#METABOLOMICS WORKBENCH Lxu_20240904_094400 DATATRACK_ID:5164 STUDY_ID:ST003459 ANALYSIS_ID:AN005684 PROJECT_ID:PR002122 VERSION 1 CREATED_ON September 4, 2024, 7:33 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 liver to support understanding of early metabolic shift in ST:STUDY_TITLE presymptomatic sepsis patients. (Part5 mice liver 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 liver tissues from mice model revealed early septic ST:STUDY_SUMMARY biomarkers in liver. 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_0194 SA020910 Sample source:Liver | Group:Sham / 13C-Serin RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-026-SA020910_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-026-SA020910_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0200 SA020942 Sample source:Liver | Group:CLP (24h) / 13C-Serin RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-025-SA020942_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-025-SA020942_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0204 SA020970 Sample source:Liver | Group:CLP (24h) / Vehicle RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-029-SA020970_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-029-SA020970_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0271 SA021005 Sample source:Liver | Group:Sham / 12C-Serin RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-033-SA021005_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-033-SA021005_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0190 SA020889 Sample source:Liver | Group:CLP (24h) / 13C-Serin RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-015-SA020889_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-015-SA020889_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0196 SA020917 Sample source:Liver | Group:Sham / Vehicle RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-020-SA020917_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-020-SA020917_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0272 SA021012 Sample source:Liver | Group:Sham / 12C-Serin RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-012-SA021012_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-012-SA021012_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0269 SA020991 Sample source:Liver | Group:Sham / 12C-Serin RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-022-SA020991_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-022-SA020991_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0198 SA020929 Sample source:Liver | Group:Sham / Vehicle RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-013-SA020929_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-013-SA020929_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0193 SA020903 Sample source:Liver | Group:Sham / 13C-Serin RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-021-SA020903_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-021-SA020903_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0189 SA020882 Sample source:Liver | Group:CLP (24h) / 13C-Serin RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-031-SA020882_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-031-SA020882_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0197 SA020924 Sample source:Liver | Group:CLP (24h) / Vehicle RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-010-SA020924_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-010-SA020924_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0270 SA020998 Sample source:Liver | Group:Sham / 12C-Serin RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-032-SA020998_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-032-SA020998_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0273 SA021019 Sample source:Liver | Group:CLP (24h) / 12C-Serin RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-011-SA021019_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-011-SA021019_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0202 SA020956 Sample source:Liver | Group:Sham / 13C-Serin RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-027-SA020956_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-027-SA020956_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0268 SA020984 Sample source:Liver | Group:CLP (24h) / 12C-Serin RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-017-SA020984_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-017-SA020984_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0192 SA020896 Sample source:Liver | Group:CLP (24h) / Vehicle RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-018-SA020896_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-018-SA020896_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0199 SA020935 Sample source:Liver | Group:CLP (24h) / 13C-Serin RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-034-SA020935_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-034-SA020935_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0203 SA020963 Sample source:Liver | Group:Sham / 13C-Serin RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-028-SA020963_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-028-SA020963_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0201 SA020949 Sample source:Liver | Group:Sham / Vehicle RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-014-SA020949_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-014-SA020949_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0267 SA020977 Sample source:Liver | Group:CLP (24h) / 12C-Serin RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-019-SA020977_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-019-SA020977_pos.mzML SUBJECT_SAMPLE_FACTORS Bl6J_23_0274 SA021026 Sample source:Liver | Group:CLP (24h) / 12C-Serin RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-024-SA021026_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-024-SA021026_pos.mzML SUBJECT_SAMPLE_FACTORS NA Blank_PB Sample source:blank | Group:Blank RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-045-Blank_PB_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-045-Blank_PB_pos.mzML SUBJECT_SAMPLE_FACTORS NA Blank_004 Sample source:blank | Group:Blank RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-044-Blank_004_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-044-Blank_004_pos.mzML SUBJECT_SAMPLE_FACTORS NA QC_001 Sample source:Liver | Group:QC RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-006-QC_001_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-006-QC_001_pos.mzML SUBJECT_SAMPLE_FACTORS NA QC_005 Sample source:Liver | Group:QC RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-016-QC_005_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-016-QC_005_pos.mzML SUBJECT_SAMPLE_FACTORS NA Blank_001 Sample source:blank | Group:Blank RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-003-Blank_001_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-003-Blank_001_pos.mzML SUBJECT_SAMPLE_FACTORS NA QC_007 Sample source:Liver | Group:QC RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-030-QC_007_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-030-QC_007_pos.mzML SUBJECT_SAMPLE_FACTORS NA Blank_002 Sample source:blank | Group:Blank RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-005-Blank_002_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-005-Blank_002_pos.mzML SUBJECT_SAMPLE_FACTORS NA Blank_003 Sample source:blank | Group:Blank RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-039-Blank_003_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-039-Blank_003_pos.mzML SUBJECT_SAMPLE_FACTORS NA QC_006 Sample source:Liver | Group:QC RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-023-QC_006_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-023-QC_006_pos.mzML SUBJECT_SAMPLE_FACTORS NA QC_008 Sample source:Liver | Group:QC RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-035-QC_008_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-035-QC_008_pos.mzML SUBJECT_SAMPLE_FACTORS NA QC_004 Sample source:Liver | Group:QC RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-009-QC_004_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-009-QC_004_pos.mzML SUBJECT_SAMPLE_FACTORS NA QC_009 Sample source:Liver | Group:QC RAW_FILE_NAME(Raw file name negative ion)=230704-EXP-PAN-1-041-QC_009_neg.mzML; RAW_FILE_NAME(Raw file name positive ion)=230704-EXP-PAN-1-041-QC_009_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 Liver #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 #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 ST003459_AN005684_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 SA020882 SA020889 SA020896 SA020903 SA020910 SA020917 SA020924 SA020929 SA020935 SA020942 SA020949 SA020956 