Urine has become one of the most attractive biofluids in clinical proteomics as it can be obtained noninvasively, in large quantities and is stable compared to other biofluids. The urinary proteome has been studied by almost any proteomic technology, but mass spectrometry based urinary protein and peptide profiling has emerged as most suitable for clinical application. In this study, SDS-PAGE was used to separate urine proteome after purification by ProteoSpinTM Urine Protein Concentration Micro Kit. The peptide mixtures extracted from each gel slice were analyzed by two-dimensional nano liquid chromatography (2D nanoLC) coupled online with tandem mass spectrometry analysis (NanoESI-Q-TOF-MS/MS). In total, 132 proteins were identified, 11 hight abundant concentration proteins, 72 low albudant concentration proteins and several isoforms of immunoglobulin.