An exploratory approach for an oriented development of an untargeted hydrophilic interaction liquid chromatography-mass spectrometry platform for polar metabolites in biological matrices

The analysis of polar metabolites based on liquid chromatography-mass spectrometry (LC-MS) methods should take into consideration the complexity interactions in LC columns to be able cover a broad range key biological pathways. Therefore, this study, different chromatographic were tested for including reversed-phase and hydrophilic interaction chromatography (HILIC) columns. Based column screening, two new generations zwitterionic HILIC selected further evaluation. A tree-based method optimization was applied investigate factors affecting retention mechanisms with stationary phases. results evaluated scoring system which more than 80 high coverage human metabolic final optimized showed complementarity analyze wide classes amino acids, small peptides, sugars, phosphorylated organic nucleobases, nucleosides, nucleotides acylcarnitines. Optimized matrices, urine, plasma liver cell extracts using an untargeted approach. number high-quality features (< 30% median relative standard deviation) ranged from 3,755 urine 5,402 intracellular metabolome cells, showing potential purposes.