Ultraperformance ion-pair liquid chromatography coupled to electrospray time-of-flight mass spectrometry for compositional profiling and quantification of heparin and heparan sulfate
Ultraperformance ion-pair liquid chromatography coupled to electrospray time-of-flight mass spectrometry for compositional profiling and quantification of heparin and heparan sulfate
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DOI：
10.1021/ac702235u
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发表时间：
2008-02-15
影响因子：
7.4
通讯作者：
Larive, Cynthia K.
中科院分区：
文献类型：
--
作者：
Korir, Albert K.;Lirntiaco, John F. K.;Larive, Cynthia K.
肝素和硫酸乙酰肝素（HS）是重要的药物靶标，因为它们结合大量蛋白质，包括生长因子和细胞因子，介导许多生物过程。由于其生物学意义和复杂性，需要开发快速和灵敏的分析技术，用于在二糖水平上对肝素和HS进行表征和成分分析，以及用于对蛋白质结合重要的较大糖胺聚糖（GAG）序列的结构阐明。在这项工作中，我们提出了一种快速分析二糖组成的方法，使用反相离子对超高效液相色谱-电喷雾飞行时间质谱（（RPIP）-UPLC-MS）。肝素二糖标准品在不到5分钟的洗脱。该方法被用来确定从普通肝素/HS从各种牛和猪组织的GAG的成分，并与文献值的结果进行了比较。
Heparin and heparan sulfate (HS) are important pharmaceutical targets because they bind a large number of proteins, including growth factors and cytokines, mediating many biological processes. Because of their biological significance and complexity, there is a need for development of rapid and sensitive analytical techniques for the characterization and compositional analysis of heparin and HS at the disaccharide level, as well as for the structure elucidation of larger glycosaminoglycan (GAG) sequences important for protein binding. In this work, we present a rapid method for analysis of disaccharide composition using reversed-phase ion-pairing ultraperformance liquid chromatography coupled with electrospray time-of-flight mass spectrometry ((RPIP)-UPLC-MS). Heparin disaccharide standards were eluted in less than 5 min. The method was used to determine the constituents of GAGs from unfractionated heparin/HS from various bovine and porcine tissues, and the results were compared with literature values.