MASLD biomarker discovery: evaluating lipidomics techniques across disease progression.

Metabolic dysfunction-associated steatotic liver disease (MASLD) spans a spectrum from benign steatosis to aggressive steatohepatitis (MASH) and fibrosis, with lipid dysregulation as a core pathophysiological driver. Lipidomics is pivotal for identifying stage-specific biomarkers, yet the optimal analytical platform remains unresolved due to diverse and rapidly evolving methodological approaches. This review comprehensively compares key lipidomics techniques including targeted and untargeted liquid chromatography-mass spectrometry (LC-MS), matrix-assisted laser desorption/ionization imaging (MALDI-IMS), gas chromatography-MS (GC-MS), and nuclear magnetic resonance (NMR) spectroscopy for MASLD research applications. We discuss in this study the comparative sensitivity of these technique, their sensitivity in detecting low-abundance pathogenic lipids during critical disease transitions, coverage of lipid classes across disease stages, throughput for clinical screening versus deep mechanistic studies, and compatibility with various sample types (serum, biopsy, organoids). By critically assessing trade-offs in quantification accuracy, standardization, and translational feasibility, we identify technique-specific niches. We ultimately provide a pragmatic framework for platform selection to accelerate the validation of lipidomic biomarkers and advance precision diagnostics for MASLD.