Size Exclusion Coupled to Reversed Phase Liquid Chromatography for the Characterization of Cranberry Products

A combination of solvent extraction, size exclusion purification, and reversed-phase liquid chromatography with UV absorption and fluorescence detection was here used for the determination of flavan-3-ols and related oligomeric compounds in cranberry products. The method relied on robust, simple, and inexpensive instrumental equipment present in most of routine analytical laboratories. Sample extracts in methanol/water/hydrochloric acid solution (60:39:1, v/v/v) were subjected to size-exclusion chromatography (SEC) using Sephadex LH-20 as the sorbent to separate simple phenolic and oligomeric components. As a novel aspect of this work, the implementation of the SEC process in a reusable packed column was practically and economically highly advantageous. Fractions were analyzed by reversed-phase chromatography using a C18 core-shell analytical column and 0.1% formic acid aqueous solution and methanol as the components of the mobile phase. Chromatograms were recorded by UV absorption spectroscopy at 280nm and by fluorescence spectroscopy at (ex) 280nm/(em) 347nm. The method was applied to the determination of flavan-3-ols in various cranberry samples. Catechin, epicatechin, and procyanidins A2 and B2 were found in different amounts depending on the cranberry varieties.