SA020963 SA020970 SA020977 SA020984 SA020991 SA020998 SA021005 SA021012 SA021019 SA021026 Factors Sample source:Liver | Group:CLP (24h) / 13C-Serin Sample source:Liver | Group:CLP (24h) / 13C-Serin Sample source:Liver | Group:CLP (24h) / Vehicle Sample source:Liver | Group:Sham / 13C-Serin Sample source:Liver | Group:Sham / 13C-Serin Sample source:Liver | Group:Sham / Vehicle Sample source:Liver | Group:CLP (24h) / Vehicle Sample source:Liver | Group:Sham / Vehicle Sample source:Liver | Group:CLP (24h) / 13C-Serin Sample source:Liver | Group:CLP (24h) / 13C-Serin Sample source:Liver | Group:Sham / Vehicle Sample source:Liver | Group:Sham / 13C-Serin Sample source:Liver | Group:Sham / 13C-Serin Sample source:Liver | Group:CLP (24h) / Vehicle Sample source:Liver | Group:CLP (24h) / 12C-Serin Sample source:Liver | Group:CLP (24h) / 12C-Serin Sample source:Liver | Group:Sham / 12C-Serin Sample source:Liver | Group:Sham / 12C-Serin Sample source:Liver | Group:Sham / 12C-Serin Sample source:Liver | Group:Sham / 12C-Serin Sample source:Liver | Group:CLP (24h) / 12C-Serin Sample source:Liver | Group:CLP (24h) / 12C-Serin 4-pyridoxic acid 8520662 11083929 3990243 7268478 6049368 3230164 5754732 4246016 4853707 4369167 4646352 3309774 5493144 2628080 1186119 1567795 1655699 5998305 10335635 4816973 6670238 4124301 Adenosine diphosphate ribose 256473.98853073 847440.672966591 288514.73981317 614909.373387669 411560.914223903 131946.149878378 576102.000411253 605735.775667523 165883.64420843 535135.407112802 104401.544866289 223417.546255266 183988.873596676 214461.321247344 82373.8731416313 63035.5974020289 195330.544402355 723359.349439442 517437.468220596 741885.308885853 745479.955217952 267232.39520672 Allantoin 3700363 3069834 2693449 2654806 3795617 1935360 2726482 2383568 1681075 2258298 2767102 1656406 1691667 2205798 1322341 1666438 990674 1764716 2075531 2352069 2318228 1759903 Glucose-6-phosphate 3980215.27425253 2427176.35874618 1226306.22881231 2737473.79537461 18026552.5837681 3069033.48420595 4361537.89197177 2079260.591607 2457716.35493014 2492862.40432309 10081787.6932105 2125731.51709509 1880821.16250383 9125422.67663737 630115.335588051 2190129.17452224 4561028.12127022 6765001.50473486 11757731.6853141 6425011.60828673 18338756.3984483 2685027.09335227 Glycylleucine 3572181 4123209 1303560 3533390 1808126 1192979 1636720 767256 830050 1867557 2475841 1118494 3102496 415441 156697 132147 339344 2456536 3293213 1656878 1483383 502204 Hippuric acid 4247269 3273351 482844 2104407 2098980 1729236 3256653 731247 1582989 2912294 1422857 921754 1667746 875836 877269 854416 380087 2422713 2340917 2315646 2716282 788722 Leucylalanine 5330587 5167933 1727152 3938178 1622427 1303366 2284243 889283 1216074 2538360 4216200 1657363 4636841 554238 210607 204627 437961 4085034 5260952 2659137 2614690 757432 Malic acid 26696958 15051698 6648804 7962003 7868033 3736155 12910183 3673883 7789418 7592167 17354792 15773245 10451891 4137245 2526982 2822326 2877370 8626757 10527163 6953026 16817824 15797919 Oxidized glutathione 39034398.6152194 72346733.937395 40511186.2692987 56961862.2875296 32638655.8333294 48730229.9742085 72308400.7749902 54900779.632421 38264625.2827347 46727596.1028281 32150175.783958 31260713.9037825 49959105.1282091 15614367.0159942 5155073.69150182 5396682.36299971 13765533.7737284 33336264.7559947 33291492.6804989 39350239.3834286 37819420.7999877 27564241.6779554 Pantothenic acid 28982428 29635002 7505347 33025024 10413821 9780834 10507076 10929060 6884845 11005016 16038823 14308448 23299100 6047114 1954458 2131558 3947911 15209427 22853570 13289150 13071786 5883900 Phenylalanine 5973410 6483761 2049664 5554284 3147735 2267547 2854413 1963502 1757060 2350098 3141679 1998338 3952654 1037671 487261 560538 753755 3378764 4167104 2372474 2919185 1225228 Threonic acid 1379920 761453 282726 583626 305469 649977 1323478 234303 186670 378130 920744 325849 604279 121851 61168 97167 194065 883570 1100579 702635 423257 162830 Uracil 2888729 2750579 2108077 1966591 1129185 1040646 2678820 1879281 2468514 1834070 1124526 1114494 1280725 1130719 1157074 1022004 704300 1399902 1436511 1373302 2292748 2885030 Xanthine 18999428 27324674 15316705 19901630 15241896 11108686 17834370 13768664 15777373 14681808 11742093 10163939 14721478 9573974 4874099 4733690 3574405 12042758 14504030 9912125 9477068 13090894 Xanthosine 22170616 32837900 11902932 