Metabolic profiling is widely used as a probe in diagnosing diseases. In this study, the metabolic profiling of urinary carbohydrates was investigated using gas chromatography/mass spectrometry (GC/MS) and multivariate statistical analysis. The kernel-based orthogonal projections to latent structures (K-OPLS) model were established and validated to distinguish between subjects with and without diabetes mellitus (DM). The model was combined with subwindow permutation analysis (SPA) in order to extract novel biomarker information. Furthermore, the K-OPLS model visually represented the alterations in urinary carbohydrate profiles of excess and deficiency syndromes in patients with diabetes. The combination of GC/MS and K-OPLS/SPA analysis allowed the urinary carbohydrate metabolic characterization of DM patients with different traditional Chinese medicine (TCM) syndromes, including biomarkers different from non-DM patients. The method presented in this study might be a complement or an alternative to TCM syndrome research.