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This work was designed to develop the chitosan-based melatonin layer-by-layer assembly (CMLLA) via the inclusion method. The structural characterizations and interaction present in CMLLA were investigated by the scanning electron microscope (SEM), X-ray diffraction (XRD) and Fourier Transform-Infrared spectroscopy (FTIR). The ratio of chitosan (CH) to carboxymethylcellulose (CMC) greatly influenced the mechanical properties, including the tensile strength, moisture content and color performance. Results showed that both antioxidant and a