#METABOLOMICS WORKBENCH VeronicaGhini_20240123_113226 DATATRACK_ID:4608 STUDY_ID:ST003054 ANALYSIS_ID:AN005008 PROJECT_ID:PR001903 VERSION 1 CREATED_ON January 24, 2024, 10:53 am #PROJECT PR:PROJECT_TITLE Integrated metabolomics and proteomics of symptomatic and early pre-symptomatic PR:PROJECT_TITLE states of colitis PR:PROJECT_TYPE NMR-based metabolomics PR:PROJECT_SUMMARY Colitis has a multifactorial pathogenesis with a strong cross-talk among PR:PROJECT_SUMMARY microbiota, hypoxia and tissue metabolism. Here, we aimed to characterize the PR:PROJECT_SUMMARY molecular signature of the disease in symptomatic and pre-symptomatic stages of PR:PROJECT_SUMMARY the inflammatory process at the tissue and fecal level. The study is based on PR:PROJECT_SUMMARY two different murine models for colitis. High-resolution Magic Angle Spinning PR:PROJECT_SUMMARY NMR on cryopreserved “intact” colon tissues and LC-MS/MS on colon tissue PR:PROJECT_SUMMARY extracts were used to derive untargeted metabolomics and proteomics information, PR:PROJECT_SUMMARY respectively. Solution NMR was used to derive metabolomic profiles of fecal PR:PROJECT_SUMMARY extracts. By combining metabolomic and proteomic analyses of the tissues, we PR:PROJECT_SUMMARY found increased anaerobic glycolysis, accompanied by altered citric acid cycle PR:PROJECT_SUMMARY and oxidative phosphorylation in inflamed colons; these changes associate with PR:PROJECT_SUMMARY inflammation-induced hypoxia taking place in colon tissues. Pre-symptomatic PR:PROJECT_SUMMARY states can be discriminated from healthy samples before macroscopic inflammation PR:PROJECT_SUMMARY is observed. Different colitis states are characterized by significantly PR:PROJECT_SUMMARY different metabolomic profiles of fecal extracts, attributable to both the PR:PROJECT_SUMMARY dysbiosis characteristic of colitis, as well as the dysregulated tissue PR:PROJECT_SUMMARY metabolism. Strong and distinctive fecal metabolomic signatures can be detected PR:PROJECT_SUMMARY before onset of symptoms. Therefore, untargeted metabolomics of tissues and PR:PROJECT_SUMMARY fecal extracts provides a comprehensive picture of the changes accompanying the PR:PROJECT_SUMMARY disease onset already at pre-clinical stages, highlighting the diagnostic PR:PROJECT_SUMMARY potential of global metabolomics for inflammatory diseases. PR:INSTITUTE University of Florence PR:DEPARTMENT Department of Chemistry PR:LABORATORY Metabolomics PR:LAST_NAME Ghini PR:FIRST_NAME Veronica PR:ADDRESS via Luigi Sacconi, 6, Sesto Fiorentino, Firenze, 50019, Italy PR:EMAIL ghini@cerm.unifi.it PR:PHONE +390554574266 #STUDY ST:STUDY_TITLE Integrated metabolomics and proteomics of symptomatic and early pre-symptomatic ST:STUDY_TITLE states of colitis ST:STUDY_SUMMARY Colitis has a multifactorial pathogenesis with a strong cross-talk among ST:STUDY_SUMMARY microbiota, hypoxia and tissue metabolism. Here, we aimed to characterize the ST:STUDY_SUMMARY molecular signature of the disease in symptomatic and pre-symptomatic stages of ST:STUDY_SUMMARY the inflammatory process at the tissue and fecal level. The study is based on ST:STUDY_SUMMARY two different murine models for colitis. High-resolution Magic Angle Spinning ST:STUDY_SUMMARY NMR on cryopreserved “intact” colon