#METABOLOMICS WORKBENCH amitch_20180414_103204 DATATRACK_ID:1378 STUDY_ID:ST000955 ANALYSIS_ID:AN001567 PROJECT_ID:PR000656 VERSION 1 CREATED_ON April 19, 2018, 11:02 am #PROJECT PR:PROJECT_TITLE Cereblon suppression offers novel approach for anti-tumor immunity in melanoma PR:PROJECT_TYPE Exploratory PR:PROJECT_SUMMARY The functional suppression of T cells in the tumor are intimately linked to PR:PROJECT_SUMMARY impared cellular metabolism. Glucose acts as the primary fuel source for the PR:PROJECT_SUMMARY generation of ATP in activated T cells and both oxidative phosphorylation and PR:PROJECT_SUMMARY glycolysis are critical for full effector functions and tumor erradication. We PR:PROJECT_SUMMARY have identified a checkpoint protein that regulates the metabolic suppression in PR:PROJECT_SUMMARY the tumor. Identification of the precise function of different metabolic events PR:PROJECT_SUMMARY regulated by this checkpoint protein will be informative. This project will PR:PROJECT_SUMMARY define the role of a glutamine/arginine mediated pathway in CD8+ T-cells. PR:INSTITUTE Moffitt Cancer Center PR:DEPARTMENT Immunology PR:LABORATORY Burnette PR:LAST_NAME Burnette PR:FIRST_NAME Pearlie PR:ADDRESS 12902 Magnolia Drive, Tampa, FL 33612 PR:EMAIL Pearlie.Burnette@moffitt.org PR:PHONE 813-270-9181 PR:FUNDING_SOURCE Moffitt Cancer Center #STUDY ST:STUDY_TITLE Effects of ODC inhibition on T cell metabolism ST:STUDY_TYPE drug treatment at two time ST:STUDY_SUMMARY Isolated T cells were activated for 48-72 hours in the presence and absence of ST:STUDY_SUMMARY 5mM DFMO, an ODC inhibitor ST:INSTITUTE University of Florida ST:DEPARTMENT SECIM ST:LAST_NAME Hesterberg ST:FIRST_NAME Rebecca ST:ADDRESS 12902 Magnolia Dr ST:EMAIL rebecca.hesterberg@moffitt.org ST:PHONE 813-270-9181 ST:NUM_GROUPS 4 ST:TOTAL_SUBJECTS 16 ST:STUDY_COMMENTS OT-1 mice express a transgenic T cell receptor in all CD8+ T cells that binds ST:STUDY_COMMENTS with high affinity to a peptide derived from the protein ovalbumin (SIINFEKL). #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:GENOTYPE_STRAIN C57Bl6 OT-1 SU:AGE_OR_AGE_RANGE 6-8 weeks SU:ANIMAL_ANIMAL_SUPPLIER Breeding at Moffitt for several generations, originally from Jax SU:ANIMAL_LIGHT_CYCLE 12 hour #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS WT1 WT1_48h_0mM_1 Time activated with peptide:48 | DFMO treatment (mM):- SUBJECT_SAMPLE_FACTORS WT1 WT1_48h_0mM_2 Time activated with peptide:48 | DFMO treatment (mM):- SUBJECT_SAMPLE_FACTORS WT1 WT1_48h_0mM_3 Time activated with peptide:48 | DFMO treatment (mM):- SUBJECT_SAMPLE_FACTORS WT1 WT1_48h_5mM_1 Time activated with peptide:48 | DFMO treatment (mM):5 SUBJECT_SAMPLE_FACTORS WT1 WT1_48h_5mM_2 Time activated with peptide:48 | DFMO treatment (mM):5 SUBJECT_SAMPLE_FACTORS WT1 WT1_48h_5mM_3 Time activated with peptide:48 | DFMO treatment (mM):5 SUBJECT_SAMPLE_FACTORS WT1 WT1_72h_0mM_1 Time activated with peptide:72 | DFMO treatment (mM):- SUBJECT_SAMPLE_FACTORS WT1 WT1_72h_0mM_2 Time activated with peptide:72 | DFMO treatment (mM):- SUBJECT_SAMPLE_FACTORS WT1 WT1_72h_0mM_3 Time activated with peptide:72 | DFMO treatment (mM):- SUBJECT_SAMPLE_FACTORS WT1 WT1_72h_5mM_1 Time activated with peptide:72 | DFMO treatment (mM):5 SUBJECT_SAMPLE_FACTORS WT1 WT1_72h_5mM_2 Time activated with peptide:72 | DFMO treatment (mM):5 SUBJECT_SAMPLE_FACTORS WT1 WT1_72h_5mM_3 Time activated with peptide:72 | DFMO treatment (mM):5 SUBJECT_SAMPLE_FACTORS WT3 WT3_48h_0mM_1 Time activated with peptide:48 | DFMO treatment (mM):- SUBJECT_SAMPLE_FACTORS WT3 WT3_48h_0mM_2 Time activated with peptide:48 | DFMO treatment (mM):- SUBJECT_SAMPLE_FACTORS WT3 WT3_48h_0mM_3 Time activated with peptide:48 | DFMO treatment (mM):- SUBJECT_SAMPLE_FACTORS WT3 WT3_48h_5mM_1 Time activated with peptide:48 | DFMO treatment (mM):5 SUBJECT_SAMPLE_FACTORS WT3 WT3_48h_5mM_2 Time activated with peptide:48 | DFMO treatment (mM):5 SUBJECT_SAMPLE_FACTORS WT3 WT3_48h_5mM_3 Time activated with peptide:48 | DFMO treatment (mM):5 SUBJECT_SAMPLE_FACTORS WT3 WT3_72h_0mM_1 Time activated with peptide:72 | DFMO treatment (mM):- SUBJECT_SAMPLE_FACTORS WT3 WT3_72h_0mM_2 Time activated with peptide:72 | DFMO treatment (mM):- SUBJECT_SAMPLE_FACTORS WT3 WT3_72h_0mM_3 Time activated with peptide:72 | DFMO treatment (mM):- SUBJECT_SAMPLE_FACTORS WT3 WT3_72h_5mM_1 Time activated with peptide:72 | DFMO treatment (mM):5 SUBJECT_SAMPLE_FACTORS WT3 WT3_72h_5mM_2 Time activated with peptide:72 | DFMO treatment (mM):5 SUBJECT_SAMPLE_FACTORS WT3 WT3_72h_5mM_3 Time