Metabolomics and lipidomics of human serum in fasting and re-feeding
Metabolomics and lipidomics analyses performed on serum collected from normal human volunteers after a 24-hour fast and 3 hours after re-feeding to identify serum mediators of fasting-associated immunomodulation.
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Please refer to the published paper for the full methods. In summary: A clinical study (NCT02719899) was conducted to assess the immunologic effects of a 24-hour fast and a re-feeding period on normal volunteers. Study participants (10 males and 11 females, aged 22-29, with no acute or chronic disease and a normal-weight BMI) consumed a fixed 500-calorie morning meal before 8 a.m. and began a 24-hour total fast before drawing fasting blood. Study participants then consumed another 500-calorie morning meal, and after 3 hours, the re-feeding samples were collected (t = 27 hours). Serum samples were extracted using a standard methanol-chloroform-water extraction and reconstituted with internal standards. Targeted detection of polar metabolites was performed with a Thermo Scientific Vanquish UHPLC+ coupled to a TSQ Quantiva mass spectrometer, and data processing and analysis were performed using Thermo Xcalibur software. Open profile lipidomics was performed using an Orbitrap Velos Elite Mass Spectrometer, and analysis was performed using XCMS. Peak intensity was normalized to internal standards. Univariate analysis of serum metabolomics and lipidomics data were performed in MetaboAnalyst (https://www.metaboanalyst.ca/). Multivariate analysis was performed with orthogonal partial least squares-discriminant analysis (OPLS-DA) using Simca® (Umetrics), generating the variables important for prediction (VIP) score.