Enhancing Sustainable Extraction of Anthocyanins from Apulian Pigmented Wheats (Triticum spp.) by Design of Experiment

Unlike conventional wheat varieties, Apulian pigmented landraces ( Triticum aestivum L., and T. durum Desf.) from Granomischio® are valuable anthocyanin sources. Such bioactive compounds were profiled via reversed-phase liquid chromatography based on a C18 stationary phase coupled with UV diode-array detector and high resolution/accuracy mass spectrometry to develop a sustainable and effective extraction method. A screening D-optimal design allowed to identify solvent composition and matrix-to-solvent ratio as the key factors to enhance the extraction efficiency, while response surface methodology provided a suboptimal point (SP) at EtOH/H 2 O 70:30 (v/v), 0.3 M HCl, and 1:40 g/mL matrix-to-solvent ratio. SP outperformed the classical MeOH-based method in terms of extraction yield (79–89% versus less than 50% in a single cycle), total phenolic content (3.00 vs 1.36 mg GAE /g), and ABTS antioxidant activity (0.88 vs 0.26 mg TE /g). Anti-inflammatory assays revealed that SP extracts significantly reduced nitric oxide production, with dose-dependent effects. In addition, SP proved to be a more sustainable alternative than MeOH, with AGREE and AGREEprep scores increasing from 0.48 to 0.72 and 0.32 to 0.73, respectively. These findings encourage further investigation of pigmented wheat brans for potential nutraceutical applications. • Anthocyanins from Apulian coloured wheat landraces were investigated by LC-DAD/MS. • Experimental design guided a green and effective method development. • Higher extraction yields (79-89%) than MeOH-based protocols (ca. 45%). • AGREE (+28%) and AGREEprep scores (+64%) showed a remarkable greenness improvement. • In vitro antioxidant and anti-inflammatory properties indicate nutraceutical potential.