#METABOLOMICS WORKBENCH neo_009_20210802_113751 DATATRACK_ID:2785 STUDY_ID:ST001897 ANALYSIS_ID:AN003079 PROJECT_ID:PR001193 VERSION 1 CREATED_ON August 3, 2021, 2:19 pm #PROJECT PR:PROJECT_TITLE Untargeted metabolomics to understand metabolism of local source of insulin in PR:PROJECT_TITLE the eye governed by phagocytosis and starvation PR:PROJECT_TYPE Untargeted Metabolomics (UPLC-MS/MS) PR:PROJECT_SUMMARY Untargeted metabolite analysis was performed on a Thermo Orbitrap IDX Tribrid MS PR:PROJECT_SUMMARY to understand changes in metabolites in eye retina and RPE tissue during PR:PROJECT_SUMMARY starvation. This study also probes the role of certain metabolites during PR:PROJECT_SUMMARY phagocytosis. PR:INSTITUTE University of Virginia PR:DEPARTMENT Center of Cell Clearance , Microbiology, Immunology, and Cancer Biology PR:LABORATORY Kodi Ravichandraan Lab PR:LAST_NAME Etchegaray PR:FIRST_NAME Iker PR:ADDRESS Department of Microbiology, Immunology, and Cancer Biology, University of PR:ADDRESS Virginia PR:EMAIL jie3c@virginia.edu; kr4h@virginia.edu; nw5es@virginia.edu PR:PHONE 4023109931 #STUDY ST:STUDY_TITLE A local source of insulin in the eye governed by phagocytosis and starvation ST:STUDY_SUMMARY Untargeted metabolite analysis was performed on a Thermo Orbitrap IDX Tribrid MS ST:STUDY_SUMMARY to understand changes in metabolites in eye retina and RPE tissue during ST:STUDY_SUMMARY starvation. This study also probes the role of certain metabolites during ST:STUDY_SUMMARY phagocytosis. ST:INSTITUTE University of Virginia ST:DEPARTMENT Center of Cell Clearance , Microbiology, Immunology, and Cancer Biology ST:LABORATORY Kodi Ravichandraan Lab ST:LAST_NAME Etchegaray ST:FIRST_NAME Iker ST:ADDRESS Center of Cell Clearance , Microbiology, Immunology, and Cancer Biology ST:EMAIL kr4h@virginia.edu; jie3c@virginia.edu; nw5es@virginia.edu; ST:NUM_GROUPS 4 ST:TOTAL_SUBJECTS 2 ST:STUDY_TYPE Untargeted Metabolomics ST:PHONE 4023109931 #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 - Sample_n_001 group:WT_Retina RAW_FILE_NAME=x03042021x_neg_001; ionizationMode=Negative SUBJECT_SAMPLE_FACTORS - Sample_n_002 group:WT_RPE RAW_FILE_NAME=x03042021x_neg_002; ionizationMode=Negative SUBJECT_SAMPLE_FACTORS - Sample_n_003 group:KO_Retina RAW_FILE_NAME=x03042021x_neg_003; ionizationMode=Negative SUBJECT_SAMPLE_FACTORS - Sample_n_004 group:KO_RPE RAW_FILE_NAME=x03042021x_neg_004; ionizationMode=Negative SUBJECT_SAMPLE_FACTORS - Sample_n_005 group:WT_Retina RAW_FILE_NAME=x03042021x_neg_005; ionizationMode=Negative SUBJECT_SAMPLE_FACTORS - Sample_n_006 group:WT_RPE RAW_FILE_NAME=x03042021x_neg_006; ionizationMode=Negative SUBJECT_SAMPLE_FACTORS - Sample_n_007 group:KO_Retina RAW_FILE_NAME=x03042021x_neg_007; ionizationMode=Negative SUBJECT_SAMPLE_FACTORS - Sample_n_008 group:KO_RPE RAW_FILE_NAME=x03042021x_neg_008; ionizationMode=Negative SUBJECT_SAMPLE_FACTORS - Sample_n_009 group:WT_Retina RAW_FILE_NAME=x03042021x_neg_009; ionizationMode=Negative SUBJECT_SAMPLE_FACTORS - Sample_n_010 