Multidimensional Liquid Chromatography-Mass Spectrometry for Metabolomics and Lipidomics: Advances in Techniques and Applications

Metabolomics and lipidomics seek to achieve comprehensive profiling of metabolites and lipids in biological and plant systems. However, the wide diversity in their physicochemical properties, such as polarity and optical activity, poses considerable challenges to conventional one-dimensional liquid chromatography (1DLC). Multidimensional liquid chromatography (MDLC) addresses these limitations by combining orthogonal separation mechanisms, offering a powerful approach for resolving complex metabolome and lipidome. This review outlines the fundamental configurations of MDLC—including heart-cutting, stop-flow, and comprehensive modes—along with key modulation techniques such as loop-based and trapping-column-based interfaces. It further summarizes applications in metabolomics and lipidomics focusing on biological and plant samples over the past decade. Owing to its high sensitivity and rich information, mass spectrometry operated in both untargeted and targeted modes has become the dominant detection platform coupled with MDLC. Overall, the integration of MDLC with MS has enhanced sensitivity and separation resolution for targeted analytes and entire sample compositions, while significantly expanding the coverage of metabolites and lipids across diverse polarity ranges.