This study showed the impact of fouling and ageing of reverse osmosis (RO) membranes on the rejection of different compounds to improve our understanding on virus rejection mechanisms. Dissolved organic matter (DOM) is generally used as an indicator of water quality. Thus, in combination with electrical conductivity profiling, the potential monitoring of DOM by three-dimensional fluorescence to detect membrane breaches was firstly investigated. It was demonstrated that DOM could be used as new membrane integrity indicator. Then, the rejections of one virus surrogate (MS2 phage) and four membrane integrity indicators (DOM, rhodamine WT, sulphate and salt) were studied with intact and impaired membranes using lab-scale set-ups. It was concluded that the presence of organic foulants on the membrane surface caused a decrease of the water permeability and an increase of compounds rejection by improving size exclusion mechanism. On the other hand, scaling did not have an impact on their rejection even if the water permeability decreased. Moreover, a chlorine exposure of 9000 ppm•h NaOCl at pH 7 caused a drop of the water permeability and compounds rejection. However, the exact modifications of the membrane surface chemistry caused by chlorine exposure are still not well understood. To conclude, statistical analysis of the data obtained during this study permitted to propose a new combination of testing techniques to monitor RO membrane.