group:WT_RPE RAW_FILE_NAME=x03042021x_neg_010; ionizationMode=Negative SUBJECT_SAMPLE_FACTORS - Sample_n_011 group:KO_Retina RAW_FILE_NAME=x03042021x_neg_011; ionizationMode=Negative SUBJECT_SAMPLE_FACTORS - Sample_n_012 group:KO_RPE RAW_FILE_NAME=x03042021x_neg_012; ionizationMode=Negative SUBJECT_SAMPLE_FACTORS - Sample_n_013 group:WT_Retina RAW_FILE_NAME=x03042021x_neg_013; ionizationMode=Negative SUBJECT_SAMPLE_FACTORS - Sample_n_014 group:WT_RPE RAW_FILE_NAME=x03042021x_neg_014; ionizationMode=Negative SUBJECT_SAMPLE_FACTORS - Sample_n_015 group:KO_Retina RAW_FILE_NAME=x03042021x_neg_015; ionizationMode=Negative SUBJECT_SAMPLE_FACTORS - Sample_n_016 group:KO_RPE RAW_FILE_NAME=x03042021x_neg_016; ionizationMode=Negative SUBJECT_SAMPLE_FACTORS - Sample_n_017 group:WT_Retina RAW_FILE_NAME=x03042021x_neg_017; ionizationMode=Negative SUBJECT_SAMPLE_FACTORS - Sample_n_018 group:WT_RPE RAW_FILE_NAME=x03042021x_neg_018; ionizationMode=Negative SUBJECT_SAMPLE_FACTORS - Sample_n_019 group:KO_Retina RAW_FILE_NAME=x03042021x_neg_019; ionizationMode=Negative SUBJECT_SAMPLE_FACTORS - Sample_n_020 group:KO_RPE RAW_FILE_NAME=x03042021x_neg_020; ionizationMode=Negative SUBJECT_SAMPLE_FACTORS - Sample_n_QC1 group:QC RAW_FILE_NAME=x03042021x_neg_QC1; ionizationMode=Negative SUBJECT_SAMPLE_FACTORS - Sample_n_QC2 group:QC RAW_FILE_NAME=x03042021x_neg_QC2; ionizationMode=Negative SUBJECT_SAMPLE_FACTORS - Sample_n_QC3 group:QC RAW_FILE_NAME=x03042021x_neg_QC3; ionizationMode=Negative SUBJECT_SAMPLE_FACTORS - Sample_n_QC4 group:QC RAW_FILE_NAME=x03042021x_neg_QC4; ionizationMode=Negative SUBJECT_SAMPLE_FACTORS - Sample_n_QC5 group:QC RAW_FILE_NAME=x03042021x_neg_QC5; ionizationMode=Negative SUBJECT_SAMPLE_FACTORS - Sample_p_001 group:WT_Retina RAW_FILE_NAME=x03042021x_pos_001; ionizationMode=Positive SUBJECT_SAMPLE_FACTORS - Sample_p_002 group:WT_RPE RAW_FILE_NAME=x03042021x_pos_002; ionizationMode=Positive SUBJECT_SAMPLE_FACTORS - Sample_p_003 group:KO_Retina RAW_FILE_NAME=x03042021x_pos_003; ionizationMode=Positive SUBJECT_SAMPLE_FACTORS - Sample_p_004 group:KO_RPE RAW_FILE_NAME=x03042021x_pos_004; ionizationMode=Positive SUBJECT_SAMPLE_FACTORS - Sample_p_005 group:WT_Retina RAW_FILE_NAME=x03042021x_pos_005; ionizationMode=Positive SUBJECT_SAMPLE_FACTORS - Sample_p_006 group:WT_RPE RAW_FILE_NAME=x03042021x_pos_006; ionizationMode=Positive SUBJECT_SAMPLE_FACTORS - Sample_p_007 group:KO_Retina RAW_FILE_NAME=x03042021x_pos_007; ionizationMode=Positive SUBJECT_SAMPLE_FACTORS - Sample_p_008 group:KO_RPE RAW_FILE_NAME=x03042021x_pos_008; ionizationMode=Positive SUBJECT_SAMPLE_FACTORS - Sample_p_009 group:WT_Retina RAW_FILE_NAME=x03042021x_pos_009; ionizationMode=Positive SUBJECT_SAMPLE_FACTORS - Sample_p_010 group:WT_RPE RAW_FILE_NAME=x03042021x_pos_010; ionizationMode=Positive SUBJECT_SAMPLE_FACTORS - Sample_p_011 group:KO_Retina RAW_FILE_NAME=x03042021x_pos_011; ionizationMode=Positive SUBJECT_SAMPLE_FACTORS - Sample_p_012 group:KO_RPE RAW_FILE_NAME=x03042021x_pos_012; ionizationMode=Positive