Unveiling the Metabolomic, Phytochemical and Bioactive Profile of Twelve Macroalgae From the Adriatic Sea: A Comprehensive Analysis Using MSPD-UHPLC-QTOF

The present study provides an exhaustive exploration of twelve macroalgal species from the Adriatic Sea, including seven brown algae (Ericaria amentacea, Fucus virsoides, Cutleria multifida, Cystoseira compressa, Cystoseira corniculata, Gongolaria barbata and Padina pavonica), three green algae (Codium adhaerens, Codium vermilara and Ulva lactuca), and two red algae (Scinaia furcellata and Asparagopsis taxiformis). Matrix solid-phase dispersion (MSPD) was applied as the extraction technique, using solvents recognised as safe (GRAS). The bioactive profile of the extracts was assessed through the quantification of total phenolic content (TPC) and antioxidant activity. Among the three phyla, U. lactuca, F. virsoides and S. furcellata exhibited the highest TPC (0.8, 26 and 3.0 mg GAE·g DW⁻¹) and antioxidant activity (1.9, 38 and 7.5 mg TE·g DW⁻¹), respectively. Targeted HPLC-MS/MS analysis enabled the identification of nineteen phenolic compounds across all taxa. Chlorophyta showed a characteristic profile enriched in coumarins, benzaldehydes and flavanones, including the selective detection of 7-hydroxycoumarin in species with higher antioxidant potential. Additionally, compounds such as chlorogenic, rosmarinic and caffeic acids exhibited taxon-specific distributions that may explain differences in antioxidant activity. Complementary untargeted UHPLC-QToF metabolomics analysis provided broader coverage, revealing eighty metabolites spanning phenolics, sugars, organic acids, lipids, amino acids and their derivatives. Notably, the detection of FAHFA lipids represents the first report of these compounds in macroalgae, alongside a pronounced presence of sulphated phenolics. Overall, these findings provide a robust baseline on the bioactivity and chemical composition of Adriatic macroalgae, highlighting their value as a natural source of functional compounds.