Abstract
LC-MS-based metabolomics is a fast-spreading approach to study microorganisms and their metabolism. This work is focused on LC-MS metabolomics of polar compounds, which are not retained on chromatographic columns using the well-established reversed-phase mechanism.
The HILIC and multimode columns are expected to improve the retention characteristics of polar compounds. In the experimental part, four HILIC columns and two mixed-mode columns were tested.
The columns were compared with a CSH-C18 column, using a set of 258 polar, human and microbial metabolites with a high diversity of their physicochemical properties. All HILIC columns significantly extended the range of the CSH-C18 column.
In terms of the number of compounds detected, the BEH Amide PEEK column using ammonium acetate pH = 3 had the highest ability to analyze a wide spectrum of polar metabolites. The combination of the mentioned results is promising for the use of this method in untargeted metabolomics.