#METABOLOMICS WORKBENCH Lxu_20240904_100539 DATATRACK_ID:5165 STUDY_ID:ST003460 ANALYSIS_ID:AN005686 PROJECT_ID:PR002122 VERSION 1 CREATED_ON September 4, 2024, 7:35 pm #PROJECT PR:PROJECT_TITLE Early Metabolic Shifts in the Sepsis: Insights from Presymptomatic Patients and PR:PROJECT_TITLE In Vivo Models. PR:PROJECT_SUMMARY Metabolomics of human serum to support understanding of early metabolic shift in PR:PROJECT_SUMMARY presymptomatic sepsis patients and further confirmed in in vivo model. PR:INSTITUTE Leibniz Institute for Natural Product Research and Infection Biology Hans Knöll PR:INSTITUTE Institute PR:DEPARTMENT Microbiome Dynamics PR:LAST_NAME XU PR:FIRST_NAME LINLIN PR:ADDRESS Beutenbergstraße 11a, Jena, Thuringia, 07745, Germany PR:EMAIL Lin-Lin.Xu@hki-jena.de PR:PHONE +4936415321265 #STUDY ST:STUDY_TITLE Metabolomics of mice serum to support understanding of early metabolic shift in ST:STUDY_TITLE presymptomatic sepsis patients. (Part6 mice serum metabolite) ST:STUDY_SUMMARY Sepsis, a life-threatening organ dysfunction caused by a dysregulated response ST:STUDY_SUMMARY to infection, is a critical medical condition with significant global impact, ST:STUDY_SUMMARY responsible for 48.9 million cases and 11 million deaths annually. Metabolic ST:STUDY_SUMMARY dysregulation in sepsis is a critical determinant of disease outcome, yet most ST:STUDY_SUMMARY studies focus on patients after severe illness onset, comparing septic versus ST:STUDY_SUMMARY non-septic groups or survivors versus non-survivors. To truly understand this ST:STUDY_SUMMARY complex disease, it is essential to investigate early metabolic shifts, ST:STUDY_SUMMARY capturing the transition from simple infection to sepsis. Our untargeted ST:STUDY_SUMMARY metabolome analysis of serum tissues from mice model revealed early septic ST:STUDY_SUMMARY biomarkers in serum. ST:INSTITUTE Leibniz Institute for Natural Product Research and Infection Biology Hans Knöll ST:INSTITUTE Institute ST:DEPARTMENT Microbiome Dynamics ST:LAST_NAME XU ST:FIRST_NAME LINLIN ST:ADDRESS Beutenbergstraße 11a, Jena, Thuringia, 07745, Germany ST:EMAIL Lin-Lin.Xu@hki-jena.de ST:PHONE +4936415321265 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS Bl6J_23_0196 SA020922 Sample source:Serum | Group:Sham / Vehicle RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-026-SA020922_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-026-SA020922_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0268 SA020989 Sample source:Serum | Group:CLP (24h) / 12C-Serin RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-018-SA020989_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-018-SA020989_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0192 SA020901 Sample source:Serum | Group:CLP (24h) / Vehicle RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-028-SA020901_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-028-SA020901_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0194 SA020915 Sample source:Serum | Group:Sham / 13C-Serin RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-033-SA020915_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-033-SA020915_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0271 SA021010 Sample source:Serum | Group:Sham / 12C-Serin RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-032-SA021010_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-032-SA021010_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0199 SA020940 Sample source:Serum | Group:CLP (24h) / 13C-Serin RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-019-SA020940_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-019-SA020940_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0273 SA021024 Sample source:Serum | Group:CLP (24h) / 12C-Serin RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-010-SA021024_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-010-SA021024_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0193 