#METABOLOMICS WORKBENCH xiaohong_20231127_144535 DATATRACK_ID:4477 STUDY_ID:ST002991 ANALYSIS_ID:AN004911 PROJECT_ID:PR001862 VERSION 1 CREATED_ON November 29, 2023, 1:55 pm #PROJECT PR:PROJECT_TITLE DNA replication stress underpins the vulnerability to oxidative phosphorylation PR:PROJECT_TITLE inhibition in colorectal cancer PR:PROJECT_SUMMARY Although targeting oxidative phosphorylation (OXPHOS) for cancer treatment is PR:PROJECT_SUMMARY currently impeded due to dose-limiting toxicities, there remain opportunities PR:PROJECT_SUMMARY through combinations that provide therapeutic benefits at doses attainable in PR:PROJECT_SUMMARY patients. On the other hand, while glycolysis-deficient cancers are generally PR:PROJECT_SUMMARY vulnerable to OXPHOS inhibition in preclinical models, the full extent of PR:PROJECT_SUMMARY phenotypical and mechanistic consequences of inhibiting OXPHOS in cancers PR:PROJECT_SUMMARY capable of glycolysis is not yet well understood. We aimed to clarify the PR:PROJECT_SUMMARY response and underlying mechanisms of colorectal cancer (CRC) that commonly PR:PROJECT_SUMMARY exhibit the glycolytic phenotype to OXPHOS inhibition and to identify potential PR:PROJECT_SUMMARY approaches to render such cells more sensitive to OXPHOS inhibitors. PR:INSTITUTE The University of Newcastle PR:LAST_NAME Zhao PR:FIRST_NAME Xiaohong PR:ADDRESS University Drive, Callaghan, NSW, 2308, Australia PR:EMAIL xiaohong.zhao@newcastle.edu.au PR:PHONE 61 0466219528 #STUDY ST:STUDY_TITLE Metabolomics studies on human colorectal cancer cell lines ST:STUDY_SUMMARY Although targeting oxidative phosphorylation (OXPHOS) for cancer treatment is ST:STUDY_SUMMARY currently impeded due to dose-limiting toxicities, there remain opportunities ST:STUDY_SUMMARY through combinations that provide therapeutic benefits at doses attainable in ST:STUDY_SUMMARY patients. On the other hand, while glycolysis-deficient cancers are generally ST:STUDY_SUMMARY vulnerable to OXPHOS inhibition in preclinical models, the full extent of ST:STUDY_SUMMARY phenotypical and mechanistic consequences of inhibiting OXPHOS in cancers ST:STUDY_SUMMARY capable of glycolysis is not yet well understood. We aimed to clarify the ST:STUDY_SUMMARY response and underlying mechanisms of colorectal cancer (CRC) that commonly ST:STUDY_SUMMARY exhibit the glycolytic phenotype to OXPHOS inhibition and to identify potential ST:STUDY_SUMMARY approaches to render such cells more sensitive to OXPHOS inhibitors. The ST:STUDY_SUMMARY responses of glycolysis-competent CRC cells to targeting OXPHOS were tested ST:STUDY_SUMMARY using targeted meatbolomics. ST:INSTITUTE The University of Newcastle ST:LAST_NAME Zhao ST:FIRST_NAME Xiaohong ST:ADDRESS University Drive, Callaghan, NSW, 2308, Australia ST:EMAIL xiaohong.zhao@newcastle.edu.au ST:PHONE 61 0466219528 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 #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 Colo205_DMSO1 LJ_001 Factor:Control RAW_FILE_NAME=LJ_001.d SUBJECT_SAMPLE_FACTORS Colo205_DMSO2 LJ_002 Factor:Control RAW_FILE_NAME=LJ_002.d SUBJECT_SAMPLE_FACTORS Colo205_DMSO3 LJ_006 Factor:Control RAW_FILE_NAME=LJ_006.d SUBJECT_SAMPLE_FACTORS Colo205_ASP1 LJ_008 Factor:10 mM Aspartate RAW_FILE_NAME=LJ_008.d SUBJECT_SAMPLE_FACTORS Colo205_ASP2 LJ_009 Factor:10 mM Aspartate RAW_FILE_NAME=LJ_009.d SUBJECT_SAMPLE_FACTORS Colo205_ASP3 LJ_010 Factor:10 mM Aspartate RAW_FILE_NAME=LJ_010.d SUBJECT_SAMPLE_FACTORS Colo205_IACS1 LJ_026 Factor:100 nM IACS-010759 RAW_FILE_NAME=LJ_026.d SUBJECT_SAMPLE_FACTORS Colo205_IACS2 LJ_027 Factor:100 nM IACS-010759 RAW_FILE_NAME=LJ_027.d SUBJECT_SAMPLE_FACTORS Colo205_IACS3 LJ_029 Factor:100 nM IACS-010759 RAW_FILE_NAME=LJ_029.d SUBJECT_SAMPLE_FACTORS Colo205_IACS+ASP1 LJ_031 Factor:100 nM IACS-010759+10 mM Aspartate RAW_FILE_NAME=LJ_031.d SUBJECT_SAMPLE_FACTORS Colo205_IACS+ASP2 LJ_032 Factor:100 nM IACS-010759+10 mM Aspartate RAW_FILE_NAME=LJ_032.d SUBJECT_SAMPLE_FACTORS Colo205_IACS+ASP3 LJ_034 Factor:100 nM IACS-010759+10 mM Aspartate RAW_FILE_NAME=LJ_034.d SUBJECT_SAMPLE_FACTORS LIM1215_DMSO1 LJ_014 Factor:Control RAW_FILE_NAME=LJ_014.d SUBJECT_SAMPLE_FACTORS LIM1215_DMSO2 