Chitin is a ubiquitous renewable biopolymer that is significantly distributed in the natural world. Biopolymeric nanoparticles (Nps) have been developed for various biomedical applications by researchers. Here, chitin derivative, dibutyrylchitin Nps (DBC) was synthesized as a nanocarrier for drug delivery using butyric anhydride and perchloric acid as a catalyst under heterogeneous conditions. The structural characterization was analyzed by FT-IR and FE SEM study showed spherical particles in a size range of 80–90nm. The physiochemical evaluation involves swelling behavior and in vitro biodegradation studies. The results of in vitro hemolytic assay validate the blood compatibility of the prepared system. Drug release profiles indicate that 5-flourouracil (Fu) loaded dibutyrylchitin Nps (DBC-Fu) gives the enhanced drug release in acidic pH when compared to neutral pH. The encapsulation efficiency of DBC-Fu was found to be 90%. The confocal analysis also confirmed the uptake of both DBC and DBC-Fu Nps by A549 cell lines. Hence, this study shows that the DBC have the potential to be used as a drug carrier and also for other biomedical applications.