SA020908 Sample source:Serum | Group:Sham / 13C-Serin RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-020-SA020908_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-020-SA020908_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0189 SA020887 Sample source:Serum | Group:CLP (24h) / 13C-Serin RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-025-SA020887_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-025-SA020887_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0267 SA020982 Sample source:Serum | Group:CLP (24h) / 12C-Serin RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-014-SA020982_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-014-SA020982_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0272 SA021017 Sample source:Serum | Group:Sham / 12C-Serin RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-017-SA021017_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-017-SA021017_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0190 SA020894 Sample source:Serum | Group:CLP (24h) / 13C-Serin RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-031-SA020894_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-031-SA020894_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0274 SA021031 Sample source:Serum | Group:CLP (24h) / 12C-Serin RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-021-SA021031_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-021-SA021031_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0203 SA020968 Sample source:Serum | Group:Sham / 13C-Serin RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-029-SA020968_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-029-SA020968_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0204 SA020975 Sample source:Serum | Group:CLP (24h) / Vehicle RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-022-SA020975_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-022-SA020975_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0198 SA021033 Sample source:Serum | Group:Sham / Vehicle RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-011-SA021033_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-011-SA021033_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0270 SA021003 Sample source:Serum | Group:Sham / 12C-Serin RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-024-SA021003_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-024-SA021003_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0200 SA020947 Sample source:Serum | Group:CLP (24h) / 13C-Serin RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-013-SA020947_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-013-SA020947_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0202 SA020961 Sample source:Serum | Group:Sham / 13C-Serin RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-012-SA020961_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-012-SA020961_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0201 SA020954 Sample source:Serum | Group:Sham / Vehicle RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-015-SA020954_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-015-SA020954_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0269 SA020996 Sample source:Serum | Group:Sham / 12C-Serin RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-027-SA020996_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-027-SA020996_pos.mzML SUBJECT_SAMPLE_FACTORS NA QC_004 Sample source:Serum | Group:QC RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-009-QC_004_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-009-QC_004_pos.mzML SUBJECT_SAMPLE_FACTORS NA QC_009 Sample source:Serum | Group:QC RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-040-QC_009_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-040-QC_009_pos.mzML SUBJECT_SAMPLE_FACTORS NA QC_006 Sample source:Serum | Group:QC RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-023-QC_006_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-023-QC_006_pos.mzML SUBJECT_SAMPLE_FACTORS NA QC_005 Sample source:Serum | Group:QC RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-016-QC_005_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-016-QC_005_pos.mzML SUBJECT_SAMPLE_FACTORS NA QC_008 Sample source:Serum | Group:QC RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-034-QC_008_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-034-QC_008_pos.mzML SUBJECT_SAMPLE_FACTORS NA Blank_002 Sample source:blank | Group:Blank RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-005-Blank_002_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-005-Blank_002_pos.mzML SUBJECT_SAMPLE_FACTORS NA Blank_001 Sample source:blank | Group:Blank RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-003-Blank_001_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-003-Blank_001_pos.mzML SUBJECT_SAMPLE_FACTORS NA QC_001 Sample source:Serum | Group:QC RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-006-QC_001_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-006-QC_001_pos.mzML SUBJECT_SAMPLE_FACTORS NA Blank_003 Sample source:blank | Group:Blank RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-038-Blank_003_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-038-Blank_003_pos.mzML SUBJECT_SAMPLE_FACTORS NA QC_007 Sample source:Serum | Group:QC RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-030-QC_007_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-030-QC_007_pos.mzML SUBJECT_SAMPLE_FACTORS NA Blank_PB Sample source:blank | Group:Blank RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-044-Blank_PB_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-044-Blank_PB_pos.mzML SUBJECT_SAMPLE_FACTORS NA Blank_004 Sample source:blank | Group:Blank RAW_FILE_NAME(Raw file name negative ion)=230707-EXP-PAN-1-043-Blank_004_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230707-EXP-PAN-1-043-Blank_004_pos.mzML #COLLECTION CO:COLLECTION_SUMMARY Mice tissue samples were collected from our in vivo mice model and frozen at -80 CO:COLLECTION_SUMMARY °C. Samples were shipped to MS-OMICS for metabolomic analysis. CO:SAMPLE_TYPE Blood (serum) #TREATMENT TR:TREATMENT_SUMMARY Approximately 30% of the cecum was ligated and punctured twice with a 23 Gauge TR:TREATMENT_SUMMARY (G) needle. A small amount of faeces was extruded and the cecum was placed back TR:TREATMENT_SUMMARY into the abdominal cavity. All animals received 40 mL/kg saline s.c. directly TR:TREATMENT_SUMMARY after the procedure and once 25mg/kg s.c. Imipenem/Cilastatin after 6 hours. TR:TREATMENT_SUMMARY This model is associated with a 14-day mortality of 30%, closely mimicking the TR:TREATMENT_SUMMARY clinical situation. Animals were orally gavaged with 500mg/kg 12C-Serin (Sigma) TR:TREATMENT_SUMMARY or 13C-Serin (CLM-1574-H-0.25 Serine-L (13C3 99%13C), Euroisotope, Germany) in TR:TREATMENT_SUMMARY 10mL/kg drinking water or drinking water only (vehicle) 6 and 18 hours after TR:TREATMENT_SUMMARY CLP98,99. After 24 hours, the animals are then perfused with ice cold PBS and TR:TREATMENT_SUMMARY organs were harvested, snap frozen and stored at -80° until further analysis. TR:TREATMENT_SUMMARY Sham animals underwent exactly to the same procedures except for the CLP. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY To prepare samples for semi-polar metabolites extraction, the different tissue SP:SAMPLEPREP_SUMMARY samples were weighted and transferred to Eppendorf tubes. Ceramic beads and SP:SAMPLEPREP_SUMMARY precooled methanol/water (1:2) were added. All tubes were placed in a pre-cooled SP:SAMPLEPREP_SUMMARY (-20°C) bead beater and homogenized (4 x 30 sec., 30 Hz) followed by SP:SAMPLEPREP_SUMMARY ultrasonication (5 min). After centrifugation (18000 RCF, 5 min, 4°C), the SP:SAMPLEPREP_SUMMARY supernatant was collected. The sample pellet was reextracted as