LC-MS(n) analysis of isomeric chondroitin sulfate oligosaccharides using a chemical derivatization strategy.
LC-MS(n) analysis of isomeric chondroitin sulfate oligosaccharides using a chemical derivatization strategy.
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DOI：
10.1007/s13361-011-0174-0
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发表时间：
2011-09
影响因子：
3.2
通讯作者：
Sharp, Joshua S.
中科院分区：
文献类型：
--
作者：
Huang, Rongrong;Pomin, Vitor H.;Sharp, Joshua S.
需要改进的糖胺多聚糖(GAG)结构分析方法来了解它们在各种生物过程中的功能作用。用液相色谱-质谱法(LC-MS)表征复杂的GaG寡糖的结构面临的主要挑战包括准确确定硫酸盐基团在碰撞活化时的气相损失所导致的硫酸盐模式，以及在MS/MS分析之前在线分离位置硫酸盐异构体的效率低下。在这里，包括全甲基化、脱硫和乙酰化在内的连续化学衍生化过程能够在线分离硫酸软骨素(CS)低聚糖的异构体混合物，并通过MSN准确地确定硫酸盐的硫化位置。衍生的低聚糖具有被乙酰基取代的硫酸盐基团，这些基团足够稳定，可以在MSN碎裂中存活，并反映原始的硫化模式。采用标准反相液质联用系统，以C18毛细管柱为分离柱，采用碰撞诱导解离(CID)法进行MSN实验。我们的结果表明，这种衍生化策略和MSN方法的结合可以准确地识别具有饱和或不饱和非还原末端的CS六糖的硫化异构体。此外，由于乙酰基修饰的位置不同，衍生的CS六糖异构体混合物可以通过LC-MS方法进行分离。
Improved methods for structural analyses of glycosaminoglycans (GAGs) are required to understand their functional roles in various biological processes. Major challenges in structural characterization of complex GAG oligosaccharides using liquid chromatography-mass spectrometry (LC-MS) include the accurate determination of the patterns of sulfation due to gas-phase losses of the sulfate groups upon collisional activation and inefficient on-line separation of positional sulfation isomers prior to MS/MS analyses. Here, a sequential chemical derivatization procedure including permethylation, desulfation, and acetylation was demonstrated to enable both on-line LC separation of isomeric mixtures of chondroitin sulfate (CS) oligosaccharides and accurate determination of sites of sulfation by MSn. The derivatized oligosaccharides have sulfate groups replaced with acetyl groups, which are sufficiently stable to survive MSn fragmentation and reflect the original sulfation patterns. A standard reversed-phase LC-MS system with a capillary C18 column was used for separation, and MSn experiments using collision-induced dissociation (CID) were performed. Our results indicate that the combination of this derivatization strategy and MSn methodology enables accurate identification of the sulfation isomers of CS hexasaccharides with either saturated or unsaturated nonreducing ends. Moreover, derivatized CS hexasaccharide isomer mixtures become separable by LC-MS method due to different positions of acetyl modifications.
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影响因子：
7.4
作者：
Saad, OM;Leary, JA
通讯作者：
Leary, JA
影响因子：
3.1
作者：
JUHASZ, P;BIEMANN, K
通讯作者：
BIEMANN, K
DOI：
10.1016/j.jchromb.2005.07.014
发表时间：
2005-09-25
影响因子：
3
作者：
Karlsson, NG;Schulz, BL;Whitelock, JM
通讯作者：
Whitelock, JM
影响因子：
7.4
作者：
Estrella, Ruby P.;Whitelock, John M.;Karlsson, Niclas G.
通讯作者：
Karlsson, Niclas G.
影响因子：
4.3
作者：
Saad, OM;Ebel, H;Leary, JA
通讯作者：
Leary, JA