tissues and LC-MS/MS on colon tissue ST:STUDY_SUMMARY extracts were used to derive untargeted metabolomics and proteomics information, ST:STUDY_SUMMARY respectively. Solution NMR was used to derive metabolomic profiles of fecal ST:STUDY_SUMMARY extracts. By combining metabolomic and proteomic analyses of the tissues, we ST:STUDY_SUMMARY found increased anaerobic glycolysis, accompanied by altered citric acid cycle ST:STUDY_SUMMARY and oxidative phosphorylation in inflamed colons; these changes associate with ST:STUDY_SUMMARY inflammation-induced hypoxia taking place in colon tissues. Pre-symptomatic ST:STUDY_SUMMARY states can be discriminated from healthy samples before macroscopic inflammation ST:STUDY_SUMMARY is observed. Different colitis states are characterized by significantly ST:STUDY_SUMMARY different metabolomic profiles of fecal extracts, attributable to both the ST:STUDY_SUMMARY dysbiosis characteristic of colitis, as well as the dysregulated tissue ST:STUDY_SUMMARY metabolism. Strong and distinctive fecal metabolomic signatures can be detected ST:STUDY_SUMMARY before onset of symptoms. Therefore, untargeted metabolomics of tissues and ST:STUDY_SUMMARY fecal extracts provides a comprehensive picture of the changes accompanying the ST:STUDY_SUMMARY disease onset already at pre-clinical stages, highlighting the diagnostic ST:STUDY_SUMMARY potential of global metabolomics for inflammatory diseases. ST:INSTITUTE University of Florence ST:LAST_NAME Ghini ST:FIRST_NAME Veronica ST:ADDRESS via Luigi Sacconi, 6, Sesto Fiorentino, Firenze, 50019, Italy ST:EMAIL ghini@cerm.unifi.it ST:PHONE +390554574266 ST:NUM_GROUPS 5 ST:TOTAL_SUBJECTS 50 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:GENDER Male and female #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 - IL10_KO_HC1 Group:Healthy_IL10-/- RAW_FILE_NAME=IL10_KO_HC1 SUBJECT_SAMPLE_FACTORS - IL10_KO_HC10 Group:Healthy_IL10-/- RAW_FILE_NAME=IL10_KO_HC10 SUBJECT_SAMPLE_FACTORS - IL10_KO_HC2 Group:Healthy_IL10-/- RAW_FILE_NAME=IL10_KO_HC2 SUBJECT_SAMPLE_FACTORS - IL10_KO_HC3 Group:Healthy_IL10-/- RAW_FILE_NAME=IL10_KO_HC3 SUBJECT_SAMPLE_FACTORS - IL10_KO_HC4 Group:Healthy_IL10-/- RAW_FILE_NAME=IL10_KO_HC4 SUBJECT_SAMPLE_FACTORS - IL10_KO_HC5 Group:Healthy_IL10-/- RAW_FILE_NAME=IL10_KO_HC5 SUBJECT_SAMPLE_FACTORS - IL10_KO_HC6 Group:Healthy_IL10-/- RAW_FILE_NAME=IL10_KO_HC6 SUBJECT_SAMPLE_FACTORS - IL10_KO_HC7 Group:Healthy_IL10-/- RAW_FILE_NAME=IL10_KO_HC7 SUBJECT_SAMPLE_FACTORS - IL10_KO_HC8 Group:Healthy_IL10-/- RAW_FILE_NAME=IL10_KO_HC8 SUBJECT_SAMPLE_FACTORS - IL10_KO_HC9 Group:Healthy_IL10-/- RAW_FILE_NAME=IL10_KO_HC9 SUBJECT_SAMPLE_FACTORS - IL10_KO_ILL1 Group:Ill_IL10-/- RAW_FILE_NAME=IL10_KO_ILL1 SUBJECT_SAMPLE_FACTORS - IL10_KO_ILL10 Group:Ill_IL10-/- RAW_FILE_NAME=IL10_KO_ILL10 SUBJECT_SAMPLE_FACTORS - IL10_KO_ILL2 Group:Ill_IL10-/- RAW_FILE_NAME=IL10_KO_ILL2 SUBJECT_SAMPLE_FACTORS - IL10_KO_ILL3 Group:Ill_IL10-/- RAW_FILE_NAME=IL10_KO_ILL3 SUBJECT_SAMPLE_FACTORS - IL10_KO_ILL4 Group:Ill_IL10-/- RAW_FILE_NAME=IL10_KO_ILL4 SUBJECT_SAMPLE_FACTORS - IL10_KO_ILL5 Group:Ill_IL10-/- RAW_FILE_NAME=IL10_KO_ILL5 SUBJECT_SAMPLE_FACTORS - IL10_KO_ILL6 Group:Ill_IL10-/- RAW_FILE_NAME=IL10_KO_ILL6 SUBJECT_SAMPLE_FACTORS - IL10_KO_ILL7 