SUBJECT_SAMPLE_FACTORS - Sample_p_013 group:WT_Retina RAW_FILE_NAME=x03042021x_pos_013; ionizationMode=Positive SUBJECT_SAMPLE_FACTORS - Sample_p_014 group:WT_RPE RAW_FILE_NAME=x03042021x_pos_014; ionizationMode=Positive SUBJECT_SAMPLE_FACTORS - Sample_p_015 group:KO_Retina RAW_FILE_NAME=x03042021x_pos_015; ionizationMode=Positive SUBJECT_SAMPLE_FACTORS - Sample_p_016 group:KO_RPE RAW_FILE_NAME=x03042021x_pos_016; ionizationMode=Positive SUBJECT_SAMPLE_FACTORS - Sample_p_017 group:WT_Retina RAW_FILE_NAME=x03042021x_pos_017; ionizationMode=Positive SUBJECT_SAMPLE_FACTORS - Sample_p_018 group:WT_RPE RAW_FILE_NAME=x03042021x_pos_018; ionizationMode=Positive SUBJECT_SAMPLE_FACTORS - Sample_p_019 group:KO_Retina RAW_FILE_NAME=x03042021x_pos_019; ionizationMode=Positive SUBJECT_SAMPLE_FACTORS - Sample_p_020 group:KO_RPE RAW_FILE_NAME=x03042021x_pos_020; ionizationMode=Positive SUBJECT_SAMPLE_FACTORS - Sample_p_QC1 group:QC RAW_FILE_NAME=x03042021x_pos_QC1; ionizationMode=Positive SUBJECT_SAMPLE_FACTORS - Sample_p_QC2 group:QC RAW_FILE_NAME=x03042021x_pos_QC2; ionizationMode=Positive SUBJECT_SAMPLE_FACTORS - Sample_p_QC3 group:QC RAW_FILE_NAME=x03042021x_pos_QC3; ionizationMode=Positive SUBJECT_SAMPLE_FACTORS - Sample_p_QC4 group:QC RAW_FILE_NAME=x03042021x_pos_QC4; ionizationMode=Positive SUBJECT_SAMPLE_FACTORS - Sample_p_QC5 group:QC RAW_FILE_NAME=x03042021x_pos_QC5; ionizationMode=Positive #COLLECTION CO:COLLECTION_SUMMARY Overnight fasted mice were euthanized with avertin after light onset. Five CO:COLLECTION_SUMMARY replicates each of both Retinal and RPE tissues (from wild type and knock out CO:COLLECTION_SUMMARY mice) were harvested/dissected, flash frozen and stored in -80 ºC until CO:COLLECTION_SUMMARY extraction. Tissues were thawed on ice and extracted using methanol:chloroform CO:COLLECTION_SUMMARY (2:1) at 4C and phase separated using water. Top aq. phase was recovered as CO:COLLECTION_SUMMARY metabolite mixture and dried overnight in speedVac and reconstituted in 100 µL CO:COLLECTION_SUMMARY of 0.1% Formic acid in water. Ten µl from each tube was removed to create a CO:COLLECTION_SUMMARY pooled QC sample that was injected at the beginning and end of the MS sequence CO:COLLECTION_SUMMARY run and additional QC samples were injected after every 5 sample injection. CO:SAMPLE_TYPE Eye tissue #TREATMENT TR:TREATMENT_SUMMARY NA #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Overnight fasted mice were euthanized with avertin after light onset. Five SP:SAMPLEPREP_SUMMARY replicates each of both Retinal and RPE tissues (from wild type and knock out SP:SAMPLEPREP_SUMMARY mice) were harvested/dissected, flash frozen and stored in -80 ºC until SP:SAMPLEPREP_SUMMARY extraction. Tissues were thawed on ice and extracted using methanol:chloroform SP:SAMPLEPREP_SUMMARY (2:1) at 4C and phase separated using water. Top aq. phase was recovered as SP:SAMPLEPREP_SUMMARY metabolite mixture and dried overnight in speedVac and reconstituted in 100 µL SP:SAMPLEPREP_SUMMARY of 0.1% Formic