LJ_015 Factor:Control RAW_FILE_NAME=LJ_015.d SUBJECT_SAMPLE_FACTORS LIM1215_DMSO3 LJ_018 Factor:Control RAW_FILE_NAME=LJ_018.d SUBJECT_SAMPLE_FACTORS LIM1215_ASP1 LJ_020 Factor:10 mM Aspartate RAW_FILE_NAME=LJ_020.d SUBJECT_SAMPLE_FACTORS LIM1215_ASP2 LJ_023 Factor:10 mM Aspartate RAW_FILE_NAME=LJ_023.d SUBJECT_SAMPLE_FACTORS LIM1215_ASP3 LJ_024 Factor:10 mM Aspartate RAW_FILE_NAME=LJ_024.d SUBJECT_SAMPLE_FACTORS LIM1215_IACS1 LJ_037 Factor:100 nM IACS-010759 RAW_FILE_NAME=LJ_037.d SUBJECT_SAMPLE_FACTORS LIM1215_IACS2 LJ_038 Factor:100 nM IACS-010759 RAW_FILE_NAME=LJ_038.d SUBJECT_SAMPLE_FACTORS LIM1215_IACS3 LJ_041 Factor:100 nM IACS-010759 RAW_FILE_NAME=LJ_041.d SUBJECT_SAMPLE_FACTORS LIM1215_IACS+ASP1 LJ_045 Factor:100 nM IACS-010759+10 mM Aspartate RAW_FILE_NAME=LJ_045.d SUBJECT_SAMPLE_FACTORS LIM1215_IACS+ASP2 LJ_046 Factor:100 nM IACS-010759+10 mM Aspartate RAW_FILE_NAME=LJ_046.d SUBJECT_SAMPLE_FACTORS LIM1215_IACS+ASP3 LJ_048 Factor:100 nM IACS-010759+10 mM Aspartate RAW_FILE_NAME=LJ_048.d #COLLECTION CO:COLLECTION_SUMMARY 1. Metabolic Arrest 1.1 Remove culture from incubator 1.2 Aspirate media from CO:COLLECTION_SUMMARY wells using vacuum sipper 1.3 Dispense about 1mL of 37oC MilliQ water to the CO:COLLECTION_SUMMARY cell surface 1.4 Briefly ‘rock’ the plate for 2 secs 1.5 Aspirate the water CO:COLLECTION_SUMMARY off the cells 1.6 Wait 3 more secs then add ~ enough LN2 to cover surface of CO:COLLECTION_SUMMARY well/plate 1.7 Briefly store on dry ice, and transfer to a -80oC freezer if not CO:COLLECTION_SUMMARY extracting immediately. 1.8 Extract/assay within 7 days 1.9 Time to complete CO:COLLECTION_SUMMARY steps 1.3 – 1.5 should be minimised to prevent changes in metabolic state due CO:COLLECTION_SUMMARY to cells being stressed 2. Metabolic Extraction 2.1 Transfer plates from the -80 CO:COLLECTION_SUMMARY deg C freezer to a 4 deg C cold room 2.2 Add 500μL of ice cold 9:1 MeOH:CHCl3, CO:COLLECTION_SUMMARY v/v (+ internal standards, allow to incubate for 10 minutes 2.3 Scrape/suspend CO:COLLECTION_SUMMARY with a cell lifter 2.4 Transfer to 1.5mL microcentrifuge tubes, Vortex for 30s CO:COLLECTION_SUMMARY 2.5 Thermomix at 950rpm for 5 mins 2.6 Centrifuge at 15,000rpm for 5 mins CO:SAMPLE_TYPE Cultured cells #TREATMENT TR:TREATMENT_SUMMARY After seeding cells in 6-well plates for 24 hours, Colo205 and LIM1215 cells TR:TREATMENT_SUMMARY were treated with Vehicle control DMSO, 100 nM IACS-010759, 10 mM Aspartate, 100 TR:TREATMENT_SUMMARY nM IACS-010759 combined with 10 mM Aspartate, for 24 hours. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Add 500uL of ice cold 1:9 chloroform:methanol (+ internal standards) and SP:SAMPLEPREP_SUMMARY scrape/suspend with a cell lifter Transfer to 1.5 mL microcentrifuge tubes SP:SAMPLEPREP_SUMMARY Vortex for 30s Thermomix at 950rpm for 5 mins Centrifuge at 15,000rpm for 5 mins #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Agilent 1200 CH:COLUMN_NAME Merck Millipore SeQuant ZIC-pHILIC column (150 mm × 4.6 mm, 5 μm) CH:SOLVENT_A 20 mM (NH4)2CO3, pH 9.0, Sigma-Aldrich CH:SOLVENT_B 100% acetonitrile, Hypergrade for LCMS LiChrosolv, Merck CH:FLOW_GRADIENT time (t) =0 min, 80% B; t =0.5 min, 80% B; t =15.5 min, 50% B; t =17.5 min, 30% CH:FLOW_GRADIENT B; t =18.5 min, 5% B, t =21.0 min, 5% B; t =23 min, 80% B, t =33 min, 80% B. CH:FLOW_RATE 300 μL/min CH:COLUMN_TEMPERATURE 30 ◦C #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Agilent 6545 QTOF MS:INSTRUMENT_TYPE Single quadrupole MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS Peak intensity data was analyzed using MetaboAnalyst. Briefly, data was MS:MS_COMMENTS normalized using median, log2FC, and p-value was generated using one-factor MS:MS_COMMENTS statistical analysis. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS peak intensity MS_METABOLITE_DATA_START Samples LJ_001 LJ_002 LJ_006 LJ_008 LJ_009 LJ_010 LJ_014 LJ_015 LJ_018 LJ_020 LJ_023 LJ_024 LJ_026 LJ_027 LJ_029 LJ_031 LJ_032 LJ_034 LJ_037 LJ_038 LJ_041 LJ_045 LJ_046 LJ_048 Factors Factor:Control Factor:Control Factor:Control Factor:10 mM Aspartate Factor:10 mM Aspartate Factor:10 mM Aspartate Factor:Control Factor:Control Factor:Control Factor:10 mM Aspartate Factor:10 mM Aspartate Factor:10 mM Aspartate Factor:100 nM IACS-010759 Factor:100 nM IACS-010759 Factor:100 nM IACS-010759 Factor:100 nM IACS-010759+10 mM Aspartate Factor:100 nM IACS-010759+10 mM Aspartate Factor:100 nM IACS-010759+10 mM Aspartate Factor:100 nM IACS-010759 Factor:100 nM IACS-010759 Factor:100 nM IACS-010759 Factor:100 nM IACS-010759+10 mM Aspartate Factor:100 nM IACS-010759+10 mM Aspartate Factor:100 nM IACS-010759+10 mM Aspartate ISTD_13C Sorbitol 44197 34178 31413 44601 56437 33122 34270 32892 31533 28186 31423 29020 35993 39245 33456 28669 44744 32536 31693 35251 27853 29759 25871 28656 ISTD_13C Valine 27466 26403 23367 25813 25858 25559 21878 20244 21689 22515 22828 24827 25068 24059 22952 23014 27597 25101 22824 23394 22529 23187 24443 22222 ISTD_13C Leucine 36627 32442 32975 32366 34439 32281 30718 27555 30034 30800 32392 33005 30183 30184 28587 29792 34942 31457 30517 31065 31418 30104 32620 30033 (R)-Lipoic acid 307 287 351 348 341 307 1190 1932 1298 2444 1917 803 1283 1571 1448 1577 1441 1509 660 557 346 992 1311 890 2-Hydroxyethanesulfonate 13066 13795 12332 14038 12451 14041 9915 8287 7330 8469 10738 10866 11099 12305 13871 9344 12348 13893 6250 6806 8688 9209 8561 9471 2-Methylcitric acid 592 296 351 348 341 307 1190 1932 1815 2444 1917 803 1283 1571 1448 1577 1441 1509 374 361 346 992 1311 890 2-Phenylglycine 1222 1289 643 633 1194 1284 2984 2462 1734 2285 2226 2490 617 1744 986 585 1099 887 1596 1574 1124 1844 2166 2710 3,3-Dimethylglutaric acid 8510 11870 8974 8241 789 7632 1534 5942 7670 12502 7630 8420 9170 16937 9481 2325 14092 8200 8715 7757 8686 898 6069 958 3,4-Dihydroxybenzeneacetic acid 1871 3654 7378 2795 3132 4538 55251 49419 50916 57872 66503 62053 9927 3880 4676 2955 4166 3515 20087 18589 22185 21734 20826 23451 4-Hydroxyproline 1794 1561 1374 1200 1710 1059 2644 3116 2722 1959 2055 2030 687 1573 1921 1333 2005 619 2029 992 1609 1577 2459 1534 5-Aminopentanoic acid 5957 5954 5247 3912 3616 3376 809 815 836 584 347 557 4777 5456 4577 3165 3934 3520 1362 1222 1486 706 974 1592 Acetoacetic acid 2469 3289 3476 2284 2010 2096 5769 4501 5760 5664 7936 4980 12795 14492 14660 2486 3469 4619 11321 12576 14540 4316 5888 6539 Acetyl-CoA 842 599 281 536 233 530 2443 2094 1895 1492 2458 2352 454 535 229 267 477 258 1518 1693 1640 959 1082 1413 Adenine 918 1581 1735 1022 1279 1516 1765 1739 1969 3111 3454 3722 867 593 1805 1507 1481 1658 1066 1136 1141 1579 1830 2070 Adenosine diphosphate 25891 18994 19682 20557 19084 21554 58334 49417 64839 71187 69212 64413 10968 12963 12674 17851 20437 19375 47331 58331 54670 34269 52238 57414 Adenosine monophosphate 22466 13310 12359 24469 19582 33069 216301 261130 208424 350960 409694 373469 5169 6242 5875 11403 12222 11638 79378 88457 92879 92593 132975 145410 Adenosine triphosphate 26730 24765 34015 23803 26057 20433 25278 17934 28487 34338 46046 41645 33960 34657 40293 26872 37331 36803 30777 52077 42877 18612 32374 38071 Alanine 26704 27304 26538 29163 31156 28221 48910 53334 51263 48490 57525 57135 19374 21361 21095 23598 25964 22410 32341 35330 35437 41544 46105 44672 Allantoin 2479 2404 2682 3241 2737 3292 4777 3822 3751 4653 5077 4822 2494 2923 3776 2383 3303 1087 4042 4551 5004 3749 3854 4344 Aminoadipic acid 735 757 668 799 882 592 814 1324 1347 810 1275 1328 446 1191 1020 896 1097 608 2393 2159 2570 1858 1906 1981 Arabinose 10836 9653 9569 9732 11508 9717 10287 10312 7303 9626 10145 10227 14275 13783 14777 9553 13692 11487 5905 6417 6600 4741 5855 6204 Arachidic acid 908 905 1277 1196 1114 1039 3793 3494 3049 4346 4429 4013 2192 1618 1906 2026 2089 1109 3743 4010 3128 3392 4079 3061 Arginine 2053 2679 3592 2134 2228 3629 3983 4204 4914 6727 7237 7723 3083 3480 4849 3509 3465 3054 4547 5303 6045 4326 5461 5072 Asparagine 4097 4286 3678 3959 4355 3219 16159 17357 17883 13500 12818 13601 3385 5523 4739 3579 4817 3765 9681 9721 10955 10831 10786 13714 Aspartic acid 118552 155204 144154 936285 770864 1165542 184617 178316 179650 1175722 1338366 1143538 14320 19767 17552 417771 704019 647696 33109 30782 32168 428288 434174 575647 Azelaic acid 3802 5414 7426 3521 1408 2589 1334 7356 