#METABOLOMICS WORKBENCH jravilap_20171002_140622 DATATRACK_ID:1248 STUDY_ID:ST000879 ANALYSIS_ID:AN001431 PROJECT_ID:PR000610 VERSION 1 CREATED_ON October 25, 2017, 9:35 am #PROJECT PR:PROJECT_TITLE Diet, genetics and gut microbiome drive dynamic changes in plasma metabolites PR:PROJECT_TITLE [plasma] PR:PROJECT_TYPE Metabolite profiling of stool and cecal contents in mice of different genotypes, PR:PROJECT_TYPE vendors, diets, and antibiotic treatments. PR:PROJECT_SUMMARY Diet, genetics and the gut microbiome are determinants of metabolic status, in PR:PROJECT_SUMMARY part through production of metabolites by gut microbiota. To understand PR:PROJECT_SUMMARY mechanisms linking these factors, we performed LC-MS-based metabolomic analysis PR:PROJECT_SUMMARY of cecal contents and plasma from C57BL/6J, 129S1/SvImJ and 129S6/SvEvTac mice PR:PROJECT_SUMMARY on chow, high-fat diet (HFD), and HFD-treated with vancomycin or metronidazole. PR:PROJECT_SUMMARY Prediction of the functional metagenome of gut bacteria by PICRUSt analysis of PR:PROJECT_SUMMARY 16S sequences revealed dramatic differences in microbial metabolism. Cecal and PR:PROJECT_SUMMARY plasma metabolites showed multifold differences reflecting the combined and PR:PROJECT_SUMMARY integrated effects of diet, antibiotics, host background, and the gut PR:PROJECT_SUMMARY microbiome. Eighteen plasma metabolites correlated positively or negatively with PR:PROJECT_SUMMARY host insulin resistance across strains and diets. Over 1,000 PR:PROJECT_SUMMARY as-of-yet-unidentified metabolite peaks were also highly regulated by diet, PR:PROJECT_SUMMARY antibiotics and genetic background. Thus diet, host genetics and gut microbiota PR:PROJECT_SUMMARY interact to create unique responses in plasma metabolites, which can contribute PR:PROJECT_SUMMARY to regulation of metabolism and insulin resistance. PR:INSTITUTE Broad Institute of MIT and Harvard PR:DEPARTMENT Metabolomics Platform PR:LAST_NAME Avila PR:FIRST_NAME Julian PR:ADDRESS 415 Main Street, Rm 7175, Cambridge, MA, 02142, USA PR:EMAIL jravilap@broadinstitute.org PR:PHONE 6177148264 PR:CONTRIBUTORS Shiho Fujisaka, Julian Avila-Pacheco, Marion Soto, Aleksandar Kostic, Jonathan PR:CONTRIBUTORS M. Dreyfuss, Hui Pan, Siegfried Ussar, Emrah Altindis, Ning Li, Lynn Bry, Clary PR:CONTRIBUTORS B. Clish, and C. Ronald Kahn #STUDY ST:STUDY_TITLE Diet, genetics and gut microbiome drive dynamic changes in plasma metabolites ST:STUDY_TITLE [plasma] ST:STUDY_TYPE Plasma data ST:STUDY_SUMMARY C57BL/6J mice (B6) and 129S1 mice (129J) were purchased from Jackson Laboratory ST:STUDY_SUMMARY (Bar Harbor, ME) and 129S6 mice (129T) were purchased from Taconic Farms ST:STUDY_SUMMARY (Germantown, NY). Mice were maintained on normal chow containing 22% calories ST:STUDY_SUMMARY from fat, 23% from protein and 55% from carbohydrates (Mouse Diet 9F 5020, ST:STUDY_SUMMARY PharmaServ, Framingham, MA) or a high fat diet (Open Source Diet, D12492, ST:STUDY_SUMMARY Research Diets, New Brunswick, NJ) containing 60% calories from fat, 20% from ST:STUDY_SUMMARY protein and 20% from carbohydrates. For antibiotic treatment, 6-week old mice ST:STUDY_SUMMARY were treated with either placebo, vancomycin (1g/L) or metronidazole (1g/L) ST:STUDY_SUMMARY (Sigma-Aldrich, St. Louis, MO) in drinking water then started on HFD from age 7 ST:STUDY_SUMMARY to 11 weeks. The mice were fasted for 2 hours and anesthetized with isoflurane ST:STUDY_SUMMARY before collecting cecum and plasma. ST:INSTITUTE Broad Institute of MIT and Harvard ST:LAST_NAME Avila ST:FIRST_NAME Julian ST:ADDRESS 415 Main Street ST:EMAIL jravilap@broadinstitute.org ST:PHONE 617-714-8264 ST:NUM_GROUPS 12 ST:TOTAL_SUBJECTS 47 #SUBJECT SU:SUBJECT_TYPE Animal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - 1_Chow_B6Jax Diet:Chow | Treatment:none | Strain:B6Jax Sample #=1; Run order=0005 SUBJECT_SAMPLE_FACTORS - 2_Chow_B6Jax