Nuclear magnetic resonance (NMR)-based metabolomics profile comparisons of embryogenic and non-embryogenic calli of sugarcane were performed using principal component analysis (PCA) to determine a possible relationship between certain metabolites and somatic embryogenesis. Mahalanobis distance (DM) analysis showed significant metabolic profile differences between the embryogenic and non-embryogenic callus groups. Significantly different spectral buckets and their corresponding metabolites have been identified using volcano- and loading-plot analyses, where glucose, fructose, sucrose, and alanine were observed at higher concentrations and asparagine, glutamine, lysine, 2-hydroxyisobutyrate, and choline were observed at lower concentrations in embryogenic calli than in non-embryogenic calli. The results of this research indicate possible roles of different sugars, amino acids, and aliphatic compounds during sugarcane somatic embryogenesis.