3396 4062 3616 4196 11634 4836 7116 2322 3192 2903 3575 2276 2560 1757 2497 1789 Behenic acid 3689 2866 2392 3289 2713 2856 2836 3319 2561 3469 3409 3343 3777 3179 2976 3210 3907 3479 3877 4404 3259 3306 3687 3521 beta-Alanine 29668 32986 30485 122017 107287 157872 37535 39326 34824 170411 206982 186454 7868 9730 8561 88084 110673 87031 7819 7795 7830 89661 88872 96512 Caffeic acid 13597 12096 7230 5470 323 5785 615 18031 5420 9534 17373 27805 49462 13232 46074 5987 11758 7440 24072 5140 6676 284 13019 762 Capric acid 1872 2745 2283 1666 2633 2689 3065 1106 1748 1760 3392 2250 2506 2197 2514 7155 3368 2451 2117 2145 2248 2083 1657 3247 Caprylic acid 1917 1620 1762 581 2948 2030 2269 1464 2041 1157 1945 1154 1636 1648 1578 4110 1651 1832 1110 1060 1680 1794 499 2582 cis-Aconitic acid 2379 2526 1641 2605 3642 2145 5083 3918 5146 8285 8360 7269 840 509 438 786 956 1353 1775 3751 3012 1597 2815 2505 Citicoline 524 562 494 569 500 318 3080 4229 5103 4785 6130 6618 209 199 195 500 745 257 2159 2442 2610 4066 3641 4382 Citric acid 35417 34669 32613 44279 53929 35876 79331 73116 77265 123122 136817 115337 9280 9904 8768 12151 10480 12526 20238 34815 35935 27970 35934 34509 Coenzyme A 5609 4986 5739 5109 4660 5073 36711 36010 38422 37413 39071 39196 3563 3747 4159 3379 4003 3983 15116 16466 16686 19287 22371 20832 D-2-Hydroxyglutaric acid 9706 13301 10281 10705 7296 12548 10806 14560 17895 20934 22789 13608 19670 24976 22332 14749 22703 23455 31967 29084 31209 28104 35366 31954 Deoxycytidine 300 278 337 291 338 313 376 316 630 291 311 466 1713 2260 1781 1742 2214 2320 2436 2398 2827 3278 3710 4210 D-Lactic acid 46195 52230 47098 52221 57449 59078 49486 43204 39577 55711 63294 63841 125877 156910 165156 128602 176855 152992 145987 162795 205391 203413 216978 216016 Dodecanoic acid 3924 3688 2385 3098 3824 4449 2010 2960 3683 2715 5435 3715 4156 3785 4648 5689 4353 4432 3057 3771 4035 1919 2923 711 Ethylmalonic acid 2462 2395 2170 1811 1071 1571 260 204 202 247 339 249 5204 5267 5346 4007 6607 4539 277 500 449 329 349 214 FAD 1987 1952 2033 2244 2207 2109 3180 3166 2685 3224 3837 3874 2154 2615 2104 1735 2347 2234 2596 3031 3065 2460 3055 3526 Fructose 6324 8050 8049 8556 7588 9382 6292 9664 8113 11819 15934 15499 10201 8797 11104 6726 9695 7243 12402 11541 14679 9820 12344 9932 Fructose 1,6-bisphosphate 1547 924 1369 1292 1473 631 1257 668 1206 1133 1805 1942 7918 8251 7245 5285 6451 6223 4490 6286 4994 1758 2710 3788 Fructose 6-phosphate 1759 1331 1666 1549 1135 2334 7203 7994 7232 9257 11264 9791 1672 2363 1992 2553 2443 2779 6879 8461 8272 7622 10801 8588 Fumaric acid 88980 101525 90481 99520 89228 93153 83984 75496 79348 129924 141147 107282 252347 276833 269945 224936 314782 290514 169188 182871 191102 218072 234835 244049 Galactonolactone 22592 19182 19438 11175 1315 16477 855 18039 15718 21879 20866 19316 21686 20037 23418 1728 22378 17022 17658 14735 17152 728 13666 698 Galacturonic acid 3945 4158 3384 4349 4270 4599 2017 1696 1930 2000 2355 1656 3240 3149 4068 2739 3980 3784 2062 1949 2253 2369 1967 2113 gamma-Aminobutyric acid 132788 138869 135890 145686 148996 141969 257897 260097 247484 280434 305172 304205 114276 131615 132819 95609 139171 125666 218012 215321 225120 202667 205700 207189 gamma-Butyrolactone 15972 12065 7842 10018 1524 11903 2816 10338 13180 6718 19956 16380 15260 14485 14438 2914 16918 13416 13020 13146 14912 2052 11956 2400 Gamma-Caprolactone 7973 9421 6570 6837 987 8628 1558 5818 8572 7644 10940 9917 7166 9989 6487 938 11087 8901 7267 8000 10309 566 6602 1531 Gluconic acid 5674 6646 6070 6872 7440 6854 3175 2793 2431 2864 3083 3638 29326 32027 32741 25503 35713 30715 9010 9678 11260 9626 7794 9865 Glucose 29400 30888 30932 38325 34701 48071 14073 16254 13186 23320 26715 22728 38465 33475 43779 26882 33242 21011 15212 17906 23962 14363 10006 14353 Glucose 6-phosphate 5299 4006 5426 3673 2495 6838 26578 28790 23808 28533 35964 31863 1606 1327 988 5902 6118 7176 17161 18600 27396 30777 36448 33390 Glucuronic acid 2707 2555 2476 3016 2887 3050 969 1133 767 568 1260 1132 2252 3038 2830 2023 2549 2601 767 1246 1204 1018 629 717 Glutaconic acid 28826 25694 14286 20970 2695 24385 