Metabolites in the colonic contents were extracted with methanol and quantified using an ultra-high performance liquid chromatography/mass spectrometry (UHPLC-MS) system (25 (link)) on a Thermo Vanquish (Thermo Fisher Scientific, United States) as previously detailed (26 (link)) and a Thermo Orbitrap Exploris 120 mass spectrometer (Thermo Fisher) using data collection from both positive and negative ion modes as previously described (27 (link)).
PCA and orthogonal partial least squares discriminant analysis (OPLS-DA) were conducted in R software to analyze and screen metabolites. Potential biomarkers were selected based on VIP scores >1 in OPLS-DA. Simultaneously, metabolites were screened further and those with FC >2 and p-value <0.05 in student’s t-test were selected.
Differential metabolites from optimized data underwent substance enrichment analysis in R software with reference to the KEGG database to determine host pig metabolic pathways.
PCA and orthogonal partial least squares discriminant analysis (OPLS-DA) were conducted in R software to analyze and screen metabolites. Potential biomarkers were selected based on VIP scores >1 in OPLS-DA. Simultaneously, metabolites were screened further and those with FC >2 and p-value <0.05 in student’s t-test were selected.
Differential metabolites from optimized data underwent substance enrichment analysis in R software with reference to the KEGG database to determine host pig metabolic pathways.
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