조준우 (서울대학교)

이도엽 (서울대학교)

조준우 (서울대학교)

최지은 (서울대학교)

김유미 (서울대학교)

이도엽 (서울대학교)

In this study, LC-Orbitrap mass spectrometry was used to examine the influence of analytical parameters on untargeted metabolomics results with pooled human fecal samples. Key variables such as isolation width, minimum automatic gain control (AGC) target, and apex trigger were investigated while baseline exclusion was also conducted to enhance metabolite feature detection, spectral fidelity, and background noise suppression.

Additionally, the effect of reconstitution solvent composition was examined by comparing single solvents and mixed solvent systems across diverse conditions. Criteria to compare were comprised of the quantity and diversity of annotated features, peak shape integrity, and the fraction of features exhibiting a coefficient of variation below 30%. As a result, we confirmed that mixed solvents demonstrated better reproducibility and expanded feature coverage compared to single solvents.

These results show the importance of careful optimization of both MS acquisition parameters and solvent composition to achieve robust and reproducible untargeted metabolomics workflows. This study would offer valuable insights for establishing reliable LC-Orbitrap MS methodologies in metabolomics research.