4421 19119 23672 12370 40364 31575 28764 27206 30159 5325 28683 24812 25260 24042 26966 3184 24569 4646 Glutamic acid 1657264 1853262 1780915 1928982 1934953 1948510 2780899 2666054 2734005 3048984 3325441 3172745 1429480 1705240 1764707 1344186 1705976 1625478 2612495 2543827 2695172 2353397 2315210 2430524 Glutamine 989539 864138 841365 916275 952618 838678 670035 696207 429575 315295 396027 515979 1036425 1103748 1097243 884045 1207105 817755 992256 948346 1090522 809412 682494 794573 Glyceraldehyde 3-phosphate 1337 1263 1343 872 1518 1116 239 261 225 537 309 790 4971 6384 6376 4203 6097 5994 2145 2100 1926 2630 2792 2885 Glyceric acid 1313 1480 1633 1509 1369 1616 1584 1452 1647 2162 1595 2417 2428 2602 1785 1842 2434 1912 1753 2145 2354 1729 1952 2125 Glycerol 3-phosphate 1863 1268 1619 705 2090 659 951 1106 821 1225 1160 517 5386 6123 5756 5993 6123 5527 1521 1102 1610 956 1295 1245 Glycerophosphocholine 3785 3767 4951 5226 5541 4823 6065 6068 5884 6253 7129 7813 1385 1581 1404 1779 2106 2150 2034 2211 2514 2768 3221 3156 Glycine 21260 21671 22286 27141 29484 25205 34293 32125 28666 31363 30617 32772 20016 25473 24768 20492 24133 19806 22816 25864 26938 25486 21619 27626 Guanosine diphosphate 928 573 566 1184 965 546 1813 1469 1740 2725 2943 3034 868 694 884 1001 1046 1104 1287 3067 2408 1040 1635 1543 Guanosine monophosphate 1640 1044 1012 1871 1880 2538 6316 7542 6664 10258 13783 11639 708 983 918 967 1145 991 3266 3045 3105 3541 5162 5218 Guanosine triphosphate 817 610 1123 836 684 440 620 548 608 1321 1496 1903 1718 1369 2005 1012 1567 1237 785 2473 1227 425 732 982 Heptadecanoic acid 3202 2113 2175 2117 1627 1790 2080 3318 2310 2643 2848 2991 1480 1479 1024 2241 2149 1733 2878 2272 1805 2324 2148 2508 Hypotaurine 13542 12573 11272 13179 10831 12109 2332 3418 3191 2559 3629 3896 13765 12020 13749 10686 14689 8357 4956 3042 3691 3580 2612 3314 Hypoxanthine 5108 4043 2414 7257 7823 5950 4636 6867 5717 17198 18804 15584 2545 3494 2189 2839 2402 2232 345 1087 1313 1940 1939 2907 Isoleucine 114672 119575 130182 128957 131219 137036 260116 276164 258345 277308 292662 314631 112579 127026 130527 112267 134375 123388 186179 193860 211190 217839 222909 237977 Isomaltose 1313 653 750 500 175 202 220 197 1022 216 638 207 548 270 711 253 561 222 237 283 850 1985 727 525 Itaconic acid 2542 3319 2819 2658 2453 3259 3410 3849 4547 5327 5176 3643 4737 5805 5680 3349 5124 6056 6090 7386 8118 7323 8724 7553 Ketoleucine 9986 9053 6920 5639 3833 9087 4860 6998 7442 8233 10545 10715 9160 11152 10043 8189 10746 9685 9938 8873 10909 4280 7720 4392 Leucine 110655 118484 126357 122921 124328 124986 256402 266702 254642 272276 297692 321946 101888 121161 126199 109392 129850 118754 180619 193267 205744 222630 233233 237387 Linoleic acid 3117 3481 2932 3430 3732 3503 3991 4400 4102 4111 4887 5058 3924 4009 4438 3837 3775 3847 3455 3554 3627 4280 4361 4287 Lysine 2547 3768 3723 3544 2970 4569 6063 6438 5952 8553 10503 10921 4317 4205 6910 4470 5471 5260 6294 7016 8959 4963 7180 7725 Maleic acid 44776 42199 23836 28433 4810 30415 4553 40059 33320 23963 56240 46765 66035 41602 52501 9691 34164 33183 43181 30707 35183 3948 40242 5684 Malic acid 396256 651551 385658 640367 433603 427678 375925 334054 367593 808901 947250 509792 1671770 1872633 1819758 1445383 2140028 1972421 1182144 1209947 1278984 1443626 1494744 1619597 Malonic acid 1818 1728 1519 2148 2008 1026 5412 4955 5278 8938 9195 6575 2088 1722 2047 1930 1200 1582 3548 3895 4170 3296 3889 4215 Methionine 42115 47849 46413 54743 59609 53440 106352 116295 101458 117247 122941 130938 48393 54232 53713 44121 55282 48428 83040 80572 89248 91010 81400 97263 Methionine sulfoxide 3707 4041 3718 3761 3831 3948 7170 6905 6465 6367 6137 6727 4197 5083 4354 3686 4954 3090 6230 6003 5742 6285 6010 6444 myo-Inositol 346444 322019 296058 362627 399924 350435 2994826 2780683 2695710 2962383 3298732 3133006 328331 380182 347458 294861 406193 341865 2425176 2506868 2667032 2533956 2523615 2762397 Myristic acid 8571 6807 6312 6236 6654 7237 2533 5158 4348 3894 4871 5624 9588 6638 9482 7458 7511 7036 4120 3802 3988 4156 3847 3434 N-Acetylalanine 10640 12791 10601 14100 13385 13053 