described above. SP:SAMPLEPREP_SUMMARY The two extract supernatants were pooled and passed through a phosphor removal SP:SAMPLEPREP_SUMMARY cartridge (Phree filter plates). A precise aliquot of the extract was evaporated SP:SAMPLEPREP_SUMMARY to dryness under a gentle stream of nitrogen, before reconstitution with 10% SP:SAMPLEPREP_SUMMARY Eluent B in Eluent A. All Samples were diluted 5 times in mobile phase eluent A SP:SAMPLEPREP_SUMMARY and stable isotope labelled standards were added before analysis. Mouse serum SP:SAMPLEPREP_SUMMARY (50 μl) was diluted with a mixture of acetonitrile and methanol (200 μl 1:1 SP:SAMPLEPREP_SUMMARY v/v). The extract was then passed through a phosphor removal cartridge (Phree, SP:SAMPLEPREP_SUMMARY Phenomenex). An aliquot (100 μl) was transferred into a high recovery HPLC vial SP:SAMPLEPREP_SUMMARY and the solvent was removed under a gentle flow of nitrogen. Extracts were SP:SAMPLEPREP_SUMMARY reconstituted in a mobile phase mixture (100 μl, 10%B in 90%A). To ensure SP:SAMPLEPREP_SUMMARY high-quality sample preparation, a quality control sample (QC sample) was SP:SAMPLEPREP_SUMMARY prepared by pooling small equal aliquots from each sample, to create a SP:SAMPLEPREP_SUMMARY representative average of the entire set. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Thermo Vanquish CH:COLUMN_NAME Waters ACQUITY UPLC HSS T3 (150 x 2.1mm,1.8um) CH:SOLVENT_A 100% water; 10 mM ammonium formate; 0.1% formic acid CH:SOLVENT_B 100% methanol; 10 mM ammonium formate; 0.1% formic acid CH:FLOW_GRADIENT 0-2 min: 0 % B, 4 min 35% B, 6-14 min 90% B, 14.1-15 min 0% B. CH:FLOW_RATE 0.3mL/min CH:COLUMN_TEMPERATURE 30 CH:METHODS_FILENAME Doneanu_etal.pdf #ANALYSIS AN:ANALYSIS_TYPE MS AN:LABORATORY_NAME MS-Omics, Denmark AN:ANALYSIS_PROTOCOL_FILE Doneanu_etal.pdf #MS MS:INSTRUMENT_NAME Orbitrap Exploris 240 MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS Semi-polar metabolite profiling was performed by MS-Omics (Vedbæk, Denmark). MS:MS_COMMENTS The analysis was carried out using a Vanquish LC (Thermo Scientific) coupled to MS:MS_COMMENTS a Orbitrap Exploris 240 MS (Thermo Scientific). The UHPLC was performed using an MS:MS_COMMENTS adapted version of the protocol described by Doneanu et al . An electrospray MS:MS_COMMENTS ionization interface was used as ionization source. Analysis was performed in MS:MS_COMMENTS positive and negative ionization mode under polarity switching. Metabolomics MS:MS_COMMENTS processing was performed untargeted using Compound Discoverer 3.3 (Thermo MS:MS_COMMENTS Scientific) and Skyline 22.2 (MacCoss Lab Software) for peak picking and feature MS:MS_COMMENTS grouping, followed by an in-house annotation and curation pipeline written in MS:MS_COMMENTS MatLab (2022b, MathWorks). Identification of compounds were performed at four MS:MS_COMMENTS levels; Level 1: identification by retention times (compared against in-house MS:MS_COMMENTS authentic standards), accurate mass (with an accepted deviation of 3ppm), and MS:MS_COMMENTS MS/MS spectra, Level 2a: identification by retention times (compared against MS:MS_COMMENTS in-house authentic standards), accurate mass (with an accepted deviation of MS:MS_COMMENTS 3ppm). Level 2b: identification by accurate mass (with an accepted deviation of MS:MS_COMMENTS 3ppm), and MS/MS spectra, Level 3: identification by accurate mass alone (with MS:MS_COMMENTS an accepted deviation of 3ppm). The data have been normalized to the QC samples MS:MS_COMMENTS to remove systematic drift over the analysis sequence. The normalization is MS:MS_COMMENTS performed using linear regression for the QC’s within each analytical batch MS:MS_COMMENTS with signal = a*injection number – b. Each value is then normalized with: MS:MS_COMMENTS normSignal = average(all QC samples)*(signal – injection number*a)/b. This is MS:MS_COMMENTS done independently for each compound. MS:MS_RESULTS_FILE ST003460_AN005686_Results.txt UNITS:Normalized