Diet:Chow | Treatment:none | Strain:B6Jax Sample #=2; Run order=0007 SUBJECT_SAMPLE_FACTORS - 3_Chow_B6Jax Diet:Chow | Treatment:none | Strain:B6Jax Sample #=3; Run order=0004 SUBJECT_SAMPLE_FACTORS - 5_HFD Placebo_B6Jax Diet:HFD | Treatment:placebo | Strain:B6Jax Sample #=5; Run order=0006 SUBJECT_SAMPLE_FACTORS - 6_HFD Placebo_B6Jax Diet:HFD | Treatment:placebo | Strain:B6Jax Sample #=6; Run order=0009 SUBJECT_SAMPLE_FACTORS - 7_HFD Placebo_B6Jax Diet:HFD | Treatment:placebo | Strain:B6Jax Sample #=7; Run order=0013 SUBJECT_SAMPLE_FACTORS - 8_HFD Placebo_B6Jax Diet:HFD | Treatment:placebo | Strain:B6Jax Sample #=8; Run order=0011 SUBJECT_SAMPLE_FACTORS - 9_HFD Vancomycin_B6Jax Diet:HFD | Treatment:vancomycin | Strain:B6Jax Sample #=9; Run order=0008 SUBJECT_SAMPLE_FACTORS - 10_HFD Vancomycin_B6Jax Diet:HFD | Treatment:vancomycin | Strain:B6Jax Sample #=10; Run order=0010 SUBJECT_SAMPLE_FACTORS - 11_HFD Vancomycin_B6Jax Diet:HFD | Treatment:vancomycin | Strain:B6Jax Sample #=11; Run order=0003 SUBJECT_SAMPLE_FACTORS - 12_HFD Vancomycin_B6Jax Diet:HFD | Treatment:vancomycin | Strain:B6Jax Sample #=12; Run order=0001 SUBJECT_SAMPLE_FACTORS - 13_HFD Metronidazole_B6Jax Diet:HFD | Treatment:metronidazole | Strain:B6Jax Sample #=13; Run order=0002 SUBJECT_SAMPLE_FACTORS - 14_HFD Metronidazole_B6Jax Diet:HFD | Treatment:metronidazole | Strain:B6Jax Sample #=14; Run order=0015 SUBJECT_SAMPLE_FACTORS - 15_HFD Metronidazole_B6Jax Diet:HFD | Treatment:metronidazole | Strain:B6Jax Sample #=15; Run order=0014 SUBJECT_SAMPLE_FACTORS - 16_HFD Metronidazole_B6Jax Diet:HFD | Treatment:metronidazole | Strain:B6Jax Sample #=16; Run order=0012 SUBJECT_SAMPLE_FACTORS - 17_Chow_129Tac Diet:Chow | Treatment:none | Strain:129Tac Sample #=17; Run order=0017 SUBJECT_SAMPLE_FACTORS - 18_Chow_129Tac Diet:Chow | Treatment:none | Strain:129Tac Sample #=18; Run order=0027 SUBJECT_SAMPLE_FACTORS - 19_Chow_129Tac Diet:Chow | Treatment:none | Strain:129Tac Sample #=19; Run order=0016 SUBJECT_SAMPLE_FACTORS - 20_Chow_129Tac Diet:Chow | Treatment:none | Strain:129Tac Sample #=20; Run order=0020 SUBJECT_SAMPLE_FACTORS - 21_HFD Placebo_129Tac Diet:HFD | Treatment:placebo | Strain:129Tac Sample #=21; Run order=0019 SUBJECT_SAMPLE_FACTORS - 22_HFD Placebo_129Tac Diet:HFD | Treatment:placebo | Strain:129Tac Sample #=22; Run order=0028 SUBJECT_SAMPLE_FACTORS - 23_HFD Placebo_129Tac Diet:HFD | Treatment:placebo | Strain:129Tac Sample #=23; Run order=0018 SUBJECT_SAMPLE_FACTORS - 24_HFD Placebo_129Tac Diet:HFD | Treatment:placebo | Strain:129Tac Sample #=24; Run order=0023 SUBJECT_SAMPLE_FACTORS - 25_HFD Vancomycin_129Tac Diet:HFD | Treatment:vancomycin | Strain:129Tac Sample #=25; Run order=0024 SUBJECT_SAMPLE_FACTORS - 26_HFD Vancomycin_129Tac Diet:HFD | Treatment:vancomycin | Strain:129Tac Sample #=26; Run order=0030 SUBJECT_SAMPLE_FACTORS - 27_HFD Vancomycin_129Tac Diet:HFD | Treatment:vancomycin | Strain:129Tac Sample #=27; Run order=0025 SUBJECT_SAMPLE_FACTORS - 28_HFD Vancomycin_129Tac Diet:HFD | Treatment:vancomycin | Strain:129Tac Sample #=28; Run order=0029 SUBJECT_SAMPLE_FACTORS - 29_HFD Metronidazole_129Tac Diet:HFD | Treatment:metronidazole | Strain:129Tac Sample #=29; Run order=0022 SUBJECT_SAMPLE_FACTORS - 30_HFD Metronidazole_129Tac Diet:HFD | Treatment:metronidazole | Strain:129Tac Sample #=30; Run order=0021 SUBJECT_SAMPLE_FACTORS - 31_HFD Metronidazole_129Tac Diet:HFD | Treatment:metronidazole | Strain:129Tac Sample #=31; Run order=0031 SUBJECT_SAMPLE_FACTORS - 32_HFD Metronidazole_129Tac Diet:HFD | Treatment:metronidazole | Strain:129Tac Sample #=32; Run order=0026 SUBJECT_SAMPLE_FACTORS - 33_Chow_129Jax Diet:Chow | Treatment:none | Strain:129Jax Sample #=33; Run order=0036 SUBJECT_SAMPLE_FACTORS - 34_Chow_129Jax Diet:Chow | Treatment:none | Strain:129Jax Sample #=34; Run order=0033 SUBJECT_SAMPLE_FACTORS - 35_Chow_129Jax Diet:Chow | Treatment:none | Strain:129Jax Sample #=35; Run order=0040 SUBJECT_SAMPLE_FACTORS - 36_Chow_129Jax Diet:Chow | Treatment:none | Strain:129Jax Sample #=36; Run order=0042 SUBJECT_SAMPLE_FACTORS - 37_HFD Placebo_129Jax Diet:HFD | Treatment:placebo | Strain:129Jax Sample #=37; Run order=0038 SUBJECT_SAMPLE_FACTORS - 38_HFD Placebo_129Jax Diet:HFD | Treatment:placebo | Strain:129Jax Sample #=38; Run order=0034 SUBJECT_SAMPLE_FACTORS - 39_HFD Placebo_129Jax Diet:HFD | Treatment:placebo | Strain:129Jax Sample #=39; Run order=0041 SUBJECT_SAMPLE_FACTORS - 40_HFD Placebo_129Jax Diet:HFD | Treatment:placebo | Strain:129Jax Sample #=40; Run order=0035 SUBJECT_SAMPLE_FACTORS - 41_HFD Vancomycin_129Jax Diet:HFD | Treatment:vancomycin | Strain:129Jax Sample #=41; Run order=0047 SUBJECT_SAMPLE_FACTORS - 42_HFD Vancomycin_129Jax Diet:HFD | Treatment:vancomycin | Strain:129Jax Sample #=42; Run order=0039 SUBJECT_SAMPLE_FACTORS - 43_HFD Vancomycin_129Jax Diet:HFD | Treatment:vancomycin | Strain:129Jax Sample #=43; Run order=0037 SUBJECT_SAMPLE_FACTORS - 44_HFD Vancomycin_129Jax Diet:HFD | Treatment:vancomycin | Strain:129Jax Sample #=44; Run order=0043 SUBJECT_SAMPLE_FACTORS - 45_HFD Metronidazole_129Jax Diet:HFD | Treatment:metronidazole | Strain:129Jax Sample #=45; Run order=0032 SUBJECT_SAMPLE_FACTORS - 46_HFD Metronidazole_129Jax Diet:HFD | Treatment:metronidazole | Strain:129Jax Sample #=46; Run order=0046 SUBJECT_SAMPLE_FACTORS - 47_HFD Metronidazole_129Jax Diet:HFD | Treatment:metronidazole | Strain:129Jax Sample #=47; Run order=0044 SUBJECT_SAMPLE_FACTORS - 48_HFD Metronidazole_129Jax Diet:HFD | Treatment:metronidazole | Strain:129Jax Sample #=48; Run order=0045 #COLLECTION CO:COLLECTION_SUMMARY The mice were fasted for 2 hours and anesthetized with isoflurane before CO:COLLECTION_SUMMARY collecting cecum and plasma. CO:BLOOD_SERUM_OR_PLASMA Plasma #TREATMENT TR:TREATMENT_SUMMARY Mice were maintained on normal chow containing 22% calories from fat, 23% from TR:TREATMENT_SUMMARY protein and 55% from carbohydrates (Mouse Diet 9F 5020, PharmaServ, Framingham, TR:TREATMENT_SUMMARY MA) or a high fat diet (Open Source Diet, D12492, Research Diets, New Brunswick, TR:TREATMENT_SUMMARY NJ) containing 60% calories from fat, 20% from protein and 20% from TR:TREATMENT_SUMMARY carbohydrates. For antibiotic treatment, 6-week old mice were treated with TR:TREATMENT_SUMMARY either placebo, vancomycin (1g/L) or metronidazole (1g/L) (Sigma-Aldrich, St. TR:TREATMENT_SUMMARY Louis, MO) in drinking water then started on HFD from age 7 to 11 weeks. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY HILIC-pos: LC-MS samples were prepared from plasma (10 μL) via protein SP:SAMPLEPREP_SUMMARY precipitation with the addition of nine volumes of 74.9:24.9:0.2 v/v/v SP:SAMPLEPREP_SUMMARY acetonitrile/methanol/formic acid containing stable isotope-labeled internal SP:SAMPLEPREP_SUMMARY standards (valine-d8, Isotec; and phenylalanine-d8, Cambridge Isotope SP:SAMPLEPREP_SUMMARY Laboratories; Andover, MA). The samples are centrifuged (10 min, 9,000 x g, SP:SAMPLEPREP_SUMMARY 4°C), and the supernatants were injected directly. HILIC-neg:Plasma samples SP:SAMPLEPREP_SUMMARY (30μL) were extracted using 120 μL of 80% methanol (VWR) containing 0.05 SP:SAMPLEPREP_SUMMARY ng/μL inosine-15N4, 0.05 ng/μL thymine-d4, and 0.1 ng/μL glycocholate-d4 as SP:SAMPLEPREP_SUMMARY internal standards (Cambridge Isotope Laboratories). The samples were SP:SAMPLEPREP_SUMMARY centrifuged (10 min, 9,000 x g, 4ºC) and the supernatants (10 μL) were SP:SAMPLEPREP_SUMMARY injected directly. C18-neg: Plasma samples (20 μL) were extracted using 60 μL SP:SAMPLEPREP_SUMMARY of methanol containing PGE2-d4 as an internal standard (Cayman Chemical Co.; Ann SP:SAMPLEPREP_SUMMARY Arbor, MI) and centrifuged (10 min, 9,000 x g, 4°C). C8-pos: Lipids were SP:SAMPLEPREP_SUMMARY extracted from plasma (10 μL) using 190 μL of isopropanol containing SP:SAMPLEPREP_SUMMARY 