Polyaniline (PANI) nanoparticles were chemically synthesized in the presence of a cross-linked carboxymethyl chitin (CM-chitin) acting as a template. The reaction was performed under acidic conditions and the template was removed after the polymerization of aniline was completed. The morphology of the synthesized PANI was globular with a diameter in the nanometer range. The degree of cross-linking of the CM-chitin played an important role in determining the size of the obtained PANI nanoparticles, which decreased from approximately 392 to 160 nm with increase in concentration of the cross-linking agent, glutaraldehyde, from 0 to 9 μmol, respectively. At a higher glutaraldehyde concentration (18 μmol), an aggregated PANI network was observed due to the incomplete removal of the more highly cross-linked CM-chitin. Molecular characterization (including UV-Visible, FTIR, TGA, and XRD techniques) revealed that the structure of the synthesized PANI nanoparticles is identical to that of conventional PANI. A mechanism is proposed for the formation of PANI nanoparticles in the presence of the cross-linked CM-chitin template.