Bio-oil from straw is a complex mixture that contains different organic compounds formed by the thermal degradation of cellulose, hemicelluloses, lignin, and other biomolecules which are presented in sugarcane straw biomass. Biofuel can be obtained from pyrolysis method from sugarcane straw. Pyrolysis is one kind of thermo-degradation technique that can be used for transformation of the biomass in bio-oil. The biomass has been converted into a liquid product such as bio-oil through pyrolysis which can be used as feedstock for fuels and valuable chemicals (Maiti et al. 2006; Li et al. 2011). Thermal pyrolytic conversion is the promising method that can be used for biomass conversion (Strezov et al. 2008). The medium tempera­ture (550 °C) leads to the production of a pyrolysis gas composed by H2 and CO (Morf et al. 2002) . Mesa-Perez et al. (2013) investigated on biofuel production through pyrolysis using sugarcane straw. They obtained 35.5 % bio-oil at 470 °C temperature that has low oxygen content and high heating value. The pyrolysis of sugarcane straw is an environment-friendly method for the production of bio-oil because it does not have another important use (Cortez and Lora 2006). The develop­ment of biorenewable agro-industry needs to integrate the coproducts obtained through pyrolysis process (Arbex et al. 2004). Bio-oil is a good coproduct of pyroly­sis and an energy carrier and feedstock for biodiesel and petrochemical production. Bio-oil is a complex nature that requires the usage of high-resolution chromato­graphic techniques. The multidimensional chromatography specially GC x GC/ TOFMS is a powerful technique for analysis of bio-oil. This technique has some advantages for separation capacity, resolution, sensitivity, and selectivity. Sfetsas et al. (2011) have used the GC x GC/FID and GC x GC/TOFMS for quantification of bio-oils. Fullana et al. (2005) reported that 70 % chromatogram could be obtained using GC x GC.