This study deals with the impact of spatio-temporal heterogeneities on the assessment of lake ecological status according to the European water framework directive (WFD). A method, based on three-dimensional coupled hydrodynamic and ecological modeling, is presented to assess the variability of lake ecological status, and to locate the most representative sampling station of Lake Geneva (France/Switzerland). Five variables used in the lake ecological status evaluation were simulated by using the free software Delft3D. The numerical simulation results showed that the simulated ecological status based on chlorophyll a and total phosphorus concentrations measured at the regulatory monitoring station depend on the choice of the sampling date. Results also indicated a strong spatial heterogeneity in ecological status that varies from “poor” to “good” along an East-West gradient. Finally, the numerical simulation results showed that the most representative point of a mean theoretical ecological quality for Lake Geneva would be located in the center of the upper basin, close to the historical sampling station.