activated with peptide:72 | DFMO treatment (mM):5 SUBJECT_SAMPLE_FACTORS WT7 WT7_48h_0mM_1 Time activated with peptide:48 | DFMO treatment (mM):- SUBJECT_SAMPLE_FACTORS WT7 WT7_48h_0mM_2 Time activated with peptide:48 | DFMO treatment (mM):- SUBJECT_SAMPLE_FACTORS WT7 WT7_48h_0mM_3 Time activated with peptide:48 | DFMO treatment (mM):- SUBJECT_SAMPLE_FACTORS WT7 WT7_48h_5mM_1 Time activated with peptide:48 | DFMO treatment (mM):5 SUBJECT_SAMPLE_FACTORS WT7 WT7_48h_5mM_2 Time activated with peptide:48 | DFMO treatment (mM):5 SUBJECT_SAMPLE_FACTORS WT7 WT7_48h_5mM_3 Time activated with peptide:48 | DFMO treatment (mM):5 SUBJECT_SAMPLE_FACTORS WT7 WT7_72h_0mM_1 Time activated with peptide:72 | DFMO treatment (mM):- SUBJECT_SAMPLE_FACTORS WT7 WT7_72h_0mM_2 Time activated with peptide:72 | DFMO treatment (mM):- SUBJECT_SAMPLE_FACTORS WT7 WT7_72h_0mM_3 Time activated with peptide:72 | DFMO treatment (mM):- SUBJECT_SAMPLE_FACTORS WT7 WT7_72h_5mM_1 Time activated with peptide:72 | DFMO treatment (mM):5 SUBJECT_SAMPLE_FACTORS WT7 WT7_72h_5mM_2 Time activated with peptide:72 | DFMO treatment (mM):5 SUBJECT_SAMPLE_FACTORS WT7 WT7_72h_5mM_3 Time activated with peptide:72 | DFMO treatment (mM):5 SUBJECT_SAMPLE_FACTORS WT8 WT8_48h_0mM_1 Time activated with peptide:48 | DFMO treatment (mM):- SUBJECT_SAMPLE_FACTORS WT8 WT8_48h_0mM_2 Time activated with peptide:48 | DFMO treatment (mM):- SUBJECT_SAMPLE_FACTORS WT8 WT8_48h_0mM_3 Time activated with peptide:48 | DFMO treatment (mM):- SUBJECT_SAMPLE_FACTORS WT8 WT8_48h_5mM_1 Time activated with peptide:48 | DFMO treatment (mM):5 SUBJECT_SAMPLE_FACTORS WT8 WT8_48h_5mM_2 Time activated with peptide:48 | DFMO treatment (mM):5 SUBJECT_SAMPLE_FACTORS WT8 WT8_48h_5mM_3 Time activated with peptide:48 | DFMO treatment (mM):5 SUBJECT_SAMPLE_FACTORS WT8 WT8_72h_0mM_1 Time activated with peptide:72 | DFMO treatment (mM):- SUBJECT_SAMPLE_FACTORS WT8 WT8_72h_0mM_2 Time activated with peptide:72 | DFMO treatment (mM):- SUBJECT_SAMPLE_FACTORS WT8 WT8_72h_0mM_3 Time activated with peptide:72 | DFMO treatment (mM):- SUBJECT_SAMPLE_FACTORS WT8 WT8_72h_5mM_1 Time activated with peptide:72 | DFMO treatment (mM):5 SUBJECT_SAMPLE_FACTORS WT8 WT8_72h_5mM_2 Time activated with peptide:72 | DFMO treatment (mM):5 SUBJECT_SAMPLE_FACTORS WT8 WT8_72h_5mM_3 Time activated with peptide:72 | DFMO treatment (mM):5 #COLLECTION CO:COLLECTION_SUMMARY At 48 or 72 hours cells in culture medium were centrifuged on a Ficoll gradient CO:COLLECTION_SUMMARY to enrich viable cells. Cells were washed 1x with PBS, counted, and frozen as CO:COLLECTION_SUMMARY cell pellets at -80. CO:SAMPLE_TYPE Cultured cells #TREATMENT TR:TREATMENT_SUMMARY Splenic CD8+ T cells were isolated from 4 OT-1 mice (WT1, 3, 7, 8) by negative TR:TREATMENT_SUMMARY immunomagnetic selection. T cells stimulated with ova-derived peptide (SIINFEKL) TR:TREATMENT_SUMMARY that binds with high affinity to the OT-1 transgenic TCR and 10ng/mL IL-2 with TR:TREATMENT_SUMMARY and without 5mM DFMO for 48-72 hours. Cell stoarge: 37 degrees, Cell growth TR:TREATMENT_SUMMARY container: 24 well plate, cell media: RPMI1640 with glutamine. Added components TR:TREATMENT_SUMMARY were: NEAA, Sodium Pyruvate, Fetal Bovine Serum, Fungizone, Gentimycin, TR:TREATMENT_SUMMARY Pen/strep. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY None SP:SAMPLEPREP_PROTOCOL_FILENAME GMetabolomics_LCMS_Protocol_092117.pdf;Appendix_A_Internal_Standard_Prep_GLCMS.pdf #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Thermo Dionex Ultimate 3000 CH:COLUMN_NAME ACE Excel 2 C18-PFP (100 x 2.1mm, 2um) CH:FLOW_RATE 350 ul/min CH:METHODS_FILENAME GMetabolomics_LCMS_Protocol_092117.pdf CH:SOLVENT_A 0.1% Formic Acid in Water CH:SOLVENT_B Acetonitrile #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:MS_COMMENTS - MS:INSTRUMENT_NAME Thermo Q Exactive Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_RESULTS_FILE ST000955_AN001567_Results.txt UNITS:peak height #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS peak height MS_METABOLITE_DATA_START Samples WT1_72h_5mM_1 WT1_72h_5mM_3 WT1_72h_5mM_2 WT3_48h_0mM_1 WT3_48h_0mM_3 WT3_48h_5mM_2 WT3_48h_5mM_1 WT3_48h_0mM_2 WT1_48h_0mM_1 WT3_72h_0mM_1 WT3_48h_5mM_3 WT3_72h_0mM_2 WT3_72h_5mM_1 WT3_72h_0mM_3 WT3_72h_5mM_3 WT7_48h_0mM_1 WT7_48h_5mM_1 WT3_72h_5mM_2 WT7_48h_0mM_3 WT7_48h_0mM_2 WT7_48h_5mM_2 WT7_48h_5mM_3 WT1_48h_0mM_2 WT7_72h_0mM_2 WT7_72h_0mM_1 WT7_72h_0mM_3 WT7_72h_5mM_2 WT7_72h_5mM_3 WT8_48h_0mM_1 WT7_72h_5mM_1 WT8_48h_0mM_3 WT1_48h_0mM_3 WT8_48h_0mM_2 WT8_72h_0mM_1 WT8_72h_0mM_3 