peak area Has m/z:Neutral masses Has RT:Yes RT units:Minutes #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Normalized peak area MS_METABOLITE_DATA_START Samples SA020887 SA020894 SA020901 SA020908 SA020915 SA020922 SA020940 SA020947 SA020954 SA020961 SA020968 SA020975 SA020982 SA020989 SA020996 SA021003 SA021010 SA021017 SA021024 SA021031 SA021033 Factors Sample source:Serum | Group:CLP (24h) / 13C-Serin Sample source:Serum | Group:CLP (24h) / 13C-Serin Sample source:Serum | Group:CLP (24h) / Vehicle Sample source:Serum | Group:Sham / 13C-Serin Sample source:Serum | Group:Sham / 13C-Serin Sample source:Serum | Group:Sham / Vehicle Sample source:Serum | Group:CLP (24h) / 13C-Serin Sample source:Serum | Group:CLP (24h) / 13C-Serin Sample source:Serum | Group:Sham / Vehicle Sample source:Serum | Group:Sham / 13C-Serin Sample source:Serum | Group:Sham / 13C-Serin Sample source:Serum | Group:CLP (24h) / Vehicle Sample source:Serum | Group:CLP (24h) / 12C-Serin Sample source:Serum | Group:CLP (24h) / 12C-Serin Sample source:Serum | Group:Sham / 12C-Serin Sample source:Serum | Group:Sham / 12C-Serin Sample source:Serum | Group:Sham / 12C-Serin Sample source:Serum | Group:Sham / 12C-Serin Sample source:Serum | Group:CLP (24h) / 12C-Serin Sample source:Serum | Group:CLP (24h) / 12C-Serin Sample source:Serum | Group:Sham / Vehicle 2-Hydroxybutyric acid 31981064 2556251 1344936 42560144 17316232 2488793 3091032 1857219 5364947 27587664 3487105 86675752 60802352 41252848 2020442 2242434 1515777 2606480 22810260 23439394 43436256 3-Indoxylsulfuric acid 37629366.4771385 12798361.7161717 21757454.9698525 36391279.4515678 23567280.9242232 1206702.67928426 21124098.5287495 2697961.22090459 48570569.0133715 16022759.904096 47101568.1375174 39787199.9229965 29829048.2438819 56785691.9385094 18471122.1695952 4225964.41942994 14177454.9033306 52392528.924582 16135861.1509347 15057796.3399926 46108770.7833762 Arginine 147803289.003676 72769400.5530445 55837949.8393954 167098709.90858 188548587.922978 306294524.063055 103575071.486083 296045107.787307 375384621.322725 323840962.700662 229095370.348843 103405624.339679 223345668.100384 218792966.592723 316578024.905812 298416825.125702 306778829.169437 260067801.259394 188488830.573601 116960953.049355 160042731.427067 Cholic acid 912668.088740718 6049386.56206442 3033.34851889136 3983001.62644137 71369426.3494689 3142.56236710363 134676.427586365 271671.312805145 10144923.3336095 1574063.47936631 2922812.44377728 883120.019256936 923159.841055286 270306.184754364 951834.944132085 440690.353267785 1057243.9479047 329023.157495753 5626461.28044935 1828965.80896548 1772177.36678659 Citric acid 1601004.15685984 353863.64680557 202512.729030264 2042831.75383439 734297.423638323 1256370.59602704 387892.813215952 363810.420084885 2349256.57048918 612573.340232112 744721.549386427 1687459.54125102 4322118.98605311 4131075.05509992 2109024.79606963 1233443.71542328 2506048.85229261 6215673.02584354 1251726.14936199 489632.606586221 3929876.48667447 Fumaric acid 108721 156768 35338 234396 280934 198885 83337 127725 327383 332361 290116 126435 137877 122956 212919 241875 232501 163673 138347 144077 343475 Glutathione 227006.23546327 977492.473678464 2854.35723647011 1204931.67510438 1619309.56359814 1740137.92237019 2007.36580873683 577751.051910176 2463898.73264692 1405.73887646827 3302777.70669846 412064.462685346 752307.005449058 570364.379461106 2755568.05858438 2328967.6844905 3695878.06730113 2357884.41789086 50421.2166425801 1605188.07922026 2556157.72087855 Malic acid 1757053.8030448 2100224.7787853 448775.441779112 3051140.70559592 3751832.84158487 2391904.58174658 740832.212383699 1581841.38318746 5159707.12826326 4294683.22800323 3601142.95930391 1607188.75377446 1900751.1905824 1932375.51702977 2278882.30052269 3394362.73395096 4297578.93388847 2745925.17025472 1335069.91559218 2027144.51715983 4986485.38583836 Succinic acid 23058072.7656105 62407192.1126047 