Miniaturized chromatography-based lipidomics methods towards single-cell analysis

Lipidomics provides detailed insight into lipid metabolism and cellular function, but conventional workflows typically rely on bulk samples that mask cellular heterogeneity. Advances in analytical chemistry are enabling lipid analysis from extremely limited material, driving the development of miniaturized chromatography-mass spectrometry (LC-MS) workflows for low-input and single-cell studies. This review summarizes recent progress in miniaturized chromatography-based lipidomics. Reducing chromatographic scale improves ionization efficiency, sensitivity, and separation performance while minimizing sample consumption. We discuss key enabling technologies, such as low-flow electrospray interfaces and the integration of ion mobility (IM) spectrometry as an orthogonal separation dimension for lipid identification. Methodological considerations for low-input lipidomics are also addressed, particularly sample preparation and quantitative challenges at picogram-scale analyte levels. Finally, we highlight future directions in automation, microfluidics, and multidimensional separations. Together, these developments position miniaturized chromatography as a critical platform for advancing single-cell lipidomics and high-resolution studies of lipid metabolism.