HPLC determination of folates in raw and processed beetroots

A sensitive HPLC method with fluorescence detection and gradient elution has been developed for the determination of folates in vegetables. The method involved extraction of folates from food matrix by heat treatment, deconjugation of folate polyglutamates to monoglutamates by incubation with hog kidney conjugase and purification of food extracts by solid-phase extraction with strong-anion exchange cartridges. The chromatographic separation of folates was achieved on Zorbax SB C8 column, which was found to be superior over conventional C18 column in terms of selectivity and sensitivity. Validation of the method included linearity tests, the addition of standard folates for the determination of recovery, repeatability and stability tests. The method developed was applied to analysis of raw and processed beetroots; 5-methyltetrahydrofolate was found to be the main folate form in beetroots. Cultivar differences and growing conditions were found to have a pronounced effect on the folate content in beetroots. Processing resulted in considerable losses of folates, whereas losses during storage appeared to be moderate.