This study explored the effects of spinach flour (SF) enrichment on pasta, focusing on chemical, nutritional and sensory properties, cooking performance, and microbiological stability. SF was added at 12.5% (PSP12) and 25% (PSP25). The enriched pasta had a lower pH than the control (CP), due to spinach-derived organic acids, with PSP25 showing the highest fiber content. Enrichment increased B vitamins and minerals, especially calcium, magnesium, sodium, and potassium. PSP25 had a shorter cooking time, higher water absorption, and greater cooking loss. Enriched pasta showed lower starch hydrolysis index and predicted glycemic index, suggesting potential benefits for managing postprandial blood sugar levels. SF significantly altered the free amino acid (FAA) profile, with PSP25 showing the highest concentration of total FAAs. Antioxidant assays demonstrated that spinach-enriched pasta retained higher levels of phenols and flavonoids, after cooking also, compared to CP. Sensory analysis indicated that while PSP12 had higher overall acceptability, PSP25 exhibited stronger herbaceous flavors, which could affect consumer preference. Microbiologically, all samples were stable for 110 days. The findings suggest that SF enrichment enhances the nutritional value, antioxidant potential, and sensory qualities of pasta, with potential for commercial applications, although consumer acceptance could be influenced by its non-traditional taste and texture.

Pasta made from durum wheat semolina has a medium–high glycemic index score, high starch digestibility, and limited nutritional value due to its low fiber, vitamin, and bioactive compound content. This study aimed to enhance pasta’s nutritional and functional qualities by incorporating Pleurotus eryngii (PE) powder at various substitution levels to achieve one nutritional claim at least. This research involved two phases: evaluating the chemical/physical, nutritional, functional, and sensory properties of laboratory-scale samples and validating the selected formulations through industrial-scale production and shelf-life analyses. The pasta sample with 8.62% PE substitution (SPE8-P) demonstrated significantly improved nutritional qualities, including high fiber content sufficient for a “high fiber content” claim, and potential prebiotic activity indicated by increased bifidobacterial density during simulated fecal microbiota fermentation. Despite its enhanced riboflavin and antioxidant content, regulatory constraints limited the inclusion of claims for vitamin B2 richness and antioxidant activity. Although significantly affecting the color, taste, and odor profiles, the sensory analysis revealed high overall acceptability, supporting the product’s potential for consumer acceptance. This study confirms the feasibility of producing innovative, nutritionally enriched pasta with PE powder as a functional ingredient. Future research will focus on in vivo evaluation to establish the potential for classifying this pasta prototype as a functional food.