7700 7491 7332 7555 7519 8852 12400 14184 14354 11111 14032 13613 7962 7439 7726 8308 8970 10968 N-Acetylglutamic acid 66261 69406 72040 72061 71890 71840 104862 104521 99313 100112 108105 109024 69508 87817 87556 67718 85397 82411 119751 125015 131075 127885 121440 132485 N-Acetylglutamine 13677 14244 13788 15397 17452 15185 17352 18455 15952 16144 17327 19034 18378 21709 19698 16170 23437 20084 20683 22608 24070 24552 23771 27776 N-Acetylneuraminic acid 34075 33571 33566 38524 36862 40054 29919 31046 25233 34140 35204 35356 19939 24897 26059 21936 26350 25185 13379 15615 18018 14749 19790 20308 NAD 19026 15331 17116 16639 17899 16836 41177 35073 39704 38546 45761 44011 17199 17636 17290 15490 21154 17585 37512 38191 40712 39115 46073 46008 NADP 1233 959 1392 774 689 723 919 335 985 842 886 765 990 1366 1042 1231 1628 1326 1198 1345 1317 703 932 1051 N-alpha-Acetyllysine 22031 17399 15218 23172 28083 16609 17732 17932 16037 14048 16141 14517 18113 13658 17380 14092 22394 10505 16046 17037 14417 14620 13234 15331 Octadecanedioic acid 1901 1079 773 1064 973 528 889 2164 663 779 2989 1621 1521 802 3298 1018 950 776 869 783 905 964 669 427 Oleic acid 70021 80549 75769 76033 74137 84401 33416 35916 35956 38976 40625 42194 60662 68327 63462 59161 63437 68329 25499 31829 27180 30280 34774 35163 Orotic acid 8222 6676 5603 11932 10226 12307 26104 23898 23426 28048 32225 26183 15012 17191 12661 11815 14290 12156 16357 17413 16952 23505 25531 21161 Orotidine 16407 15952 15181 20059 18544 15984 64357 60000 57620 69471 76180 73499 15241 16908 17902 13776 19012 16266 37489 43116 42852 38891 40382 41439 Oxidized glutathione 3523 4733 2876 2108 1993 1940 3681 4227 3646 6138 4656 5439 6414 6608 7643 2264 2935 3106 2489 3231 3684 4064 6606 4246 Oxoglutaric acid 1867 2658 2401 1819 1865 1862 3789 3642 4410 3809 5995 3239 9566 11362 10214 2090 2591 2734 8356 9786 10577 2695 4691 4532 PA(16:0/16:0) 1202 1465 1356 1299 1492 1374 3140 2989 2876 2739 2910 2644 1878 1876 1867 1584 1878 1690 2078 2282 2528 2368 2348 2109 Palmitic acid 69297 70754 48254 44510 45539 40806 31822 37238 27553 41153 37165 39760 50918 39386 41069 49391 44442 40789 55544 44672 32444 31945 32169 35417 Pantothenic acid 278530 299723 256547 321053 296632 300419 812312 744715 743898 810677 922730 829543 237705 243280 252173 188825 257686 225022 691293 822482 805077 787349 809661 818658 Pelargonic acid 8092 8556 5560 5520 20100 8936 17769 4801 6794 6152 9033 6770 6032 8333 7155 45260 8090 8392 6289 7367 8313 15334 5339 18869 Pentadecanoic acid 2878 3201 2030 2336 1030 2302 936 4242 2007 2494 2724 3066 5120 2587 3928 1644 2419 2173 2191 2132 2198 1732 1607 1705 Phenylalanine 113568 125472 131308 136599 140659 138518 306355 318333 295695 321117 336371 361057 118955 131399 135203 116451 136995 129427 217413 227885 236075 250987 267459 275301 Phthalic acid 77993 69441 40267 31044 7904 30212 7916 111981 33343 53282 104481 138439 212914 61401 209850 22658 58161 37322 123381 27377 46520 7363 66672 8650 Proline 61134 65119 60425 66737 74814 67313 183000 183269 183805 178855 201837 211971 46553 58167 51974 48297 56293 51748 202973 224991 226453 248707 258657 262211 Pyridoxine 5590 7917 6567 8364 7714 8723 10143 10915 10340 12849 15597 12264 6266 8247 9840 4970 8556 6642 10457 11810 13898 9352 9338 10848 Pyroglutamic acid 714913 866886 840508 876561 849784 940891 962457 1002690 953084 1002617 1061256 1073797 962644 1160538 1211934 893741 1189585 1002936 992928 1093063 1198852 1118273 1068802 1080835 Pyruvic acid 2186 2205 2369 2403 1554 2406 2194 1862 2112 2795 3001 2707 4644 5291 5086 4147 6200 6070 3601 3042 3486 4082 4572 4458 Ribose 5-phosphate 1537 1003 1432 1530 930 1605 1216 1213 1424 1468 1643 1630 791 886 1299 1660 1650 1703 1511 1732 1006 1808 2193 1544 S-Adenosylhomocysteine 2687 3031 1699 6735 5428 7016 33153 33008 26882 23249 21403 38294 3229 5437 6341 3170 4980 3918 17561 21609 16527 15092 12873 9299 Serine 56199 70680 64252 72948 79339 72203 111396 116877 99985 127500 126209 114161 67411 87859 81254 63663 84762 82597 135630 139991 147303 139728 136069 159754 Shikimic acid 10082 13378 9619 4737 1818 8291 2120 7610 8370 11822 12406 12248 13161 14216 