On-line nano-HPLC/ESI QTOF MS and tandem MS for separation, detection, and structural elucidation of human erythrocytes neutral glycosphingolipid mixture
On-line nano-HPLC/ESI QTOF MS and tandem MS for separation, detection, and structural elucidation of human erythrocytes neutral glycosphingolipid mixture
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DOI：
10.1021/ac702175f
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发表时间：
2008-06-15
影响因子：
7.4
通讯作者：
Peter-Katalinle, Jasna
中科院分区：
文献类型：
--
作者：
Kirsch, Stephan;Zarei, Mostafa;Peter-Katalinle, Jasna
本文介绍了一种上级方法，包括纳米高效液相色谱（nano-HPLC）在线结合电喷雾离子化四极杆飞行时间质谱（ESI QTOF MS）和串联质谱，用于中性鞘糖脂（GSU）复杂混合物的筛选和结构表征。从人红细胞中纯化的中性GSL有效地分离，根据糖链长度的差异，通过优化的纳米HPLC方案和流动通过ESI QTOF MS检测在低飞摩尔水平。此外，GSL物质与混合物中的伴随脂质准确区分，从而允许通过数据依赖性分析确定详细的结构特征，以在单个实验中鉴定GSL构成。设计了一种替代的nano-HPLC/ESI QTOF MS方法，用于解剖定义GSL疏水部分的神经酰胺部分的不饱和度/饱和度，并随后通过nano-HPLC/ESI QTOF MS/MS对神经酰胺区域内的-CH= CH-进行定位。该方法是快速，高灵敏度，高通量的顺从，并强调糖脂组学作为一个新的和有价值的分析维度。
A superior approach involving nano-high-performance liquid chromatography (nano-HPLC) in on-line conjunction to electrospray ionization quadrupole time-of-flight mass spectrometry (ESI QTOF MS) and tandem MS for screening and structural characterization of complex mixtures of neutral glycosphingolipids (GSU) is here described. Neutral GSLs purified from human erythrocytes were efficiently separated according to the differences in carbohydrate chain length by an optimized nano-HPLC protocol and flow-through detected by ESI QTOF MS at the low femtomole level. Additionally, GSL species were accurately distinguished from the accompanying lipids in the mixture, thus permitting the determination of detailed structural characteristics by data-dependent analysis for identification of GSL constitution within single experiments. An alternative nano-HPLC/ESI QTOF MS approach was designed for dissection of unsaturation/saturation degree of the ceramide moieties defining the hydrophobic portion of GSLs and subsequent localization by nano-HPLC/ESI QTOF MS/MS of the -CH=CH-within the ceramide regions. The method is fast, highly sensitive, and high-throughput amenable and is highlighted as a new and valuable analytical dimension in glycolipidomics.