Abstract: Traditional methods of pesticide residue detection are time-consuming, complicated, and require a lot of pretreatment processes. In this study, hyperspectral imaging technology combined with fluorescence analysis was used for determining trace chemicals in vegetable samples. Hyperspectral fluorescence images of chlorpyrifos in the range of 400～1100nm for vegetable samples were acquired, and the fluorescence emission spectral curves from the hyperspectral images were extracted by ENVI 4.3 analysis software. The results show that chlorpyrifos have strong fluorescence characteristic when methanol is used as solvent. The emission spectrum of chlorpyrifos indicates that the peak emission wavelength is 437 nm, and hlorpyrifos samples with different concentration have different fluorescence emission spectral intensity at the peak, and fluorescence emission peak value reduces when the concentration of chlorpyrifos decreases. The results can provide theoretical basis for developing rapid detection instrument for vegetable pesticide residue.