Determination of folates in seaweeds by high-performance liquid chromatography

A reversed-phase high-performance liquid chromatography (HPLC) method, with a combination of fluorescence and ultraviolet detectors, to determine the folate forms present in commercial macroalgae products form northwest Spain as part of nutritional studies in dehydrated and canned seaweeds is reported. The method includes extraction of folates from seaweed by heat treatment, deconjugation of folate polyglutamates by incubation with hog kidney conjugase and purification by solid-phase extraction (SPE) with strong anion-exchange (SAX) cartridges. Separation was achieved with a Tracer Extrasil ODS 5 μm 25 cm×0.4 cm column using acetonitrile and potassium phosphate buffer (pH 2.2) as mobile phase. Good results were obtained with respect to repeatability (relative standard deviation (R.S.D.) ≤4.12%) and recovery (≥90.80%). The amount of folate (as folic acid) in the six species ranged from 61.4 to 161.6 μg per 100 g dry mass. In all the seaweeds studied (Himanthalia elongata, Laminaria ochroleuca, Palmaria spp., Undaria pinnatifida and Porphyra spp. and Saccorhiza polychides) the single most abundant form is 5-CH3H4-folate, except Porphyra and Himanthalia.