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Chitosan is one kind of good biocompatible polymer and is suitble for drug carriers. Preparation of 5-fluorouracil (5-Fu) loaded chitosan (CS) particles and in vitro release experiment were performed using ionic crosslinking method with sodium tripolyphosphate (TPP) as crosslinker. The optimal preparing parameters were verified by 5-Fu release experiments. The drug loading, and release behavior of drug loaded microparticles in vitro were investigated. The optimal preparation conditions were: the temperature 25°C, the ratio of CS to TPP 5:1, the CS concentration 1.5g/L, stirring speed 650rpm. Under these conditions, the drug loading of particles was up to 45%.