Skip to main content
. 2021 Oct 19;12:6000. doi: 10.1038/s41467-021-26209-8

Fig. 1. Overview of experimental approach and multi-metabolomics analyses.

Fig. 1

a Experimental workflow started with sample harvest and metabolite extraction of feces, blood sera, and cerebral cortical brain tissues from 8-week age-matched germ-free (GF, N = 12) and conventionally raised (CONV-R) specific-pathogen-free C57BL/6 mice (N = 12). A novel high-coverage metabolomics approach was used featuring orbitrap high-resolution mass spectrometry, targeted annotation based on an in-house mass spectral library, untargeted annotation using a streamlined cheminformatic pipeline for de novo structural dereplication, univariate and multivariate statistics, and data visualization. b Number of total and significant ion features (p-value < 0.01, fold change ≥ 1.5, two-sided Welch’s t-test) detected for feces, blood sera, and cortical brain tissues under HESI positive and negative modes of analysis. c Trend distribution of significantly altered ion features. d Venn diagram of all identified metabolites of GF/CONV-R difference among the three sample matrices. Chroma chromatography, HESI heated electrospray ionization, PRM parallel reaction monitoring, QA/QC quality assurance/quality control, ISD internal standard, PCA principal component analysis, RT/mz library retention time and mass-to-charge ratio pair library, ExpDB experimental database, MoNA MassBank of North America, GNPS The Global Natural Product Social Molecular Networking.