Hydrophilic interaction liquid chromatography coupled to mass spectrometry for the characterization of prebiotic galactooligosaccharides
Hydrophilic interaction liquid chromatography coupled to mass spectrometry for the characterization of prebiotic galactooligosaccharides
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DOI：
10.1016/j.chroma.2011.11.047
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发表时间：
2012-01-13
影响因子：
4.1
通讯作者：
Sanz, M. L.
中科院分区：
文献类型：
--
作者：
Hernandez-Hernandez, O.;Calvillo, I.;Sanz, M. L.
三种不同的固定相（磺烷基甜菜碱两性离子，聚羟乙基甜菜酰胺和乙烯桥杂化（BEH）与三官能键合酰胺），在亲水相互作用液相色谱（HILIC）模式下操作，已被测定和比较分析的复杂混合物的低聚半乳糖（GOS）。已优化色谱方法，以获得两个连续的含半乳糖的标准品和麦芽糖糊精之间的最佳分离，根据分辨率进行测量。研究了化学修饰剂（甲酸、乙酸铵和氢氧化铵）、有机溶剂和移动的梯度等因素对寡糖分离的影响。在BEH酰胺固定相上，以乙腈：水+0.1%氢氧化铵为移动的流动相，大多数寡糖被成功地分离。评价了含有半乳糖、葡萄糖和/或果糖单元的具有不同键合的二糖的特征性MS 2断裂谱，并用于通过HILIC-MS”表征三种商业益生元GOS混合物（GOS-1、GOS-2和GOS-3）的二糖、三糖和四糖。观察到GOS-1和GOS-3的定性和定量组成相似，而GOS-2的连接和丰度不同。一般来说，（1 -> 4）和（1 -> 6）糖苷键是在GOS中发现的主要结构，尽管也鉴定出（1 -> 2）和（1 -> 3）糖苷键。关于分子量，在这些样品中检测到高达五糖，双糖是最丰富的碳水化合物。(C)2011爱思唯尔有限公司版权所有。
Three different stationary phases (sulfoalkylbetaine zwitterionic, polyhydroxyethyl aspartamide and ethylene bridge hybrid (BEH) with trifunctionally bonded amide), operating at hydrophilic interaction liquid chromatographic (HILIC) mode, have been assayed and compared for the analysis of complex mixtures of galactooligosaccharides (GOS). Chromatographic methods have been optimized to obtain the best separation between two consecutive galactose containing standards and maltodextrins, measured on the basis of resolution. Influence of several factors such as chemical modifiers (formic acid, ammonium acetate and ammonium hydroxide), organic solvent and gradients of the mobile phases in the separation of oligosaccharides have been studied. The best results were achieved on the BEH amide stationary phase, using acetonitrile:water with 0.1% ammonium hydroxide as mobile phase, where the most of oligosaccharides were successfully resolved. Characteristic MS2 fragmentation profiles of disaccharides containing galactose, glucose and/or fructose units with different linkages were evaluated and used for the characterization of di-, tri- and tetrasaccharides of three commercial prebiotic GOS mixtures (GOS-1, GOS-2 and GOS-3) by HILIC-MS". Similar qualitative and quantitative composition was observed for GOS-1 and GOS-3, whereas different linkages and abundances were detected for GOS-2. In general, (1 -> 4) and (1 -> 6) glycosidic linkages were the main structures found in GOS, although (1 -> 2) and (1 -> 3) linkages were also identified. Regarding molecular weight, up to pentasaccharides were detected in these samples, disaccharicles being the most abundant carbohydrates. (C) 2011 Elsevier B.V. All rights reserved.