WT8_48h_5mM_1 WT8_48h_5mM_3 WT8_72h_5mM_1 WT8_72h_0mM_2 WT8_48h_5mM_2 WT1_48h_5mM_1 WT8_72h_5mM_2 WT8_72h_5mM_3 WT1_48h_5mM_3 WT1_72h_0mM_2 WT1_48h_5mM_2 WT1_72h_0mM_1 WT1_72h_0mM_3 Factors Time activated with peptide:72 | DFMO treatment (mM):5 Time activated with peptide:72 | DFMO treatment (mM):5 Time activated with peptide:72 | DFMO treatment (mM):5 Time activated with peptide:48 | DFMO treatment (mM):- Time activated with peptide:48 | DFMO treatment (mM):- Time activated with peptide:48 | DFMO treatment (mM):5 Time activated with peptide:48 | DFMO treatment (mM):5 Time activated with peptide:48 | DFMO treatment (mM):- Time activated with peptide:48 | DFMO treatment (mM):- Time activated with peptide:72 | DFMO treatment (mM):- Time activated with peptide:48 | DFMO treatment (mM):5 Time activated with peptide:72 | DFMO treatment (mM):- Time activated with peptide:72 | DFMO treatment (mM):5 Time activated with peptide:72 | DFMO treatment (mM):- Time activated with peptide:72 | DFMO treatment (mM):5 Time activated with peptide:48 | DFMO treatment (mM):- Time activated with peptide:48 | DFMO treatment (mM):5 Time activated with peptide:72 | DFMO treatment (mM):5 Time activated with peptide:48 | DFMO treatment (mM):- Time activated with peptide:48 | DFMO treatment (mM):- Time activated with peptide:48 | DFMO treatment (mM):5 Time activated with peptide:48 | DFMO treatment (mM):5 Time activated with peptide:48 | DFMO treatment (mM):- Time activated with peptide:72 | DFMO treatment (mM):- Time activated with peptide:72 | DFMO treatment (mM):- Time activated with peptide:72 | DFMO treatment (mM):- Time activated with peptide:72 | DFMO treatment (mM):5 Time activated with peptide:72 | DFMO treatment (mM):5 Time activated with peptide:48 | DFMO treatment (mM):- Time activated with peptide:72 | DFMO treatment (mM):5 Time activated with peptide:48 | DFMO treatment (mM):- Time activated with peptide:48 | DFMO treatment (mM):- Time activated with peptide:48 | DFMO treatment (mM):- Time activated with peptide:72 | DFMO treatment (mM):- Time activated with peptide:72 | DFMO treatment (mM):- Time activated with peptide:48 | DFMO treatment (mM):5 Time activated with peptide:48 | DFMO treatment (mM):5 Time activated with peptide:72 | DFMO treatment (mM):5 Time activated with peptide:72 | DFMO treatment (mM):- Time activated with peptide:48 | DFMO treatment (mM):5 Time activated with peptide:48 | DFMO treatment (mM):5 Time activated with peptide:72 | DFMO treatment (mM):5 Time activated with peptide:72 | DFMO treatment (mM):5 Time activated with peptide:48 | DFMO treatment (mM):5 Time activated with peptide:72 | DFMO treatment (mM):- Time activated with peptide:48 | DFMO treatment (mM):5 Time activated with peptide:72 | DFMO treatment (mM):- Time activated with peptide:72 | DFMO treatment (mM):- 2-hydroxyglutarate 165221 170670 169271 203473 199178 125860 136789 182542 151147 100413 126033 49597 93436 96224 100647 147359 152840 97526 150871 155479 151621 49444 158614 84879 85709 91037 179041 168912 116584 180882 118778 142270 131710 122724 125290 135147 146287 131119 124857 152265 118460 131819 120412 133410 106328 125355 116478 111380 3-methyladipic acid 1129386 1159935 1153941 1229522 1255598 867538 858936 1233534 1248094 1246218 879272 1273499 818411 1209917 769197 1097806 1201543 812028 1095979 1044897 1108594 1184563 1070944 604719 636721 647375 821462 851783 1031246 840646 974938 1138120 1015199 1194685 1196490 1264944 1266652 1101411 1162582 1508791 1061251 1084697 1041329 1098134 1114865 1056668 1167408 1208497 3-SULFINO-L-ALANINE 174459 209819 185946 260729 233533 236854 231525 226012 139166 171370 200487 176025 183945 157976 191263 275313 383402 217323 256501 206332 293253 329725 152454 167181 199622 175025 174215 204514 195792 175228 213112 155490 250692 95866 69531 141175 138357 419456 76483 165057 120523 398288 394048 142843 122754 122882 121151 135514 4-oxoproline 4817743 5390465 5368582 4365186 4324931 3961648 3875061 3842722 4626359 3268631 3361269 3598159 4378860 3463840 4592379 2762407 3737771 4512174 2704330 2255110 3178915 3870938 4342512 2771207 2928347 2881950 3084346 3518770 2810435 3148731 2901075 4465666 3022118 7598823 7471094 4926154 4832789 4352454 6280685 5758682 1962987 4348097 4163411 2366895 3072492 1985275 3545621 3536396 6-Hydroxycaproic acid 266458 268021 275086 214080 215080 243408 232415 209598 215456 267069 220778 259847 368974 265140 369498 689090 409862 385746 683065 622183 383064 413330 216906 257060 283148 275262 239557 246144 235569 