Residue analysis of emamectin benzoate in tobacco and soil by Reversed-Phase high performance liquid chromatography was assessed. Residues were first extracted with acetone and water, filtrated with Florisil-SPE columns, and then determined with UV detection. Lowest detection limit with this method were 2.0×10^-10g（B1a）and 1.0×10^-9g（B1b）. The average spiked recoveries were 94.92%-102.18% in tobacco leaves, and 96.76%-99.02% in the soil. The precision of the method is acceptable with relative standard deviation (RSD) of 2.66%-3.69% for tobacco leaves and 2.23%-4.48% for the soil, respectively.