Organic thin films usually feature vertical phase segregation, and film-depth-dependent light absorption spectroscopy is an emerging characterization method to study the vertical phase separation of active layer films in organic electronics field. However, the interference effects on thin films can lead to optical errors in their characterization results. In this work, the interference effects on fluctuations of peak intensity and peak position of film-depth-dependent light absorption spectroscopy are investigated. Subsequently, a numerical method based on inverse transfer matrix is proposed to obtain the optical constants of the active layer through the film-depth-dependent light absorption spectroscopy. The extinction coefficient error in the non-absorbing wavelength range caused by interference effect is reduced by ∼95% compared with the traditional film-depth-dependent light absorption spectroscopy measurement. Thus, the optical properties of the thin film and quantitative spectrographic analysis based on these optical constants largely avoid the effects of interference including fluctuations of peak intensity and peak position. It is concluded that for many morphologically homogenously films, the spatial (film-depth) resolution of this film-depth-dependent light absorption spectroscopy can be optimized to be <1 nm. Subsequently, this modified film-depth-dependent light absorption spectroscopy approach is employed to simulate the local optical properties within devices with a multilayer architecture.