STRUCTURAL CHARACTERIZATION OF OLIGOSACCHARIDES BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY, FAST-ATOM BOMBARDMENT MASS-SPECTROMETRY, AND EXOGLYCOSIDASE DIGESTION
STRUCTURAL CHARACTERIZATION OF OLIGOSACCHARIDES BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY, FAST-ATOM BOMBARDMENT MASS-SPECTROMETRY, AND EXOGLYCOSIDASE DIGESTION
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DOI：
10.1016/0003-2697(84)90057-5
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发表时间：
1984-01-01
影响因子：
2.9
通讯作者：
SWEELEY, CC
中科院分区：
文献类型：
--
作者：
WANG, WT;LEDONNE, NC;SWEELEY, CC
描述了制备紫外吸收衍生物后对寡糖进行结构表征的方法。这些衍生物可以通过高效液相色谱进行快速分析和纯化，主要根据尺寸和糖组成来分离各种结构。衍生化需要低至 0.5-1.0 nmol 的寡糖，通过监测 229 nm 处的吸光度可以检测低至 50 pmol 的寡糖。此外，发现衍生物的碳水化合物部分保留了其对外切糖苷酶的敏感性，允许进行连续的酶消化以确定糖序列和异头性。这些衍生物还具有潜在正电荷位点，使其易于通过快原子轰击质谱法进行分析。衍生物的全甲基化允许通过毛细管气相色谱法分离它们，从而可以通过气相色谱-质谱法研究它们的结构。这些技术的结合将能够确定少量寡糖的几乎完整结构。
A method for structural characterization of oligosaccharides after prepararing UV-absorbing derivatives was described. The derivatives can be rapidly analyzed and purified by high-performance liquid chromatography, with separation of various structures determined primarily by size and sugar composition. Derivatization requires as little as 0.5-1.0 nmol of oligosaccharide and detection of down to 50 pmol of oligosaccharide is possible by monitoring absorbance at 229 nm. In addition, the carbohydrate portion of the derivative was found to retain its sensitivity to exoglycosidases, allowing sequential enzymatic digestions for determination of sugar sequence and anomerity to be performed. The derivatives also possessed a site of potential positive charge, making them amenable to analysis by fast-atom bombardment-mass spectrometry. Permethylation of the derivatives permitted their separation by capillary gas chromatography, thus allowing investigation of their structures by gas chromatography-mass spectrometry. The combination of these techniques will allow almost the complete structure of small amounts of oligosaccharides to be determined.