12816 1473 12892 8672 11486 7315 8391 2290 6675 3344 Sorbitol 3857 4376 4298 5131 5569 4884 5978 5953 5149 6445 8503 7574 6185 6729 6706 4453 6562 4659 8507 9061 9635 8531 7175 8837 Stearic acid 71499 75170 42129 37626 32168 28902 44130 44684 39385 58794 46543 49667 41704 30226 31353 57680 38186 32056 69339 55061 45152 37335 40444 43734 Succinic acid 28786 36383 26503 33344 20504 30380 11709 24179 20955 28926 32765 23919 26579 25916 25882 6766 16459 16765 26115 17542 18950 7009 20000 7277 Succinic anhydride 3200 3283 2686 2733 1949 3111 1774 2544 1933 3289 3457 2682 2861 2472 1960 806 1503 1639 2333 1771 2307 1118 1731 713 Taurine 1125358 1150884 1128326 780470 822530 676707 942265 922340 864937 249553 300227 357915 1109107 1225230 1322443 786800 1054136 882791 1035243 1045972 1112476 754870 747580 712450 Thiamine pyrophosphate 1017 560 257 1456 931 1881 20021 21736 17182 34394 39555 33582 270 262 240 271 595 488 2591 4574 5163 4745 8852 9781 Threonic acid 426512 422940 398643 503358 671998 420499 231052 211688 211396 232604 260728 248660 631123 687920 672679 375235 710254 651109 240725 264827 275225 295910 295868 332539 Threonine 297472 338521 336857 392820 412052 387890 432507 565776 411403 645448 501436 569752 306146 360821 337831 288776 334404 308419 332916 357425 392845 385785 305730 413410 Tryptophan 24348 27000 29385 27940 28522 28632 71770 74492 69451 71927 78645 86115 24205 27814 28649 22354 29804 27083 46492 50661 54664 60060 59842 62889 Tyrosine 101495 118192 116440 129160 133667 126178 314049 321474 289168 311201 341691 356083 111304 129791 128542 107697 130847 119102 220397 224264 244563 257653 250813 280674 Uracil 10209 8796 9030 9357 8727 9364 32927 31978 30172 35527 39317 37558 9471 10549 9745 8580 10351 9498 21394 21960 21399 20644 22458 22893 Ureidosuccinic acid 1464 1277 1307 1341 1679 1528 765 404 1008 339 1011 1062 1913 2521 2371 1618 2318 1817 845 1231 1335 915 938 543 Uridine 5'-diphosphate 3324 2414 2218 2286 2403 2895 6998 5756 7388 7282 7724 7302 1400 1467 1656 2718 2683 2589 5290 6960 6351 3828 5134 6943 Uridine 5'-monophosphate 3402 2915 2417 4002 3119 5617 19682 21828 20324 41168 45951 29285 928 1474 1390 2075 2335 2047 7213 8203 8795 9081 12901 14000 Uridine diphosphate glucose 9885 9585 8413 8300 8538 8708 13036 11472 13433 16312 19397 17507 8561 8731 9086 9008 10304 9412 8800 9739 9849 9640 11429 10405 Uridine diphosphate-N-acetylglucosamine 93267 85006 86472 73160 73995 75030 380188 362982 384816 392033 597732 399895 21913 23043 23689 20788 27349 25057 63016 71373 73602 69721 78133 70985 Urocanic acid 1783 2093 1477 1003 1486 1273 1275 1563 903 1446 1177 644 1393 1101 1177 753 969 763 1279 1394 1344 1125 794 1034 Valine 74433 82050 81863 80623 89319 90934 172018 181918 170990 182566 199491 208448 80540 85615 87533 77679 88186 85202 125555 132907 145608 147397 147446 161181 Xanthine 12371 13837 13905 18497 17665 19184 29048 35743 29631 53991 71375 61754 14648 17377 20190 13481 18182 15124 24411 26746 30185 31078 29037 34696 Xylitol 2588 2519 2305 2640 3324 3620 1787 1666 1639 1715 1941 1594 9779 12072 9588 7872 10339 8812 2254 2245 2564 2289 1782 2336 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name HMDB PubChem KEGG ISTD_13C Sorbitol NA NA NA ISTD_13C Valine NA NA NA ISTD_13C Leucine NA NA NA (R)-Lipoic acid HMDB0001451 6112 C16241 2-Hydroxyethanesulfonate HMDB0003903 7866 C05123 2-Methylcitric acid HMDB0000379 5290 C02225 2-Phenylglycine HMDB0002210 99291 NA 3,3-Dimethylglutaric acid HMDB0002441 20984 NA 3,4-Dihydroxybenzeneacetic acid HMDB0001336 547 C01161 4-Hydroxyproline HMDB0000725 5810 C01157 5-Aminopentanoic acid HMDB0003355 138 C00431 Acetoacetic acid HMDB0000060 96 C00164 Acetyl-CoA HMDB0001206 444493 C00024 Adenine HMDB0000034 190 C00147 Adenosine diphosphate HMDB0001341 6022 C00008 Adenosine monophosphate HMDB0000045 6083 C00020 Adenosine triphosphate HMDB0000538 5957 C00002 Alanine HMDB0000161 5950 C00041 Allantoin HMDB0000462 204 C01551 Aminoadipic acid HMDB0000510 469 C00956 Arabinose HMDB0000646 439195 C02479 Arachidic acid HMDB0002212 10467 C06425 Arginine HMDB0000517 6322 C00062 Asparagine HMDB0000168 6267 C00152 Aspartic acid HMDB0000191 