Group:Ill_IL10-/- RAW_FILE_NAME=IL10_KO_ILL7 SUBJECT_SAMPLE_FACTORS - IL10_KO_ILL8 Group:Ill_IL10-/- RAW_FILE_NAME=IL10_KO_ILL8 SUBJECT_SAMPLE_FACTORS - IL10_KO_ILL9 Group:Ill_IL10-/- RAW_FILE_NAME=IL10_KO_ILL9 SUBJECT_SAMPLE_FACTORS - WT_DSS4_1 Group:Day_4_DSS RAW_FILE_NAME=HRMAS-Mice_colon-WT_DSS4_1 SUBJECT_SAMPLE_FACTORS - WT_DSS4_10 Group:Day_4_DSS RAW_FILE_NAME=HRMAS-Mice_colon-WT_DSS4_10 SUBJECT_SAMPLE_FACTORS - WT_DSS4_2 Group:Day_4_DSS RAW_FILE_NAME=HRMAS-Mice_colon-WT_DSS4_2 SUBJECT_SAMPLE_FACTORS - WT_DSS4_3 Group:Day_4_DSS RAW_FILE_NAME=HRMAS-Mice_colon-WT_DSS4_3 SUBJECT_SAMPLE_FACTORS - WT_DSS4_4 Group:Day_4_DSS RAW_FILE_NAME=HRMAS-Mice_colon-WT_DSS4_4 SUBJECT_SAMPLE_FACTORS - WT_DSS4_5 Group:Day_4_DSS RAW_FILE_NAME=HRMAS-Mice_colon-WT_DSS4_5 SUBJECT_SAMPLE_FACTORS - WT_DSS4_6 Group:Day_4_DSS RAW_FILE_NAME=HRMAS-Mice_colon-WT_DSS4_6 SUBJECT_SAMPLE_FACTORS - WT_DSS4_7 Group:Day_4_DSS RAW_FILE_NAME=HRMAS-Mice_colon-WT_DSS4_7 SUBJECT_SAMPLE_FACTORS - WT_DSS4_8 Group:Day_4_DSS RAW_FILE_NAME=HRMAS-Mice_colon-WT_DSS4_8 SUBJECT_SAMPLE_FACTORS - WT_DSS4_9 Group:Day_4_DSS RAW_FILE_NAME=HRMAS-Mice_colon-WT_DSS4_9 SUBJECT_SAMPLE_FACTORS - WT_HC_1 Group:WT RAW_FILE_NAME=WT_HC_1 SUBJECT_SAMPLE_FACTORS - WT_HC_10 Group:WT RAW_FILE_NAME=WT_HC_10 SUBJECT_SAMPLE_FACTORS - WT_HC_2 Group:WT RAW_FILE_NAME=WT_HC_2 SUBJECT_SAMPLE_FACTORS - WT_HC_3 Group:WT RAW_FILE_NAME=WT_HC_3 SUBJECT_SAMPLE_FACTORS - WT_HC_4 Group:WT RAW_FILE_NAME=WT_HC_4 SUBJECT_SAMPLE_FACTORS - WT_HC_5 Group:WT RAW_FILE_NAME=WT_HC_5 SUBJECT_SAMPLE_FACTORS - WT_HC_6 Group:WT RAW_FILE_NAME=WT_HC_6 SUBJECT_SAMPLE_FACTORS - WT_HC_7 Group:WT RAW_FILE_NAME=WT_HC_7 SUBJECT_SAMPLE_FACTORS - WT_HC_8 Group:WT RAW_FILE_NAME=WT_HC_8 SUBJECT_SAMPLE_FACTORS - WT_HC_9 Group:WT RAW_FILE_NAME=WT_HC_9 SUBJECT_SAMPLE_FACTORS - WT_DSS_1 Group:Day_10_DSS RAW_FILE_NAME=WT_PSS_1 SUBJECT_SAMPLE_FACTORS - WT_DSS_10 Group:Day_10_DSS RAW_FILE_NAME=WT_PSS_10 SUBJECT_SAMPLE_FACTORS - WT_DSS_2 Group:Day_10_DSS RAW_FILE_NAME=WT_PSS_2 SUBJECT_SAMPLE_FACTORS - WT_DSS_3 Group:Day_10_DSS RAW_FILE_NAME=WT_PSS_3 SUBJECT_SAMPLE_FACTORS - WT_DSS_4 Group:Day_10_DSS RAW_FILE_NAME=WT_PSS_4 SUBJECT_SAMPLE_FACTORS - WT_DSS_5 Group:Day_10_DSS RAW_FILE_NAME=WT_PSS_5 SUBJECT_SAMPLE_FACTORS - WT_DSS_6 Group:Day_10_DSS RAW_FILE_NAME=WT_PSS_6 SUBJECT_SAMPLE_FACTORS - WT_DSS_7 Group:Day_10_DSS RAW_FILE_NAME=WT_PSS_7 SUBJECT_SAMPLE_FACTORS - WT_DSS_8 Group:Day_10_DSS RAW_FILE_NAME=WT_PSS_8 SUBJECT_SAMPLE_FACTORS - WT_DSS_9 Group:Day_10_DSS RAW_FILE_NAME=WT_PSS_9 #COLLECTION CO:COLLECTION_SUMMARY Colons were harvested from euthanized mice and flash-frozen in liquid nitrogen. CO:COLLECTION_SUMMARY For all downstream analyses, the distal region of the colon (0.5-2 cm from CO:COLLECTION_SUMMARY rectum) was used, since it is where colitis is most pronounced in these models. CO:SAMPLE_TYPE Colon CO:STORAGE_CONDITIONS -80℃ #TREATMENT TR:TREATMENT_SUMMARY Acute colitis was induced in wild type (WT) C57BL/6 mice via addition of 1.25% TR:TREATMENT_SUMMARY DSS into the drinking water for seven days. To gain insight on chronic TR:TREATMENT_SUMMARY intestinal inflammation, we used C57BL/6 IL-10-/- mice that spontaneously TR:TREATMENT_SUMMARY develop chronic colon inflammation. This inflammation is not chemically induced, TR:TREATMENT_SUMMARY