1-dodecanoyl-2-tridecanoyl-sn-glycero-3-phosphocholine as an internal standard SP:SAMPLEPREP_SUMMARY (Avanti Polar Lipids; Alabaster, AL). After centrifugation (10 min, 9,000 x g, SP:SAMPLEPREP_SUMMARY ambient temperature), supernatants (10 μL) were injected directly #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Shimadzu Nexera X2 CH:COLUMN_NAME Waters Acquity BEH C18 (150 x 2.1mm, 1.7um) CH:SOLVENT_A 0.1% formic acid in water CH:SOLVENT_B acetonitrile with 0.1% formic acid #ANALYSIS AN:ANALYSIS_TYPE MS AN:OPERATOR_NAME Kevin Bullock #MS MS:MS_COMMENTS - MS:INSTRUMENT_NAME Thermo Q Exactive Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE NEGATIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Peak area MS_METABOLITE_DATA_START Samples 1_Chow_B6Jax 2_Chow_B6Jax 3_Chow_B6Jax 5_HFD Placebo_B6Jax 6_HFD Placebo_B6Jax 7_HFD Placebo_B6Jax 8_HFD Placebo_B6Jax 9_HFD Vancomycin_B6Jax 10_HFD Vancomycin_B6Jax 11_HFD Vancomycin_B6Jax 12_HFD Vancomycin_B6Jax 13_HFD Metronidazole_B6Jax 14_HFD Metronidazole_B6Jax 15_HFD Metronidazole_B6Jax 16_HFD Metronidazole_B6Jax 17_Chow_129Tac 18_Chow_129Tac 19_Chow_129Tac 20_Chow_129Tac 21_HFD Placebo_129Tac 22_HFD Placebo_129Tac 23_HFD Placebo_129Tac 24_HFD Placebo_129Tac 25_HFD Vancomycin_129Tac 26_HFD Vancomycin_129Tac 27_HFD Vancomycin_129Tac 28_HFD Vancomycin_129Tac 29_HFD Metronidazole_129Tac 30_HFD Metronidazole_129Tac 31_HFD Metronidazole_129Tac 32_HFD Metronidazole_129Tac 33_Chow_129Jax 34_Chow_129Jax 35_Chow_129Jax 36_Chow_129Jax 37_HFD Placebo_129Jax 38_HFD Placebo_129Jax 39_HFD Placebo_129Jax 40_HFD Placebo_129Jax 41_HFD Vancomycin_129Jax 42_HFD Vancomycin_129Jax 43_HFD Vancomycin_129Jax 44_HFD Vancomycin_129Jax 45_HFD Metronidazole_129Jax 46_HFD Metronidazole_129Jax 47_HFD Metronidazole_129Jax 48_HFD Metronidazole_129Jax Factors Diet:Chow | Treatment:none | Strain:B6Jax Diet:Chow | Treatment:none | Strain:B6Jax Diet:Chow | Treatment:none | Strain:B6Jax Diet:HFD | Treatment:placebo | Strain:B6Jax Diet:HFD | Treatment:placebo | Strain:B6Jax Diet:HFD | Treatment:placebo | Strain:B6Jax Diet:HFD | Treatment:placebo | Strain:B6Jax Diet:HFD | Treatment:vancomycin | Strain:B6Jax Diet:HFD | Treatment:vancomycin | Strain:B6Jax Diet:HFD | Treatment:vancomycin | Strain:B6Jax Diet:HFD | Treatment:vancomycin | Strain:B6Jax Diet:HFD | Treatment:metronidazole | Strain:B6Jax Diet:HFD | Treatment:metronidazole | Strain:B6Jax Diet:HFD | Treatment:metronidazole | Strain:B6Jax Diet:HFD | Treatment:metronidazole | Strain:B6Jax Diet:Chow | Treatment:none | Strain:129Tac Diet:Chow | Treatment:none | Strain:129Tac Diet:Chow | Treatment:none | Strain:129Tac Diet:Chow | Treatment:none | Strain:129Tac Diet:HFD | Treatment:placebo | Strain:129Tac Diet:HFD | Treatment:placebo | Strain:129Tac Diet:HFD | Treatment:placebo | Strain:129Tac Diet:HFD | Treatment:placebo | Strain:129Tac Diet:HFD | Treatment:vancomycin | Strain:129Tac Diet:HFD | Treatment:vancomycin | Strain:129Tac Diet:HFD | Treatment:vancomycin | Strain:129Tac Diet:HFD | Treatment:vancomycin | Strain:129Tac Diet:HFD | Treatment:metronidazole | Strain:129Tac Diet:HFD | Treatment:metronidazole | Strain:129Tac Diet:HFD | Treatment:metronidazole | Strain:129Tac Diet:HFD | Treatment:metronidazole | Strain:129Tac Diet:Chow | Treatment:none | Strain:129Jax Diet:Chow | Treatment:none | Strain:129Jax Diet:Chow | Treatment:none | Strain:129Jax Diet:Chow | Treatment:none | Strain:129Jax Diet:HFD | Treatment:placebo | Strain:129Jax Diet:HFD | Treatment:placebo | Strain:129Jax Diet:HFD | Treatment:placebo | Strain:129Jax Diet:HFD | Treatment:placebo | Strain:129Jax Diet:HFD | Treatment:vancomycin | Strain:129Jax Diet:HFD | Treatment:vancomycin | Strain:129Jax Diet:HFD | Treatment:vancomycin | Strain:129Jax Diet:HFD | Treatment:vancomycin | Strain:129Jax Diet:HFD | Treatment:metronidazole | Strain:129Jax Diet:HFD | Treatment:metronidazole | Strain:129Jax Diet:HFD | Treatment:metronidazole | Strain:129Jax Diet:HFD | Treatment:metronidazole | Strain:129Jax PGE2-d4 (internal standard) 16834311 16945553 16362296 16276339 16806618 16870704 16296022 17072993 17426951 16583185 17124604 17092263 17771230 17374649 17116229 17120099 16440535 16858903 17027513 16511522 16989796 16751763 16598525 17195131 17060171 16725093 16414249 16809785 16802097 16317985 17193897 17262997 15444442 16587806 16951711 17009406 16789567 16011094 16811558 15948061 15803665 16731128 16191099 15648700 16966826 16285059 16027708 Alpha-muricholic acid 613553 3763 1808 191791 2845 3550 21273 50575 4787 359588 3464 9049 95561 Beta-muricholic acid 230490 9898 38576 4340351 533524 55397 75485 23194 56227 16658 299190 83156 44560 105208 22337 29520 1215065 28107 209664 178569 2822 6627 10181 9161 2507 217360 246131 35734 4377 217219 4129052 11487 118790 421986 234373 17237 2032607 29648 407477 123325 400781 27987 4133082 951628 tauromuricholic acid 1789359849 11610136 29008376 18987848233 4698480 4670507 2914074 8651590 4085954 60651711 54073599 150683514 2643981 106351996 43804416 1619816 28843341 1886691 1582336 2769127 2435327 5560182 13470862 35732031 4708215 4301495 3330962 38318688 2045549 4711558 5327825 155891743 19945904616 3650027 73357947 187187016 116341955 33811546 6865331961 1928363 1503263917 28597039 2285029986 62162521 28339519713 51631492 2252419 Myristoleic acid 62176 119991 196904 73732 24727 117803 131818 107091 99156 118988 45361 71635 74553 61699 26909 123546 91162 18475 19200 83064 46102 129231 57911 20870 68548 55363 47826 35962 80790 64957 65686 94585 171846 84911 25257 92087 85887 41631 64398 70112 111163 54953 208292 80647 93904 177411 200443 Myristic acid 95352 129392 114009 105034 59961 132132 120674 134105 124969 156593 59208 86471 80709 193622 115655 57922 96790 35227 21028 81837 33706 58702 83494 36122 75855 41123 39036 28999 77078 68834 72653 127265 352082 157894 100363 121158 94877 98121 66946 90277 102145 128341 243635 75506 104118 207366 206519 Palmitoleic acid 519968 1005745 633018 409465 332892 751850 613384 746943 826886 742802 302400 340594 409153 973541 650679 376703 630065 165326 90255 328382 208496 571958 578149 106275 432330 133702 174811 93973 440787 195758 314272 991334 4551448 1163415 435721 637845 955685 482744 404343 524367 1222218 524667 2944065 646942 627849 1766894 1444404 Palmitic acid 1187426 1870581 1457277 1477694 992178 1233306 1613802 1660845 1780752 1766997 1070397 1262158 1308539 2323895 1392937 941899 1171712 566223 410814 1125995 1239126 1196887 1097236 741258 1399449 730387 828065 601897 1019644 1213824 992952 1818272 3649851 1463291 1037570 1637033 1237183 1661428 1341993 1236817 2451841 1621020 3971942 1289510 1288912 2506226 2685192 Gamma-linolenic acid 333969 532520 348409 437703 342089 427208 450997 454021 473371 591614 296854 333665 355569 631472 426506 263359 344063 194661 123796 323660 273244 406734 318922 181873 393709 236564 246454 177814 291593 311967 247827 527459 962742 461716 240748 524813 478859 342096 307549 315742 639904 479665 1189881 382940 399257 935468 1499476 Linoleic acid 4672289 10704648 10644728 6986202 4902443 9427793 11597708 10921420 10942562 12642298 5546179 6867223 7172062 14849820 10105007 3083721 5248226 2033756 1177805 6281212 6219067 7854526 6276448 2969364 9224875 4413859 3913304 2370900 6541831 7323506 4872841 10084280 32223916 6872214 3340030 10861666 11186859 9758680 7078093 6974999 20159394 12493921 35619881 8890260 7731327 24411668 46749014 Oleic acid 4331555 10171347 6368258 6839919 4319781 9853190 10093893 9530415 9186728 11555060 5846507 6858923 6135472 12378979 9644153 2264133 4040450 913183 524340 6521451 5074803 6366973 6161305 2673889 8371945 3846994 3367982 1960488 5765501 5931077 4328841 7060206 24513216 5230729 2217663 8736830 8030273 7681177 5811258 6009452 15411733 10031119 31056200 6390628 6228662 18741659 26702026 Stearic