Svennerholm L, Fredman P
Biochim Biophys Acta. 1980 Jan 18;617(1):97-109. doi: 10.1016/0005-2760(80)90227-1.
In a systematic study of the optimal conditions for the quantitative isolation of gangliosides from brain tissue and their further purification the yield of gangliosides obtained by extraction of the tissue twice with twenty volumes of chloroform/methanol/water (4 : 8 : 3, v/v) was larger than that obtained with all other solvents tested, including tetrahydrofuran/phosphate buffer. The gangliosides were separated from other lipids by phase partition, water was added to the total lipid extract to give a final chloroform/methanol/water volume ratio of 4 : 8 : 5.6. Isolation of gangliosides from the total lipid extract with the aid of anion-exchange resins was not practical as a routine procedure on a large scale. The crude gangliosides extract was freed from low molecular weight contaminants by dialysis against water. This method was superior to the purification on gel filtration media or on anion-exchange resins, which required large columns with selective losses of gangliosides as a result. The present method has been applied to human brain, and the concentration and distribution of gangliosides in the human forebrain in infancy and old age are given.
在一项关于从脑组织中定量分离神经节苷脂及其进一步纯化的最佳条件的系统研究中,用二十倍体积的氯仿/甲醇/水(4∶8∶3,v/v)对组织进行两次提取所获得的神经节苷脂产量,高于用包括四氢呋喃/磷酸盐缓冲液在内的所有其他测试溶剂所获得的产量。通过相分配将神经节苷脂与其他脂质分离,向总脂质提取物中加水,使氯仿/甲醇/水的最终体积比为4∶8∶5.6。借助阴离子交换树脂从总脂质提取物中分离神经节苷脂,作为一种大规模常规方法并不实用。粗制神经节苷脂提取物通过对水透析除去低分子量污染物。该方法优于在凝胶过滤介质或阴离子交换树脂上的纯化方法,后两种方法需要大柱子,结果会导致神经节苷脂有选择性损失。本方法已应用于人类大脑,并给出了婴儿期和老年期人类前脑中神经节苷脂的浓度和分布情况。