Merging metabolomics and lipidomics into one analytical run

A novel integrated metabolomics/lipidomics workflow is introduced enabling high coverage of polar metabolites and non-polar lipids within one analytical run. Dual HILIC and RP chromatography were combined to high-resolution mass spectrometry. As a major advantage, only one data file per sample was o...

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Bibliographic Details
Published in:Analyst (London) 2019-01, Vol.144 (1), p.220-229
Main Authors: Schwaiger, Michaela, Schoeny, Harald, El Abiead, Yasin, Hermann, Gerrit, Rampler, Evelyn, Koellensperger, Gunda
Format: Article
Language:English
Subjects:
Automation
Blood plasma
Chromatography
Chromatography, Liquid - methods
Chromatography, Reverse-Phase - methods
Data acquisition
Elution
Humans
Hydrophobic and Hydrophilic Interactions
Lipids
Lipids - blood
Mass spectrometry
Mass Spectrometry - methods
Mathematical analysis
Metabolites
Metabolomics - methods
Organic acids
Orthogonality
Polarity
Sugar
Triglycerides
Workflow
Yeast
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Summary:A novel integrated metabolomics/lipidomics workflow is introduced enabling high coverage of polar metabolites and non-polar lipids within one analytical run. Dual HILIC and RP chromatography were combined to high-resolution mass spectrometry. As a major advantage, only one data file per sample was obtained by fully automated simultaneous analysis of two extracts per sample. Hence, the unprecedented high coverage without compromise on analytical throughput was not only obtained by the orthogonality of the chromatographic separations, but also by the implementation of dedicated sample preparation procedures resulting in optimum extraction efficiency for both sub-omes. Thus, the method addressed completely hydrophilic sugars and organic acids next to water-insoluble triglycerides. As for the timing of the dual chromatography setup, HILIC and RP separation were performed consecutively. However, re-equilibration of the HILIC column during elution of RP compounds and vice versa reduced the overall analysis time by one third to 32 min. Application to the Standard Reference Material SRM 1950 - Metabolites in Frozen Human Plasma resulted in >100 metabolite and >380 lipid identifications based on accurate mass implementing fast polarity switching and acquiring data dependent MS2 spectra with the use of automated exclusion lists. Targeted quantification based on external calibrations and 13C labeled yeast internal standards was successfully accomplished for 59 metabolites. Moreover, the potential for lipid quantification was shown integrating non-endogenous lipids as internal standards. In human plasma, concentrations ranging over 4 orders of magnitude (low nM to high μM) were assessed.
ISSN:0003-2654
1364-5528
DOI:10.1039/c8an01219a