acid in water. Ten µl from each tube was removed to create a SP:SAMPLEPREP_SUMMARY pooled QC sample that was injected at the beginning and end of the MS sequence SP:SAMPLEPREP_SUMMARY run and additional QC samples were injected after every 5 sample injection. SP:PROCESSING_STORAGE_CONDITIONS -80℃ SP:EXTRACT_STORAGE -80℃ #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Low pH polar CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Thermo Vanquish CH:COLUMN_NAME Waters Acquity BEH C18 (100 x 2mm, 1.7um) CH:FLOW_GRADIENT 0-8 minutes 50% B, 8 – 13 minutes held at 98% B, 13.1 to 15.0 minutes revert CH:FLOW_GRADIENT to 0% B to re-equilibration for next injection CH:FLOW_RATE 0.25 mL/min CH:COLUMN_TEMPERATURE 30C CH:SOLVENT_A 0.1% Formic acid in water CH:SOLVENT_B 0.1% Formic acid in Methanol #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Orbitrap IDX MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE Other MS:ION_MODE NEGATIVE MS:MS_COMMENTS Conventional DDA mode was used for data acquisition and samples were randomized MS:MS_COMMENTS during extraction and analysis on the mass spectrometer. The injection volume MS:MS_COMMENTS was 5 µL. ESI source condition were set as follows: Ion source type: H-ESI; MS:MS_COMMENTS positive ion voltage set at 3500 and negative at 2500. Sheath gas was set at 35 MS:MS_COMMENTS and auxiliary gas at 7. Ion transfer tube temperature was set at 275 °C while MS:MS_COMMENTS the vaporizer temperature was set at 320 °C. Data was acquired in full MS mode MS:MS_COMMENTS (1 µscan) at a resolution of 120,000 with a scan range of 67-1000 m/z at a MS:MS_COMMENTS normalized AGC Target of 25% and 50 millisecond of maximum injection time was MS:MS_COMMENTS allowed. RF lens amplitude was set at 35%. Tandem MS/MS performed by applying MS:MS_COMMENTS quadrupole isolation with an isolation window of 1.6 m/z. Activation type was MS:MS_COMMENTS set at HCD and masses were fragmented with HCD Assisted Collision Energy (%) of MS:MS_COMMENTS 15,35,50. Fragment masses were detected by Orbitrap at a resolution of 30,000. MS:MS_COMMENTS For data analysis, Thermo *.RAW files were converted to open source *.mzML MS:MS_COMMENTS format using msConvert software MS:MS_COMMENTS (http://proteowizard.sourceforge.net/download.html) and brought into MS-DIAL MS:MS_COMMENTS software for analysis (http://prime.psc.riken.jp/compms/msdial/main.html). MS:MS_COMMENTS MS-DIAL analysis parameters were as follows: MS1 tolerance was set at 0.01 Da MS:MS_COMMENTS and Ms2 tolerance at 0.025 Da. Data was collected from 0-15 min at a mass range MS:MS_COMMENTS from 50-1000 Da. Peak detection was performed with a minimum peak height of MS:MS_COMMENTS 1x105 and mass slice width of 0.1 Da. Metabolites were identified by searching MS:MS_COMMENTS the MS2 spectra in MS-DIAL public database (290,915 records for positive and MS:MS_COMMENTS 36,848 entries for negative mode; Version 14) using a mass tolerance of 0.01 Da MS:MS_COMMENTS for MS1 and 0.05 Da for MS/MS with identification score cutoff of 60%. MS:MS_COMMENTS Additionally, peaks were also searched against in-house IROA library (in both MS:MS_COMMENTS positive and negative mode) with a mass tolerance of 0.01, identification cutoff MS:MS_COMMENTS of 90% and RT tolerance of 0.5 min (Mass Spectrometry Metabolite Library of MS:MS_COMMENTS Standards (MSMLS); http://iroatech.com/). For peak alignment maximum retention MS:MS_COMMENTS tolerance was set at 0.2 min. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS intensity MS_METABOLITE_DATA_START Samples Sample_n_001 Sample_n_002 Sample_n_003 Sample_n_004 Sample_n_005 Sample_n_006 Sample_n_007 Sample_n_008 Sample_n_009 Sample_n_010 Sample_n_011 Sample_n_012 Sample_n_013 Sample_n_014 Sample_n_015 Sample_n_016 Sample_n_017 Sample_n_018 Sample_n_019 Sample_n_020 Sample_n_QC1 Sample_n_QC2 Sample_n_QC3 Sample_n_QC4 Sample_n_QC5 Factors group:WT_Retina group:WT_RPE group:KO_Retina group:KO_RPE group:WT_Retina group:WT_RPE group:KO_Retina group:KO_RPE group:WT_Retina group:WT_RPE group:KO_Retina group:KO_RPE group:WT_Retina group:WT_RPE group:KO_Retina group:KO_RPE group:WT_Retina group:WT_RPE group:KO_Retina group:KO_RPE group:QC group:QC group:QC group:QC group:QC Phosphoric acid 18124160 1711928 7006856 3727045 19712202 2263301 16905086 9918485 17437012 4685478 8273816 3138585 9766460 5639184 21729812 9514229 17046168 6383538 13324464 5445334 3547407 7051249 8144062 896333 1643433 SUCCINIC ACID 9852003 974518 14844274 952629 11314920 776669 7404718 922438 13633024 688717 14719687 781369 17347996 915946 15824038 919744 15863357 814940 9836488 867042 5186544 5005670 1811924 5791 5979 Glycerophosphate(2) 543618 103841 1066755 131244 868503 131667 783497 125817 626649 96949 773198 52639 909883 131894 828031 102096 368126 157656 880476 112659 577540 603144 368311 718 0 FUMARATE (IORA) 2594814 327027 2044056 315670 2469493 346585 1619283 300546 1370783 294286 1610532 202522 2138498 206868 2272390 135813 1830172 315298 2097338 175264 846018 859660 222333 0 729 ADENINE 4068211 81497 3143200 51384 3629959 32209 3544886 71647 1792144 41782 2741309 78545 2316671 62687 4371718 44003 2562328 67665 4003549 74924 1294870 1286713 668875 18040 14584 Lactic acid 223979376 14129601 158785936 12857257 240881872 10425335 170303216 10275928 159591680 9272425 131934936 6703748 224487424 11079046 220114816 7344698 161706384 9659015 175064896 9630021 91001472 94471208 35337296 8392391 2145718 HYPOXANTHINE (IORA) 2098616 2504366 1783153 1995084 2388155 2246398 1738416 2131960 1331365 1769065 1581440 1632509 2275390 2419788 2777110 1800069 2524977 2810524 2464489 1940719 2172608 1934119 765280 25037 949 OROTATE (IORA) 22374 71129 22311 51455 21638 46938 12254 54614 36425 46574 18975 52665 23399 65544 18384 63216 18182 38106 20598 55914 32424 38825 34051 726 750 Creatine 5825492 2149128 5345718 1977689 5430068 1564029 4989568 1822685 4930047 1476094 4777441 1442916 5588368 1952685 5120202 1524408 5158930 1548121 5328636 1530250 4356360 3962616 2327574 19052 1036 DIHYDROOROTATE (IORA) 167943 