5960 C00049 Azelaic acid HMDB0000784 2266 C08261 Behenic acid HMDB0000944 8215 C08281 beta-Alanine HMDB0000056 239 C00099 Caffeic acid HMDB0001964 1549111 C01481 Capric acid HMDB0000511 2969 C01571 Caprylic acid HMDB0000482 379 C06423 cis-Aconitic acid HMDB0000072 643757 C00417 Citicoline HMDB0001413 13804 C00307 Citric acid HMDB0000094 311 C00158 Coenzyme A HMDB0001423 87642 C00010 D-2-Hydroxyglutaric acid HMDB0000606 439391 C01087 Deoxycytidine HMDB0000014 13711 C00881 D-Lactic acid HMDB0001311 61503 C00256 Dodecanoic acid HMDB0000638 3893 C02679 Ethylmalonic acid HMDB0000622 11756 NA FAD HMDB0001248 643975 C00016 Fructose HMDB0000660 439709 C02336 Fructose 1,6-bisphosphate HMDB0001058 445557 C05378 Fructose 6-phosphate HMDB0000124 69507 C00085 Fumaric acid HMDB0000134 444972 C00122 Galactonolactone HMDB0002541 5640 C03383 Galacturonic acid HMDB0002545 84740 C08348 gamma-Aminobutyric acid HMDB0000112 223130 C00334 gamma-Butyrolactone HMDB0000549 7302 C01770 Gamma-Caprolactone HMDB0003843 12756 NA Gluconic acid HMDB0000625 10690 C00257 Glucose HMDB0000122 5793 C00221 Glucose 6-phosphate HMDB0001401 5958 C00092 Glucuronic acid HMDB0000127 94715 C00191 Glutaconic acid HMDB0000620 5280498 C02214 Glutamic acid HMDB0000148 33032 C00025 Glutamine HMDB0000641 5961 C00064 Glyceraldehyde 3-phosphate NA NA NA Glyceric acid HMDB0000139 439194 C00258 Glycerol 3-phosphate HMDB0000126 439162 C00093 Glycerophosphocholine HMDB0000086 71920 C00670 Glycine HMDB0000123 750 C00037 Guanosine diphosphate HMDB0001201 8977 C00035 Guanosine monophosphate HMDB0001397 6804 C00144 Guanosine triphosphate HMDB0001273 6830 C00044 Heptadecanoic acid HMDB0002259 10465 NA Hypotaurine HMDB0000965 107812 C00519 Hypoxanthine HMDB0000157 790 C00262 Isoleucine HMDB0000172 6306 C00407 Isomaltose HMDB0002923 440656 C05400 Itaconic acid HMDB0002092 811 C00490 Ketoleucine HMDB0000695 70 C00233 Leucine HMDB0000687 6106 C00123 Linoleic acid HMDB0000673 5280450 C01595 Lysine HMDB0000182 5962 C00047 Maleic acid HMDB0000176 444972 C01384 Malic acid HMDB0000744 525 C03668 Malonic acid HMDB0000691 867 C04025 Methionine HMDB0000696 6137 C00073 Methionine sulfoxide HMDB0002005 847 C02989 myo-Inositol HMDB0000211 NA C00137 Myristic acid HMDB0000806 11005 C06424 N-Acetylalanine HMDB0000766 88064 NA N-Acetylglutamic acid HMDB0001138 185 C00624 N-Acetylglutamine HMDB0006029 25561 NA N-Acetylneuraminic acid HMDB0000230 445063 C19910 NAD HMDB0000902 5893 C00003 NADP HMDB0000217 5886 C00006 N-alpha-Acetyllysine HMDB0000446 192590 C12989 Octadecanedioic acid HMDB0000782 70095 NA Oleic acid HMDB0000207 445639 C00712 Orotic acid HMDB0000226 967 C00295 Orotidine HMDB0000788 92751 C01103 Oxidized glutathione HMDB0003337 975 C00127 Oxoglutaric acid HMDB0000208 51 C00026 PA(16:0/16:0) HMDB0000674 446066 C00416 Palmitic acid HMDB0000220 985 C00249 Pantothenic acid HMDB0000210 6613 C00864 Pelargonic acid HMDB0000847 8158 C01601 Pentadecanoic acid HMDB0000826 13849 C16537 Phenylalanine HMDB0000159 6140 C00079 Phthalic acid HMDB0002107 1017 C01606 Proline HMDB0000162 145742 C00148 Pyridoxine HMDB0000239 1054 C00314 Pyroglutamic acid HMDB0000267 7405 C01879 Pyruvic acid HMDB0000243 1060 C00022 Ribose 5-phosphate HMDB0001548 439167 C00117 S-Adenosylhomocysteine HMDB0000939 439155 C00021 Serine HMDB0062263 5951 C00716 Shikimic acid HMDB0003070 8742 C00493 Sorbitol HMDB0000247 5780 C00794 Stearic acid HMDB0000827 5281 C01530 Succinic acid HMDB0000254 1110 C00042 Succinic anhydride HMDB0032523 7922 C19524 Taurine HMDB0000251 1123 C00245 Thiamine pyrophosphate HMDB0001372 1132 C00068 Threonic acid HMDB0000943 151152 C01620 Threonine HMDB0000167 6288 C00188 Tryptophan HMDB0000929 6305 C00078 Tyrosine HMDB0000158 6057 C00082 Uracil HMDB0000300 1174 C00106 Ureidosuccinic acid HMDB0000828 93072 C00438 Uridine 5'-diphosphate HMDB0000295 6031 C00015 Uridine 5'-monophosphate HMDB0000288 6030 C00105 Uridine diphosphate glucose HMDB0000286 53477679 C00029 Uridine diphosphate-N-acetylglucosamine HMDB0000290 9547196 C00043 Urocanic acid HMDB0000301 736715 C00785 Valine HMDB0000883 6287 C00183 Xanthine HMDB0000292 1188 C00385 Xylitol HMDB0002917 6912 C00379 METABOLITES_END #END