Chromatographic behavior of oxygenated derivatives of cholesterol
Chromatographic behavior of oxygenated derivatives of cholesterol
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DOI：
10.1016/s0039-128x(02)00185-x
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发表时间：
2003-03-01
期刊：
影响因子：
2.7
通讯作者：
Schroepfer, GJ
中科院分区：
文献类型：
--
作者：
Shan, H;Pang, JH;Schroepfer, GJ
胆固醇的氧化衍生物在许多生物化学过程中具有重要的功能。这些氧化固醇难以研究，因为它们的生理浓度低，容易形成胆固醇自氧化伪影，并且缺乏关于其色谱行为的信息。专注于胆固醇的代谢产物和自氧化产物，我们已经记录了35氧化甾醇在各种条件下的色谱迁移率：8个溶剂系统的硅胶薄层色谱，几个移动的相的反相高效液相色谱（HPLC），和两种类型的固定相的毛细管气相色谱（GC）使用三甲基甲硅烷基衍生物。通过改变固定相或移动的相可以获得选择性的显著差异。在正相、反相、手性或银离子HPLC柱上实现了侧链碳或C-7处的氧甾醇对异构体的分离。色谱行为也描述了侧链hexadeutated和heptafluorescent氧化甾醇，这是有用的同位素稀释分析和自氧化研究，分别作为标准。总体结果与氧化甾醇分析的许多问题有关，包括氧化甾醇与胆固醇的初始分离、高极性和非极性氧化甾醇的测定、异构体对的分离、GC分析的衍生化条件的选择以及胆固醇自氧化程度的定量。(C)2002年爱思唯尔科学公司All rights reserved.
Oxygenated derivatives of cholesterol have important functions in many biochemical processes. These oxysterols are difficult to study because of their low physiological concentrations, the facile formation of cholesterol autoxidation artifacts, and lack of information on their chromatographic behavior. Focusing on metabolites and autoxidation products of cholesterol, we have documented the chromatographic mobilities of 35 oxysterols under a variety of conditions: eight solvent systems for thin-layer chromatography on silica gel, several mobile phases for reversed-phase high-performance liquid chromatography (HPLC), and two types of stationary phase for capillary gas chromatography (GC) using trimethylsilyl derivatives. Notable differences in selectivity could be obtained by modifying the stationary or mobile phases. Separations of oxysterol pairs isomeric at side-chain carbons or C-7 were achieved on normal-phase, reversed-phase, chiral, or silver-ion HPLC columns. Chromatographic behavior is also described for side-chain hexadeuterated and heptafluorinated oxysterols, which are useful as standards in isotope dilution analyses and autoxidation studies, respectively. The overall results are relevant to many problems of oxysterol analysis, including the initial separation of oxysterols from cholesterol, determination of highly polar and nonpolar oxysterols, separation of isomeric pairs, selection of derivatization conditions for GC analysis, and quantitation of the extent of cholesterol autoxidation. (C) 2002 Elsevier Science Inc. All rights reserved.