通过自动多次展开实现GM1b型神经节苷脂的高效薄层色谱分离

Enhanced thin-layer chromatographic separation of GM1b-type gangliosides by automated multiple development.

作者信息

Müthing J, Ziehr H

机构信息

Institut für Zellkulturtechnik, Universität Bielefeld, Germany.

出版信息

J Chromatogr B Biomed Appl. 1996 Dec 13;687(2):357-62. doi: 10.1016/s0378-4347(96)00258-7.

DOI:10.1016/s0378-4347(96)00258-7

PMID:9017458

Abstract

Enhancement in separation of gangliosides on silica gel precoated high-performance TLC plates has been obtained by automated multiple development chromatography. A less polar mixture of the standard solvent chloroform-methanol-20 mM aqueous CaCl2 (120:85:20, v/v) was used. Lowering the water content achieved separation of two complex monosialoganglioside fractions, isolated from murine YAC 1 T lymphoma and MDAY-D2 lymphoreticular cells. Three-fold chromatography in the solvent chloroform-methanol-20 mM aqueous CaCl2 (120:85:14, v/v) resulted in TLC separation of GM1b-type gangliosides, substituted with C24 and C16 fatty acids and with Neu5Ac and Neu5Gc as well, which could not be achieved by unidirectional standard chromatography. Compared to conventional single chromatography, the technique described allows high-resolution separation of extremely heterogenous ganglioside mixtures and offers a convenient tool for both analytical and preparative TLC.

摘要

通过自动多次展开色谱法，已实现硅胶预涂高效薄层色谱板上神经节苷脂分离的增强。使用了标准溶剂氯仿 - 甲醇 - 20 mM 氯化钙水溶液（120:85:20，v/v）的极性较小的混合物。降低水含量实现了从鼠 YAC 1 T 淋巴瘤和 MDAY - D2 淋巴网状细胞中分离出的两种复杂单唾液酸神经节苷脂组分的分离。在氯仿 - 甲醇 - 20 mM 氯化钙水溶液（120:85:14，v/v）中进行三次展开色谱，实现了 GM1b 型神经节苷脂的薄层色谱分离，这些神经节苷脂被 C24 和 C16 脂肪酸取代，并且还含有 Neu5Ac 和 Neu5Gc，而单向标准色谱法无法实现这种分离。与传统的单次色谱法相比，所述技术能够对极其异质的神经节苷脂混合物进行高分辨率分离，并为分析型和制备型薄层色谱提供了一种便捷工具。