Decoding Membrane Lipids: Analytical Barriers and Technological Advances in Modern Lipidomics.

Biological membranes are dynamic, information-rich platforms whose structural and functional properties are dictated by lipid composition rather than acting as passive barriers. Recent advances in lipidomics have revealed that variations in lipid headgroups, acyl-chain length and saturation, sn-positional architecture, and oxidative modifications profoundly influence membrane mechanics, lateral organization, and protein-lipid interactions. These features collectively regulate fundamental cellular processes, including signaling, trafficking, curvature generation, and transbilayer asymmetry. In parallel, a wide range of pathological conditions-including cancer, metabolic disorders, neurodegeneration, and inflammatory diseases-are increasingly associated with coordinated lipid remodeling that reshapes membrane material properties and electrostatic landscapes. In this review, we integrate biophysical principles with lipidomics-based evidence to elucidate how lipid chemical diversity translates into membrane-level behavior. We discuss the roles of major membrane lipid classes, the functional consequences of acyl-chain and sn-positional remodeling, and the biological significance of lipid asymmetry and lateral heterogeneity. Particular attention is given to disease-associated lipid reprogramming and extracellular vesicle lipidomes as functional extensions of cellular membranes. Finally, we examine key analytical barriers in modern lipidomics and outline strategies required to connect lipid structural information with biological function. Together, this framework highlights membrane lipid architecture as a central determinant of cellular physiology and a promising axis for mechanistic insight and translational biomarker discovery.