acid 248375 668658 447220 965864 328439 836245 1052671 695365 1064017 1069505 528181 774495 467654 1385129 726049 218935 321844 228259 220489 1436620 1103529 980348 853617 1052272 1160773 528763 862536 447822 653161 1025293 751435 896721 954623 628200 388766 2481640 998896 1692602 981352 1361100 2134073 2061432 2808403 1374501 846815 2996241 3516189 Eicosapentaenoic acid 4675733 5283945 11410276 1067136 1063922 1805896 713418 1052761 1537604 1040659 639285 1015992 1153126 1365063 1093690 5621243 5137506 4686837 4201336 732710 683180 793517 746083 679521 483643 583296 431953 379327 387501 529171 439301 6426163 6853123 6443192 5407558 983780 663079 896456 805410 670707 954551 1110657 953109 864496 849449 1226594 2852099 Arachidonic acid 6761145 8468823 27346265 10564342 11760526 18972923 11163021 15251379 19873271 14699001 11192313 12671104 14193148 22975138 16550424 5798294 6802239 5945564 5287168 13391253 14937740 17045438 14706588 13889641 11103893 11491557 10404900 11065844 10597818 11040631 11288194 12564650 19113177 7374052 7707815 15957231 13203268 25188603 18469462 13673424 24522542 19902418 15463084 22653288 14684721 19749406 92911631 8.11.14-Eicosatrienoic acid 74875 129788 152111 128436 94285 163832 95173 143718 176299 134841 54574 102466 106132 164536 104892 69158 68571 55023 46729 102694 117852 141253 111330 133719 127963 84605 90186 68301 84226 105280 112594 157865 165598 112282 88797 173840 104599 194794 140422 125748 236501 207994 457412 176083 107838 218924 1343929 DHA 41191369 62741607 63311176 25915449 28567080 42966101 25135670 34088640 44149078 32893849 25352284 28457471 33501851 35006910 31497079 52861242 52566811 46768832 37936000 25085968 25201377 28215437 25787106 20651925 23305608 17730911 19333859 16117582 16628312 19762678 21173508 53209554 45588834 54948824 42098150 30030216 20847862 30390477 26506497 21689233 29679772 34760170 35206792 30508231 26214059 37556264 57699998 DPA 770985 1460076 1233952 641424 592497 1004547 694225 853475 1165978 937835 473775 657892 699772 912054 529172 794854 896861 653971 484641 460162 440611 578654 502904 386019 455678 359643 328632 258994 308009 489692 379614 1446463 2039362 1219016 806394 937697 571601 740772 622764 522870 1123547 936119 1654464 713603 550709 1780623 2365975 Adrenic Acid 71192 137179 92980 248189 191970 365387 233947 268384 371318 316092 197324 244695 255075 268442 182247 64916 72259 55783 45297 297630 236165 247165 203522 209142 258483 194317 180594 160070 172582 201347 210853 141664 136291 94161 84895 366419 257926 291059 295678 230268 456858 424724 373549 235330 215994 455463 487640 5-HETE 25226 38568 42992 41913 35436 51272 40530 56074 46874 72056 44769 50688 25674 81781 197592 34459 40170 39989 31685 48583 48614 51521 21850 74862 30566 77790 37532 29562 22988 27699 205674 133796 238105 71794 27885 62464 130871 570968 374009 57226 176113 69051 93174 130270 370367 99383 94323 11-HETE 4153 14937 45963 12045 23572 15237 28937 22959 21322 6390 209119 455022 11876 19299 10625 3881 24091 38465 9948 6277 14614 328293 5741 5880 11540 227379 846811 774518 45601 19796 4688 226721 1389424 1161649 17275 1348083 12861 185275 817931 514898 53471 34399 12S-HETE 70795 365306 17171048 2408152 156467 423174 3122495 1122223 810065 1971540 1907703 1105202 1501206 15080092 14189593 382147 73507 117192 148622 1447379 3191890 3937795 374433 619287 297602 34596590 203556 478547 409701 680228 745559 68327966 21274159 610775 782936 139764 1972557 49938931 33960185 968137 50882214 1027930 1103706 40725099 4404586 822880 2718217 15S-HETE 11001 239240 34898 15486 89334 22808 17987 31766 19911 22089 8777 401330 851650 13354 10728 8362 17012 47890 11431 10330 6443 305390 7395 5226 386460 733809 5189570 15155 21981 384457 2032787 1504047 11639 1764535 30268 231261 965542 891113 45722 32969 LTB4 10411 25656 48374 38449 22264 90618 49086 79053 9779 36835 38782 23970 PGE2 63140 71559 107940 80897 