240112 249002 219086 255620 286004 273765 528248 502732 692087 263792 539682 366816 708007 695422 382757 338733 377930 364069 364177 ADENINE 108154 139239 140706 373080 399812 144353 159757 381285 381522 87515 117345 89149 184248 96652 198868 217397 74463 204202 218520 165203 70351 74542 366332 126024 144059 141393 328824 261069 286852 346991 290046 348726 345291 89718 76759 151008 162850 225053 83159 175597 129290 208945 225365 125850 98730 111525 99242 100102 ADP 33999 40048 34668 105428 119598 80182 73622 107755 102587 36554 71476 37261 56971 36873 58561 15658 44806 45519 16522 16109 38150 36657 97809 41002 41030 36589 113850 113978 131795 124193 119559 107356 132474 42211 43428 31557 31988 18900 47434 34684 30555 19668 20834 28264 31384 23936 35352 30500 ANTHRANILATE 59994 68425 64033 46764 55321 52888 47819 42542 48353 51221 43083 57684 35880 50941 35891 74296 48282 42173 70887 64814 38819 52387 51149 48336 63545 50523 44400 38354 31393 49868 36210 50634 38776 52704 51562 56182 56686 57117 48103 67430 98779 52554 60850 108609 48219 94596 50833 53401 ASPARAGINE 949171 1083949 1063366 2073407 1996288 1284418 1310697 1728264 2167622 1299900 1093279 1266370 1168205 1253536 1231178 911463 1408462 1271829 855689 763579 1181281 1516895 2368345 1254387 1422152 1353027 2213451 2412849 2114140 2190669 2072319 2330338 2515008 1438771 1475935 607381 633291 868272 1180290 662526 462977 850193 898675 535874 1067633 443665 1271411 1235073 ASPARTATE 3437637 3869858 3930722 9128155 9077043 5570541 5581917 8041060 9438098 4606698 4843050 4316671 5216239 4327949 5599610 5062349 5521260 5529791 4964137 4123698 4561159 5657210 9253395 4677544 5335457 5257711 10300000 11000000 8821930 10500000 9098659 9236186 9888345 5796844 5771573 4584025 4655335 6804471 5029918 4597504 2491209 6733864 7236014 3019121 4526518 2632688 5075182 5179491 BOC-L-Tyrosine_R1 140000000 150000000 147000000 150000000 152000000 148000000 143000000 147000000 148000000 151000000 142000000 147000000 152000000 148000000 148000000 148000000 152000000 153000000 153000000 145000000 148000000 148000000 142000000 142000000 140000000 147000000 150000000 148000000 147000000 147000000 142000000 147000000 146000000 153000000 150000000 156000000 151000000 157000000 155000000 148000000 151000000 150000000 149000000 153000000 142000000 155000000 150000000 151000000 BOC-L-Tyrosine_R2 112000000 120000000 117000000 119000000 120000000 118000000 116000000 117000000 117000000 121000000 112000000 116000000 119000000 119000000 116000000 118000000 122000000 120000000 121000000 114000000 118000000 118000000 111000000 114000000 113000000 118000000 119000000 118000000 117000000 115000000 113000000 118000000 115000000 118000000 118000000 123000000 123000000 124000000 117000000 116000000 119000000 121000000 115000000 118000000 114000000 119000000 118000000 120000000 C12H22O11-DISACCHARIDE-(6C/6C; GLC-GLC/GLC-FRC/GAL-GLC) 237621 281028 277026 345143 353502 296543 293680 302784 428991 570125 256553 579881 310647 586338 314287 429083 528505 318698 418535 341654 441201 521937 448009 138124 140390 139403 249600 252738 152215 252913 154155 436049 166125 284955 289187 268716 315598 497474 245155 311267 175200 495249 481759 225082 221967 207632 280531 277535 C6H12O6-HEXOSE/KETOSE/INOSITOL 746850 794898 849199 1169992 1185343 851112 851858 1019342 1447473 723123 777238 713305 698950 751153 785101 634576 1085613 707061 580958 519362 965055 1079766 1587290 863634 932680 978173 1019155 1127820 1158517 1030323 1166326 1642976 1268778 688090 740392 1366494 1355268 709588 626992 1330416 433517 696797 679196 500251 549109 446201 662387 659200 CITRULLINE 335984 354814 353720 187042 196395 206879 213275 167506 276906 2499276 179800 2416653 822420 2463563 788641 512225 4422617 807856 520595 490350 3927888 4389760 278482 262010 279646 255245 193439 179321 151029 176128 150031 286284 183382 3091823 3034015 729583 724852 368405 2774916 733229 173983 369224 391205 205503 1420950 181921 1644846 1635819 CMP 595655 699896 683824 1514634 1483023 803844 824051 1367635 1767954 1019771 738423 1079071 1026051 1041180 1006580 567120 993316 938234 568809 469542 834034 983233 1728189 573993 594795 589435 1750932 1700318 1591908 1630555 1610041 1849443 1652043 1147807 1069471 362946 352119 604709 1002339 354777 383872 558274 565300 448254 1114336 400626 1292894 1323917 Creatine-D3 1467923 1708688 1660939 1742464 1731187 1781747 1801244 1546421 1577597 1772060 1635725 1786803 1906636 1824892 1858918 1710463 1810641 1937816 1724191 1480140 1553194 1753341 1534880 1592056 