239408 185698 185276 201208 206662 259062 280355 160033 180641 236529 231600 325077 305708 261660 241468 135297 170174 300411 218328 221228 201127 125975 0 0 Xanthine 3260069 1666078 2756605 1618622 3972349 1823502 2684192 1632800 1688121 1271058 2347618 1171368 2651126 1942100 3545681 1311433 3042642 2560268 3532138 1781660 2265527 2139910 725133 1471 0 Itaconic acid (Not validated) 2770723 1252094 1825179 943683 1461225 845224 1082830 884065 2479725 708187 1791371 867963 3018289 654512 3186632 657312 980052 582687 2541307 814448 2464715 2097823 1064817 8011 5217 3-HYDROXYBENZOATE (IORA) 4427 5391 9662 40456 10039 5794 11100 6818 4879 10832 36629 13154 10175 6193 37847 20135 10564 11794 8180 8237 11693 8945 3778 1228 3059 UROCANATE (IORA) 4609 2298 6347 1872 39373 3528 6425 836 1233 1024 1335 913 1740 675 31360 906 2469 1112 2531 0 1373 1450 1963 0 0 D-PANTOTHENIC ACID 235856 108624 588286 307697 150996 72509 552775 331629 234350 139755 473439 213965 492071 190785 674288 167052 485059 225042 539646 210594 300769 305981 103672 0 0 4-Nitrophenol 478010 418352 312608 585468 412980 312777 493649 413527 233779 347020 555238 477132 311055 481852 698902 569289 457495 357039 407792 462976 500898 516065 277412 110677 57885 MALEATE (IORA) 13715 14181 20972 5780 16922 15238 18200 20077 12108 10826 11223 11132 13834 31120 28993 29613 13134 7547 14038 13051 12492 18631 16293 685 0 OXOPROLINE (IORA) 2813364 320952 2973829 218356 2727592 249732 2237132 290164 2453741 221822 2564179 201644 2710727 378074 3030642 289946 1983898 201059 2693902 247002 1774725 1562956 965935 9394 5353 2-METHYLMALEATE (IORA) 283162 15380 223945 21057 291903 26886 275398 27801 197322 30637 324560 33219 281734 34813 374278 26503 273615 26376 261801 18001 151671 136541 68448 8369 20492 UREIDOPROPIONATE (IORA) 42417 89221 25127 39320 20491 82607 21813 46640 19962 66632 23373 68853 21332 91468 38500 30160 45629 45859 38502 72515 28036 35077 21950 0 0 Phenylphosphoric acid 5495745 793560 3947435 554276 3682807 486930 2677719 550278 4103378 445621 2640880 504481 5343583 455409 4383684 450990 1962496 199236 4198484 217723 2909271 2446858 1110150 1014 1436 METHIONINE SULFOXIMINE (IORA) 352649 1465057 283757 1294342 388769 1243684 347335 1370554 304786 925254 332473 958804 251813 1312588 239755 1012541 587795 827804 238602 1173018 468761 388271 116841 996 0 Meglutol 255428 100648 259195 125272 261316 117166 261147 108090 163686 111883 237153 96263 258547 107845 307395 74058 248674 127019 212923 47697 166790 164098 53907 1623 823 HydroxyButyric acid 2041878 963594 1763079 2788452 1394911 2812691 1404963 1829909 1770148 1403000 1863661 1100401 2008144 1173612 1573717 1833402 1473956 1842224 1980940 1898118 2508451 2560404 1798362 602048 897941 Azelaic acid 1471306 1054474 1270796 1074932 4048572 1057291 1785391 1490505 1137760 1010755 1186346 904765 1038364 1444416 1354673 972910 1115518 1056824 955648 946234 1639191 1534919 989387 607664 681201 2-Aminoadipic acid 1826496 224132 1102876 113461 1224390 255092 1036842 284165 