Characterization of Major Anthocyanins and the Color of Red-Fleshed Budd Blood Orange (Citrus sinensis)

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Abstract
High-performance liquid chromatography (HPLC) with photodiode array detection was applied for the characterization of anthocyanins in red-fleshed Budd Blood (Citrus sinensis) orange. More than seven anthocyanin pigments were separated within 30 min by using a binary gradient (0.1% H3P04 in water and 0.1% H3PO4 in acetonitrile) elution on a Prodigy ODS column. Separations by reversed-phase HPLC and semipreparative HPLC on a Prodigy 10-μm ODS Prep column, and acid and alkali hydrolyses were used for identification of anthocyanins. The primary anthocyanins in Budd Blood orange grown in Florida were cyanidin-3-(6‘ ‘-malonylglucoside) (44.8%) followed by cyanidin-3-glucoside (33.6%). Two other minor pigments were also acylated with malonic acid. Malonated anthocyanins represented the major proportion (>51%) of anthocyanins in Budd Blood orange. Total anthocyanin contents and juice color parameters (CIE L*,a*,b*) were compared with six other Florida-grown blood oranges. Keywords: Budd Blood orange; HPLC; color; pigment; anthocyanin; malonated anthocyanins