MetaboLights

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ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15098/FILES/DERIVED_FILES/metabolite_identification_quantification_table.xlsxftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15098/m_MTBLS15098_LC-MS_alternating_normal-phase_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15098/a_MTBLS15098_LC-MS_alternating_normal-phase.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15098/s_MTBLS15098.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15098/i_Investigation.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15098/FILES/RAW_FILES/f_24h_1.rawftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15098/FILES/RAW_FILES/f_24h_3.rawftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15098/FILES/RAW_FILES/f_24h_5.rawftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15098/FILES/RAW_FILES/food_3.rawftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15098/FILES/RAW_FILES/food_4.rawftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15098/FILES/RAW_FILES/f_12h_4.rawftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15098/FILES/RAW_FILES/f_12h_1.rawftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15098/FILES/RAW_FILES/food_1.rawftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15098/FILES/RAW_FILES/f_24h_4.rawftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15098/FILES/RAW_FILES/f_12h_2.rawftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15098/FILES/RAW_FILES/f_12h_3.rawftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15098/FILES/RAW_FILES/food_5.rawftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15098/FILES/RAW_FILES/food_2.rawftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15098/FILES/RAW_FILES/f_24h_2.rawftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15098/FILES/RAW_FILES/f_12h_5.rawprimaryOK200ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15098The metabolites were identified by accuracy mass and MS/MS data which were matched with HMDB (http://www.hmdb.ca), massbank (http://www.massbank.jp/), KEGG (https://www.genome.jp/kegg/), LipidMaps (http://www.lipidmaps.org), mzcloud (https://www.mzcloud.org) and the metabolite database bulid by Panomix Biomedical Tech Co., Ltd. (Shuzhou, China). The molecular weight of metabolites was determined according to the m/z (mass-to-charge ratio) of parent ions in MS data. Molecular formula was predicted by ppm (parts per million) and adduct ion, and then matched with the database. At the same time, the MS/MS data from quantitative table of MS/MS data, were matched with the fragment ions and other information of each metabolite in the database, so as to realize the MS/MS identification of metabolites.MetaboLightsPublicLiquid Chromatography MS - alternating - normal-phaseThe LC analysis was performed on a Vanquish UHPLC System (Thermo Fisher Scientific, USA). Chromatography was carried out with an ACQUITY UPLC ® HSS T3 (150×2.1 mm, 1.8 μm) (Waters, Milford, MA, USA). The column maintained at 40 ℃. The flow rate and injection volume were set at 0.3 mL/min and 2 μL, respectively. For LC-ESI (+)-MS analysis, the mobile phases consisted of (B2) 0.1% formic acid in acetonitrile (v/v) and (A2) 0.1% formic acid in water (v/v). Separation was conducted under the following gradient: 0~1 min, 10% B2; 1~5 min, 10%~98% B2; 5~6.5 min, 98% B2; 6.5~6.6 min, 98%~10% B2; 6.6~8 min, 10% B2. For LC-ESI (-)-MS analysis, the analytes was carried out with (B3) acetonitrile and (A3) ammonium formate (5mM). Separation was conducted under the following gradient: 0~1 min, 10%B3; 1~5 min, 10%~98% B3; 5~6.5 min, 98% B3; 6.5~6.6 min, 98%~10% B3; 6.6~8 min, 10% B3.Dual Effects of Fasting on Gut Microbiota and Metabolism: Protective Adaptation versus Barrier Dysfunction Risks.Sen YeGuangzhou University of Chinese Medicinefecesmass spectrometry assaySample preparation:1. Accurately weigh an appropriate amount of sample into a 2 mL centrifuge tube,add 600 μL MeOH (stored at -20 ℃) (Containing 2-Amino-3-(2-chloro-phenyl)-propionic acid(4 ppm), vortex for 30 s;2. Add 100 mg glass bead, placed in a tissue grinder for 120 s at 50 Hz;3. Room temperature ultrasound for 10 min;4. Centrifuge for 10 min at 12,000 rpm and 4 °C,filter the supernatant by 0.22 μm membrane and transfer into the detection bottle for LC-MS detection.Mus musculushttps://www.ebi.ac.uk/metabolights/MTBLS15098Sen Ye. Guangzhou University of Chinese Medicine. yesen@gzucm.edu.cn.The raw data were firstly converted to mzXML format by MSConvert in ProteoWizard software package (v3.0.8789) and processed using R XCMS(v3.12.0) for feature detection, retention time correction and alignment.Feeding conditionyesen@gzucm.edu.cnIntestinal content samples were obtained from male C57BL/6J mice in three groups: ad libitum feeding, 12 h fasting, and 24 h fasting, with five mice per group. After one week of acclimatization, mice underwent the assigned feeding or fasting intervention with free access to water. At the endpoint, mice were anesthetized with 3% isoflurane and euthanized by cervical dislocation. Intestinal contents were collected under sterile conditions, immediately flash-frozen in liquid nitrogen, and stored at −80°C until LC-MS/MS-based untargeted metabolomic analysis.MetabolomicsMetabolomicsMus musculusVanquishcolitisuntargeted analysisOrbitrap Exploris 120fecessepsisobeseexperimental sampleMetabolomicsMus musculusVanquishcolitisuntargeted analysisOrbitrap Exploris 120fecessepsisobeseexperimental sampleMass spectrometric detection of metabolites was performed on Orbitrap Exploris 120 (Thermo Fisher Scientific, USA) with ESI ion source. Simultaneous MS1 and MS/MS (Full MS-ddMS2 mode, data-dependent MS/MS) acquisition was used. The parameters were as follows: sheath gas pressure, 40 arb; aux gas flow, 10 arb; spray voltage, 3.50 kV and -2.50 kV for ESI(+) and ESI(-), respectively; capillary temperature, 325 ℃; MS1 range, m/z 100-1000; MS1 resolving power, 60000 FWHM; number of data dependant scans per cycle, 4; MS/MS resolving power, 15000 FWHM; normalized collision energy, 30%; dynamic exclusion time, automatic.falseDual Effects of Fasting on Gut Microbiota and Metabolism: Protective Adaptation versus Barrier Dysfunction RisksFasting is increasingly recognized as a metabolic intervention that reshapes gut microbiota and host metabolism, but its duration-dependent effects on intestinal epithelial integrity remain incompletely understood. In this study, we compared ad libitum feeding, 12 h fasting, and 24 h fasting in male C57BL/6J mice using 16S rRNA gene sequencing, untargeted metabolomics, histological assessment, Occludin immunofluorescence, and a cecal ligation and puncture inflammatory challenge model. Fasting induced time-dependent microbial and metabolic remodeling. The 12 h fasting condition was characterized by enrichment of Akkermansia muciniphila and fasting-responsive metabolites associated with mucosal homeostasis and immune modulation. However, early epithelial structural changes, including reduced crypt/gland depth and decreased Occludin signal, were already detectable after fasting and became more evident after 24 h. In the CLP model, 24 h fasting was associated with increased mortality, suggesting reduced resilience to severe polymicrobial inflammatory stress. Correlation analyses further linked fasting-responsive microbial taxa with metabolites involved in inflammatory and metabolic pathways, although these associations do not establish causality. Collectively, our findings suggest that acute fasting induces concurrent microbial-metabolic adaptation and epithelial vulnerability, with prolonged fasting shifting the balance toward impaired stress resilience. These results highlight the importance of defining fasting duration carefully and warrant future studies using direct permeability assays and mechanistic microbiota-intervention models.2026-07-202026-07-20MTBLS15098