Metabolomic profiling using Orbitrap Fourier transform mass spectrometry with hydrophilic interaction chromatography: a method with wide applicability to analysis of biomolecules

Abstract

It was shown that coupling hydrophilic interaction chromatography (HILIC) to Orbitrap Fourier transform mass spectrometery (FT-MS) provided an excellent tool for metabolic profiling, principally due to rapid elution of lipids in advance of most metabolites entering the mass spectrometer. We used in vitro cultivated procyclic forms of the protozoan parasite Trypanosoma brucei as a source of metabolites to test the performance of the HILIC column and the mass accuracy of MS. The mass accuracy achieved fell within 2 ppm for all the metabolites identified within samples. It was, for example, possible to identify the signature metabolite of the trypanosome, trypanothione, and also glutathione which were well retained by the HILIC column. By comparing trypanosomes grown in two different media we were able to clearly distinguish the samples in terms of the relative abundance of a number of metabolites using Sieve 1.1 software.

Metabolomics: Moving to the Clinic

“…However, the drawback of RP chromatography is that polar metabolites have poor retention and will thus elute in the void volume, making correct identification and quantification of these substances difficult. To address this, investigators have started to employ hydrophilic interaction chromatography (Tolstikov and Fiehn 2002;Appelblad et al 2008;Kamleh et al 2008;Kim et al 2009;T'Kindt et al 2008), which results in almost orthogonal retention order to RP (most polar metabolites elutes last). Multidimensional chromatographic setups using two or more LC separation phases for metabolic profiling are also described (Sabatine et al 2005;Lewis et al 2008).…”

Mass Spectrometry for Metabolomics

“…Hydrophilic interaction liquid chromatography (HIlIC) has also gained in popularity over the last few years [55,56], and may provide an excellent orthogonal separation approach to traditional reversed-phase chromatography. Alternatives to lC-MS metabolomics include gas chromatography (gC) [33,34,45,46], capillary electrophoresis (CE) [23,47], or supercritical fluid chromatography (SFC) [57,58].…”

Metabolomics: a valuable tool for stem cell monitoring in regenerative medicine

“…This gives broad retention characteristics for small molecules [17]. Another benefit is that their selectivity allows organic solvents to be used for metabolite extraction without further processing prior to loading on the column.…”