The treatment of colon cancer has been aimed by approaches of oral drug administration. 5-Fluorouracil is the standard treatment still nowadays and would be a candidate to be delivered orally to the colon. A pH-sensitive polymer Eudragit P-4135F was used to prepare microspheres by a simple oil/water emulsification process. Process parameters were analyzed in order to optimize the drug loading and release profiles. In further attempts mixtures with Eudragit RS100 were prepared to prolong drug release. Scanning electron microscopy and confocal laser scanning microscopy permitted a structural analysis. The solvent extraction was preferable over solvent evaporation with a view to the encapsulation rate (extraction: 37%; evaporation: 19%) due to the hydrophilic character of the drug while release pattern were nearly unchanged. Eudragit P-4135F, pure or in mixture, was found to retain drug release at pH 6.8 lower than 35% within 6 h. At pH 7.4, nearly immediate release (within 30 min) was observed for pure P-4135F, while mixtures enabled to prolong the release slightly. Analysis of the morphology led to an inhomogeneous polymer distribution of P-4135F and RS100 throughout the particle core. A capsule-like structure was concluded which allowed only slight changes of the release kinetics by the addition of RS100. However, the formulation proved its applicability in-vitro as a promising device for pH-dependent colon delivery of 5-fluorouracil.