71764 76620 87585 67460 92425 87232 64344 76478 79044 119964 100169 70694 61801 72540 75206 76073 91143 41877 57759 63398 58180 174587 47481 68243 61815 75631 107306 237467 461763 75663 65122 64765 78052 157437 176544 62374 351871 78465 85983 168832 73554 70121 73357 PGF2alpha 5509 6436 21505 179720 18288 84766 17141 Sphingosine-1-phosphate 24572046 28118538 22889875 21491290 26822636 24681322 27803157 24175682 37876548 28399934 22116848 24269994 32629178 26329544 28356374 32431593 38441603 34835472 35464227 29809931 39698542 38812797 43083875 48536410 37796787 34890819 37396669 43707529 28223848 34239186 35526956 29629152 31376134 31674467 31964816 26738749 33008589 27180960 29257996 27347219 37159483 34370238 28718563 35596928 38346010 34575627 35762247 Cholic acid 785126 128283 14518475 18942 5299 3725 208858 2846 30037 12167 25225 7323938 2907673 871889 2013521 1610513 701721 216520 3942 21468 491243 9768249 18875 192254 65361 4195 104905 3745 26812 38971 394868 Deoxycholic acid 1016031 603761 552725 1346053 145420 440263 1437536 87386 37233 346264 149817 1101049 702578 152629 707092 1078331 582177 506940 791834 346029 577517 832362 98653 25986 113073 402074 18962 11371 11094 275664 577501 523194 462733 717385 112683 730688 467262 2075575 2928 117584 38928 565895 150385 1404577 6028 Hyodeoxycholic acid/Ursodeoxycholic acid 106743 18932 66983 52113 10512 8294 11373 14167 3159 311714 208052 32760 79379 69182 22752 2619 25057 24875 16453 26005 22550 101714 7947 12157 35640 428933 Chenodeoxycholic Acid 7675 43740 13710 32993 9738 Lithocholic acid glycine conjugate 2897 Glycochenodeoxycholic acid 21397 25992 1556289 17694 25132 10975 11327 16961 11499 24245 42955 45250 11666 8370 17975 13517 2821 3924 8017 4859 2729 3146 5960 2431 6691 6676 6893 2748 502724 3271 3487 3518 98898 Glycodeoxycholic acid 23811 18619 167546 24681 16635 15975 25963 24654 8278 34966 39046 43948 11218 11213 10223 7737 14072 8397 3286 13302 3880 8722 3566 9506 3753 3804 146845 3239 3424 2742 30800 Glycoursodeoxycholic acid 4017 1947106 2786 2611 11681 3946 20892 5545 3534 1568747 2617 9559 289382 Glycocholic Acid 24791 2636 28990251 2108 2297 161961 61983 2913 20395 3277 87231 26030 14402 29547 8936 112183 40758 2284 17097 16859176 288206 499228 2358 154341 2540 2585 4518262 Tauroursodeoxycholic acid 1633967 899410 1803698828 981615 1185464 1092457 6435727 1814624 3857752 2543955 74088391 1835957 2850349 7707235 7049348 3747770 1057850 694203 1395896 385431 4099156 322387 1578048 2890620 364569 327769 245703 595540 205904 604360 353325 13110050 2162185019 271315 2124764 3891739 393374 156929319 6065588 192095 430687227 5788968 117007902 10392469 4012483 4159678 2550507745 Taurocholic acid 6117014 2233391 2894306 2112960 1924965 46965443 4622432 17045342 7581619 457352094 5324235 12491537 41164140 44716671 36757838 4031071 5395508 6963147 3025391 45564950 4528428 32009702 54026857 7963204 9023235 4472587 14129601 2860572 18076775 11014386 141495742 378843 3071004 30534155 63974249 4471141 3077615714 103637692 3173949 6183001089 108557914 2008942890 185832535 88595549 94049400 Taurodeoxycholic acid 955883 693734 320278735 382175 344742 474236 7367532 3267 158570 130776 1170675 597557 672491 1082544 892102 2958074 748812 537607 4445 1888640 603591636 452612 1001525 537875 144052 11544824 2130239 192303 75860 6832 27889 2805 1515635 Taurohyodeoxycholic acid 1633967 899410 1803698828 981615 1185464 1092457 6435727 1814624 3857752 2543955 74088391 1835957 2850349 7707235 7049348 3747770 1057850 694203 1395896 385431 4099156 322387 1578048 2890620 364569 327769 245703 595540 205904 604360 353325 13110050 2162185019 271315 2124764 3891739 393374 156929319 6065588 192095 430687227 5788968 117007902 10392469 4012483 4159678 2550507745 Taurolithocholic acid 38128 38958 8935752 19300 15813 28540 48099 22404 16535 22271 57395 24779 35682 36576 26775 58966 29918 34512 195926 92083 37244 