Chimie Biologie des Membranes et Nanoobjets CBMN-UMR 5248 Centre de Génomique Fonctionnelle Université Bordeaux 2, Université de Bordeaux, 146, rue Léo Saignat, 33076 Bordeaux cedex, France.
Anal Bioanal Chem. 2013 Jan;405(1):203-13. doi: 10.1007/s00216-012-6466-9. Epub 2012 Oct 13.
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.
液相色谱-串联质谱法已被用于与 shotgun 分析进行比较，旨在寻找单次全细胞磷脂分析的最佳折衷方案。亲水相互作用液相色谱 (HILIC)、正相 (NP) 和反相 (RP) 液相色谱分别参照属于磷脂酸 (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) 类的磷脂进行了评估。NP-HPLC- 和 RP-HPLC-ESI-MS/MS 被应用于酵母磷脂组学分析，使用野生型菌株和两种已知参与从头合成或磷脂重塑的酰基转移酶缺陷菌株。MRM 模式用于基于含有奇数碳原子脂肪酸链的参考磷脂对单个化合物进行相对定量。组合 LC-MS/MS 优于 shotgun 分析，比 shotgun 分析鉴定出更多的定量物质。最后，RP-HPLC-MS/MS 因其更高的选择性、稳健性和更好的重现性而成为首选方法。
