Tomas Cajka, Jiri Hricko, Stanislava Rakusanova, Kristyna Brejchova, Michaela Novakova, Lucie Rudl Kulhava, Veronika Hola, Michaela Paucova, Oliver Fiehn, Ondrej Kuda
International journal of molecular sciences 2024 Mar 01Liquid chromatography with mass spectrometry (LC-MS)-based metabolomics detects thousands of molecular features (retention time-m/z pairs) in biological samples per analysis, yet the metabolite annotation rate remains low, with 90% of signals classified as unknowns. To enhance the metabolite annotation rates, researchers employ tandem mass spectral libraries and challenging in silico fragmentation software. Hydrogen/deuterium exchange mass spectrometry (HDX-MS) may offer an additional layer of structural information in untargeted metabolomics, especially for identifying specific unidentified metabolites that are revealed to be statistically significant. Here, we investigate the potential of hydrophilic interaction liquid chromatography (HILIC)-HDX-MS in untargeted metabolomics. Specifically, we evaluate the effectiveness of two approaches using hypothetical targets: the post-column addition of deuterium oxide (D2O) and the on-column HILIC-HDX-MS method. To illustrate the practical application of HILIC-HDX-MS, we apply this methodology using the in silico fragmentation software MS-FINDER to an unknown compound detected in various biological samples, including plasma, serum, tissues, and feces during HILIC-MS profiling, subsequently identified as N1-acetylspermidine.
Tomas Cajka, Jiri Hricko, Stanislava Rakusanova, Kristyna Brejchova, Michaela Novakova, Lucie Rudl Kulhava, Veronika Hola, Michaela Paucova, Oliver Fiehn, Ondrej Kuda. Hydrophilic Interaction Liquid Chromatography-Hydrogen/Deuterium Exchange-Mass Spectrometry (HILIC-HDX-MS) for Untargeted Metabolomics. International journal of molecular sciences. 2024 Mar 01;25(5)
PMID: 38474147
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