23864234.6808543 50706885.3874026 59520926.3237064 46189448.4059515 12843460.5730193 69467792.6792847 83336357.4846811 33100684.4767722 49296142.7647998 20698790.3325445 1843876.18838056 2954711.79641099 38456549.3668167 41088960.7372125 35085281.8604514 19182388.8738213 9282368.89480995 15949747.3335548 50003351.0072599 Taurocholic acid 943.971323398175 25321 2195.3193623668 2884437 117733504 1523434 2854.7995670444802 3362.98726737802 3591368 1336080 1986253 1193.56003252463 17602 31353 408085 402162 998501 435921 27499 2384.09393741703 1606448 Threonic acid 2761717.20784382 146585.294502766 69162.7138529706 243716.734849564 134886.282554646 806221.576062534 150392.905878403 206645.976813002 677611.889265147 800993.497516214 650125.717991693 781904.605964224 3435970.9577892 4581917.83778367 514442.092561084 691162.047028694 842966.598962583 1653245.61451484 1107452.77141249 344801.133044383 3200.47956031055 Uric acid 16048559 39019396 24457080 34096440 32876848 34549012 33497646 55857028 48749900 26925976 32513614 21285220 10284635 12511656 37058976 33653788 34445600 35844888 25119796 23622958 38251332 Uridine 3358672.49723959 69060555.6419486 20324822.1310668 52833122.3610141 29392504.7004696 33291415.3113328 6267706.68809175 35518776.5044784 36223323.524619 25791846.3681348 30223057.5503387 8422712.87569102 3599613.76760302 2971066.17682942 20306744.20787 18814729.0705836 19352702.2881652 15446025.6600649 4106747.61171713 6267210.85151367 38837714.9138164 Xanthine 3051.15643790551 8250063 2543258 4676944 28904 4005236 3240.76172656985 119475 10748223 22.3139561449643 126648 3612 12404 481.221891945461 3932609 5219346 5207236 3000863 2734.67741290224 2288.35648736171 2958304 2-(hydroxymethyl)butanoic acid 935166 230173 137373 443067 295814 122332 437904 149553 269584 279441 199215 535289 704303 663936 154831 198895 125556 208319 781973 538827 381056 2-C-Methyl-erythritol-4-phosphoric acid 160968564.11044 8261871.30552882 9825135.87129658 92672186.559434 103218861.258899 210886297.554568 9915606.21072779 141129864.801621 267741245.111117 145454661.208206 137399211.599685 19890098.583392 137028758.894915 139002484.455687 226331754.988896 179062602.59078 194911032.157404 181192635.860268 100913779.869213 49852540.2137534 85270954.8400127 2-Hydroxyisocaproic acid 2867023 552033 295805 2751443 1296446 1081470 507253 527091 779478 1526403 618395 2642644 2107945 2443171 648551 962685 499490 796707 1642302 974991 1827037 2-Oxoglutaric acid 342670 200470 116451 735156 653971 371347 308126 209716 637162 687071 468533 189418 265863 231824 620220 1077884 825254 931506 295433 361519 1248412 3-Methyl-2-oxovaleric acid/ 4-Methyl-2-oxovaleric acid 1542742 742807 259677 2074856 3468828 1277394 1290745 782293 3442001 1963359 1260470 1241906 1156075 955417 1849364 2387705 1646306 2253185 1470649 1377615 4952590 cis-Aconitic acid 1909094 1788013 502181 1907656 1363007 1205672 867664 1159737 1579840 791028 1313131 2468726 2733299 2474993 984782 974514 1107140 1318261 1952270 2359097 1865998 Deoxyuridine 731985 792062 306815 1366953 1110287 386427 861910 871478 538609 676454 801452 982961 1109941 1010881 433706 381421 620133 653649 1092134 681413 1219867 Galactonic acid 30734216 2175271 1500210 5127119 3853465 6424657 1477716 5041259 10199331 5240917 5747241 5329244 25939994 31564700 3433000 4668414 6801482 8042267 7697178 2344926 7006562 Glucuronic acid 10795925 1741583 1112141 2815075 1151535 2078516 1476724 1023328 2345511 3163933 2106322 2213256 24351030 25279250 583494 1300936 1357848 1890324 5699049 1947260 2263807 Glutamine 17260430 13777886 7869863 18165404 19394300 19627480 11518182 17104232 20108842 21250964 15760951 21765058 11064191 10643353 17979770 20350634 20200182 18210654 13500772 13170659 14998478 Glutaric acid 913582 2891196 505704 1394710 1378743 166765 776285 773678 1091862 184767 1144907 898253 238045 630678 402461 671341 448492 315104 454173 204127 1004946 Glycolic acid 379682 