1776079 1700541 1709936 1697875 1531906 1761428 1558822 1726605 1695887 1842674 1874308 1739934 1741959 1810857 1627937 1780600 1631681 1807774 1729961 1841154 1590308 1660155 1885904 1888257 CYTIDINE 2',3'-CYCLIC MONO-PHOS-PHATE 61146 67090 72443 26544 24238 51545 49522 18446 63874 15314 38029 7395 33770 7455 28620 34165 8549 31922 32391 24627 8643 8864 60995 47414 55121 49439 36858 36197 55882 42008 51847 57641 43601 12789 17185 236935 226603 142759 12162 243873 485252 128190 142284 514762 23549 494890 25270 23078 D-GLUCURONIC ACID/D-GLUCURONOLACTONE/D-(+)-GALACTURONIC ACID_R1 276921 260215 282213 337543 324052 298412 329570 271854 319113 345206 313766 322230 401892 307144 401501 363149 341567 432507 366139 312093 267157 337123 301210 373850 407478 440708 381531 373275 348013 365366 329806 318155 366122 343946 365838 302085 342208 350035 325809 258915 342784 337322 337452 388136 322248 372566 375419 406520 D-GLUCURONIC ACID/D-GLUCURONOLACTONE/D-(+)-GALACTURONIC ACID_R2 107945 69738 71006 311500 69270 98589 84052 83101 72437 68848 76145 105059 61820 71272 61678 57837 80662 70517 65808 53404 53012 78396 51836 38030 56560 63302 63802 81450 60179 79961 63685 66373 65968 66840 56701 54285 61282 146896 70727 57341 54140 93516 116998 52201 57271 50865 56859 60949 DIHYDROXYACETONE PHOSPHATE 237255 240157 254737 4211236 3839990 626134 635688 3603405 2799398 577772 559823 521059 418926 581966 446272 36679 453646 472894 39929 35552 443945 446052 2842170 177389 180584 183548 4063943 4508390 2534305 4188887 2586478 2888890 2877419 677597 683737 37951 38292 75366 615579 39062 38117 79711 64500 45359 400710 56770 471660 475768 D-RIBOSE 5-PHOSPHATE/RIBULOSE 5-PHOSPHATE 253683 272932 268122 722044 678649 358481 340960 635465 638442 292722 323374 284197 261571 283304 284552 413496 295826 281949 440136 409725 285054 292650 669602 253168 272899 275524 635043 684204 628291 623245 629767 667763 661097 314305 287084 451789 421714 516507 294782 397157 457540 543723 464748 463865 307101 445757 317845 315380 GLUCONIC ACID/D-GULONIC ACID GAMA-LACTONE 213374 226629 237129 467478 475128 260670 248427 425958 289948 342016 265992 373321 163953 381172 192405 255802 238696 184851 233340 217422 250703 244944 300703 149151 153290 149737 237519 229893 226893 245996 224445 294424 232814 198449 195835 359903 356089 218687 183450 353258 157239 209402 197917 162937 182945 137569 203241 214057 GLYCEROL 2-PHOSPHATE/SN-GLYCEROL 3-PHOSPHATE 232365 238984 245705 338324 340110 227363 233337 292322 338855 225930 227929 232012 240017 203184 228831 206205 196374 235955 206536 187632 185769 201351 374986 146933 161213 149620 544761 558411 393102 540227 360114 377899 401205 192812 191768 132603 120524 222872 160750 146444 121906 230302 221617 115544 244862 111451 229228 241762 GLYCOLATE 676040 584375 594841 512309 548030 476897 484201 612261 754596 567948 678742 519612 548942 532718 494424 627116 581415 513142 600350 844378 783862 568006 657188 672299 603157 558688 633675 626943 661000 614894 613627 620013 589551 962443 999217 587726 561163 549897 1033582 565343 674233 539522 556171 526818 683119 647889 519056 597201 LACTATE-dimer 503678 753919 704227 1767961 1842868 446492 447307 1284119 1046596 507831 307190 587928 453725 578228 465831 1007946 1131003 462885 1030642 631446 717305 1096036 927661 234773 286114 282500 1221134 1494795 648690 1246718 710081 1094183 798091 767671 816647 1070841 983823 2079939 506319 1230355 305430 1954898 1950465 484107 791009 365476 1156589 1274735 L-ARGININE 1462486 1631584 1574131 2519579 2364576 1854906 1845998 2208239 2771186 1142568 1650216 1084474 1772517 1079884 1770303 2233868 850061 1837521 2127679 2019764 737797 880338 2867871 1954520 2191779 2149856 2573141 2655248 2802767 2556715 2772439 2912232 2918657 1365521 1363229 1083628 1125133 1620869 1127099 1152712 847375 1660689 1685284 973729 1308704 829224 1492313 1478923 L-CYSTEIC ACID 264869 301309 287479 590019 551306 194077 194304 493998 91955 56247 180091 63938 198039 50397 199528 535703 240867 199038 481327 395112 212463 255947 116452 137833 164318 152522 219318 205912 343628 220184 337078 110098 386087 68944 53308 369450 393337 630701 46010 384222 226726 581500 632856 225138 142189 219875 180175 168866 L-GLUTAMIC ACID 6752003 7409422 7610813 18800000 19100000 10900000 10800000 17300000 19100000 6822404 9849334 7086648 10700000 6663567 10900000 8542193 8731435 10600000 8583206 7392964 7627964 9229230 18600000 8213428 9283572 9214163 20000000 19900000 16900000 20000000 17100000 19300000 18000000 9192105 9178687 