Seyfried T N, Ando S, Yu R K
J Lipid Res. 1978 Jul;19(5):538-43.
The monosialoganglioside hematoside (GM3) is an important precursor in the synthesis of the more complex gangliosides. To obtain large quantities of GM3 for use in ganglioside biosynthetic experiments, we have devised a column chromatographic procedure for the isolation and purification of GM3 from human liver. A total ganglioside mixture was obtained from a 550 g sample of normal human liver. Quantitative analysis by gas-liquid chromatography indicated about 66 microgram of lipid-bound N-acetylneuraminic acid per gram of fresh tissue. Hematoside appeared as double bands on thin-layer plates and represented 86.4% of the total sialic acid content. Additional ganglioside species, notably GD3, were also seen. Hematoside was separated from the other gangliosides by GM3 obtained was 120 mg, which represented a 90% recovery. Although the long-chain base and sugar compositions of the upper and lower GM3 fractions were similar, striking fatty acid differences were detected. The upper fraction contained predominantly unsubstituted fatty acids, while the lower fraction showed a preponderance of alpha-hydroxy fatty acids. The unsubstituted fatty acid and hydroxy fatty acid composition of the total GM3 fraction was calculated to be 56.9% and 43.1%, respectively.
单唾液酸神经节苷脂血型糖苷（GM3）是合成更复杂神经节苷脂的重要前体。为了获得大量GM3用于神经节苷脂生物合成实验，我们设计了一种柱色谱方法，用于从人肝脏中分离和纯化GM3。从550克正常人肝脏样本中获得了总神经节苷脂混合物。气液色谱法定量分析表明，每克新鲜组织中约含66微克脂质结合的N-乙酰神经氨酸。血型糖苷在薄层板上呈现为两条带，占总唾液酸含量的86.4%。还观察到了其他神经节苷脂种类，特别是GD3。通过硅胶柱色谱法将血型糖苷与其他神经节苷脂分离，得到的GM3为120毫克，回收率为90%。尽管上层和下层GM3组分的长链碱和糖组成相似，但检测到了显著的脂肪酸差异。上层组分主要含有未取代的脂肪酸，而下层组分则以α-羟基脂肪酸为主。总GM3组分的未取代脂肪酸和羟基脂肪酸组成分别计算为56.9%和43.1%。
