Characterization of bioactive cranberry fractions by mass spectrometry

Abstract

Increasing evidence indicates that fruits contain functional bio-active compounds that have several preventive and therapeutic health benefits. Our group has recently conducted studies to assess the potential effects of cranberry polyphenolic fractions on intestinal Caco-2/15 epithelial cells and a substantial reduction in oxidative stress and inflammation was observed. The aim of the present work was to determine the polyphenolic species most likely responsible for the observed biological activity. Low, medium, and high molecular weight cranberry fractions were generated with a Sephadex LH-20 column by elution with 60% MeOH, 100% MeOH, and 70% acetone, respectively. The total phenolic content in these fractions was determined by the Folin–Ciocalteu method. A combination of LC–MS and MALDI-TOF methods were used to characterize the nature of polyphenolic compounds in the cranberry extracts. High resolution mass spectrometry was used to generate empirical formulae for the detected species. The low molecular weight fraction was essentially constituted of small phenolic acids (hydroxycinnamic and hydroxybenzoic acids). The medium fraction was mostly composed of anthocyanin, flavonols (quercetin, myricetin, isorhamnetin, kaempferol), procyanidins monomers (epicatechin, catechin), dimers, and few small oligomers. The heavy fraction was devoid of phenolic acids and anthocyanins, and it contained for the most part oligomers and polymers of procyanidins. Proanthocyanidins oligomers up to n = 22 were detected, which is, to our knowledge, the largest individual polymers reported to date. With this approach, it was also possible to distinguish between the A-type and B-type linkages.