In the present work, we report the chemical characterization of asphaltenes from a light Colombian crude oil, 36° API, Colorado Crude Oil (CCO). Asphaltenes of CCO were extracted using the standard ASTM procedure, ASTM D6560-12. The approach to the chemical composition of asphaltenes was carried out by using elemental analysis (C, H, N, O, S), Fourier transform infrared spectroscopy (FT-IR), Raman Spectroscopy (RS), Matrix Assisted Laser Desorption Ionization - Time of Flight (MALDI-TOF) mass spectrometry, and proton and carbon nuclear magnetic resonance spectroscopy (1H- and 13C-NMR). Finally, the crystallite parameters of nanoaggregates of CCO asphaltenes in solid phase were calculated using X-ray powder diffraction data. The average diameters of aromatic layers La calculated from XRD and Raman were compared together and proved to be in good agreement. Average molecular parameters AMPs were calculated from elemental analysis, NMR and Raman spectroscopies, and X-ray diffractometry. &nbsp