An exploratory approach for an oriented development of an untargeted hydrophilic interaction liquid chromatography-mass spectrometry platform for polar metabolites in biological matrices
Iturrospe, Elias; Da Silva, Katyeny Manuela; Talavera Andujar, Begonaet al.
[en] The analysis of polar metabolites based on liquid chromatography-mass spectrometry (LC-MS) methods should take into consideration the complexity of interactions in LC columns to be able to cover a broad range of metabolites of key biological pathways. Therefore, in this study, different chromatographic columns were tested for polar metabolites including reversed-phase and hydrophilic interaction liquid chromatography (HILIC) columns. Based on a column screening, two new generations of zwitterionic HILIC columns were selected for further evaluation. A tree-based method optimization was applied to investigate the chromatographic factors affecting the retention mechanisms of polar metabolites with zwitterionic stationary phases. The results were evaluated based on a scoring system which was applied for more than 80 polar metabolites with a high coverage of key human metabolic pathways. The final optimized methods showed high complementarity to analyze a wide range of metabolic classes including amino acids, small peptides, sugars, amino sugars, phosphorylated sugars, organic acids, nucleobases, nucleosides, nucleotides and acylcarnitines. Optimized methods were applied to analyze different biological matrices, including human urine, plasma and liver cell extracts using an untargeted approach. The number of high-quality features ( < 30% median relative standard deviation) ranged from 3,755 for urine to 5,402 for the intracellular metabolome of liver cells, showing the potential of the methods for untargeted purposes.
Disciplines :
Pharmacy, pharmacology & toxicology
Author, co-author :
Iturrospe, Elias; Universiteit Antwerpen - UA > Toxicological Centre
Da Silva, Katyeny Manuela; Universiteit Antwerpen - UA > Toxicological Centre
Talavera Andujar, Begona ; University of Luxembourg > Luxembourg Centre for Systems Biomedicine (LCSB) > Environmental Cheminformatics
Cuykx, Matthias; Universiteit Antwerpen - UA > Toxicological Centre
Boeckmans, Joost; Vrije Universiteit Brussel - VUB > Department of In Vitro Toxicology and Dermato- Cosmetology
Vanhaecke, Tamara; Vrije Universiteit Brussel - VUB > Department of In Vitro Toxicology and Dermato-cosmetology
Covaci, Adrian; Universiteit Antwerpen - UA > Toxicological Centre
Van Nuijs, Alexander L.N.; Universiteit Antwerpen - UA > Toxicological Centre
External co-authors :
yes
Language :
English
Title :
An exploratory approach for an oriented development of an untargeted hydrophilic interaction liquid chromatography-mass spectrometry platform for polar metabolites in biological matrices