Rezanka T, Mares P
Institute of Microbiology, Czechoslovak Academy of Sciences, Prague.
J Chromatogr. 1990 Jun 22;509(2):333-46. doi: 10.1016/s0021-9673(01)93091-2.
Six fractions containing tri- to pentaglycosylceramides were isolated from the green, fresh water alga Chlorella kessleri, grown heterotrophically, by using preparative high-performance liquid chromatography (HPLC). Up to twelve fractions were obtained by further reversed-phase HPLC of each glycosylceramide. The use of a polar capillary column with Supelcowax 10 as the stationary phase allowed an excellent separation of the individual molecular species of ceramides, even though the separation did not occur when the ceramides differed only in the position of the amide bond. The individual molecular species (even if present in mixtures) were identified by gas chromatography-chemical ionization mass spectrometry. The evidence for a complete structure was obtained by enzyme splitting with alpha- and beta-galactosidases (the sequence of monosaccharides) and by negative ionization fast atom bombardment mass spectrometry. More than 400 molecular species of glycosylceramides were identified.
通过制备型高效液相色谱（HPLC），从异养生长的绿色淡水藻类克氏小球藻中分离出了六个含有三至五糖基神经酰胺的组分。对每个糖基神经酰胺进一步进行反相HPLC操作，最多可得到十二个组分。使用以Supelcowax 10为固定相的极性毛细管柱，能够出色地分离神经酰胺的各个分子种类，不过当神经酰胺仅在酰胺键位置不同时，无法实现分离。通过气相色谱 - 化学电离质谱法鉴定了各个分子种类（即使存在于混合物中）。借助α - 和β - 半乳糖苷酶进行酶解（单糖序列）以及负离子快速原子轰击质谱法，获得了完整结构的证据。鉴定出了400多种糖基神经酰胺的分子种类。
