High-pressure liquid chromatographic separation of 3-glucosides, 3,5-diglucosides, 3-(6-O- p-coumaryl)glucosides and 3-(6-O- p-coumarylglucoside)-5-glucosides of anthocyanidins

Abstract

A high-pressure liquid chromatomagraphic method has been developed for separation of anthocyanins from fruits and their products on a μBondapak c 18 analytical column. Separation of 3- and 3,5-diglucosides f anthocyanidins can be achieved with aqueous acetic acid. That of the p-coumaryl-3 and 3,5-diglucosides requires an aqueous methanolic acetic acid solution. For the separation of complex mixtures of anthocyanins containing members of all four above mentioned pigment groups, a programmed non-linear-gradient elution between aqueous acetic acid and aqueous methanolic acetic solution is required. This technique enables a relatively fast separation and identification of twenty anthocyains in one run without prior treatment of the plant extract, or derivatization of the piments.