Optimization of normal phase chromatographic conditions for lipid analysis and comparison of associated detection techniques
Optimization of normal phase chromatographic conditions for lipid analysis and comparison of associated detection techniques
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DOI：
10.1016/j.chroma.2017.07.063
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发表时间：
2017-09-08
影响因子：
4.1
通讯作者：
Chaminade, Pierre
中科院分区：
文献类型：
--
作者：
Abreu, Sonia;Solgadi, Audrey;Chaminade, Pierre
脂质类分析中的一个重要挑战是开发一种适合或至少可适应各种样品的方法。在目前的研究中，一种改进的正相液相色谱（NPLC）方法允许分析哺乳动物，植物以及微生物（酵母和细菌）脂质样品中存在的脂质类。该方法有效地分离了30种脂质类或亚类，特别关注中等极性脂质。分离采用裸二氧化硅固定相进行，并与蒸发光散射检测（ELSD）、荷电气溶胶检测（Corona-CAD（R））和质谱联用。提供解决方案以规避技术问题（例如泵送低粘度溶剂、溶剂纯度、冲洗步骤）。记录了移动的相组成和添加离子改性剂对特定脂质类色谱行为的影响。ELSD和Corona-CAD（R）之间的比较证实了这款后来的检测器对具有广泛浓度的不同脂质的样本的兴趣。使用三个常见的大气压电离接口将NPLC分离耦合到LTQ Velos Pro（R）质谱仪。色谱图的比较表明，大气压化学电离（APCI）和大气压光电离（APPI）都适合于检测不同的脂质类，而APPI允许在低浓度的脂质更好的灵敏度。(C)2017爱思唯尔B. V.保留所有权利。
One important challenge in lipid class analysis is to develop a method suitable or, at least adaptable, for a vast diversity of samples. In the current study, an improved normal-phase liquid chromatography (NPLC) method allowed analyzing the lipid classes present in mammalian, vegetable as well as microorganism (yeast and bacteria) lipid samples. The method effectively separated 30 lipid classes or subclasses with a special focus on medium polarity lipids. The separation was carried out with bare silica stationary phase and was coupled to evaporative light scattering detection (ELSD), charged aerosol detection (Corona-CAD (R)) and mass spectrometry. Solutions are provided to circumvent technical issues (such as pumping solvents of low viscosity, solvent purity, rinsing step). The influence of mobile phase composition and addition of ionic modifiers on the chromatographic behavior of particular lipid classes is documented. A comparison between ELSD and Corona-CAD (R) confirmed the interest of this later detector for samples with a wide range of concentration of different lipids. Three common atmospheric pressure ionization interfaces were used for coupling the NPLC separation to a LTQ Velos Pro (R) mass spectrometer. The comparison of the chromatographic profiles showed that atmospheric pressure chemical ionization (APCI) and atmospheric pressure photoionization (APPI) are both suitable to detect the different lipid classes whereas APPI allows a better sensitivity for lipids at low-concentration. (C) 2017 Elsevier B.V. All rights reserved.