用于从酵母中分离磷脂和中性脂质的薄层色谱法。

Thin-Layer Chromatography to Separate Phospholipids and Neutral Lipids from Yeast.

作者信息

Knittelfelder Oskar L, Kohlwein Sepp D

机构信息

Institute of Molecular Biosciences, University of Graz, BioTechMed Graz, 8010 Graz, Austria.

Institute of Molecular Biosciences, University of Graz, BioTechMed Graz, 8010 Graz, Austria

出版信息

Cold Spring Harb Protoc. 2017 May 1;2017(5):2017/5/pdb.prot085456. doi: 10.1101/pdb.prot085456.

DOI:10.1101/pdb.prot085456

PMID:28461652

Abstract

Thin-layer chromatography (TLC) is a versatile technique for the separation of lipid classes. It provides excellent resolution, can be used for both preparative and analytical applications, and does not require expensive equipment. Here we describe the use of different solvent systems to separate yeast phospholipids and neutral lipids by TLC in one dimension. Resolved lipid species are visualized by iodine vapor or by charring after treatment with sulfuric acid and manganese chloride. Neither of these staining techniques yields a quantitative readout because the mixture of various lipids in yeast affects iodine absorption and charring efficiency; standard curves are required to obtain semiquantitative estimates of the relative lipid composition.

摘要

薄层色谱法（TLC）是一种用于分离脂质类别的通用技术。它具有出色的分辨率，可用于制备和分析应用，且不需要昂贵的设备。在此，我们描述了使用不同的溶剂系统通过一维TLC分离酵母磷脂和中性脂质的方法。分离出的脂质种类通过碘蒸气或用硫酸和氯化锰处理后炭化来可视化。这两种染色技术都无法产生定量读数，因为酵母中各种脂质的混合物会影响碘吸收和炭化效率；需要标准曲线来获得脂质相对组成的半定量估计。