Untargeted metabolomics of human plasma with mass spectrometry is of particular interest in medical research to explore pathophysiology, find disease biomarkers or for the understanding of the response to pharmacotherapy. Since analytical performances may be impacted by the laboratory environment and the acquisition method settings, the objectives of this study were to assess the role of interfering compounds and to propose an acquisition method to maximize the metabolome coverage for human plasma metabolomic analysis. Human plasma samples were processed with liquid/liquid extraction then analysed with HILIC-high resolution mass spectrometry. A method with a single m/z range was compared to four methods with different split acquisition ranges and four sets of ionization source parameters were compared. The data were analysed with the R software and on the Worklow4Metabolomics online platform. The major interfering compounds were identified in blank samples where they accounted for up to 86% of the signal intensity. Splitting the acquisition range into 3 m/z ranges improved the number of detected features, the number of features with proposed annotation in the Human Metabolome Database, as well as signal intensity throughout the whole m/z range. The method performing best was the one using three m/z ranges of approximatively the same extent. Ionization source parameters also strongly affected the number of detected features. Splitting the acquisition range into 3 m/z ranges with optimized ionization source parameters allows a comprehensive analysis of the human plasma metabolome with perspectives for applications to pathophysiological studies.
Keywords: High-resolution mass spectrometry; Human plasma; Hydrophilic interaction liquid chromatography; Metabolomics.
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