Bio-oils usually contain many types of compounds with various chemical properties. A bio-oil sample derived from rice husk through rapid pyrolysis was fractioned using solvent- or solid-extraction techniques based on their various properties. Ultraviolet-visible spectroscopy, three-dimensional excitation-emission matrix (EEM) fluorescence spectroscopy and Fourier transform infrared spectroscopy were used to characterize their various spectral properties for further understanding the characteristics of the bio-oil. Bio-oil mostly contains many aromatic ring components, acidic polar fractions, few weak- and non-polar components. The results all show that the main compounds and functional groups in the various bio-oil fractions were different and depended on the fractionation methods. The compositions of the bio-oil fractions were also analyzed with a gas chromatography/mass spectrometry (GC/MS) method. The consistency of the results obtained from the spectrometric methods with the GC/MS method indicates that the spectrometric methods have a good potential for rapid and effective characterization of bio-oils. (C) 2010 Elsevier Ltd. All rights reserved.