Phospholipids in liquid chromatography/mass spectrometry bioanalysis: comparison of three tandem mass spectrometric techniques for monitoring plasma phospholipids, the effect of mobile phase composition on phospholipids elution and the association of phospholipids with matrix effects
Phospholipids in liquid chromatography/mass spectrometry bioanalysis: comparison of three tandem mass spectrometric techniques for monitoring plasma phospholipids, the effect of mobile phase composition on phospholipids elution and the association of phospholipids with matrix effects
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DOI：
10.1002/rcm.4121
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发表时间：
2009-07-01
影响因子：
2
通讯作者：
Jemal, Mohammed
中科院分区：
文献类型：
--
作者：
Xia, Yuan-Qing;Jemal, Mohammed
由于血浆磷脂可能会导致生物分析液相色谱/串联质谱（LC/MS/MS）方法中的基质效应，因此在方法开发和应用过程中建立最佳质谱技术以监测磷脂的去向非常重要。我们评估了三种MS/MS技术，使用正和负电喷雾电离（ESI）监测磷脂。第一种技术基于使用m/z 184的正前体离子扫描、141 Da的正中性损失扫描和m/z 153的负前体离子扫描。第二种技术是基于使用类别特异性阳性和阴性选择反应监测（SRM）转换来监测类别代表性磷脂。先前报道的第三种技术利用基于源内碰撞诱导解离（CID）的m/z 184 -> 184的正SRM。我们建议将全包技术1用于所有类别磷脂的定性评估，将技术2用于类别代表性磷脂的定量评估。其次，我们评价了血浆磷脂在不同反相移动的条件下的洗脱行为。移动的相对磷脂的洗脱强度主要取决于有机洗脱剂的种类和用量M，洗脱强度按甲醇、乙腈和异丙醇的顺序增大。在LC/MS/MS生物分析中常用的梯度和等度洗脱方案下，并非所有磷脂都从色谱柱上洗脱下来。第三，我们使用碱性、酸性和中性分析物研究了磷脂和基质效应在阳性和阴性EST中的关联。虽然磷脂引起的基质效应，在积极和消极的EST，电离抑制的程度是分析依赖性的，并呈负相关的保留因子和磷脂峰的宽度。通常在反相色谱中较早洗脱的溶血磷脂与较晚洗脱的磷脂相比更可能引起基质效应，尽管后者在血浆中的浓度较大。版权所有（C）2009约翰威利父子有限公司
Because plasma phospholipids may cause matrix effects in bioanalytical liquid chromatography/tandem mass spectrometry (LC/MS/MS) methods, it is important to establish optimal mass spectrometric techniques to monitor the fate of phospholipids during method development and application. We evaluated three MS/MS techniques to monitor phospholipids using positive and negative electrospray ionization (ESI). The first technique is based on using positive precursor ion scan of m/z 184, positive neutral loss scan of 141 Da and negative precursor ion scan of m/z 153. The second technique is based on using class-specific positive and negative selected reaction monitoring (SRM) transitions to monitor class-representative phospholipids. The third technique, previously reported, utilizes in-source collision-induced dissociation (CID)-based positive SRM of m/z 184 -> 184. We recommend the all-inclusive technique 1 for use in qualitative assessment of all classes of phospholipids and technique 2 for use in quantitative assessment of class-representative phospholipids. Secondly, we evaluated the elution behaviors of the plasma phospholipids under different reversed-phase mobile phase conditions. The phospholipid-eluting strength of a mobile phase was mainly dependent on the type and amount M of the organic eluent and the strength increased in the order of methanol, acetonitrile and isopropyl alcohol. Under the commonly used gradient and isocratic elution schemes in LC/MS/MS bioanalysis, not all the phospholipids are eluted off the column. Thirdly, we investigated the association between phospholipids and matrix effects in positive and negative EST using basic, acidic and neutral analytes. While the phospholipids caused matrix effects in both positive and negative EST, the extent of ionization suppression was analyte-dependent and was inversely related to the retention factor and broadness of the phospholipids peaks. The lysophospholipids which normally elute earlier in reversed-phase chromatography are more likely to cause matrix effects compared to the later-eluting phospholipids in spite of the larger concentrations of the latter in plasma. Copyright (C) 2009 John Wiley & Sons, Ltd.