Miller-Podraza H, Månsson J E, Svennerholm L
Department of Psychiatry and Neurochemistry, University of Göteborg, St. Jörgen Hospital, Hisings Backa, Sweden.
Biochim Biophys Acta. 1992 Feb 20;1124(1):45-51. doi: 10.1016/0005-2760(92)90124-e.
A simple procedure that enables the isolation of ganglioside GQ1b and other complex gangliosides from the human brain is described. The tissue was extracted with a mixture of chloroform, methanol, and water, and the extract purified twice by means of silica gel column chromatography and preparative HPTLC. Phase partition and ion exchange chromatography were omitted. The silica gel chromatography was based on a two step developing system, which provided an efficient separation of oligosialogangliosides. The yields of chromatographically homogenous fractions of ganglioside GQ1b isolated from the whole cerebrum, cerebellar cortex and occipital grey matter of a 60-year-old woman were 62, 138 and 110 nmol SA per g of fresh tissue. The problem of co-extraction of protein-positive material with gangliosides into the organic solvents is discussed. Chromatographic search of gangliosides in different regions of the human brain revealed the presence of small quantities of more complex gangliosides than GQ1b.
本文描述了一种从人脑中分离神经节苷脂GQ1b和其他复杂神经节苷脂的简单方法。用氯仿、甲醇和水的混合物提取组织,提取物通过硅胶柱色谱和制备型高效薄层层析纯化两次。省略了相分配和离子交换色谱步骤。硅胶色谱基于两步展开系统,可有效分离低唾液酸神经节苷脂。从一名60岁女性的全脑、小脑皮质和枕叶灰质中分离出的色谱纯神经节苷脂GQ1b级分,每克新鲜组织的唾液酸产量分别为62、138和110 nmol。讨论了蛋白质阳性物质与神经节苷脂共萃取到有机溶剂中的问题。对人脑不同区域神经节苷脂的色谱研究表明,存在少量比GQ1b更复杂的神经节苷脂。