An overview of domestic and foreign recent status on bio-oil analysis and characterization was presented,which focused on the applications of some spectroscopy and chromatography techniques in chemical structure characterization of the small organic molecules,such as,Fourier transform infrared spectroscopy（FTIR）,nuclear magnetic resonance（NMR）,mass spectrometry（MS）,gas chromatography（GC）,high performance liquid chromatography（HPLC） techniques,as well as the one and two-dimensional chromatography coupling technology.The evaluations on the role and value of these techniques in structure characterization were afforded,and the detailed descriptions of the quality standards and some typical analysis methods of bio-oil were involved,and then the effects of physical and chemical properties of bio-oil on its separation and analysis were discussed.Moreover,the structure characterizations of oligosaccharide and pyrolytic lignin remained in bio-oil were introduced.Finally,the main problems existed in bio-oil separation and analysis were summarized,and the prospect on the development trend for bio-oil separation and analysis was presented.