Groupe de Chimie Analytique de Paris-Sud, Faculty of Pharmacy, University Paris-Sud, EA 4041, IFR 141, 92296 Châtenay-Malabry, France.
Molecules. 2010 Jan 18;15(1):352-73. doi: 10.3390/molecules15010352.
Phospholipids are essential constituents of all living cell membranes. There are many analytical methods available for the quantitative and qualitative determination of phospholipids, but since these molecules lack chromophores, common absorbance based methods are of limited use. Beside mass spectrometry, some less specific approaches that are routinely used are evaporative light scattering detection or fluorescence, which exhibit sufficient sensitivity. Here, we focus on fluorescence, which remains an interesting way to quantify phospholipids. Two ways of detecting phospholipids by fluorescence are possible coupled with separation techniques such as thin layer chromatography (TLC), high performance liquid chromatography (HPLC) and capillary electrophoresis (CE): firstly, pre-column derivatization procedures and secondly, probe assisted post-column detection with suitable fluorescence reagents. In both cases, the common purpose is to increase the detection sensitivity. It is shown that, whereas pre-column derivatization is characterized by selectivity due to the chemical functionality of the analyte involved in the derivatization process, in supramolecular post-column derivatization, the selectivity only proceeds from the capacity of the lipid to involve supramolecular assemblies with a fluorescence probe. The aim of this review is to summarize available experiments concerning fluorescence detection of phospholipids. The interest and limitation of such detection approaches are discussed.
磷脂是所有活细胞膜的重要组成部分。有许多分析方法可用于定量和定性测定磷脂,但由于这些分子缺乏发色团,常见的基于吸光度的方法的用途有限。除了质谱法外,一些常规使用的不太特定的方法是蒸发光散射检测或荧光,它们具有足够的灵敏度。在这里,我们专注于荧光,它仍然是定量磷脂的一种有趣方法。通过与分离技术(如薄层色谱(TLC)、高效液相色谱(HPLC)和毛细管电泳(CE))结合,可以通过两种荧光检测方式来检测磷脂:首先,柱前衍生程序,其次,使用合适的荧光试剂进行柱后探针辅助检测。在这两种情况下,共同的目的是提高检测灵敏度。结果表明,尽管柱前衍生由于涉及衍生化过程的分析物的化学官能团而具有选择性,但在超分子柱后衍生中,选择性仅来自脂质与荧光探针形成超分子组装的能力。本文综述了有关磷脂荧光检测的现有实验。讨论了这种检测方法的优点和局限性。