Bouhours J F, Guignard H
J Lipid Res. 1979 Sep;20(7):879-907.
Free ceramide, glucosylceramide, and sphingomyelin were isolated from mature cells of adult rat small intestine. Free ceramide and ceramide cleaved from sphingomyelin by enzymatic hydrolysis were fractionated by thin-layer chromatography on borate-impregnated silica gel plates. Sphingoid bases were characterized by gas-liquid chromatography of aldehydes formed upon periodate oxidation. Fatty acids were quantified as methyl esters. Ceramide structures were confirmed by direct-inlet mass spectrometry. Free ceramide was found to contain two major long-chain bases in nearly equal quantity: sphingosine, mainly linked to palmitic acid, and 4D-hydroxysphinganine associated with C20 to C24 fatty acids, 22% being hydroxylated. Sphinganine occurred as a minor component linked to nonhydroxy fatty acids. Sphingomyelin contained the three long-chain bases and 63% of its ceramide was N-palmitoyl-sphingosine. Mass spectrometry of glucosylceramide confirmed 4D-hydroxyshingamine as the major sphingoid base associated preferentially with longer chain hydroxy fatty acids.
从成年大鼠小肠的成熟细胞中分离出游离神经酰胺、葡萄糖神经酰胺和鞘磷脂。通过硼酸盐浸渍硅胶板上的薄层色谱法对游离神经酰胺和通过酶水解从鞘磷脂中裂解得到的神经酰胺进行分离。通过对高碘酸盐氧化后形成的醛进行气液色谱分析来鉴定鞘氨醇碱。脂肪酸以甲酯形式进行定量。通过直接进样质谱法确认神经酰胺结构。发现游离神经酰胺含有两种数量几乎相等的主要长链碱：主要与棕榈酸相连的鞘氨醇，以及与C20至C24脂肪酸相关的4D-羟基鞘氨醇，其中22%被羟基化。鞘氨醇以与非羟基脂肪酸相连的次要成分形式存在。鞘磷脂含有这三种长链碱，其63%的神经酰胺为N-棕榈酰鞘氨醇。葡萄糖神经酰胺的质谱分析证实4D-羟基鞘氨胺是主要的鞘氨醇碱，优先与较长链的羟基脂肪酸相关。
