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Hydrophilic interaction liquid chromatography-mass spectrometry of (lyso)phosphatidic acids, (lyso)phosphatidylserines and other lipid classes
Authors: Cífková Eva | Hájek Roman | Lísa Miroslav | Holčapek Michal
Year: 2016
Type of publication: článek v odborném periodiku
Name of source: Journal of Chromatography A
Publisher name: Elsevier Science BV
Place: Amsterdam
Page from-to: 65-73
Titles:
Language Name Abstract Keywords
cze Hydrofilní interakční chromatografie ve spojen s hmotnostní spektrometrií pro stanovení kyseliny fosfatidové, fosfatidylserinu a jejich lysoderivátů. Cílem této práce byla optimalizace separace polárních lipidů zejména kyseliny fosfatidové, fosfatidylserinu a jejich lysoderivátů. Optimalizace byla provedena pomocí kapalinové chromatografie (HILIC) ve spojení s hmotnostní spektrometrií (ESI). Byly testovány kolony a vliv pH a koncentrace aditiv. Vyvinutá metoda byla použita pro analýzu mozku a ledvin prasete. Hydrofilní interakční chromatografie; hmotnostní spektrometrie; kyselina fosfatidová; fosfatidylserin
eng Hydrophilic interaction liquid chromatography-mass spectrometry of (lyso)phosphatidic acids, (lyso)phosphatidylserines and other lipid classes 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. Phosphatidic acid; Phosphatidylserine; Phospholipids; Lysophospholipids; HILIC; LC/MS

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