Hydrophilic Interaction Chromatography on Silica: Group Analysis of Grape Anthocyanins

Abstract

A group control procedure is developed, in which the anthocyanin complex of a grape extract is divided into six groups without differentiation by aglycons: 3-glucosides (Zone III); 3-glucosides acylated with p-coumaric acid (Zone I); 3-glucosides acylated with acetic acid (Zone II); 3,5-diglucosides (Zone IV); 3,5-diglucosides acylated with p-coumaric acid (Zone IV); and 3,5-diglucosides acylated with acetic acid (Zone V). The anthocyanins were separated by hydrophilic interaction chromatography in a column packed with a silica using the mobile phases of acetonitrile–water (20–8 vol %) containing 0.5 vol % of orthophosphoric acid. Due to the low viscosity of the mobile phase and the complete separation of anthocyanin groups, the procedure suitable for determining the inheritance of genetic traits in the selection of grapes is transferred to a format of microcolumn chromatography using MiLiChrome chromatographs.