Use of graphitised carbon negative ion LC-MS to analyse enzymatically digested glycosaminoglycans
Use of graphitised carbon negative ion LC-MS to analyse enzymatically digested glycosaminoglycans
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DOI：
10.1016/j.jchromb.2005.07.014
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发表时间：
2005-09-25
影响因子：
3
通讯作者：
Whitelock, JM
中科院分区：
文献类型：
--
作者：
Karlsson, NG;Schulz, BL;Whitelock, JM
使用石墨化碳固定相和离子阱质谱的毛细管液相色谱-质谱法被证明是一种强大的技术，用于分析糖胺聚糖内切酶消化。通常发现的二糖从肝素/硫酸乙酰肝素酶在亚纳摩尔水平被发现分离的质量和/或保留时间和检测的负离子电喷雾质谱法主要作为[M - H]（-）离子使用标准电喷雾接口和6-10 μ L/min之间的流速。石墨化碳液相色谱-裂解质谱法提供的序列数据的二糖和寡糖。从[M-H]（-）离子（低硫酸化二糖）或[M + Na -2 H]（-）离子（高硫酸化二糖）的碰撞中获得序列信息。使用阳离子交换和石墨化碳的组合的糖苷内切酶消化和样品制备的这种分离和鉴定方法被用于成功地分析来自滑液样品的硫酸角质素（角质素酶）和肝素（肝素酶）标准品以及透明质酸（透明质酸酶）。(c)2005 Elsevier B. V.保留所有权利。
Capillary liquid chromatography-mass spectrometry using graphitised carbon stationary phase and ion trap mass spectrometry was shown to be a powerful technique for analysing glycosaminoglycans digested with endoglycosidases. Commonly found disaccharides from heparin/heparan sulphate digests at sub nanomole levels were found to be separated by mass and/or retention time and detected by negative ion electrospray mass spectrometry predominantly as [M - H](-) ions using a standard electrospray interface and flow rate between 6-10 mu L/min. Graphitised carbon liquid chromatography-fragmentation mass spectrometry provided sequence data of disaccharides and oligosaccharides. Sequence information was obtained from either collision of the [M - H](-) ions (low sulphated disaccharides) or of the [M + Na - 2H](-) ions (highly sulphated disaccharides). This separation and identification method of endoglycosidase digestion and sample preparation using a combination of cation exchange and graphitised carbon, was used to successfully analyse digests of keratan sulphate (keratanase) and heparin (heparinase) standards, and hyaluronic acid (hyaluronidase) from synovial fluid samples. (c) 2005 Elsevier B.V. All rights reserved.