title: Comprehensive Two-Dimensional Hydrophilic Interaction Chromatography (HILIC) × Reversed-Phase Liquid Chromatography Coupled to High-Resolution Mass Spectrometry (RP-LC-UV-MS) Analysis of Anthocyanins and Derived Pigments in Red Wine.
abstract: Changes in anthocyanin chemistry represent some of the most important transformations involved in red wine aging. However, accurate analysis of the derived pigments, as required to study the evolution of anthocyanins and tannins during aging, is hampered by their extreme structural diversity, low levels, and the fact that many of these compounds have identical mass spectral characteristics. In this context, chromatographic separation is critical. In this contribution, the application of online hydrophilic interaction chromatography (HILIC) × reversed-phase liquid chromatography (RP-LC) separation coupled to high-resolution mass spectrometry (MS) is described for the detailed characterization of anthocyanins and their derived pigments in aged red wine. A systematic approach was followed for the optimization of HILIC × RP-LC separation parameters using a capillary liquid chromatography (LC) system in the first dimension and an ultrahigh-pressure LC system in the second dimension to ensure maximum sensitivity and performance. Ninety four (94) anthocyanin-derived pigments were tentatively identified in one- and six-year-old Pinotage wines using accurate mass and fragmentation information obtained using quadrupole-time-of-flight mass spectrometry (Q-TOF-MS). Online HILIC × RP-LC-MS was found to offer high-resolution separation, because of the combination of two different separation modes, while the structured elution order observed improved the certainty in compound identification. Therefore, this approach shows promise for the detailed elucidation of the chemical alteration of anthocyanins during wine aging.
authors.affiliation: Department of Chemistry and Polymer Science, Stellenbosch University , Private Bag X1, Matieland 7602, South Africa.
authors.forename: Chandré M
authors.initials: CM
authors.lastname: Willemse
authors.affiliation: Central Analytical Facility, Stellenbosch University , Private Bag X1, Matieland 7602, South Africa.|Department of Biochemistry, Stellenbosch University , Private Bag X1, Matieland 7602, South Africa.
authors.forename: Maria A
authors.initials: MA
authors.lastname: Stander
authors.affiliation: Center of Analytical Chemistry and Microbiology, Department of Microbiology and Biochemistry, Hochschule Geisenheim University , 65366 Geisenheim, Germany.|Université de Bordeaux, ISVV, EA 4577, Unité de recherche Œnologie, 33882 Villenave d'Ornon, France.|INRA, ISVV, USC 1366 Œnologie, 33882 Villenave d'Ornon, France.
authors.forename: Jochen
authors.initials: J
authors.lastname: Vestner
authors.affiliation: Department of Chemistry and Polymer Science, Stellenbosch University , Private Bag X1, Matieland 7602, South Africa.
authors.forename: Andreas G J
authors.initials: AG
authors.lastname: Tredoux
authors.affiliation: Department of Chemistry and Polymer Science, Stellenbosch University , Private Bag X1, Matieland 7602, South Africa.
authors.forename: André
authors.initials: A
authors.lastname: de Villiers
mesh_terms: D000872:Anthocyanins / Q000032:analysis*; D002853:Chromatography, Liquid; D056148:Chromatography, Reverse-Phase; D005504:Food Analysis / Q000379:methods*; D057927:Hydrophobic and Hydrophilic Interactions; D013058:Mass Spectrometry; D010860:Pigments, Biological / Q000032:analysis*; D014920:Wine / Q000032:analysis*
chemical_list: D000872:Anthocyanins; D010860:Pigments, Biological