such as postnatal maturation, asphyxia/hypoxia, inborn errors of metabolism, nutrition, nephrouropathies, nephrotoxicity, cardiovascular diseases, and other conditions have been investigated using a metabolomic approach. Together with genomics and proteomics, metabolomics appears to be a promising tool in neonatology for the monitoring of postnatal metabolic maturation, the identification of biomarkers as early predictors of outcome, the diagnosis and monitoring of various diseases, and the "tailored" management of neonatal disorders.