but rather is considered to be a result of a dysregulated immune system, which TR:TREATMENT_SUMMARY more resembles the clinical manifestation of chronic UC and CD patients and TR:TREATMENT_SUMMARY mimics a monogenic form of IBD20. In order to synchronize and accelerate the TR:TREATMENT_SUMMARY onset of chronic colitis, mice were exposed to 200 ppm piroxicam (Sigma-Aldrich TR:TREATMENT_SUMMARY ltd.) via supplementation in normal murine chow for 14 days. With the described TR:TREATMENT_SUMMARY experimental procedures, we obtained five groups of mice (10 mice per group) for TR:TREATMENT_SUMMARY further analysis: 1) Healthy WT; 2) Day 4 of DSS (pre-symptomatic); 3) Day 10 of TR:TREATMENT_SUMMARY DSS (acute colitis); 4) Healthy IL-10-/- (pre-symptomatic); and 5) Ill IL-10-/- TR:TREATMENT_SUMMARY (chronic colitis). #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Frozen colon samples were trimmed (15–20 mg) to fit HR-MAS ZrO2 rotor insert SP:SAMPLEPREP_SUMMARY capacity (50 μL). Each insert was filled with 2H2O containing 5.8 mM sodium SP:SAMPLEPREP_SUMMARY trimethylsilyl [2,2,3,3-2H4]propionate (TMSP). Inserts were covered with plug SP:SAMPLEPREP_SUMMARY and plug-restraining screw and inserted into the 4 mm rotor for HR-MAS probe. #ANALYSIS AN:ANALYSIS_TYPE NMR #NMR NM:INSTRUMENT_NAME Bruker 600 MHz NM:INSTRUMENT_TYPE FT-NMR NM:NMR_EXPERIMENT_TYPE 1D-1H NM:SPECTROMETER_FREQUENCY 600 MHz NM:NMR_PROBE HR-MAS TXI 1H/13C/15N probe NM:NMR_SOLVENT H2O NM:NMR_TUBE_SIZE 4 mm rotor for HR-MAS probe NM:PULSE_SEQUENCE CPMG NM:WATER_SUPPRESSION water peak suppression-presaturation NM:TEMPERATURE 277 K NM:NUMBER_OF_SCANS 64 NM:DUMMY_SCANS 4 NM:ACQUISITION_TIME 1.36s NM:SPECTRAL_WIDTH 12 KHz NM:NUM_DATA_POINTS_ACQUIRED 32 k NM:LINE_BROADENING 1 NM:BASELINE_CORRECTION_METHOD automatic NM:CHEMICAL_SHIFT_REF_STD doublet at 5.24 ppm NM:BINNED_INCREMENT 0.02 ppm NM:BINNED_DATA_EXCLUDED_RANGE 4.5-5.5 ppm #NMR_METABOLITE_DATA NMR_METABOLITE_DATA:UNITS AU NMR_METABOLITE_DATA_START Samples IL10_KO_HC1 IL10_KO_HC10 IL10_KO_HC2 IL10_KO_HC3 IL10_KO_HC4 IL10_KO_HC5 IL10_KO_HC6 IL10_KO_HC7 IL10_KO_HC8 IL10_KO_HC9 IL10_KO_ILL1 IL10_KO_ILL10 IL10_KO_ILL2 IL10_KO_ILL3 IL10_KO_ILL4 IL10_KO_ILL5 IL10_KO_ILL6 IL10_KO_ILL7 IL10_KO_ILL8 IL10_KO_ILL9 WT_DSS4_1 WT_DSS4_10 WT_DSS4_2 WT_DSS4_3 WT_DSS4_4 WT_DSS4_5 WT_DSS4_6 WT_DSS4_7 WT_DSS4_8 WT_DSS4_9 WT_HC_1 WT_HC_10 WT_HC_2 WT_HC_3 WT_HC_4 WT_HC_5 WT_HC_6 WT_HC_7 WT_HC_8 WT_HC_9 WT_DSS_1 WT_DSS_10 WT_DSS_2 WT_DSS_3 WT_DSS_4 WT_DSS_5 WT_DSS_6 WT_DSS_7 WT_DSS_8 WT_DSS_9 Factors Group:Healthy_IL10-/- Group:Healthy_IL10-/- Group:Healthy_IL10-/- Group:Healthy_IL10-/- Group:Healthy_IL10-/- Group:Healthy_IL10-/- Group:Healthy_IL10-/- Group:Healthy_IL10-/- Group:Healthy_IL10-/- Group:Healthy_IL10-/- Group:Ill_IL10-/- Group:Ill_IL10-/- Group:Ill_IL10-/- Group:Ill_IL10-/- Group:Ill_IL10-/- Group:Ill_IL10-/- Group:Ill_IL10-/- Group:Ill_IL10-/- Group:Ill_IL10-/- Group:Ill_IL10-/- Group:Day_4_DSS Group:Day_4_DSS Group:Day_4_DSS Group:Day_4_DSS Group:Day_4_DSS Group:Day_4_DSS Group:Day_4_DSS Group:Day_4_DSS Group:Day_4_DSS Group:Day_4_DSS Group:WT Group:WT Group:WT Group:WT Group:WT Group:WT Group:WT Group:WT Group:WT Group:WT Group:Day_10_DSS Group:Day_10_DSS Group:Day_10_DSS Group:Day_10_DSS Group:Day_10_DSS