title: Hydrophilic interaction liquid chromatography-mass spectrometry of (lyso)phosphatidic acids, (lyso)phosphatidylserines and other lipid classes.
abstract: The goal of this work is a systematic optimization of hydrophilic interaction liquid chromatography (HILIC) separation of acidic lipid classes (namely phosphatidic acids-PA, lysophosphatidic acids-LPA, phosphatidylserines-PS and lysophosphatidylserines-LPS) and other lipid classes under mass spectrometry (MS) compatible conditions. The main parameters included in this optimization are the type of stationary phases used in HILIC, pH of the mobile phase, the type and concentration of mobile phase additives. Nine HILIC columns with different chemistries (unmodified silica, modified silica using diol, 2-picolylamine, diethylamine and 1-aminoanthracene and hydride silica) are compared with the emphasis on peak shapes of acidic lipid classes. The optimization of pH is correlated with the theoretical calculation of acidobasic equilibria of studied lipid classes. The final method using the hydride column, pH 4 adjusted by formic acid and the gradient of acetonitrile and 40 mmol/L of aqueous ammonium formate provides good peak shapes for all analyzed lipid classes including acidic lipids. This method is applied for the identification of lipids in real samples of porcine brain and kidney extracts.
authors: Department of Analytical Chemistry, Faculty of Chemical Technology, University of Pardubice, Studentská 573, 53210 Pardubice, Czech Republic.
authors: Eva
authors: E
authors: Cífková
authors: Department of Analytical Chemistry, Faculty of Chemical Technology, University of Pardubice, Studentská 573, 53210 Pardubice, Czech Republic.
authors: Roman
authors: R
authors: Hájek
authors: Department of Analytical Chemistry, Faculty of Chemical Technology, University of Pardubice, Studentská 573, 53210 Pardubice, Czech Republic.
authors: Miroslav
authors: M
authors: Lísa
authors: Department of Analytical Chemistry, Faculty of Chemical Technology, University of Pardubice, Studentská 573, 53210 Pardubice, Czech Republic. Electronic address: Michal.Holcapek@upce.cz.
authors: Michal
authors: M
authors: HolĿapek
mesh_terms: D000818:Animals; D001923:Brain Chemistry; D002853:Chromatography, Liquid / Q000379:methods; D006863:Hydrogen-Ion Concentration; D057927:Hydrophobic and Hydrophilic Interactions; D007668:Kidney / Q000737:chemistry; D008055:Lipids / Q000032:analysis*; D008246:Lysophospholipids / Q000032:analysis; D013058:Mass Spectrometry / Q000379:methods; D010718:Phosphatidylserines / Q000032:analysis; D012822:Silicon Dioxide; D013237:Stereoisomerism; D013552:Swine
chemical_list: D008055:Lipids; D008246:Lysophospholipids; D010718:Phosphatidylserines; C025059:lysophosphatidylserine; D012822:Silicon Dioxide
keywords: HILIC; LC/MS; Lysophospholipids; Phosphatidic acid; Phosphatidylserine; Phospholipids