423404 211348 370930 514339 747095 299474 386676 544705 896402 756412 221467 220009 178495 672071 589005 524084 551064 247915 234906 402149 Hexanoylglycine 2076319 658034 858148 3017084 1880458 704008 793320 1481188 1879510 1424201 2813609 2054007 4335969 4519170 1640527 1069468 1408009 9070396 2314133 1688707 4710735 Hexose 80989021.3117621 9682819.08011237 11581974.2100317 56271361.3702255 68623360.8979573 136868841.352771 9804170.79648405 86905265.2101262 200267818.719288 150430367.925509 87401007.0694998 14211223.7002409 92734031.2497924 92402616.4968492 158992714.053476 129234292.376752 142635468.782556 121405630.933468 65203644.7465455 39761066.443867 57452399.8315026 Isocitric acid 5625247.56618707 6686204.13196102 3608442.52909308 7668630.77984422 6005532.07062353 6079777.87700514 4419788.92521223 5023975.56105539 7659571.5278833 3736657.6343431 5814889.75754984 7529062.99274005 5862571.62796855 6021988.59110219 6329244.51944647 4689097.17525058 6842359.91756157 6386883.82576747 4899665.97868832 7195967.35196366 8648699.9375388 Itaconic acid 469283 822241 496481 131151 58522 71621 1111958 1355703 103922 124020 60742 2127723 715944 774655 107675 87603 94042 127948 725606 2018009 88381 Lactic acid 447454076.223459 606726940.840823 352783712.508011 780124691.625818 805527952.156003 1065817628.89518 545849175.133385 848904210.518648 855911504.944056 619268069.922557 767476050.658335 527783934.074136 398570024.975306 294085314.381182 669988269.697866 781003140.464453 788574624.381946 753166392.877008 530217151.544083 555636783.582574 830802118.025849 Ornithine 246681.297456406 892425.251420478 674606.608957124 735052.793042059 796519.224810712 1195550.15784448 1201465.34758758 1448413.14871599 779784.221331156 663949.522641891 926224.349228293 1107511.02993713 678057.175555095 817059.66794812 711070.443711449 895587.380182898 884250.82464777 777362.847784066 843076.503992415 388176.232213723 907964.84762999 Oxidized glutathione 8726 505574 384189 189245 404155 913412 144941 708269 1821529 248885 692726 242521 441277 237371 908897 1047068 1403175 884859 341234 595639 2032941 Phenylalanine 1915245.86739511 3283012.47240943 1527186.3453086 2141757.01055003 2242453.18392819 2538523.98781237 2911790.06139023 3493941.18180922 2262572.67389603 1607004.67681805 2479298.85100016 1742621.94961372 1803037.54406863 1505411.85354111 1352344.51030114 2158634.12257785 1353821.40272806 1536286.38300894 1890077.38489154 1603730.56881006 1961763.2097035 Pyruvic acid 4607117 4350174 1719491 8075338 7153355 8487298 3352591 4671643 6376775 6121974 7802753 5650607 6898985 6177252 4839333 5734990 6390791 6181998 4276513 7761834 5755059 Sugar Alcohol 784182 655609 327331 625334 560483 1545337 255122 935137 1276582 683766 1176972 235940 867657 692039 935062 568354 1321868 1024484 360190 536662 669657 Tauro-α-Muricholic acid/Tauro-β-Muricholic acid 695.625475692097 535260 66594 2542290 84697232 765361 1257.02424540324 36260 1860886 781538 1362690 2457.20034250151 110842 117349 370767 388973 691959 303722 992987 13743 1355994 Thymidine 3283150 4364302 2163618 7733340 5988473 2950670 4665880 6144392 3444213 3631182 4924936 4649374 5315294 5242421 2724285 2823638 3724472 3510022 4971513 3255288 7131180 Traumatic acid 955093 205475 108103 1062237 1053118 175562 256467 104321 426448 260471 600324 549624 889429 1083173 214544 293873 196773 318062 806309 267043 1547732 UMP 2108.82038959092 103545 192.34876258974 13338 3005.69691597018 16609 672.282715031877 3124.92003871943 247879 491.417539723683 187099 2308.35100926575 2299.03137634206 1596.61007895926 169549 297975 157428 186180 793.101776812226 2144.45354045811 200167 Uracil 1660960 2055577 1535112 962036 799393 825114 2156649 2338158 437140 459444 579899 1726126 490773 528996 360274 350267 329688 401301 1483548 1318386 688839 Xylose 1874661 100383 200902 1158336 1360793 2946639 189371 1857415 4472132 3136328 1905331 226355 2283986 2299766 3792362 2955948 3186623 