Group:Day_10_DSS Group:Day_10_DSS Group:Day_10_DSS Group:Day_10_DSS Group:Day_10_DSS L-Valine 0.000397167 0.000559937 0.000116455 0.000637725 0.000118624 0.000926617 0.000162498 0.00055931 0.00013983 0 0.000131425 0.000963932 0.003777783 0 0.000937922 0.002180613 0.000185717 0.000183723 0.000509685 0.000832493 9.07E-05 0.00032068 0.000245528 0.000479627 0.001033095 0.000495922 0.073196432 3.69E-05 0 0.000103932 0 0.000432907 0.000169641 0.000304852 0.000605266 7.03E-05 0.000127488 0.000158383 0.000234704 0 0.001112892 0.000263733 0.000563058 0 0.001037051 0.000465915 0.000180856 0.000438036 0.000232453 0.000159092 L-Isoleucine 0.000341781 0.000765075 0.000379442 0.000120673 0.00030102 0.000378735 0.000286301 0.000358308 0.000192155 0.000308322 0.000301503 0.000540741 0.000318012 0.000236888 0.000186569 0.000395408 0.000648768 0.000373264 0.000508746 0.000738369 0.000203923 0.00038709 0 0.000219691 0.000427421 0.000297854 0.023928979 0.000234467 0.000401597 0.000170639 0.000287986 0.000322177 0.000171885 0.000129744 0.000275312 0.000211703 0.000153428 0.000311366 0.000272832 0.000184111 0.000352307 0.000458038 0.000473442 0.000372683 0.000446992 0.000434909 0.000380355 0.000369403 0.000343775 0.000404553 Succinate 0.000187482 0.000557862 1.98E-05 0.000256055 0.000216166 7.62E-05 0 0.004306349 0.000437635 0 0.000323808 0.001153383 0.002585032 0 0.00130028 0.005762974 0 0.000146658 0.001060499 0.000272815 0 0.000240942 4.73E-05 0.000574628 0.000423944 0.000274331 0 4.90E-05 0.000125976 0 0 0.00057788 0 0.000127399 0.000397055 0 0 0.000136518 0 0 0.001994186 0.000222531 0.001069588 7.14E-05 0.002709314 0 0 0.000552409 0.000264513 0 L-Glutamic_acid 0.00335232 0.007491552 0.004088259 0.006720471 0.004455681 0.008231809 0.006658904 0.0132858 0.011887575 0.004441697 0.006345939 0.012518168 0.010749703 0.006762777 0.011964045 0.011673047 0.004472775 0.004621691 0.016477115 0.014276834 0.001996334 0.007235614 0.006346833 0.009576153 0.012327381 0.010697122 0 0.005323697 0.009714398 0.007292622 0.001957872 0.006758311 0.001472185 0.007292314 0.004855788 0.001287462 0.002071949 0.003368903 0.004629442 0.003028723 0.008807589 0.004420938 0.014969656 0.00171199 0.007009207 0.006266945 0.002767213 0.008966975 0.006170479 0.003240435 glutamine 6.30E-05 2.80E-05 0.000204724 0.00029961 0 0.001073741 0.000779494 0.001203255 0.001252485 0.000174415 0.000333048 0.00050854 0.001069701 0 0 0 3.32E-06 0.000449117 0.001814867 0.000944189 0 0.001157417 0 0.000434673 1.01E-05 0.001392821 0 0.000824607 0.000668678 0.000984643 3.13E-05 0.000424783 0 0.000877353 0.000239503 0 0.000234242 0 0.000241747 0.000115559 0.001004943 0.000574037 0.002636818 0.000243064 0.000929605 0.000136791 0.000376853 0.000999787 0.00081807 0 Glutathione 0.001401546 0.003577438 0.000968129 0.003206505 0.000870052 0.004345377 0.003210511 0.003371571 0.004198567 0.001575584 0.003473738 0.004752885 0.006372097 0.00224206 0.004484315 0.003725102 0.00185296 0.001902849 0.008850474 0.007842431 0.000948912 0.002673198 0.003637441 0.002246166 0.001292842 0.003502162 0.003170274 0.003103189 0.002784128 0.003348399 8.61E-06 0.002068014 0.000528369 0.00350904 0.000952856 0.000503655 0.001737239 0.000576621 0.002761154 0.001425114 0.004818676 0.00135659 0.007045371 0.001228201 0.004695829 