The transformation of dissolved organic matter (DOM) during various disinfection processes has raised great concerns due to the generation of carcinogenic disinfection by-products (DBPs). Ultraviolet (UV) irradiation is an effective method for drinking water disinfection, during which DOM undergoes changes in functional groups and molecular weight. In this study, the spectrophotometric titration and gel permeation chromatography (GPC) determination were employed to investigate the changes in oxygenated groups and weight-averaged molecular weight (Mw) of two typical DOM during UV irradiation. The differential absorbance spectra (DAS) of DOM could be deconvoluted into six Gaussian bands. The change of relative band intensity was attributed to the change of oxygenated groups (carboxylic and phenolic groups), which was confirmed by combining DAS data and revised Non-Ideal Competitive Adsorption -Donnan model. The GPC result demonstrated that the Mw of DOM decreased after UV disinfection. Moreover, a linear correlation between Mw and the intensity of deconvoluted Gaussian band from DAS was established, which might be served as an alternative approach to estimate Mw and predict the hydrophobicity and DBPs formation potential of DOM in drinking water treatment and monitoring.