6891338 6719798 10900000 8470978 6947323 4093809 10700000 10300000 4760994 7192909 4349050 8318576 7935882 L-GLUTAMINE 1188134 1327096 1404263 2443043 2531470 1575044 1532308 2257843 2810291 1801817 1445291 1779557 1326522 1844345 1312386 1477583 2284639 1403062 1403634 1208149 2065920 2421196 2885963 1594712 1753742 1668107 2774381 2914050 2925806 2776401 2920548 2934965 3137060 2191770 2161099 1001860 1030243 1119038 1887853 1033048 724217 1114889 1196697 839905 1679624 730203 1891311 1990341 L-HISTIDINE 599002 669971 709220 1329036 1270351 882915 793598 1070878 1746370 744358 727691 765250 989323 751077 1069576 682509 972773 1116534 709747 580325 836813 951337 1799824 744622 800248 801389 1471817 1634005 1405409 1503747 1536716 1864087 1611010 1008333 1023476 620405 688302 861817 856585 728664 349499 850993 898709 374409 676279 374404 776375 772413 L-Phenylalanine-13C6 111214 123909 133106 121472 131074 132281 135167 120094 120193 137298 121645 121117 130527 123946 140081 140049 125161 143675 124081 112025 127273 146553 122369 123385 146293 121080 129251 122121 110825 127272 116066 118217 121825 141073 150551 129199 140937 127504 133274 132993 132735 125267 129282 135092 118316 126971 132870 142180 L-SERINE 2117282 2421786 2477813 3297794 3231834 2359566 2351338 2740820 3570231 2564524 2118833 2636302 2416997 2658958 2695259 1944693 2577958 2746014 1700800 1611063 2258288 2803608 3898565 2301335 2564901 2515340 3699855 3835236 3456608 3581755 3394232 3893058 4082324 3555264 3420610 1892800 1963398 2736625 2833866 1967471 1037426 2675227 2803353 1406538 2099693 1130015 2515799 2430390 L-TYROSINE 138471 162118 170555 230665 216575 173647 187984 212137 267646 171843 150193 184368 142855 155662 160205 182452 200086 159529 163808 141768 168735 218269 270826 163158 173559 176253 240443 263620 272291 245258 253053 258294 278405 220901 216158 127285 128726 184060 178948 151485 102336 153379 168575 133312 147152 102525 190797 195680 MALATE 1553431 1499742 1586689 8610301 8663091 2319034 2292304 8958404 9142199 1022995 2385674 977524 1309027 1005174 1273866 1013356 1256029 1271248 1005561 1074553 1283193 1222794 8600792 1274505 1267663 1249638 5645434 5801871 4645893 5700015 4492154 8994764 4391257 1206090 1213667 1274552 1383389 1005896 1260200 1364839 1849113 955928 971759 1746711 1032462 1807742 1009808 1008326 Mannitol 105671 126315 106427 305871 252223 108178 96801 268819 1503802 219268 103843 197896 113360 221790 99244 140646 105097 92828 152677 128771 115006 121846 1588331 66350 75018 89051 322319 305292 145341 313747 149381 1534787 150525 95324 94008 444699 440200 94817 84953 407231 94235 99276 106658 101042 164918 87394 199064 177687 N-ACETYL-HEXOSAMINE 38030 49977 47154 108431 99944 78917 81289 89313 100641 36577 67697 34218 74542 29082 74437 20162 29135 75208 18657 16675 32423 27158 91158 48977 57112 63080 108892 121460 80293 104736 92808 103821 91722 33391 38376 22804 29231 24069 28306 18476 8642 17922 18348 9501 24647 13849 30709 25730 N-ACETYL-L-LEUCINE 254693 265416 275590 510011 532395 289818 308997 493695 367097 613706 283251 617450 864278 576172 853407 2846423 1160361 868380 2854640 2623570 1186453 1220250 346977 472798 492740 495415 547316 535750 1859624 558030 1804963 363234 1843998 534939 536001 462775 459778 472402 523632 526168 299563 447881 481486 314507 1424344 281412 1465990 1489705 N-ACETYLNEURAMINATE 190871 205439 224552 359412 403418 88288 80040 314077 488460 276114 74222 254770 173914 295899 185705 111349 246942 187543 102366 101992 249819 259935 436522 61895 62448 66737 372406 418195 408401 401046 384751 467248 382323 291307 303965 74732 86180 145447 274369 75187 82182 148163 144738 97055 271664 86814 303953 303713 N-BOC-L-Aspartic Acid_R1 18400000 19100000 18900000 18100000 18500000 18200000 18300000 17900000 17200000 18500000 18600000 18400000 19300000 18500000 18800000 18700000 19400000 20000000 18900000 17700000 18700000 18800000 16400000 18500000 18100000 18800000 18000000 18100000 18300000 17700000 17900000 17600000 17900000 19400000 19200000 18600000 18900000 19000000 18500000 18000000 18500000 19000000 18500000 18500000 19100000 18700000 19600000 19400000 N-BOC-L-Aspartic Acid_R2 2574669 2712250 2616919 2545866 2622780 2591366 2680914 2494175 2504894 2600180 2594072 2665952 2956054 2680060 2702870 2732038 2654186 2834565 2652671 2488598 2598015 2695568 2461034 2594332 2636790 2662735 2606173 2479413 2563348 2579457 2461305 2456110 2692190 2781312 2730056 2649138 2686503 2650089 2649498 2756049 2605183 2706074 2732808 2674933 2666625 