1406693 250656 1471432 209179 2222557 471723 2142228 230394 512360 339312 1798433 307506 917528 817578 373199 0 628 2-Hydroxybenzaldehyde 483686 231728 473151 341274 423976 182086 439949 234202 371061 182331 1596317 277652 323596 272575 1853176 287448 506527 143353 550196 169433 454206 460662 217753 33661 12666 2-Hydroxyisobutyric acid 28054902 5367830 21310526 4725664 29115336 4321766 16646993 2347900 28776350 5186115 10918745 2450766 38369176 4511692 24230472 2988916 24444560 6260148 21370106 3282046 13477321 12211728 7095486 2557682 935965 4-HYDROXYBENZALDEHYDE 1481196 170063 1277003 309507 939657 153866 834028 149676 1083833 214536 1206164 391835 981628 175564 1664997 310574 1151903 144796 984373 97757 538817 565034 221446 28013 67150 Aspartate 15931509 1184787 15266297 749800 15270450 606937 10879916 762505 12851909 534310 10624924 505111 14196294 1368395 18992072 1099832 9208255 572464 14186870 881158 8616846 7548002 5835738 33527 4201 DL-3-Aminoisobutyric acid 2293999 228080 2367069 155176 2257667 167721 1839553 210879 1951209 148536 2157506 127623 2307892 262913 2540755 191890 1647226 132351 2276299 184676 1347002 1263151 757808 2400 0 Glucose 155633 1465057 160829 1294342 162658 1243684 158277 1370554 214371 925254 191829 958804 131275 1312588 169386 1012541 587795 827804 192349 1173018 192764 209791 187679 895 0 Glutamine 2408114 2024545 2323807 1245855 2781816 1511588 2157496 1666742 2363819 1243132 2362051 1265750 2358605 2072146 2822474 1677586 4037836 1242336 2171442 1307336 2360248 2174120 1352699 34306 10553 GLYCERATE 188583 126060 72950 140694 75064 83395 51069 100703 94737 67761 165351 102824 115556 233190 245309 172165 55035 102826 98228 114737 157863 149118 73013 5706 1080 L-5-Oxoproline 4625214 512512 3787062 658979 11088073 588186 4600534 562254 2449547 436161 4302158 473026 3359396 527727 10912424 680849 4686730 825533 6233930 510249 2506166 2364130 867285 14335 3093 L-Glutamic acid 32477736 4131217 32654146 2828860 30618988 3030941 25355038 3611895 27094230 2573174 28197920 2250917 29221648 4280912 31887186 3317498 22179142 2271678 28177908 3086484 19674724 17436700 11292401 82117 30168 N-ACETYLALANINE (IORA) 812388 21136 518058 16250 667919 11909 599079 6260 582800 18601 557592 13075 880790 20253 899185 9397 746945 18829 707782 12708 277245 244285 140717 765 634 N-Acetylaspartate 33875632 1159076 27733098 1250915 35621064 1065426 23426362 861322 22942100 969167 20718786 848270 32050596 1039721 33534344 777101 25869846 1220530 25289856 655873 13803025 12414599 4983057 12418 0 O-Phosphoethanolamine 861319 1491692 966754 1021653 726560 982792 865837 1050731 867347 804834 803645 921730 845835 1357619 776248 1282084 773299 760659 734026 1012739 1050809 1420538 1316294 19297 5171 Salicylic acid 922211 672570 1196457 774393 1463956 957681 1564544 1009466 691961 847146 1586058 1080878 2012682 1150685 1535572 896919 1301705 987442 1292750 576230 914384 967734 504667 41788 58624 Taurine 314238784 65528336 283512864 39386604 286551392 38783576 242573056 40701632 