IDENTIFICATION AND FRACTIONATION OF HUMAN-MILK OLIGOSACCHARIDES BY PROTON-NUCLEAR MAGNETIC-RESONANCE SPECTROSCOPY AND REVERSE-PHASE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY
IDENTIFICATION AND FRACTIONATION OF HUMAN-MILK OLIGOSACCHARIDES BY PROTON-NUCLEAR MAGNETIC-RESONANCE SPECTROSCOPY AND REVERSE-PHASE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY
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DOI：
10.1016/0003-2697(83)90215-4
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发表时间：
1983-01-01
影响因子：
2.9
通讯作者：
BUSH, CA
中科院分区：
文献类型：
--
作者：
DUA, VK;BUSH, CA
含n -乙酰基氨基糖的低聚糖在反相高效液相色谱中的保留时间对链长和立体化学差异都很敏感。结果表明，在C-18色谱柱上，以水为洗脱剂，可以对母乳中的低聚糖进行立体化学分离。在纳摩尔水平的检测是可能的，使用紫外线吸收在190nm的对乙酰氨基基团的结果。通过分离碳水化合物组成相同的两种四糖，即乳酸-n -四糖和乳酸-n -新四糖，以及通过分离两种异构体五糖，即乳酸-n -fucopentaose I和II，可以分离出连锁异构体。在某些情况下，有可能从复杂混合物中制备分离纯低聚糖。在300 MHz的质子核磁共振波谱下，对不同的异构体寡糖进行了结构报告质子的鉴定和纯度鉴定。除了分配异构质子共振外，还可以分配连接在O3的半乳糖的H4，并聚焦H5和H6共振。
The retention times of oligosaccharides containing N-acetyl amino sugars in reverse-phase high-performance liquid chromatography are sensitive both to chain length and to stereochemical differences. Data showing that oligosaccharides of human milk can be fractionated according to stereochemistry on C-18 columns using water as eluant is presented. Detection at the nanomolar level is possible using UV absorption at 190 nm as a result of the acetamido group. The separation of linkage isomers by fractionation of 2 tetrasaccharides of identical carbohydrate composition, lacto-N-tetraose and lacto-N-neotetraose, and by the separation of 2 isomeric pentasaccharides, lacto-N-fucopentaose I and II, is shown. In some cases it was possible to preparatively fractionate pure oligosaccharides from complex mixtures. The identity and purity of the oligosaccharides was determined by proton-NMR spectroscopy at 300 MHz in which structural reporter protons are identified for the different isomeric oligosaccharides. In addition to assignment of the anomeric proton resonances, it was possible to assign H4 of galactose linked at O3 and fucose H5 and H6 resonances.