0.00407532 0.001432552 0.003706155 0.002068342 0.00132468 Formate 4.23E-05 5.44E-06 0 3.16E-05 0.000104939 0 0 0.000529658 2.82E-05 2.42E-05 6.34E-05 0 0.000159906 5.55E-05 0 0.000153945 1.35E-05 0 0 0.000126932 0.000257845 3.22E-05 0.000587039 0 1.25E-05 0 2.57E-05 0.000124618 0.000407138 0 8.14E-05 3.50E-05 7.50E-05 0 0 0 5.18E-05 7.60E-06 0 3.97E-05 0.000409326 1.59E-05 0.00012478 1.52E-05 0.001531061 0 3.41E-05 0 0 2.95E-05 Ethanol 0.015672626 0.029081265 0.033177111 0.10637173 0.020273775 0.087836963 0.080659538 0.026009114 0.044467406 0.005191734 0.036557571 0.030249475 0.048647891 0.051465532 0.227648557 0.016453058 0.04121298 0.014923866 0.048826801 0.006985835 0.004630144 0.041499632 0.117066918 0.278900241 0.03494185 0.027454797 0.003041953 0.097864884 0.041401503 0.036184512 0.057996386 0.006944355 0.142535009 0.117591372 0.069360813 0.03333977 0.029016177 0.004705699 0.038096083 0.045499634 0.150786501 0.033475283 0.060501079 0.018146716 0.137392163 0.06735151 0.221526273 0.149036264 0.307000665 0.004745744 L-Alanine 0.001448339 0.003465381 0.001536119 0.001883829 0.001452656 0.004997983 0.003979753 0.003994885 0.003667206 0.003392672 0.004422302 0.005358655 0.007059317 0.003593516 0.005705492 0.006677331 0.002884607 0.001931823 0.006829643 0.009201552 0.000907527 0.002723234 0.002061747 0.00289264 0.00184502 0.002581103 0 0.002758159 0.003525615 0.004248691 0.001480667 0.003060789 0.001176852 0.005160115 0.001555104 0.000893935 0.001259178 0.001021345 0.002428649 0.001924117 0.00555694 0.00277004 0.007026947 0.001274957 0.006287479 0.0026359 0.002571326 0.003589308 0.003992057 0.003538386 Lactate 0.000637873 0.002537529 0.000742968 0.001561862 0.000284883 0.006992069 0.00275599 0.003136625 0.00509356 0.002815175 0.004689374 0.003126385 0.004367031 0.003913488 0.003460281 0.003408557 0.001114078 0.001146373 0.00800026 0.005817763 0.000858406 0.001195308 0.00201904 0.001828035 0.000217187 0.00145996 0 0.000656233 0 0.000529718 0.001051202 0.001356685 0.000624152 0.002826264 0.000185453 0.000209393 0.000886327 7.27E-05 0.000859287 0.000520179 0.009146658 0.000966428 0.006755269 0.001164221 0.006160457 0.002621107 0.002085429 0.003551188 0.003269009 0.001673487 Propionate 0.002876217 0.002928129 0.000280438 0.006328506 0.001408578 0.002161852 0.000455111 0.00991107 0.000179236 0.00029024 0.000407697 0.006627616 0.019514318 0 0.008467957 0.017480656 0.000425474 0.000378596 0.001121223 0.001674254 0.001083269 0.001486086 0.005074962 0.004042093 0.006232883 0.001295124 0.108991851 0.000751238 0 0.000476935 5.43E-05 0.004127181 0.001281865 0.001015552 0.002815761 0.000261795 0.000303254 0.002536103 0.000476077 0.00022505 0.005795521 0.000649844 0.00104258 0 0.01322174 0.001101864 0.000774419 0.001024597 0.000613288 0.000384818 Taurine 0.017340889 0.027775865 0.014882817 0.022024777 0.011272029 0.06114288 0.027736216 0.045615449 0.046113421 0.021690612 0.038541692 0.037890312 0.05435555 0.027167586 0.033539963 0.036589396 0.022360366 0.025706684 0.102568893 0.084204945 0.01395956 0.040479361 0.038213446 0.027930335 0.01447274 0.04409412 0.007785657 0.046388596 0.037730911 0.045441539 0.014353933 0.024923354 0.009998298 0.046430938 0.019144096 0.009714939 0.022779233 0.009737254 0.026163929 0.020670626 0.048424967 