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Non-Targeted Quantitation of Lipid Classes Using Hydrophilic Interaction Liquid Chromatography - Electrospray Ionization Mass Spectrometry Using Single Internal Standard and Response Factor Approach
Authors: Cífková Eva | Holčapek Michal | Lísa Miroslav | Ovčačíková Magdaléna | Lyčka Antonín | Lynen Frederic | Sandra Pat
Year: 2012
Type of publication: článek v odborném periodiku
Name of source: Analytical Chemistry
Publisher name: American Chemical Society
Place: Washington
Page from-to: 10064-10070
Titles:
Language Name Abstract Keywords
cze Necílená kvantitativní analýza tříd lipidů pomocí HILIC-HPLC/ESI-MS s použitím jednoho interního standardu a odezvových faktorů V této práci byla popsána kvantitativní analýza polárních tříd lipidů pomocí kapalinové chromatografie hydrofilních interakcí a hmotnostní spektrometrie s ionizací elektrosprejem s využitím jednoho interního standardu a odezvových faktorů jednotlivých tříd. Necílená; kvantitativní analýza; lipidy; HILIC; ESI-MS; použitím
eng Non-Targeted Quantitation of Lipid Classes Using Hydrophilic Interaction Liquid Chromatography - Electrospray Ionization Mass Spectrometry Using Single Internal Standard and Response Factor Approach The identification and quantitation of a wide range of lipids in complex biological samples is an essential requirement for the lipidomic studies. High-performance liquid chromatography−mass spectrometry (HPLC/MS) has the highest potential to obtain detailed information on the whole lipidome, but the reliable quantitation of multiple lipid classes is still a challenging task. In this work, we describe a new method for the nontargeted quantitation of polar lipid classes separated by hydrophilic interaction liquid chromatography (HILIC) followed by positive-ion electrospray ionization mass spectrometry (ESI-MS) using a single internal lipid standard to which all class specific response factors (RFs) are related to. The developed method enables the nontargeted quantitation of lipid classes and molecules inside these classes in contrast to the conventional targeted quantitation, which is based on predefined selected reaction monitoring (SRM) transitions for selected lipids only. In the nontargeted quantitation method described here, concentrations of lipid classes are obtained by the peak integration in HILIC chromatograms multiplied by their RFs related to the single internal standard (i.e., sphingosyl PE, d17:1/12:0) used as common reference for all polar lipid classes. The accuracy, reproducibility and robustness of the method have been checked by various means: (1) the comparison with conventional lipidomic quantitation using SRM scans on a triple quadrupole (QqQ) mass analyzer, (2) 31P nuclear magnetic resonance (NMR) quantitation of the total lipid extract, (3) method robustness test using subsequent measurements by three different persons, (4) method transfer to different HPLC/MS systems using different chromatographic conditions, and (5) comparison with previously published results for identical samples, especially human reference plasma from the National Institute of Standards and Technology (NIST human plasma). lipidomics; quantitation; hydrophilic interaction liquid chromatography; mass spectrometry; phospholipids; response factor; nuclear magnetic resonance.

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