Because of its impact on water quality, organic matter (OM) in karst groundwater has been widely studied. The present article describes a method for monitoring OM in karst aquifers characterized by quick responses to rainfall. This method combines weekly manual sampling and continuous monitoring to provide, qualitative and quantitative information about OM flow. Weekly samples were analyzed for Total Organic Carbon (TOC) content and spectrofluorescence, while continuous monitoring was carried out at the main spring, using a field fluorimeter (310/400-700 nm and 280/300-600 nm) to quantify chromophoric organic matter (COM). The type and quantity of COM were defined by decomposing Excitation Emission Matrices (EEMs) and by applying a 2D fluorescence decomposition method. Continuous monitoring data showed that the dominant COM was humic-like (HL). We found three types of relationship between HL and discharge and between HL and TOC, showing that caution must be exercised when using field fluorimeter measurements to quantify TOC. Each relationship was characterized by global differences in OM content and by the presence of different percentages of non-chromophoric organic matter. These three relationships are associated with changes in hydrology and microorganism activity during the year. We used these relationships to estimate the annual OM flow (about 15 kg/ha/year) and thereby quantify OM flow during the year. Our results show the importance of the non-chromophoric organic matter in such estimation. That work illustrates the need to couple qualitative and quantitative monitoring of OM in karst spring to improve the global comprehension of karst system and of the sources implies in the OM flow.