Objective: The aim of the present study was to investigate the metabolic profile of preterm human breast milk (HBM) by using a metabolomic approach. Methods: NMR spectroscopy and GC/MS were used to analyze the water-soluble and lipid fractions extracted from milk samples obtained from mothers giving birth at 26-36 weeks of gestation. For the sake of comparison, preterm formula milk was also studied. Results: The multivariate statistical analysis of the data evidenced biochemical variability both between preterm HBM and commercial milk and within the group of HBM samples. Conclusions: The preliminary results of this study suggest that metabolomics may provide a promising tool to study aspects related to the nutrition and health of preterm infant.