Sonnino S, Kirschner G, Ghidoni R, Acquotti D, Tettamanti G
J Lipid Res. 1985 Feb;26(2):248-57.
A new procedure is described for preparing the molecular species of GM1 ganglioside that carry a single fatty acid (myristic (C14:0), stearic (C18:0), arachidic (C20:0) or lignoceric (C24:0) acid) and a single long chain base (C18 or C20 sphingosine, C18 or C20 sphinganine, each of them in natural 3D(+)erythro or unnatural 3L(-)threo form). The procedure consisted of the following steps: a) alkaline hydrolysis of GM1 ganglioside in the presence of tetramethylammonium hydroxide, which produces de-N-acylation of the ceramide and de-N-acetylation of the sialic acid residue; b) specific re-N-acylation at the long chain base amino group with a new fatty acid (myristic, stearic, arachidic, or lignoceric) in the presence of 1-(3-dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride; and c) final re-N-acetylation at the level of the sialic acid residue. GM1 ganglioside molecular species, completely homogeneous in the ceramide portion, were prepared by reversed phase high performance liquid chromatography. The GM1 ganglioside molecular species were analyzed for saccharide, fatty acid, and long chain base composition by chemical and spectrometric analyses. Using a combination of the two procedures, 32 different molecular species of GM1 ganglioside, over 99% homogeneous, have been prepared.
描述了一种制备携带单一脂肪酸（肉豆蔻酸（C14:0）、硬脂酸（C18:0）、花生酸（C20:0）或木蜡酸（C24:0））和单一长链碱（C18或C20鞘氨醇、C18或C20鞘氨醇胺，它们各自呈天然3D(+)赤藓糖型或非天然3L(-)苏阿糖型）的GM1神经节苷脂分子种类的新方法。该方法包括以下步骤：a）在氢氧化四甲铵存在下对GM1神经节苷脂进行碱性水解，这会导致神经酰胺的脱N-酰化和唾液酸残基的脱N-乙酰化；b）在1-(3-二甲基氨基丙基)-3-乙基碳二亚胺盐酸盐存在下，用新的脂肪酸（肉豆蔻酸、硬脂酸、花生酸或木蜡酸）对长链碱氨基进行特异性再N-酰化；以及c）在唾液酸残基水平进行最终的再N-乙酰化。通过反相高效液相色谱法制备了神经酰胺部分完全均匀的GM1神经节苷脂分子种类。通过化学和光谱分析对GM1神经节苷脂分子种类的糖类、脂肪酸和长链碱组成进行了分析。通过结合这两种方法，制备了32种不同的GM1神经节苷脂分子种类，其纯度超过99%。
