Reversed-phase ion-pairing liquid chromatography/ion trap mass spectrometry for the analysis of negatively charged, derivatized glycans
Reversed-phase ion-pairing liquid chromatography/ion trap mass spectrometry for the analysis of negatively charged, derivatized glycans
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DOI：
10.1002/rcm.1079
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发表时间：
2003-01-01
影响因子：
2
通讯作者：
Vouros, P
中科院分区：
文献类型：
--
作者：
Gennaro, LA;Harvey, DJ;Vouros, P
显著的复杂性、相似的极性和缺乏可电离位点使得聚糖的分析成为分析挑战。这些化合物通常被衍生化，并通过正相高效液相色谱法（HPLC）或毛细管电泳法（CE）分离，然后进行UV或荧光检测。由于广泛使用反相色谱耦合电喷雾质谱作为分析工具，我们的实验室已经开发了这种方法用于分析带负电荷的标签，8-氨基萘-1，3，6-三磺酸（ANTS）衍生的聚糖。可以利用这种带负电荷的标签与移动的相离子配对试剂相互作用的能力，允许保留在反相C-18柱上用于随后的在线UV或MS分析。ANTS衍生的样品，包括麦芽低聚糖梯形和从牛核糖核酸酶B、牛胎球蛋白和鸡卵清蛋白释放的聚糖，使用该方法进行分析。除了反相保留，核糖核酸酶B和卵清蛋白衍生物显示出非常理想的异构体分离。通过使用质谱检测聚糖鉴别，这允许对单个组分进行相对定量。版权所有（C）2003约翰威利父子有限公司。
The significant complexity, similar polarity and lack of ionizable sites make the analysis of glycans an analytical challenge. These compounds are often derivatized and separated by normal-phase high-performance liquid chromatography (HPLC) or capillary electrophoresis (CE) followed by UV or fluorescence detection. Due to widespread use of reversed-phase chromatography coupled to electrospray mass spectrometry as an analytical tool, our laboratory has developed this methodology for the analysis of glycans derivatized with a negatively charged tag, 8-aminonaphthalene-1,3,6-trisulfonic acid (ANTS). It is possible to exploit the ability of this negatively charged tag to interact with a mobile phase ion-pairing reagent, allowing retention on a reversed-phase C-18 column for subsequent on-line UV or MS analysis. ANTS-derivatized samples, including a maltooligosaccharide ladder and glycans released from bovine ribonuclease B, bovine fetuin, and chicken ovalbumin, were analyzed using this method. In addition to reversed-phase retention, ribonuclease B and ovalbumin derivatives displayed highly desirable isomeric separation. With the use of mass spectrometric detection for glycan identity, this allowed relative quantitation of individual components. Copyright (C) 2003 John Wiley Sons, Ltd.