Stellenbosch University, Department of Chemistry and Polymer Science, Private Bag X1, Matieland 7602, South Africa.
Stellenbosch University, Central Analytical Facility, Private Bag X1, Matieland 7602, South Africa; Stellenbosch University, Department of Biochemistry, Private Bag X1, Matieland 7602, South Africa.
J Chromatogr A. 2014 Sep 12;1359:189-201. doi: 10.1016/j.chroma.2014.07.044. Epub 2014 Jul 22.
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.
花色苷是天然存在的植物色素，由于其复杂性以及与柱上转化反应相关的独特色谱行为，对其进行准确分析受到了阻碍。本文报道了离线全面二维液相色谱（LC×LC）分析花色苷的评估。在第一维中使用亲水相互作用色谱（HILIC），在第二维中使用反相液相色谱（RP-LC）。为了选择性检测花色苷，使用二极管阵列检测，而用于化合物鉴定的则是高分辨率四极杆飞行时间质谱（Q-TOF）。作为应用实例，展示了蓝莓、红萝卜、黑豆、红葡萄皮和红甘蓝中各种花色苷的 HILIC×RP-LC 分离。离线 HILIC×RP-LC 揭示了一维 HPLC 方法无法获得的信息，而花色苷的结构化洗脱顺序简化了化合物鉴定，并通过等高线图有助于比较天然产物中花色苷的含量。
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