271037056 33966048 232903744 32034578 265546896 61975084 303919744 47278704 191954592 24244152 245952720 36757748 205765808 191963888 162744128 2973150 963296 Threonine 426011 411110 296429 235383 480946 284544 413159 331588 262137 212360 359310 239072 338260 485731 412979 392834 363799 235775 311142 281930 360161 320796 143201 1809 1403 TRYPTOPHAN 124031 152576 102462 159294 133261 161201 95310 265077 76674 144658 64353 113432 81996 123109 140783 89628 105408 261354 85274 105987 133499 114444 38930 0 0 DOPA 11455 14377 11206 7188 2330 8313 2511 10510 13697 7340 1396 13551 19937 12681 28930 13751 6844 866 31945 21051 20169 14502 8333 0 0 Gluconate 357923 257432 252941 192744 358107 210120 258829 197270 236767 157446 246095 152521 355201 227968 452193 176202 228325 174174 293555 170118 237549 225783 70689 0 0 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Average Rt(min) Average Mz Formula KEGG_Code Phosphoric acid 2.585 96.96552 H3O4P C00009 SUCCINIC ACID 2.919 117.01439 C4H6O4 C00042 Glycerophosphate(2) 1.979 170.99908 C3H9O6P C00093 FUMARATE (IORA) 1.963 114.99886 null C00122 ADENINE 2.977 134.0414 C5H5N5 C00147 Lactic acid 2.221 89.02062 C3H6O3 C00256 HYPOXANTHINE (IORA) 2.247 135.02551 null C00262 OROTATE (IORA) 1.567 155.00737 null C00295 Creatine 1.568 130.05667 C4H9N3O2 C00300 DIHYDROOROTATE (IORA) 1.584 157.02994 null C00337 Xanthine 2.544 151.01955 C5H4N4O2 C00385 Itaconic acid (Not validated) 1.708 129.01378 C5H6O4 C00490 3-HYDROXYBENZOATE (IORA) 6.101 137.01849 null C00587 UROCANATE (IORA) 1.885 137.02965 null C00785 D-PANTOTHENIC ACID 4.915 218.09381 C9H17NO5 C00864 4-Nitrophenol 9.752 138.01387 C6H5NO3 C00870 MALEATE (IORA) 1.57 115.00999 null C01384 OXOPROLINE (IORA) 1.531 128.0298 null C01879 2-METHYLMALEATE (IORA) 2.331 129.01378 null C02226 UREIDOPROPIONATE (IORA) 1.497 131.04036 null C02642 Phenylphosphoric acid 1.708 173.00182 C6H7O4P C02734 METHIONINE SULFOXIMINE (IORA) 1.48 179.04857 null C03510 Meglutol 3.279 161.03854 C6H10O5 C03761 HydroxyButyric acid 1.63 103.03561 C4H8O3 C05984 Azelaic acid 10.599 187.08942 C9H16O4 C08261 2-Aminoadipic acid 1.628 160.05479 C6H11NO4 C00956 2-Hydroxybenzaldehyde 7.406 121.02428 C7H6O2 C06202 2-Hydroxyisobutyric acid 3.614 103.03564 C4H8O3 C05984 4-HYDROXYBENZALDEHYDE 10.34 121.02428 C7H6O2 C00633 Aspartate 1.519 132.02455 C4H7NO4 C00049 DL-3-Aminoisobutyric acid 1.533 102.05167 C4H9NO2 C02356 Glucose 1.543 179.04851 C6H12O6 C00031 Glutamine 1.482 145.05574 C5H10N2O3 C00064 GLYCERATE 1.701 105.01485 C3H6O4 C00258 L-5-Oxoproline 2.583 128.0298 C5H7NO3 C01879 L-Glutamic acid 1.524 146.03943 C5H9NO4 C00025 N-ACETYLALANINE (IORA) 2.294 130.04533 null N-Acetylaspartate 2.298 174.03329 C6H9NO5 C01042 O-Phosphoethanolamine 1.486 140.00583 C2H8NO4P C00346 Salicylic acid 10.753 137.01849 C7H6O3 C00805 Taurine 1.495 124.00192 C2H7NO3S C00245 Threonine 1.495 118.04585 C4H9NO3 C00188 TRYPTOPHAN 5.84 203.0742 C11H12N2O2 C00078 DOPA 1.714 196.0659 C9H11NO4 C00355 Gluconate 1.567 195.0425 C6H12O7 C00257 METABOLITES_END #END