2740833 2828183 2738886 N-BOC-L-tert-Leucine_R1 27100000 26100000 28000000 25200000 24300000 24500000 24200000 24400000 26100000 27600000 24100000 26100000 28100000 26200000 27100000 25300000 26000000 30900000 24800000 24900000 25800000 27800000 25200000 26600000 27500000 28400000 25600000 27700000 25300000 25700000 25500000 25300000 24600000 27400000 25700000 20900000 22800000 27700000 25900000 21400000 26400000 28300000 28600000 27200000 25600000 26800000 26700000 27000000 N-BOC-L-tert-Leucine_R2 15700000 15100000 16100000 14500000 14100000 13900000 14000000 14100000 14800000 16200000 13800000 15000000 15800000 15300000 15500000 14200000 15300000 17500000 14400000 14400000 15100000 16000000 14000000 15400000 15800000 16400000 14900000 16100000 14800000 14800000 14600000 14600000 14100000 15400000 14700000 11900000 12800000 16500000 14400000 12000000 15400000 16300000 16200000 15500000 14900000 14800000 15000000 15400000 N-BOC-L-Tryptophan_R1 54900000 59400000 59100000 60000000 58700000 56300000 54100000 56600000 55800000 57600000 53200000 54400000 59200000 56800000 57200000 58600000 59600000 58400000 61500000 58400000 59000000 59300000 52400000 50600000 53400000 53200000 56200000 56200000 57600000 55500000 55100000 56100000 55600000 55300000 56700000 61800000 61900000 63900000 53300000 59200000 63100000 62200000 60300000 63700000 54800000 63000000 55700000 56100000 N-BOC-L-Tryptophan_R2 47200000 50800000 51000000 50800000 51800000 48100000 46800000 48200000 48000000 50000000 46100000 46800000 53500000 48900000 50400000 51500000 51500000 50400000 53400000 50600000 51000000 50900000 45500000 44300000 45500000 46500000 49100000 47300000 49400000 48700000 48000000 48800000 50200000 47700000 49400000 53300000 53400000 55600000 47800000 52600000 54500000 54300000 52600000 55900000 47300000 56500000 48400000 48200000 Pantothenic Acid 128358 147298 134249 336205 314492 293838 314087 316651 312751 100454 300034 95135 187688 108319 190245 86060 113912 201286 85924 80246 111101 118504 289883 179868 185714 187226 228135 244003 231850 229235 229023 306794 235426 125923 120806 83325 82601 79307 122375 90821 79647 77501 85059 79634 87859 79612 99601 100242 PROPIONATE 82698 97059 89500 93067 90526 104973 97077 67019 80363 96162 83052 98326 110299 99911 97442 77970 103808 91387 95718 59690 71482 89295 81456 80082 96682 89259 132691 90676 72930 100329 70737 100726 84729 96701 93391 97183 90714 124653 105675 103562 91136 110506 127687 106721 77610 89745 109197 99072 PYRIDOXAL 28972 30412 28490 93725 92427 43451 44339 74288 75650 30324 41112 25894 35345 28439 45056 45925 37250 41842 44892 33987 31254 47210 67977 31330 40510 37336 80193 92460 81072 71665 68868 77006 76004 33342 32603 37247 44555 48690 32492 46718 28149 54961 45251 31140 27529 28136 35150 34144 Salicylic Acid-D4 6335731 6053707 6143292 5961875 5740823 5836369 5664658 5786594 5415716 6267685 5394413 5581347 5974512 6053629 5819275 5964630 6310032 7077434 5916352 6369803 5880602 6788933 5213937 5725152 5941370 6394733 5800657 6546447 6004092 5580071 5900084 5349071 5552946 6483814 5812122 5904973 6830868 5882762 5751092 6173874 5716396 6064830 6331989 6156461 5906301 5846495 6019958 6344507 SARCOSINE/BETA-ALANINE 52469 56926 62065 99591 102928 82730 68139 103219 114748 60127 70238 55388 77031 48741 76233 79624 71601 78115 73171 69515 69028 82267 89280 56700 75773 65608 103322 112503 95357 106131 117233 104953 109960 70751 77371 54861 68163 96345 69178 61253 42871 94074 113862 50106 64724 36425 69674 80936 STACHYOSE 69270 79945 86376 15100 4672 27484 23599 0 36616 5458 36348 14144 15449 12283 16316 52362 16903 12468 55228 48113 10599 33542 33136 26626 17710 23446 25671 21937 48768 24493 46953 40011 46425 151159 132472 89128 94532 401178 114810 102695 84608 422858 425217 91889 18250 87927 17963 17869 SUCCINATE 1309606 1471499 1524363 1505488 1519561 1531379 1464962 1396997 1488082 1912296 1280821 2035607 1476379 2005055 1536314 1739661 2369227 1481253 1740938 1565256 2136642 2464599 1511272 953826 964972 1021965 1584386 1707679 1230834 1657537 1249131 1476541 1323562 2104254 2042402 1536978 1541179 1447800 1878326 1772388 1194895 1396945 1404035 1343905 1488642 1210722 1699363 1727879 Succinic-2,2,3,3-D4 Acid 5814313 6092854 6234324 6330185 6106516 6520433 6570996 5870986 5868488 6139519 5897767 6478522 6568165 6260252 7118295 6707031 6771770 7034652 6561628 6037501 5922265 6714985 5580156 5908185 5973187 6199079 5910887 6503089 6119517 5857817 5975770 5899573 6307474 6681233 6866831 6187817 6210306 6733955 6175146 