19711042 20438060 10707107 24836468 14073723 14241564 16682110 29766944 10111446 18671850 7460385 3375166 4264500 6987202 24748760 35402292 19027566 17980030 13307616 2-Aminoadipic acid 3362697 14433516 586656 7142477 3536337 88408 1379392 1680471 3345457 576806 265897 294085 1316048 4936323 359668 191111 51754 420765 198963 114674 405801 355622 2-C-Methyl-erythritol-4-phosphoric acid 52005988 13774256 14553057 34368656 36676916 92592960 61560848 17421490 12426422 40196252 96560792 60237468 61712380 8923267 12383717 21704898 65016288 71335280 81917304 78389232 17309016 15088338 Aspartic acid 10946552 12013534 4413637 9310554 6906045 5634299 7564048 4867031 3457339 4874644 6498607 4630231 12354901 2683270 972434 1066702 1944811 5915812 7095305 4025110 5499345 2781478 cis-Aconitic acid 127075 165954 1026245 81116 495420 254449 1460436 21471 292440 1388914 2330029 493077 772363 23928 27388 138.65803669807963 367161 1883174 2310620 1365323 2113993 869781 Deoxyuridine 293180 1498039 360766 970504 194567 161309 107284 240097 243131 223387 46666 336777 419666 292136 42146 60241 19128 55562 75481 87460 72190 49455 Fumaric acid 295030 270954 148849 165862 149096 107556 254174 97428 182064 202733 276938 333102 196932 111525 84448 118862 105199 163467 193064 170164 277002 292830 Galactonic acid/ Gluconic acid 108607160 66737984 60378912 100000504 44489128 90802368 96920504 56973324 27757744 44952128 120336304 56502056 122073912 18119976 6039487 5602037 14922516 94666696 142326896 59833616 73664432 26875176 Glucuronic acid 6841652.83233616 3141347.74334535 1914572.48278484 5358001.9424095 1948425.5092274 3429234.15752237 4147794.91444365 3241032.18110723 1209486.59049742 1410328.08214147 3604498.01668391 3302404.6440993 5061595.03446432 896337.713689705 817261.078920438 827876.320463061 488003.613164359 3243573.37019924 5000193.04852218 2908961.12692039 2916204.23461446 982813.063341006 Glutamic acid/ N-Methylaspartic acid 51021868 26040800 13622726 25984952 27164184 22817196 32712482 10751750 11752489 15734642 39439080 19687396 40404748 8549311 2950517 3043359 8380261 30325906 41176524 23294658 16682267 10140268 Gulonic acid γ-lactone 2154058 1254733 1201554 1971527 842803 1911565 1971094 1156567 561632 888196 2380747 1049746 2485095 346299 99768 115161 276379 1824432 2923586 1035640 1468116 558742 Hexoses 38037140 7816943 6549384 20777818 20474768 70065032 47096112 6922655 4514794 19468574 100765240 36359844 48916628 3055196 5855655 9319945 33512862 53812800 78137496 58841648 8371045 5359085 Hexoses dimer 588425 87606 71426 130917 253467 18404170 372568 74748 64508 90449 5538556 1472000 3750829 30886 53039 76911 2917903 3668164 8497057 3695884 127404 74576 Isocitric acid 781247.85811781 431176.880981529 4425251.30950803 251170.856408231 818523.2792112 1715663.62213944 4384761.92703161 199994.242643653 1596835.49222726 5620391.23696975 9407193.17138018 3596890.90266553 2574547.60133549 289873.469339614 505372.077567732 212009.528111318 3963358.82165713 5668896.19921931 6541095.46530934 4495799.16999111 6137594.38182984 4418119.95231303 Itaconic acid 1153724 377475 184407 53021 24873 30271 702455 36539 2045396 130971 56584 44217 46583 523972 461742 816870 1043 22668 20268 42395 1721632 1249629 Lactic acid 70359104 27515606 34651508 34663304 31646766 65129020 103222440 25191232 13595226 49400864 145999856 37033152 42906836 10475579 21234844 23751730 46843300 73526672 100700400 70602600 41937436 22532250 Methanesulfonic acid 4997776 2227493 422935 5324412 3545914 1755566 1378106 947983 475981 1111525 751356 1374771 2096420 304967 156209 312261 288175 971548 1323810 818814 670643 247595 Mucic acid 910361 1294243 289590 878591 424165 390781 675466 284219 399625 414978 529589 638239 528069 258805 21020 3615.711855551197 55420 453746 692392 567245 589550 260368 N-Acetylalanine 631641 535597 228802 606454 433079 319401 405587 263293 211879 295576 331150 205349 490680 191373 107123 107500 156711 372044 408129 284830 332751 163593 N-Acetylglucosamine 1278045 1235908 607915 703155 589724 387954 991054 441596 529248 602664 530100 257619 677938 343438 160572 159814 131129 541414 656225 462428 588001 413026 N-Acetylneuraminic acid 699541 747798 634559 397010 415194 315322 571033 317689 415878 498197 424170 229920 486783 265636 70741 79493 76786 209381 257602 206139 330600 257312 Pentoses 2206146 1332297 826934 1830882 1166213 2860382 2499359 991199 597682 