Lipidomics by ultrahigh performance liquid chromatography-high resolution mass spectrometry and its application to complex biological samples

Alexander Triebl, M. Trötzmüller, Jürgen Hartler, Tatjana Stojakovic, Harald C. Köfeler

    Research output: Contribution to journalArticlepeer-review

    Abstract

    An improved approach for selective and sensitive identification and quantitation of lipid molecular species using reversed phase chromatography coupled to high resolution mass spectrometry was developed. The method is applicable to a wide variety of biological matrices using a simple liquid-liquid extraction procedure. Together, this approach combines multiple selectivity criteria: Reversed phase chromatography separates lipids according to their acyl chain length and degree of unsaturation and is capable of resolving positional isomers of lysophospholipids, as well as structural isomers of diacyl phospholipids and glycerolipids. Orbitrap mass spectrometry delivers the elemental composition of both positive and negative ions with high mass accuracy. Finally, automatically generated tandem mass spectra provide structural insight into numerous glycerolipids, phospholipids, and sphingolipids within a single run. Calibration showed linearity ranges of more than four orders of magnitude, good values for accuracy and precision at biologically relevant concentration levels, and limits of quantitation of a few femtomoles on column. Hundreds of lipid molecular species were detected and quantified in three different biological matrices, which cover well the wide variety and complexity of various model organisms in lipidomic research. Together with a software package, this method is a prime choice for global lipidomic analysis of even the most complex biological samples.

    Original languageEnglish
    Pages (from-to)72-80
    Number of pages9
    JournalJournal of Chromatography B
    Volume1053
    DOIs
    Publication statusPublished - 15 May 2017

    Keywords

    • Animals
    • Caenorhabditis elegans
    • Cell Line, Tumor
    • Chromatography, High Pressure Liquid
    • Chromatography, Reverse-Phase
    • Humans
    • Isomerism
    • Lipids
    • Liquid-Liquid Extraction
    • Mass Spectrometry
    • Metabolomics
    • Mice
    • Journal Article

    Fields of Expertise

    • Human- & Biotechnology

    Treatment code (Nähere Zuordnung)

    • Experimental

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