The infiltration of parasitical water into two sewer systems in Rome (Italy) was quantified during a dry weather period. Infiltration was estimated using the hydrograph separation method with two water components and δ18O as a conservative tracer. The two water components were groundwater, the possible source of parasitical water within the sewer, and drinking water discharged into the sewer system. This method was applied at an urban catchment scale in order to test the effective water-tightness of two different sewer networks. The sampling strategy was based on an uncertainty analysis and the errors have been propagated using Monte Carlo random sampling. Our field applications showed that the method can be applied easily and quickly, but the error in the estimated infiltration rate can be up to 20%. The estimated infiltration into the recent sewer in Torraccia is 14% and can be considered negligible given the precision of the method, while the old sewer in Infernetto has an estimated infiltration of 50%.