Alpert A J, Shukla M, Shukla A K, Zieske L R, Yuen S W, Ferguson M A, Mehlert A, Pauly M, Orlando R
PolyLC Inc., Columbia, MD 21045.
J Chromatogr A. 1994 Jul 29;676(1):191-22. doi: 10.1016/0021-9673(94)00467-6.
Complex carbohydrates can frequently be separated using hydrophilic-interaction chromatography (HILIC). The mechanism was investigated using small oligosaccharides and a new column, PolyGLYCOPLEX. Some carbohydrates exhibited anomer separation, which made it possible to determine the orientation of the reducing end relative to the stationary phase. Amide sugars were consistently good contact regions. Relative to amide sugars, sialic acids and neutral hexoses were better contact regions at lower levels of organic solvents than at higher levels. HILIC readily resolved carbohydrates differing in residue composition and position of linkage. Complex carbohydrate mixtures could be resolved using volatile mobile phases. This was evaluated with native glycans and with glycans derivatized with 2-aminopyridine or a nitrobenzene derivative. Both asialo- and sialylated glycans could be resolved using the same set of conditions. With derivatized carbohydrates, detection was possible at the picomole level by UV detection or on-line electrospray mass spectrometry. Selectivity compared favorably with that of other modes of HPLC. HILIC is promising for a variety of analytical and preparative applications.
复杂碳水化合物通常可以使用亲水相互作用色谱法（HILIC）进行分离。利用小寡糖和新型色谱柱PolyGLYCOPLEX对其分离机制进行了研究。一些碳水化合物表现出端基异构体分离，这使得确定还原端相对于固定相的方向成为可能。酰胺糖始终是良好的接触区域。相对于酰胺糖，在较低有机溶剂水平下，唾液酸和中性己糖比在较高水平下是更好的接触区域。HILIC很容易分离出残基组成和连接位置不同的碳水化合物。复杂碳水化合物混合物可以使用挥发性流动相进行分离。使用天然聚糖以及用2-氨基吡啶或硝基苯衍生物衍生化的聚糖对此进行了评估。在相同的条件下，脱唾液酸聚糖和唾液酸化聚糖都可以得到分离。对于衍生化的碳水化合物，可以通过紫外检测或在线电喷雾质谱在皮摩尔水平进行检测。其选择性与其他高效液相色谱模式相比具有优势。HILIC在各种分析和制备应用中很有前景。
