박영재 (서울대학교)

이도엽 (서울대학교)

박영재 (서울대학교)

최지은 (서울대학교)

조주연 (서울대학교)

이도엽 (서울대학교)

Sample preparation can substantially affect metabolite coverage and analytical reproducibility in untargeted plasma metabolomics. In this study, pooled plasma from healthy individuals was analyzed to compare a range of reconstitution and extraction solvent conditions for reversed-phase liquid chromatography–Orbitrap mass spectrometry (RP-LC–Orbitrap MS). Reconstitution solvents included aqueous and organic mixtures of water, methanol, and acetonitrile, while extraction conditions comprised methanol-, acetonitrile-, and isopropanol-based solvent systems. Replicate samples were prepared and analyzed in randomized order. Raw data were processed using MZmine for feature detection, alignment, filtering, and metabolite annotation. The solvent conditions were comparatively assessed according to annotated feature coverage, analytical variability, and chemical-class-dependent recovery. This study characterizes solvent-dependent differences in plasma metabolite profiles and provides a methodological reference for selecting sample-preparation conditions in untargeted RP-LC–MS analysis.

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