Crude oil is a complex compound of a myriad of liquid and gaseous hydrocarbons,oxygenated, nitrogenous, and inorganic compounds. Tragacanth is a biocompatible and biodegradable polysaccharide which has attracted the attention of researchers in different fields. Zinc Oxide has attracted the attention of researchers as a semi-conductor metal oxide. Zinc Oxide has a wide range of application. In a previous study, tragacanth nanofiber layer has been used to separate water from petroleum products. This study aimed at electrospinning tragacanth nanofibers containing zinc oxide nanoparticles with the aim of separating sulphur compounds from petroleum products. Zin oxide incorporated tragacanth nanofibers were electrospun from tragacanth solutions containing zinc oxide nanoparticles with concentration of 5, 10, 15 and 20 (% w/w). The Average diameter of these nanofibers were 91.6, 94.1, 95.4 and 97.8 nm, respectively. The effect of parameters such as tragacanth concentration, zinc oxide concentration, viscosity, electrical conductivity and surface tension on the morphology of the electrospun nanofibers were investigated. SEM images showed that zinc oxide nanoparticles incorporated tragacanth nanofibers were uniformly electrospun with no beads. FTIR analysis showed some interactions between zinc oxide nanoparticles and tragacanth macrolecules. XRD studies showed a two phase microstructure structure consisting of amorphous tragacanth matrix with zinc oxide crystallites dispersed in it . Filteration experiments showed the highest filtration efficiency for Isfahan and Gachsaran crude oil to be 75.3% and 62.2%, respectively. Moreover, the highest sulphur adsorption capacity obtained for Isfahan and Gachsaran crude oil was 211.2 mg.gr-1 and 108.1 mg.gr-1, respectively. Keywords: tragacanth, zinc oxide nanoparticles, electrospinning, nanofibers, crude oil, sulphur compounds, filteratio