The combination of normal-phase and reverse-phase high-pressure liquid chromatography with NMR for the isolation and characterization of oligosaccharide alditols from ovarian cyst mucins.
The combination of normal-phase and reverse-phase high-pressure liquid chromatography with NMR for the isolation and characterization of oligosaccharide alditols from ovarian cyst mucins.
复制标题
正相和反相高压液相色谱与 NMR 相结合，用于从卵巢囊肿粘蛋白中分离和表征寡糖糖醇。
DOI：
10.1016/0304-4165(84)90030-8
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发表时间：
1984
期刊：
影响因子：
--
通讯作者：
Bush,CA
中科院分区：
文献类型：
--
作者：
Dua,VK;Dube,VE;Bush,CA
氨基键合硅胶正相高压液相色谱柱与十八烷基硅胶反相液相色谱柱的互补，尤其适用于碱性硼氢化物降解粘蛋白糖蛋白所产生的低聚糖醛糖醇的分离。前者在分子大小的基础上可以很好地分离，而后者对立体异构体表现出选择性。四糖、五糖和六糖糖醇的立体异构体对的相对保留时间在3到12之间，因此在反相色谱中具有良好的制备分离效果。从单一的卵巢囊性糖蛋白H和Lewis b活性中，已经分离出13种寡糖，基本上代表整个碳水化合物的含量。其中12个低聚糖的结构经核磁共振氢谱确证。对于那些从其他来源分离的低聚糖，其核磁共振谱以前已经报道过，直接进行明确的结构鉴定。与已知的核磁共振谱只有一个或两个残基不同的低聚糖的结构可以通过利用化学位移类比的简单算法来推断。
Normal-phase high-pressure liquid chromatography (HPLC) on amino-bonded silica with elution by aqueous acetonitrile is shown to be an especially suitable complement to reverse-phase HPLC on octadecyl silica for the fractionation of oligosaccharide alditols produced by alkaline borohydride degradation of mucin glycoproteins. The former technique separates well on the basis of molecular size, while the latter method shows selectivity for stereoisomers. Stereoisomeric pairs of tetra-, penta- and hexasaccharide alditols show relative retention times ranging from 3 to 12, resulting in excellent preparative separations in reverse-phase chromatography. From a single ovarian cyst glycoprotein, H and Lewis b active, 13 oligosaccharides, representing essentially the entire carbohydrate content, have been isolated. The structures of 12 of the oligosaccharides have been determined by1H-NMR spectroscopy. For those oligosaccharides which have been isolated from other sources and whose NMR spectra have been previously reported, unambiguous structural identification follows directly. Structures of oligosaccharides differing by only one or two residues from those whose NMR spectra are known may be deduced by a simple algorithm utilizing chemical shift analogies.