title: Characterization of complex lipid mixtures in contaminant exposed JEG-3 cells using liquid chromatography and high-resolution mass spectrometry.
abstract: The aim of this study was to develop a method based on ultra high performance liquid chromatography coupled with mass spectrometry (UHPLC-MS) for lipid profiling in human placental choriocarcinoma (JEG-3) cells. Lipids were solid-liquid extracted from JEG-3 cells using a solution of chloroform/methanol (2:1, v/v) in a simple procedure requiring minimal sample alteration. Simultaneous separation of complex lipid mixtures in their major classes was achieved with a reversed-phase (C8) UHPLC column and a mobile phase containing methanol with 1 mM ammonium formate and 0.2 % formic acid (A)/water with 2 mM ammonium formate and 0.2 % formic acid (B). Lipids were characterized using time-of-flight (TOF) and Orbitrap under full scan and positive electrospray ionization mode with both analyzers. A total of 178 species of lipids, including 37 phosphatidylcholines (PC), 32 plasmalogen PC, 9 lyso PC, 4 lyso plasmalogen PC, 30 triacylglycerols, 22 diacylglycerols, 7 cholesterol esters, 25 phosphatidylethanolamines, and 12 sphingomyelins, were identified using TOF and Orbitrap. The identification of all lipid classes was based on exact mass characterization with an error < 5 ppm. The developed methodology was applied to study lipid alterations in human placental cells against the exposure to perfluorinated chemicals (PFCs) and tributyltin (TBT).
authors: Department of Environmental Chemistry, Institute of Environmental Assessment and Water Research (IDAEA), Consejo Superior de Investigaciones Científicas (CSIC), Jordi Girona 18-26, Barcelona, 08034, Catalonia, Spain.
authors: Eva
authors: E
authors: Gorrochategui
authors: Josefina
authors: J
authors: Casas
authors: Elisabet
authors: E
authors: Pérez-Albaladejo
authors: Olga
authors: O
authors: Jáuregui
authors: Cinta
authors: C
authors: Porte
authors: Sílvia
authors: S
authors: Lacorte
mesh_terms: D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005466:Fluorocarbons / Q000633:toxicity*; D006801:Humans; D008055:Lipids / Q000032:analysis* / Q000302:isolation & purification; D013058:Mass Spectrometry / Q000379:methods*; D055442:Metabolome / Q000187:drug effects; D014220:Trialkyltin Compounds / Q000633:toxicity*
chemical_list: D005466:Fluorocarbons; D008055:Lipids; D014220:Trialkyltin Compounds; C011559:tributyltin