One of the most exciting areas of research in pharmaceutical sciences is the development of new delivery systems for controlled drug release. The main objective of this research focused on chitosan-fatty acid derivatized matrix for drug delivery. Saturated fatty acids (lauric acid and stearic acid) as well as unsaturated fatty acids (oleic acid and linoleic acid) are involved in this study which enhance the chemical properties of chitosan in drug delivery and enhance the permeability of ciprofloxacin drug compounds. The structure of the synthesized derivatives was characterized by Fourier transform infrared spectroscopy and X-ray diffraction. Intercalation and in vitro release of ciprofloxacin was investigated by UV spectrophotometrically. All the prepared chitosan derivatives showed a potent antimicrobial activity. It was observed that the release study of drug was maximum at the beginning and then released slowly. The initial burst release of drug may be due to the drug which exists on the surface of the film without being trapped efficiently. Chitosan linoleic ciprofloxacin composite (CS-Lin-CF) showed 52% drug release, chitosan stearic ciprofloxacin composite (CS-S-CF) showed 53.2% drug release and chitosan lauric ciprofloxacin composite (CS-L-CF) showed 63.1% drug release, chitosan oleic ciprofloxacin composite (CS-Ol-CF) showed 79.1% drug release at 8 hours.