2646561 1566167 838261 1235698 β-Muricholic acid 32475 1835354 23471 645044 10466964 3836.9419309427 55805 1054.116946836 1826154 317531 388062 191115 51490 2588.6781248888 163110 47114 75362 179547 622699 303356 281730 β-Pinene 613143 468907 227178 875045 998220 1014551 247143 628381 912869 707648 588612 850838 1042495 1094172 474245 764886 618139 741317 591350 370597 710956 4-Phenolsulfonic acid 212609276.189675 16539510.8900132 17173133.8061967 65019318.257635 52608066.0107662 1093788.90830755 24891793.0286446 11310667.6514983 75870021.9653109 13574921.4607334 71303088.9514627 75968526.595475107 170086543.172828 181113307.68153 1158514.01190693 4386867.46353563 5324891.47276184 8613438.13744727 86905655.0153883 38069625.1117906 82538597.2176472 L-Ascorbic acid 2-sulfate 39763512.4869485 3770689.46247931 1876427.05330612 4354288.28203995 4206527.53502338 3258609.1761628 4254287.65468196 4211395.45822933 4736650.87505588 5153625.90999587 3964047.03260227 7792920.78178354 33598450.6931169 53293004.6830079 4350860.23253487 4267156.17682704 4161966.81889313 22073535.9783953 5794155.70602388 4077263.09692957 5055793.61104135 Methyl acetoacetate 748775.582658065 372214.205656184 136046.945473824 960500.833213473 1286710.44161137 505744.047063127 510493.541194364 285093.760239173 1459744.98798568 873584.20013858 604849.261933596 645007.490220028 767613.09457098006 738386.033102335 910419.134572845 1008206.44579413 841779.146969667 994831.978914738 900359.126022781 579436.954072582 2096380.46713927 Methyl salicylate 7773687.10874645 4283502.34166866 3457823.27870323 9968068.41402485 7424690.42925696 12445231.6280098 4704789.94952791 6325516.78117894 12437563.5715127 15364254.698615 5110066.20115585 7208883.5011091 4362092.36575663 7334946.0714021 9037085.64772267 9905939.52738695 9628011.77723405 10819681.0412314 9761623.10032163 9029869.87318243 13585800.6220561 N-Glycolylneuraminic acid 49384277.9602983 22394150.9709377 10010891.8468846 702900.625849918 195922.289891535 319214.904313145 42648271.8836508 27050460.8854835 312562.69820763 318824.44317334 74406.3639957568 28480082.4618004 19425799.5196744 22401950.1971186 88793.1912926562 1992.47512403046 43887.984858327 311241.382785908 13418322.7955149 14969963.8529092 287540.577450332 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Neutral_masses Retention_time 2-Hydroxybutyric acid 104.047344 4.52 3-Indoxylsulfuric acid 213.00953 6.114 Arginine 174.11159 1.208 Cholic acid 408.28753 8.809 Citric acid 192.02695 2.321 Fumaric acid 116.010959 2.41 Glutathione 307.08368 2.48 Malic acid 134.02148 1.605 Succinic acid 118.02655 4.321 Taurocholic acid 515.291674 7.9 Threonic acid 136.03709 1.29 Uric acid 168.02834 3.79 Uridine 244.06946 4.615 Xanthine 152.033425 4.56 2-(hydroxymethyl)butanoic acid 118.062994 6 2-C-Methyl-erythritol-4-phosphoric acid 216.04005 1.228 2-Hydroxyisocaproic acid 132.078644 6.88 2-Oxoglutaric acid 146.021523 1.76 3-Methyl-2-oxovaleric acid/ 4-Methyl-2-oxovaleric acid 130.062994 6.06 cis-Aconitic acid 174.016438 1.56 Deoxyuridine 228.074622 4.94 Galactonic acid 196.058303 1.21 Glucuronic acid 194.042653 1.14 Glutamine 146.069142 1.19 Glutaric acid 132.042259 5.36 Glycolic acid 76.016044 1.43 Hexanoylglycine 173.105193 7 Hexose 180.06335 1.229 Isocitric acid 192.02694 1.616 Itaconic acid 130.026609 5.52 Lactic acid 90.03163 2.099 Ornithine 132.08987 1.203 Oxidized glutathione 612.151962 4.51 Phenylalanine 165.07899 5.571 Pyruvic acid 88.016044 1.54 Sugar Alcohol 182.079038 1.24 Tauro-α-Muricholic acid/Tauro-β-Muricholic acid 515.291674 7.54 Thymidine 242.090272 5.53 Traumatic acid 228.136159 7.69 UMP 324.035867 1.89 Uracil 112.027277 2.5 Xylose 150.052824 1.24 β-Muricholic acid 408.287575 8.16 β-Pinene 182.057909 6.18 4-Phenolsulfonic acid 173.99856 5.956 L-Ascorbic acid 2-sulfate 255.9889 1.809 Methyl acetoacetate 116.0473 4.641 Methyl salicylate 152.04732 6.528 N-Glycolylneuraminic acid 325.10096 1.184 METABOLITES_END #END