0.022608295 0.061851237 0.013374079 0.041948304 0.040039368 0.018401885 0.047330338 0.021753421 0.019765572 Creatine 0.008421682 0.017112728 0.009928453 0.012375709 0.007424879 0.030007508 0.013591338 0.010998954 0.023713359 0.007544637 0.014164082 0.015556872 0.019220393 0.009431325 0.016422446 0.02081742 0.011078162 0.012389203 0.03966031 0.037562103 0.006372884 0.021485361 0.014321527 0.013529674 0.007759589 0.014740974 0.000730646 0.024410232 0.023517041 0.027598155 0.006396442 0.009171244 0.006867267 0.016577872 0.00701386 0.006280028 0.01298948 0.004138585 0.019001469 0.016094078 0.019252749 0.008341586 0.024403563 0.005960836 0.018056571 0.018434958 0.008744485 0.020615241 0.010932843 0.007908507 Myo-inositol 0.009700988 0.022146342 0.008796264 0.015733885 0.007845975 0.028676444 0.013821083 0.015786437 0.027905389 0.013857936 0.010456965 0.012099305 0.017423488 0.010258757 0.014947709 0.018249596 0.011461278 0.009662054 0.023069493 0.030213788 0.007884769 0.018383397 0.016968025 0.017778365 0.010258495 0.016847457 0.003888421 0.020426081 0.017132442 0.028980258 0.005772477 0.010351715 0.005762709 0.012714573 0.005674699 0.005234325 0.016669237 0.00532527 0.018895747 0.015282917 0.01490679 0.006879215 0.019791947 0.004609726 0.015606658 0.015257841 0.007669169 0.014727679 0.00588817 0.006594796 Ascorbic_acid 0.000878615 0.001915934 0.000812335 0.001867568 0.000615778 0.003092271 0.001169159 0.00319224 0.001824443 0.000962091 0.002286613 0.003680665 0.00347975 0.001473822 0.002198859 0.001949133 0.001253083 0.001091978 0.005019686 0.005299802 0.000708824 0.001523764 0.002152337 0.001632205 0.0012301 0.002036351 0 0.001466809 0.002019829 0.003061376 9.80E-06 0.000788967 0.000807473 0.0031787 0.001152937 0.000562258 0.001617433 0.000320833 0.002581363 0.001359925 0.002876162 0.000945232 0.004381952 0.000808826 0.002356209 0.003120211 0.001102513 0.00332012 0.00149053 0.000765952 L-Aspartic_acid 0.00021099 0.000702681 0.00019459 0.000477328 0.000134323 0.000175165 0.000333478 0.000834549 0.000233626 0.000319276 0.000321563 0.000543251 0.000782369 0.000502418 0.00065436 0.000679855 0.000414383 0.000189014 0.000394507 0.000702686 3.52E-05 0.0001534 0.000394456 0.000495512 0.000252751 0.000292232 9.21E-05 0.000160053 0.000529251 0.000478278 0.000180607 0.000510892 3.19E-05 0.00062615 0.000258161 3.45E-05 9.35E-05 0.000192323 0.000303087 0.00029613 0.000337282 0.000133171 0.00064853 5.49E-05 0.000417791 0.000245865 0.000152513 0.000177408 0.000153525 0.000154384 Glycine 0.000980935 0.002251825 0.000774152 0.001053688 0.000943807 0.001761434 0.002539593 0.002282876 0.001140913 0.00203711 0.002651462 0.003788267 0.004629407 0.001696424 0.003851475 0.004012788 0.001858874 0.00202053 0.007127566 0.007550206 0.000429003 0.001381428 0.001438978 0.001675236 0.002123534 0.002495279 0.000164495 0.001001161 0.001753085 0.00185916 0.000819549 0.001949584 0.000360179 0.00325115 0.000797136 0.000275978 0.000524573 0.000774062 0.00123746 0.000444983 0.002613199 0.001324076 0.003337749 0.000861472 0.003170935 0.002147001 0.001493892 0.002375419 0.002062631 0.001291358 Choline 0.008007755 0.015649886 0.005997337 0.008977147 0.00584274 0.022905846 0.00933523 0.009980209 0.015675189 0.014217698 0.013451752 0.012521521 0.020487302 0.013437075 