Recent advances in liquid‐phase separations for clinical metabolomics

Abstract

Over the last decades, several technological improvements have been achieved in liquid-based separation techniques, notably, with the advent of fully porous sub-2 μm particles and superficially porous sub-3 μm particles, the comeback of supercritical fluid chromatography, and the development of alternative chromatographic modes such as hydrophilic interaction chromatography. Combined with mass spectrometry, these techniques have demonstrated their added value, substantially increasing separation efficiency, selectivity, and speed of analysis. These benefits are essential in modern clinical metabolomics typically involving the study of large-scale sample cohorts and the analysis of thousands of metabolites showing extensive differences in physicochemical properties. This review presents a brief overview of the recent developments in liquid-phase separation sciences in the context of clinical metabolomics, focusing on increased throughput as well as metabolite coverage. Relevant metabolomics applications highlighting the benefits of ultra-high performance liquid chromatography, core-shell technology, high-temperature liquid chromatography, capillary electrophoresis, supercritical fluid chromatography, and hydrophilic interaction chromatography are discussed.

Comprehensive metabonomic analysis of heart tissue from isoproterenol-induced myocardial infarction rat based on reversed-phase and hydrophilic interaction chromatography coupled to mass spectrometry

“…HILIC offers a different selectivity, with better retention of polar analytes not easily retained or indeed not retained at all using RPLC approaches . HILIC also provides high MS sensitivity because of increased ionization resulting from the use of mobile phases containing a high proportion of organic solvents . In this study, it was proved that HILIC separation method could be a suitable complementary separation method for RP analysis.…”

Capillary electrophoresis mass spectrometry based metabolomics

“…CE provides a flat solvent front compared to the convex front with HPLC separation systems. Peak spreading by CE is only due to longitudinal diffusion across the capillary column thus producing separations with better peak shape, greater dynamic range for certain metabolites over LC-MS meth- 2016 [114] Iadarola P, Fumagalli M, Bardoni AM, Salvini R, Viglio S.…”

“…Issue 37 of the Journal Electrophoresis [110] provided an entire publication to LC-MS and CE-MS, demonstrating equal status, although the CE-MS applications were not specific to metabolomics. Hankemeier's laboratory at Leiden University provided 4 separate reviews [111][112][113][114] covering different aspects of CE-MS from growing applications to technical developments. Tang et al [109] provides a discussion comparing HILIC Boizard F, Brunchault V, Moulos P, at al.…”

Data processing, multi-omic pathway mapping, and metabolite activity analysis using XCMS Online

“…Pooled samples are run throughout the mass spectrometric sequence regularly (e.g., one in ten injections) as a QC check for signal intensity and retention time drift, but are also extremely valuable as a method for including preliminary metabolite validation using data-dependent or targeted MS/MS 37 . Further details on metabolomic sampling, extraction and chromatographic methods have been discussed elsewhere 49–52 .…”