Fluorescence spectroscopy has been used to detect polycyclic aromatic hydrocarbons (PAHs) in soil. However, the soil moisture has a strong interference to the fluorescence intensity of PAHs, which is a challenge to the development of the rapid real-time detection technology of PAHs in soil. In order to analyze the effect of soil moisture on the fluorescence characteristics of PAHs, 8 phenanthrene contaminated soil samples were prepared with different moisture content (5%~40%, interval of 5%). The dynamic one-dimensional fluorescence spectra of soil samples with different moisture content were obtained by LS-55 fluorophotometer. Two-dimensional (2D) correlation fluorescence spectrum was calculated under the perturbation of soil moisture content. It was found that the fluorescence intensity of phenanthrene at 386, 408 and 432 nm enhanced with the increase of soil moisture content, while the Rayleigh scattering light intensity at 333 nm was decreased. And, it is pointed out that the effect of soil moisture on fluorescence can be corrected by establishing the relationship among fluorescence intensity, Rayleigh scattering intensity and soil moisture content. Meanwhile, the effect of soil moisture on the standard curve of phenanthrene concentration was studied. It is demonstrated that the soil moisture has a great influence on the quantitation of phenanthrene.