0.014986754 0.020452476 0.008285729 0.009384392 0.032116682 0.030875202 0.004165666 0.010013526 0.009769399 0.015332244 0.010500782 0.014614846 0.001556275 0.010259691 0.010306692 0.012743186 0.005295986 0.014863416 0.003514784 0.013685982 0.00818813 0.003346921 0.005896134 0.007096149 0.00739099 0.006621239 0.019257838 0.008518147 0.020626478 0.004603563 0.020537673 0.011367332 0.004956572 0.014593876 0.007404931 0.007384658 D-Glucose 0.001248185 0.002375468 0.001113219 0.001241359 0.00110707 0.002289411 0.000534076 0.001193373 0.001114235 0.001551172 0.001694484 0.001872017 0.002448166 0.00169111 0.001822509 0.001535787 0.00180165 0.001646733 0.005759246 0.005072187 0.000831764 0.001808277 0.00227897 0.002096498 0.001543875 0.002980287 0 0.002353373 0.002967225 0.002542253 0.000741261 0.001666351 0.00094487 0.002094816 0.00109463 0.000819111 0.000954636 0.000859364 0.0021069 0.001596877 0.005452678 0.001603333 0.00472011 0.00121514 0.003195934 0.00401824 0.001372233 0.005405731 0.001993827 0.00080586 Acetate 0.008130637 0.013150921 0.000735819 0.012904771 0.008007369 0.002294989 0.001182675 0.030803243 0.002466119 0.005090596 0.0008802 0.025102153 0.053696092 0.001399007 0.017514557 0.0439766 0.001303288 0.000790485 0.00241075 0.004125755 0.003925639 0.005122385 0.022384552 0.018942253 0.029454274 0.003504313 0.000476386 0.001871381 0.00513507 0.001845814 0.000698553 0.009570553 0.000690127 0.001140805 0.006917142 0.000540933 0.000684947 0.012714523 0.001150398 0.001094036 0.012205004 0.001587288 0.00245598 0.000763157 0.044120694 0.002089878 0.000886304 0.006214586 0.001253668 0.0016663 Hypotaurine 0.000766142 0.001682442 0.000390674 0.001243231 0.00033888 0.001229698 0.001380317 0.001603956 0.001774665 0.001692003 0.001174586 0.001767923 0.001805539 0.000851577 0.001341215 0.002294815 0.00038882 0.00094454 0.003292715 0.002720858 0.000338888 0.000838426 0.001291805 0.000697763 0.000647831 0.00095466 0.00036714 0.000667812 0.000681832 0.000816167 0.000175038 0.001503333 0.000316941 0.002306536 0.000468542 0.000147905 0.000788057 0.000495955 0.000720806 0.000444701 0.001297713 0.000298607 0.002353722 0.000386858 0.001299434 0.002104593 0.000676018 0.001779059 0.000545961 0.000519209 Fumarate 0.000103786 6.18E-05 3.50E-05 2.24E-05 6.40E-05 0.00013959 0.000104727 0.000241518 0.000107176 3.87E-05 6.72E-05 2.06E-05 0.000113324 0.000203779 0.000133648 0.000252079 7.32E-05 4.74E-05 0.000260629 0.000454073 3.90E-05 8.39E-05 9.51E-05 0.000102143 3.36E-05 0.000146479 5.19E-05 4.20E-05 2.34E-05 5.17E-05 4.55E-05 0.000117329 2.30E-05 0 8.06E-05 1.16E-05 4.04E-05 0 6.18E-06 6.51E-05 0.00016209 5.30E-05 0.000125606 3.38E-05 0.000161697 0.000115972 0.000100994 0.000185848 0.000223601 2.13E-05 NMR_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name HMDB ID Acetate HMDB00042 L-Alanine HMDB00161 Ascorbic_acid HMDB00044 L-Aspartic_acid HMDB00191 Choline HMDB00097 Creatine HMDB00064 Ethanol HMDB00108 Fumarate HMDB00134 D-Glucose HMDB00122 L-Glutamic_acid HMDB00148 Glutathione HMDB00125 Glycine HMDB00123 Hypotaurine HMDB00965 L-Isoleucine HMDB00172 Lactate HMDB00190 Myo-inositol HMDB00211 Propionate HMDB00237 Succinate HMDB00254 Taurine HMDB00251 glutamine HMDB003423 L-Valine HMDB00883 Formate HMDB0304356 METABOLITES_END #END