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Determination of the uronic acid composition of seaweed dietary fibre by HPLC
Authors:Sánchez-Machado D I  López-Cervantes J  López-Hernández J  Paseiro-Losada P  Simal-Lozano J
Affiliation:Departamento de Química Analítica, Nutrición y Bromatología, Facultad de Farmacia, Universidad de Santiago de Compostela, 15782 Santiago de Compostela, La Coru?a, Spain.
Abstract:A high-performance liquid chromatographic (HPLC) method is described for determination of the ratio of beta-d-mannuronic acid to alpha-l-guluronic acid (M/G ratio) in dietary fibre of edible seaweeds. Total dietary fibre (TDF) content was determined gravimetrically. The TDF fraction was hydrolysed with 12 m and 1 m H(2)SO(4), then neutralized with AG 4 x 4 resin. The uronic acids were separated in a Tracer Extrasil SAX 5 micro m column (25 cm x 4 mm) at 35 degrees C, with 2 mm KH(2)PO(4) containing 5% methanol as mobile phase at a fl ow rate of 1.5 mL/min. The detection wavelength was UV 210 nm. The chromatographic identifications of beta-d-mannuronic acid and alpha-l-guluronic acid were confirmed by liquid chromatography-mass spectrometry (LC-MS). The method precision was 1.4% for beta-d-mannuronic acid and 3.5% for alpha-l-guluronic acid. The method was used to determine M/G ratio in canned seaweeds (Saccorhiza polyschides and Himanthalia elongata) and in dried seaweeds (H. elongata, Laminaria ochroleuca, Undaria pinnatifida, Palmaria sp. and Porphyra sp.).
Keywords:edible seaweeds  β‐d‐mannuronic acid and α‐l‐guluronic acid ratio  ion‐exchange chromatography  LC‐MS
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