The red prickly pear fruit (Opuntia ficus-indica L. Mill), endemic from Mexico’s semi-desert regions and present in North Africa and Southern Europe, particularly Italy and Spain, is a valuable source of nutrients, bioactive compounds, and polysaccharides. This study used non-destructive techniques like microscopy and Raman and infrared (IR) spectroscopy to characterize polysaccharides extracted from two red prickly pear varieties. The polysaccharides constitute approximately 80% of the peel and 39–18% of the pulp; microscopy provided insights into its microstructural details, while Raman and IR spectroscopy enabled the identification of its specific functional groups. The results revealed distinct microstructural attributes: mucilage displays a microstructure influenced by the ratio of acidic to neutral sugar monomers; pectin exhibits a low degree of methoxylation alongside a characteristic egg-box structure facilitated by calcium ions; hemicellulose presents a delicate, porous layer; and cellulose reveals a layered microstructure supported by thin or robust fibers and calcium crystals. The functional groups identified via Raman and IR spectroscopy provided specific information that could be used to infer chemical interactions influenced by functional groups like hydroxyl, carboxyl, and methyl, suggesting potential binding, stabilization, and water retention properties that enhance their utility as functional ingredients in food products. These findings, obtained using non-destructive methods, enhance the understanding of the compositional and microstructural characteristics of polysaccharides in the red prickly pear, which, in turn, can be used to predict their promising technological applications as functional ingredients.
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