Clinical Metabolomics: Expanding the Metabolome Coverage Using Advanced Analytical Techniques

Metabolomics, the comprehensive analysis of all metabolites and intermediate products of reactions present within a biological system, is a promising field to enable precision medicine. Clinical metabolomics faces two main challenges at the bioanalytical level. The first is the need for high resolution to obtain maximum metabolome coverage. This is exemplified by the latest version of the Human Metabolome Database (HMDB), which reports more than 110,000 metabolites and endogenous compounds. The second is the high-throughput needed to enable the analysis of a large number of samples typically encountered in large-scale cohort studies. Reversed-phase liquid chromatography (LC) at regular or ultrahigh pressures combined with high-resolution mass spectrometry (HRMS) has long been considered the “gold standard” in metabolomics. However, these conventional reversed-phase LC–MS approaches are no longer sufficient to analyze the vast variety of polar compounds, as well as discriminate closely related compounds such as isomers or enantiomers. This review article discusses the novel separation and detection strategies that are considered promising in clinical metabolomics to enhance the metabolome coverage. It includes hydrophilic interaction chromatography (HILIC), supercritical fluid chromatography (SFC), multidimensional LC approaches, as well as ion-mobility mass spectrometry (IM-MS) and data-independent acquisition (DIA) analysis methods.