1190051 3470437 1507152 2343320 483582 389444 492005 1146367 2256303 3190931 2333427 709312 520277 Ribulose 5-phosphate 1712361 1636415 908461 1936605 1828681 1287498 1650547 1819883 1196711 1016805 2228922 1560794 1945045 841978 518951 607459 974638 1867164 2485961 1549122 1487676 792784 Serine 4576610 4610916 2198180 5059024 4600331 4152613 4577556 3181927 1639198 2288727 3941547 2231998 5660926 1392735 1559971 1743586 1786549 4098022 4244752 3004841 3899280 1617956 Sugar alcohol (C5) 494567 885970 566901 537544 470028 865556 751359 431459 484625 415611 618380 368516 826627 281479 127968 81581 271480 730054 727224 618095 306769 310758 Taurine 41998032 71375328 51565628 52686988 32802768 44636404 79508120 32311632 42654984 52349376 15017675 37434224 30802260 32522284 22366176 23468436 18778108 39545212 44702152 38674288 44883916 38523776 Threonine 6486534 5339995 2215330 6461757 4537460 5102517 5248285 2940446 2060431 3041851 6870424 3316543 7826760 1285019 948903 1039218 1996815 5081084 6090891 4165550 3641912 1640080 Thymidine 839039 5013484 1936201 4057685 4957421 1562869 573895 2305436 1147364 1196869 387020 2461912 1963068 1996314 505434 1191533 219280 331096 412801 791923 622870 287366 Uric acid 745569 881981 282268 135214 76400 265136 2630060 671585 3044035 1542105 1383255 680373 623810 275681 232609 106951 223211 1043605 1096221 352497 563767 2467358 Uridine 51020384 33816584 22834966 31282826 27644258 16094807 48395420 18905246 22358554 21386482 26722350 14656590 22483500 16216361 10630296 12120206 10832319 23462524 34250040 24555908 27835564 15055603 α-aminobutyric acid/ N,N-Dimethylglycine 1672719 945882 494058 922813 993999 770465 1138838 381961 447921 515043 1340910 652699 1290150 282867 89276 106388 259460 987799 1284608 732713 618021 333142 δ-Gluconolactone 207315 58229 359110 220836 129564 401378 351678 382438 44941 75646 205189 79730 176295 65688 72985 63545 103369 153407 141683 111955 71320 125774 4-chloro-1H-indazol-3-amine 1689716.84840061 1793314.46526506 1857088.4128158 1683675.57188315 471971.854327769 569363.99852577 1444518.59395096 434582.235843 1028283.38230161 877152.260631556 463752.808460942 713176.720901166 1396792.5441674 820631.474953639 582416.804858542 671902.674197926 282875.994584373 1250372.50294467 1080470.87989717 1220846.54552787 1340337.85572075 762729.165329703 N-Glycolylneuraminic acid 6291350.91008562 8233239.17622911 5316283.16059611 3436366.06579527 4658179.34941437 2453209.73542675 5760635.70378511 2686368.56709913 4710904.75047063 5042579.07131683 4519712.78613244 2714303.34910886 5240432.97233459 1486017.32967188 565431.231402537 405565.146881921 1184772.4909895 2788671.01637477 4378941.1687576 3046696.68012615 2613775.65225482 2147797.95919983 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Neutral_masses Retention_time 4-pyridoxic acid 183.053158 5.19 Adenosine diphosphate ribose 559.07221 1.405 Allantoin 158.04399 1.37 Glucose-6-phosphate 260.02974 1.1 Glycylleucine 188.116092 5.84 Hippuric acid 179.058243 6.45 Leucylalanine 202.131743 5.13 Malic acid 134.021523 1.6 Oxidized glutathione 612.15205 4.51 Pantothenic acid 219.110673 5.8 Phenylalanine 165.078979 5.57 Threonic acid 136.037173 1.24 Uracil 112.027277 2.5 Xanthine 152.033425 4.56 Xanthosine 284.075684 5.35 2-Aminoadipic acid 161.068808 1.47 2-C-Methyl-erythritol-4-phosphoric acid 216.03989 1.24 Aspartic acid 133.037508 1.14 cis-Aconitic acid 174.016438 1.56 Deoxyuridine 228.074622 4.93 Fumaric acid 116.010959 2.4 Galactonic acid/ Gluconic acid 196.058303 1.19 Glucuronic acid 194.04263 1.15 Glutamic acid/ N-Methylaspartic acid 147.053158 1.23 Gulonic acid γ-lactone 178.047738 1.2 Hexoses 180.063388 1.23 Hexoses dimer 342.116212 1.47 Isocitric acid 192.02693 1.603 Itaconic acid 130.026609 5.52 Lactic acid 90.031694 2.04 Methanesulfonic acid 95.988115 1.24 Mucic acid 210.037567 1.15 N-Acetylalanine 131.058243 4.28 N-Acetylglucosamine 221.089937 1.47 N-Acetylneuraminic acid 309.105981 1.28 Pentoses 150.052824 1.34 Ribulose 5-phosphate 230.019154 1.16 Serine 105.042593 1.13 Sugar alcohol (C5) 152.068474 1.27 Taurine 125.014664 1.14 Threonine 119.058243 1.2 Thymidine 242.090272 5.54 Uric acid 168.02834 3.79 Uridine 244.069536 4.62 α-aminobutyric acid/ N,N-Dimethylglycine 103.063329 1.22 δ-Gluconolactone 178.047738 1.33 4-chloro-1H-indazol-3-amine 167.02518 1.584 N-Glycolylneuraminic acid 325.10102 1.178 METABOLITES_END #END