Holistic Lipidomics of the Human Gut Phenotype Using Validated Ultra-High-Performance Liquid Chromatography Coupled to Hybrid Orbitrap Mass Spectrometry

L. Van Meulebroek, E. De Paepe, V. Vercruysse, B. Pomian, S. Bos, B. Lapauw, L. Vanhaecke

Research output: Contribution to journalArticlepeer-review

29 Citations (Scopus)


As lipids are assigned a plethora of biological functions, it is evident that dysregulated lipid metabolism signifies a key element in many pathological conditions. With this rationale, this study presents a validated lipidomics platform to map the fecal lipidome, which integrates unique information about host-gut microbiome interactions, gastrointestinal functionality, and dietary patterns. This particular method accomplished coverage across all eight lipid categories: fatty acyls, glycerolipids, phosphoglycerolipids, polyketides, prenols, saccharolipids, sphingolipids, and sterols. Generic extraction of freeze-dried feces was achieved by solid-liquid extraction using methanol and methyl tert-butyl ether. Extracted components were separated by liquid chromatography, whereby the selected ethylene-bridged hybrid phenyl ultra-high-performance liquid chromatography stationary phase allowed fast separation of both individual lipid species and categories. Detection was achieved by high-resolution full-scan Q-Exactive Orbitrap mass spectrometry and covered a broad m/z scan range (67-2300 Da). Method validation was performed in a targeted fashion to evaluate the analytical performance across all lipid categories, revealing excellent linearity (R2 ≥ 0.9921), acceptable repeatability (coefficients of variance ≤15.6%), and stable recovery (coefficients of variance ≤11.9%). Method suitability for untargeted fingerprinting was verified, demonstrating adequate linearity (R2 ≥ 0.90) for 75.3% and acceptable repeatability (coefficients of variance ≤30%) for 84.5% of about 9000 endogenous fecal compounds. Eventually, the potential of fecal lipidomics was exemplified within a clinical context of type 2 diabetes, thereby revealing significant perturbations [orthogonal partial least-squares discriminant analysis Q2(Y) of 0.728] in the fecal lipidome between participants with normal blood glucose levels (n = 26) and those with type 2 diabetes (n = 17). © 2017 American Chemical Society.
Original languageEnglish
Pages (from-to)12502-12510
JournalAnalytical Chemistry
Issue number22
Early online date20 Oct 2017
Publication statusPublished - 21 Nov 2017

Bibliographical note

cited By 2


  • Chromatography
  • Discriminant analysis
  • Ethylene
  • Extraction
  • High performance liquid chromatography
  • Least squares approximations
  • Lipids
  • Liquids
  • Mass spectrometry
  • Spectrometry, Analytical performance
  • Coefficients of variances
  • Methyl tert butyl ether
  • Orbitrap mass spectrometries
  • Partial least squares - discriminant analysis
  • Pathological conditions
  • Solid-liquid extraction
  • Ultra-high performance liquid chromatographies, Liquid chromatography


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