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In this paper, horseshoe crab exoskeleton and exuviae were collected on the coasts of Yucatan and physicochemical studies were carried out. Samples from different sections of the exoskeleton were taken to evaluate chitin content and properties in order to be compared with exuviae. The isolation of chitin was realized by chemical processes involving deproteinization, demineralization and discoloration. Physicochemical properties of chitin extracted from both sources were compared in terms of characterization analysis in particular achieved under EDX, FTIR and calorimetric techniques (DSC and TGA). In such comparative analysis lies the importance to establish differences or similitudes in chitin contents, structure, etc. Scanning Electron Microcopy was conducted on exuviae and exoskeleton selected parts in order to identify horseshoe crab structure and surface morphology. Differential scanning calorimetry was focused on changes in physical and chemical structures at different samples showing two endothermic peaks related to the complete elimination of hydroxyl groups attached to polysaccharide rings. TGA thermograms were characterized by endo and exotherms corresponding to water evaporation and decomposition of polymer respectively. Exoskeleton SEM images shown a tubular network and porous surface while exuviae exhibited irregular surface and diatom unicellular microalgae embedded. Calorimetric measurements confirmed that exuviae is richer in chitin than exoskeleton and that the main allotropic crystal structure corresponds to α‐chitin. FTIR analysis supported these results by exhibiting different absorption bands corresponding to α‐chitin. [ABSTRACT FROM AUTHOR]