The inner filter effect (IFE) and the absorption depth from the fluorescence material will directly lead to an artifact for intensity and spectral shape of the fluorescence spectra, and they restrict the application of fluorescence analysis. A two-component mixed solution with overlapping absorption spectra was used to develop a new method based on physical absorption model to correct the influences of IFE and the absorption depth on fluorescence intensity. The spectral investigations of terthiophene/quinquethiophene mixture solution show that a good correction result can be obtained by using the above correction method, and the error after correction is less than 5%.