Hydrophilic interaction chromatographic analysis of anthocyanins

• Published in: Journal of Chromatography A Vol. 1319; pp. 127 - 140
• Main Authors: Willemse, Chandré M., Stander, Maria A., de Villiers, André
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 06.12.2013; Elsevier
• Subjects: acylation; Anthocyanins; Anthocyanins - analysis; Arrays; Biological and medical sciences; black beans; blueberries; cabbage; Chromatography; Chromatography, High Pressure Liquid - methods; Consumption; Diodes; Food industries; Fruit and vegetable industries; Fundamental and applied biological sciences. Psychology; General pharmacology; glycosylation; grapes; HPLC; hydrophilic interaction chromatography; Hydrophilic interaction chromatography (HILIC); hydrophilicity; Hydrophobic and Hydrophilic Interactions; Liquid chromatography; mass spectrometry; Medical sciences; Pharmacognosy. Homeopathy. Health food; Pharmacology. Drug treatments; Phases; phenolic compounds; Phenolics; Quadrupole-time-of-flight mass spectrometry (Q-TOF-MS); radishes; reversed-phase liquid chromatography; Separation; silica; Spectrometry, Mass, Electrospray Ionization; Spectrophotometry, Ultraviolet; Temperature; Asia; China; Anthocyanins; Hydrophilic interaction chromatography (HILIC); Quadrupole-time-of-flight mass spectrometry (Q-TOF-MS); Phenolics; HPLC; Tandem mass spectrometry; Chemical analysis; Vegetables; Vaccinium myrtillus; Small fruits; HPLC chromatography; Flavonoid; Raphanus sativus; Electrospray; Quadrupole spectrometer; Red cabbage; Medicinal plant; Polyphenol; Folk medicine; Cruciferae; Dicotyledones; Blueberry; Angiospermae; Peel; Anthocyanin; Fruit; Coupled method; Pharmacognosy; Hydrophilic Interaction Liquid chromatography; Ultraviolet detector; Leguminosae; Phaseolus vulgaris; Grape; Plant origin; Spermatophyta; Ericaceae; Time-of-flight mass spectrometers
• ISSN: 0021-9673; 1873-3778
• DOI: 10.1016/j.chroma.2013.10.045

•The first application of HILIC for anthocyanin analysis is reported.•The choice of HILIC stationary phase and mobile phase composition were optimised.•Highly acidic mobile phases, high temperature and low flow rates improve separation.•Optimised HILIC–ESI-Q-TOF-MS methods were developed for anthocyanins in natural products.•HILIC offers an alternative separation mechanism to RP-LC for anthocyanins. Hydrophilic interaction chromatography (HILIC) provides an alternative separation mode for the analysis of phenolic compounds, in which aqueous-organic mobile phases with polar stationary phases are used. This paper reports the evaluation of HILIC for the analysis of the natural pigments anthocyanins, which are of importance because of their chromophoric properties and a range of health benefits associated with their consumption. Several HILIC stationary phases (silica, diol, amine, cyanopropyl and amide) and mobile phase combinations were evaluated, with the latter proving particularly important due to the distinctive chromatographic behaviour of anthocyanins. Diode array detection was used for selective detection of anthocyanins, while high resolution quadrupole-time-of-flight mass spectrometry (Q-TOF-MS) was used for compound identification. The potential of HILIC separation is demonstrated for a range of anthocyanins varying in glycosylation and acylation patterns found in blueberries, grape skins, black beans, red cabbage and red radish. HILIC is shown to be a complementary separation method to reversed phase liquid chromatography (RP-LC) due to the alternative retention mechanism.