Bodennec J, Koul O, Aguado I, Brichon G, Zwingelstein G, Portoukalian J
Institute Michel Pacha, University Claude Bernard-Lyon I, 83500 La Seyne sur Mer, France.
J Lipid Res. 2000 Sep;41(9):1524-31.
Solid-phase extraction (SPE) methods are easy, rapid, and reliable. Their growing popularity is in part due to their operational simplicity and cost reduction in solvents, and partly because they are easier to automate. Sphingolipids are implicated in various cellular events such as growth, differentiation, and apoptosis. However, their separation by small SPE cartridges has attracted limited attention. Here we describe an SPE procedure on aminopropyl cartridges that by sequential elution allows the separation of a lipid mixture into free ceramides, neutral glycosphingolipids, neutral phospholipids (sphingomyelin), and a fraction containing the acidic phospholipids and phosphorylated sphingoid bases, phosphoceramides and sulfatides. Individual components are obtained in high yield and purity. We applied the procedure to obtain data on separation of [(3)H]myristic acid-labeled sphingolipids from fish gills, and from human melanoma tumor tissue. Individual lipids in the SPE fractions were identified by chromatography on several high-performance thin-layer chromatography (HPTLC) systems. The chromatographic behavior of free sphingoid bases is also reported.
固相萃取（SPE）方法简便、快速且可靠。其日益普及部分归功于操作简单以及溶剂成本降低，部分原因是更易于自动化。鞘脂类参与多种细胞活动，如生长、分化和凋亡。然而，使用小型SPE柱对其进行分离受到的关注有限。在此，我们描述了一种在氨丙基柱上的SPE方法，该方法通过顺序洗脱可将脂质混合物分离为游离神经酰胺、中性糖鞘脂、中性磷脂（鞘磷脂）以及含有酸性磷脂和磷酸化鞘氨醇碱、磷酸神经酰胺和硫苷脂的组分。各组分以高收率和高纯度获得。我们应用该方法获取了从鱼鳃和人黑色素瘤肿瘤组织中分离[³H]肉豆蔻酸标记鞘脂类的数据。通过在多个高效薄层色谱（HPTLC）系统上进行色谱分析，鉴定了SPE馏分中的各个脂质。还报道了游离鞘氨醇碱的色谱行为。
