ArticleScientific reports2026
Hydrophobic interaction chromatography resolves extracellular vesicle fractions with distinct lipidomic signatures.
Article in Scientific reports, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Current extracellular vesicle (EV) isolation workflows are dominated by size- and density-based approaches, which provide limited insight into surface chemical properties of vesicular particles. Here, we report a hydrophobic interaction chromatography (HIC) workflow for resolving EV fractions along differences in membrane interfacial hydrophobicity. Using a commercially available HIC column, reproducible fractionation of EV samples was achieved, yielding discrete fractions that differed in retention behaviour and lipid composition. Transmission electron microscopy (TEM) showed vesicle-like nanoparticles with EV-consistent morphology, indicating preservation of vesicle integrity during HIC. Lipidomic profiling by reversed-phase liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (RP-LC-Q-TOF-MS) revealed fraction-specific differences in lipid composition. Importantly, we demonstrate the feasibility of direct injection of pre-cleaned biofluids onto the HIC column, enabling fractionation of nanoparticle populations containing EVs without prior ultracentrifugation. Furthermore, an operational interfacial hydrophobicity index derived from lipidomic data showed a clear correlation with HIC retention, providing an orthogonal compositional descriptor consistent with the proposed fractionation mechanism. Together, this hydrophobicity-based strategy introduces a previously unexplored physicochemical dimension to EV analysis, revealing chemically structured heterogeneity that is not accessible using conventional separation strategies. The workflow provides a practical framework for fractionating EVs in a manner directly relevant to lipidomic profiling and studies of EV membrane chemistry.
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