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Comprehensive two-dimensional liquid chromatographic analysis of anthocyanins A

Willemse, Chandré M., Stander, Maria A., Tredoux, Andreas G.J., de Villiers, André
Journal of chromatography 2014 v.1359 pp. 189-201
anthocyanins, black beans, blueberries, cabbage, comprehensive two-dimensional liquid chromatography, diodes, grapes, high performance liquid chromatography, hydrophilic interaction chromatography, radishes, reversed-phase liquid chromatography
Anthocyanins are naturally occurring plant pigments whose accurate analysis is hampered by their complexity and unique chromatographic behaviour associated with on-column conversion reactions. This paper reports the evaluation of off-line comprehensive two-dimensional liquid chromatography (LC×LC) for the analysis of anthocyanins. Hydrophilic interaction chromatography (HILIC) was used in the first dimension in combination with reversed phase liquid chromatography (RP-LC) in the second dimension. For the selective detection of anthocyanins, diode array detection was used, while high resolution quadrupole-time-of-flight mass spectrometry (Q-TOF) was used for compound identification. As application, the HILIC×RP-LC separation of diverse anthocyanins in blueberries, red radish, black beans, red grape skins and red cabbage is demonstrated. Off-line HILIC×RP-LC revealed information which could not be obtained by one-dimensional HPLC methods, while the structured elution order for the anthocyanins simplifies compound identification and facilitates the comparison of anthocyanin content of natural products by means of contour plots.