29260 54460 50560 141523 49643 7794 98842 75381 121656 17705 237031 3517239 190327 175649 205924 214666 201509 232523 121035 269001 203203 204829 162116 159052 120869 677458 Taurochenodeoxycholic acid 639423 266657 1836810158 278007 302059 457190 1220763 366459 1294678 584088 35081071 519584 967897 4234337 5143087 1144559 248133 389730 628100 150816 1049637 235894 1029031 1090323 204662 210120 210645 361657 128868 270044 151482 2874992 651687642 103633 874650 1232600 312920 38722399 2343293 183351 125473557 2086744 40548726 3748289 1822437 1330534 917648340 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name retention time quantified m/z PubChem ID KEGG ID HMDB ID Study Metabolite Name Study identifier PGE2-d4 (internal standard) 7.87 355.2417 Internal Standard PGE2-d4 (internal standard) m1 Alpha-muricholic acid 7.99 407.2792 126523331 HMDB00506 alpha-muricholic acid m2 Beta-muricholic acid 8.15 407.2792 beta-muricholic acid m3 tauromuricholic acid 6.26 514.2833 103850044 HMDB00932 tauromuricholic acid m4 Myristoleic acid 10.36 225.1849 103636868 C08322 HMDB02000 Myristoleic acid m5 Myristic acid 13.79 227.2006 103329704 C06424 HMDB00806 Myristic acid m6 Palmitoleic acid 14.01 253.2162 10299739 C08362 HMDB03229 Palmitoleic acid m7 Palmitic acid 14.95 255.2319 103045306 C00249 HMDB00220 Palmitic acid m8 Gamma-linolenic acid 13.54 277.2162 103636892 C06426 HMDB03073 Gamma-Linolenic acid m9 Linoleic acid 14.29 279.2319 103167732 C01595 HMDB00673 Linoleic acid m10 Oleic acid 15.08 281.2475 10299740 C00712 HMDB00207 Oleic acid m11 Stearic acid 15.89 283.2632 102851490 C01530 HMDB00827 Stearic acid m12 Eicosapentaenoic acid 13.43 301.2162 103582828 C06428 HMDB01999 Eicosapentaenoic acid m13 Arachidonic acid 14.11 303.2319 10299602 C00219 HMDB01043 Arachidonic acid m14 8.11.14-Eicosatrienoic acid 14.59 305.2475 103636894 C03242 HMDB02925 8.11.14-Eicosatrienoic acid m15 DHA 13.89 327.2319 10299728 C06429 HMDB02183 Docosahexaenoic acid m16 DPA 14.26 329.2475 103635881 C16513 HMDB06528 Docosapentaenoic acid m17 Adrenic Acid 14.88 331.2632 104046534 C16527 HMDB02226 Adrenic acid m18 5-HETE 12.15 319.2268 103419145 C04805 HMDB11134 5-HETE m19 11-HETE 11.76 319.2268 114160464 HMDB04682 11-HETE m20 12S-HETE 11.9 319.2268 11057217 HMDB06111 12-HETE m21 15S-HETE 11.58 319.2268 103719170 C04742 HMDB03876 15-HETE m22 LTB4 9.9 335.2217 113859175 HMDB05089 LTB4 m23 PGE2 7.87 351.2166 103176817 C00584 HMDB01220 PGE2 m24 PGF2alpha 7.73 353.2323 104046446 C02314 HMDB01483 PGF2alpha m25 Sphingosine-1-phosphate 10.49 378.2404 113909917 HMDB00277 Sphingosine-1-Phosphate m26 Cholic acid 10.54 391.2843 103051112 C00695 HMDB00619 Cholic acid m28 Deoxycholic acid 10.33 391.2843 103578732 C04483 HMDB00626 Deoxycholic acid m29 Hyodeoxycholic acid/Ursodeoxycholic acid 9.05 391.2843 126523525 HMDB00733 Hyodeoxycholic acid/Ursodeoxycholic acid m30 Chenodeoxycholic Acid 8.92 407.2792 103542516 C02528 HMDB00518 Chenodeoxycholic acid m31 Lithocholic acid glycine conjugate 10.68 432.3109 10236704 C15557 HMDB00698 Glycolithocholic acid m32 Glycochenodeoxycholic acid 9.06 448.3058 126523439 HMDB00637 Glycochenodeoxycholic acid m33 Glycodeoxycholic acid 9.31 448.3058 126523434 HMDB00631 Glycodeoxycholic acid m34 Glycoursodeoxycholic acid 7.9 448.3058 103578912 HMDB00708 Glycoursodeoxycholic acid m35 Glycocholic Acid 7.93 464.3007 126523032 HMDB00138/ Glycocholic acid m36 Tauroursodeoxycholic acid 9.24 482.2935 126523640 HMDB00874 Tauroursodeoxycholic acid m37 Taurocholic acid 8.31 498.2884 126522968 HMDB00036 Taurocholic acid m38 Taurodeoxycholic acid 8.06 498.2884 103579024 HMDB00896 Taurodeoxycholic acid m39 Taurohyodeoxycholic acid 7.05 498.2884 Taurohyodeoxycholic acid m40 Taurolithocholic acid 7.05 498.2884 126523516 HMDB00722 Taurolithocholic acid m41 Taurochenodeoxycholic acid 7.2 514.2833 103457854 C05465 HMDB00951 Taurochenodesoxycholic acid m42 METABOLITES_END #END