6683944 6144527 6710749 6525369 6655724 6033797 6114736 6910630 6854608 Sulfoacetaldehyde 79084 81627 75443 103698 83328 71763 74781 97506 85354 103570 62207 111066 93604 87407 99351 75198 70373 88846 67740 65567 68330 71933 112588 65170 60810 77869 81555 94336 80905 94691 68860 99737 95035 98534 89199 73467 72392 110592 61191 79129 77043 120096 102256 77709 59034 65748 78508 81764 TAURINE 347557 379901 423920 4690080 4848298 890729 881505 4253570 2754516 662213 775531 636796 529153 598287 581223 231538 492672 565613 213032 180127 420306 546892 2797774 313482 355450 347272 2520938 2588938 1607312 2643061 1665876 2655147 1796313 561144 526409 320764 342560 258104 499326 322502 254034 258833 264149 292442 536648 256115 559777 608984 THREONINE/HOMOSERINE 883146 933937 938068 1612999 1600652 987741 936058 1517494 1686401 1165411 906778 1227037 1390699 1168740 1430530 1317464 1362380 1336820 1171629 1122311 1248600 1380749 1758854 1339286 1452161 1448710 2093952 1999137 2036330 2123469 2068374 1736759 2220603 1342323 1395549 864232 803889 1232128 1262704 820978 600806 1144185 1171514 679510 1213340 681844 1295557 1392373 Tryptophan 157377 182107 165349 267196 257194 197768 207459 247187 292064 186151 180874 179618 198124 182146 201450 186879 230737 205586 179058 175346 211961 214372 261642 191094 199528 189177 276052 289317 291017 289159 282086 272847 284451 228342 214390 188240 169258 219874 223475 177548 138608 194200 183735 147787 194489 148307 214507 197241 UROCANATE 237152 265871 265307 255464 248459 375447 362258 256604 289948 164528 310671 173797 367028 194739 375756 226494 286313 397531 224617 193589 257625 279842 259370 209992 224067 208755 198066 215194 191594 193192 188675 294381 204756 503449 520130 454616 467628 376349 463108 535793 188306 385023 374905 203579 157394 166538 193428 179084 Xylenesulfonate 353940 390332 384768 640998 688870 313608 320790 618236 383749 1310609 311602 1286532 595559 1301725 546144 621091 4725309 603087 584285 540000 4770217 4869257 366549 234984 232595 226589 566838 517459 1270994 555976 1302273 405715 1326509 311557 328933 343087 310843 726459 315289 353108 614906 753109 706879 631273 486431 602353 500829 485630 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Retention Index Moverz Quant Local ID PubChem CID KEGG ID mass spectrum 2-hydroxyglutarate 1.51 43 147.0299 3-methyladipic acid 7.83 12292 159.0665 3-SULFINO-L-ALANINE 0.72 1549098 152.0023 4-oxoproline 1.76 107541 128.0353 6-Hydroxycaproic acid 7.78 14490 131.0714 ADENINE 1.46 190 134.0472 ADP 1.06 6022 426.0220 ANTHRANILATE 7.94 227 136.0398 ASPARAGINE 0.68 6267 131.0462 ASPARTATE 0.68 83887 132.0303 BOC-L-Tyrosine_R1 9.99 206.0458 BOC-L-Tyrosine_R2 9.99 280.1189 C12H22O11-DISACCHARIDE-(6C/6C; GLC-GLC/GLC-FRC/GAL-GLC) 0.86 341.1086 C6H12O6-HEXOSE/KETOSE/INOSITOL 0.74 179.0561 CITRULLINE 0.76 9750 174.0867 CMP 1.01 6131 322.0444 Creatine-D3 0.88 133.0810 CYTIDINE 2',3'-CYCLIC MONO-PHOS-PHATE 1.05 53481030 304.0338 D-GLUCURONIC ACID/D-GLUCURONOLACTONE/D-(+)-GALACTURONIC ACID_R1 0.74 193.0328 D-GLUCURONIC ACID/D-GLUCURONOLACTONE/D-(+)-GALACTURONIC ACID_R2 0.72 193.0362 DIHYDROXYACETONE PHOSPHATE 0.70 668 168.9901 D-RIBOSE 5-PHOSPHATE/RIBULOSE 5-PHOSPHATE 0.72 229.0118 GLUCONIC ACID/D-GULONIC ACID GAMA-LACTONE 0.75 195.0523 GLYCEROL 2-PHOSPHATE/SN-GLYCEROL 3-PHOSPHATE 0.70 171.0062 GLYCOLATE 0.86 757 75.0082 LACTATE-dimer 1.20 201.0380 L-ARGININE 0.69 6322 173.1043 L-CYSTEIC ACID 0.70 72886 167.9972 L-GLUTAMIC ACID 0.75 33032 146.0459 L-GLUTAMINE 0.72 5961 145.0619 L-HISTIDINE 0.67 6274 154.0622 L-Phenylalanine-13C6 6.25 71309396 170.0918 L-SERINE 0.67 5951 104.0351 L-TYROSINE 3.32 6057 180.0665 MALATE 1.05 92824 133.0142 Mannitol 0.72 6251 181.0719 N-ACETYL-HEXOSAMINE 0.81 220.0826 N-ACETYL-L-LEUCINE 7.54 70912 172.0980 N-ACETYLNEURAMINATE 0.79 439197 308.0987 N-BOC-L-Aspartic Acid_R1 8.49 232.0826 N-BOC-L-Aspartic Acid_R2 8.49 158.0094 N-BOC-L-tert-Leucine_R1 10.85 156.0666 N-BOC-L-tert-Leucine_R2 10.85 230.1396 N-BOC-L-Tryptophan_R1 11.43 303.1349 N-BOC-L-Tryptophan_R2 11.43 229.0617 Pantothenic Acid 6.61 6613 218.1023 PROPIONATE 0.73 1032 109.0075 PYRIDOXAL 1.84 1050 166.0507 Salicylic Acid-D4 9.94 46782907 141.0495 SARCOSINE/BETA-ALANINE 0.68 88.0400 STACHYOSE 0.89 439531 665.2140 SUCCINATE 2.24 1110 117.0193 Succinic-2,2,3,3-D4 Acid 2.21 121.0443 Sulfoacetaldehyde 0.68 160226 122.9757 TAURINE 0.68 1123 124.0071 THREONINE/HOMOSERINE 0.72 118.0510 Tryptophan 7.64 6305 203.0827 UROCANATE 1.91 736715 137.0356 Xylenesulfonate 8.05 11868 185.0277 METABOLITES_END #END