A single run LC-MS/MS method for phospholipidomics
A single run LC-MS/MS method for phospholipidomics
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DOI：
10.1007/s00216-012-6466-9
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发表时间：
2013-01-01
影响因子：
4.3
通讯作者：
Schmitter, Jean-Marie
中科院分区：
文献类型：
--
作者：
Bure, Corinne;Ayciriex, Sophie;Schmitter, Jean-Marie
液相色谱-串联质谱法已被比较，鸟枪分析的目的是找到一个完整的细胞磷脂的单次运行分析的最佳妥协。采用属于磷脂酸（PA）、磷脂酰胆碱（PC）、磷脂酰乙醇胺（PE）、磷脂酰甘油（PG）、磷脂酰肌醇（PI）和磷脂酰丝氨酸（PS）类的参比磷脂评价了亲水相互作用液相色谱（HILIC）、正相（NP）和反相（RP）液相色谱。NP-HPLC-和RP-HPLC-ESI-MS/MS应用于酵母磷脂组分析，使用野生型菌株和两种已知参与从头磷脂合成或磷脂重塑的酰基转移酶缺陷菌株。基于含有奇数碳原子脂肪酸链的参比磷脂，使用MRM模式对单个化合物进行相对定量。发现组合LC-MS/MS上级于鸟枪法分析，导致比鸟枪法分析更大数量的定量物质。最后，RP-HPLC-MS/MS因其更高的选择性、耐用性和更好的重复性而成为首选方法。
Liquid chromatography coupled to tandem mass spectrometry has been compared to shotgun analysis with the objective of finding the best compromise for a single run analysis of whole cell phospholipids. Hydrophilic interaction liquid chromatography (HILIC), normal phase (NP), and reversed phase (RP) liquid chromatography were evaluated with reference phospholipids belonging to phosphatidic acid (PA), phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylglycerol (PG), phosphatidylinositol (PI), and phosphatidylserine (PS) classes. NP-HPLC- and RP-HPLC-ESI-MS/MS were applied to yeast phospholipidome analysis, using a wild-type strain and two strains defective for acyltransferases that are known to be involved in de novo phospholipid synthesis or phospholipid remodeling. The MRM mode was used for relative quantitation of individual compounds based on reference phospholipids bearing fatty acid chains with an odd number of carbon atoms. Combined LC-MS/MS was found superior to shotgun analysis, leading to a larger number of quantified species than shotgun analysis. Finally, RP-HPLC-MS/MS was the preferred method for its higher selectivity, robustness, and better repeatability.