#METABOLOMICS WORKBENCH spinelli_lab_20240725_130516 DATATRACK_ID:5034 STUDY_ID:ST003397 ANALYSIS_ID:AN005576 PROJECT_ID:PR001988 VERSION 1 CREATED_ON August 12, 2024, 11:39 am #PROJECT PR:PROJECT_TITLE Rhodoquinone is an Electron Carrier in the Mammalian Electron Transport Chain PR:PROJECT_SUMMARY Ubiquinone (UQ), the only known electron carrier in the mammalian electron PR:PROJECT_SUMMARY transport chain (ETC), delivers electrons to both oxygen (O2) and fumarate as PR:PROJECT_SUMMARY terminal electron acceptors. As fumarate has a lower reduction potential than PR:PROJECT_SUMMARY UQ, fumarate reduction is only thermodynamically favorable when ubiquinol, the PR:PROJECT_SUMMARY reduced form of UQ, accumulates. Paradoxically, some tissues reduce fumarate PR:PROJECT_SUMMARY without ubiquinol buildup, suggesting another mechanism enables fumarate PR:PROJECT_SUMMARY reduction in mammals. Here, we identify rhodoquinone (RQ), a novel component of PR:PROJECT_SUMMARY the mammalian ETC that carries electrons to fumarate, instead of O2, as the PR:PROJECT_SUMMARY terminal electron acceptor. RQ, which is undetectable in cultured mammalian PR:PROJECT_SUMMARY cells, is enriched in mitochondria from mouse and human tissues that catalyze PR:PROJECT_SUMMARY high levels of fumarate reduction. UQ and RQ-directed ETC circuits support PR:PROJECT_SUMMARY unique programs of mitochondrial function. Through expression of a bacterial PR:PROJECT_SUMMARY enzyme that converts UQ into RQ and development of a novel RQ analog, we PR:PROJECT_SUMMARY demonstrate that reprogramming the mammalian ETC from the UQ to RQ circuit PR:PROJECT_SUMMARY renders cells highly resistant to hypoxia exposure in vitro and in vivo. Thus, PR:PROJECT_SUMMARY we establish RQ as a fundamental component of the mammalian ETC and unveil that PR:PROJECT_SUMMARY reprogramming the ETC to the RQ-circuit is a tractable strategy to ameliorate PR:PROJECT_SUMMARY hypoxia-related conditions. PR:INSTITUTE UMass Chan Medical School PR:LAST_NAME UMass Chan PR:FIRST_NAME Spinelli Lab PR:ADDRESS 55 Lake Avenue North, Worcester, Massachusetts, 01605, USA PR:EMAIL spinellilab@gmail.com PR:PHONE (508) 856-8989 ext. 68148 #STUDY ST:STUDY_TITLE Glutamine Tracing in Mice injected with HKJS001 and Malonic Acid ST:STUDY_SUMMARY Mice were injected by IP with 13C515N2-glutamine solution. Each mice was ST:STUDY_SUMMARY injected with 15uL of 50mg/mL glutamine solution per gram of mouse. Mice were ST:STUDY_SUMMARY injected with DMSO as a control, HKJS001 compound, 1M malonic acid, or a ST:STUDY_SUMMARY combination of HKJS001 and Malonic Acid. The mice were sacrificed and the liver ST:STUDY_SUMMARY was harvested. Polar metabolites extraction performed on the liver tissue. ST:INSTITUTE UMass Chan Medical School ST:LAST_NAME UMass Chan ST:FIRST_NAME Spinelli Lab ST:ADDRESS 55 Lake Avenue North, Worcester, Massachusetts, 01605, USA ST:EMAIL spinellilab@gmail.com ST:PHONE (508) 856-8989 ext. 68148 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 #FACTORS #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 - Liver 1 Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:Vehicle RAW_FILE_NAME(File Name )=20240529_JV131_INVIVO_001_MalonicAcid_01 SUBJECT_SAMPLE_FACTORS - Liver 2 Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:Vehicle RAW_FILE_NAME(File Name )=20240529_JV131_INVIVO_001_MalonicAcid_02 SUBJECT_SAMPLE_FACTORS - Liver 3 Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:Vehicle RAW_FILE_NAME(File Name )=20240529_JV131_INVIVO_001_MalonicAcid_03 SUBJECT_SAMPLE_FACTORS - Liver 4 Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:Vehicle RAW_FILE_NAME(File Name )=20240529_JV131_INVIVO_001_MalonicAcid_04 SUBJECT_SAMPLE_FACTORS - Liver 5 Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:HKJS001 Injected RAW_FILE_NAME(File Name )=20240529_JV131_INVIVO_001_MalonicAcid_05 SUBJECT_SAMPLE_FACTORS - Liver 6 Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:HKJS001 Injected RAW_FILE_NAME(File Name )=20240529_JV131_INVIVO_001_MalonicAcid_06 SUBJECT_SAMPLE_FACTORS - Liver 7 Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:HKJS001 Injected RAW_FILE_NAME(File Name )=20240529_JV131_INVIVO_001_MalonicAcid_07 SUBJECT_SAMPLE_FACTORS - Liver 8 Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:HKJS001 Injected RAW_FILE_NAME(File Name )=20240529_JV131_INVIVO_001_MalonicAcid_08 SUBJECT_SAMPLE_FACTORS - Liver 9 Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:Malonic Acid Injected RAW_FILE_NAME(File Name )=20240529_JV131_INVIVO_001_MalonicAcid_09 SUBJECT_SAMPLE_FACTORS - Liver 10 Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:Malonic Acid Injected RAW_FILE_NAME(File Name )=20240529_JV131_INVIVO_001_MalonicAcid_10 SUBJECT_SAMPLE_FACTORS - Liver 11 Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:Malonic Acid Injected RAW_FILE_NAME(File Name )=20240529_JV131_INVIVO_001_MalonicAcid_11 SUBJECT_SAMPLE_FACTORS - Liver 12 Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:Malonic Acid Injected RAW_FILE_NAME(File Name )=20240529_JV131_INVIVO_001_MalonicAcid_12 SUBJECT_SAMPLE_FACTORS - Liver 13 Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:Combo HKJS001 and Malonic Acid RAW_FILE_NAME(File Name )=20240529_JV131_INVIVO_001_MalonicAcid_13 SUBJECT_SAMPLE_FACTORS - Liver 14 Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:Combo HKJS001 and Malonic Acid RAW_FILE_NAME(File Name )=20240529_JV131_INVIVO_001_MalonicAcid_14 SUBJECT_SAMPLE_FACTORS - Liver 15 Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:Combo HKJS001 and Malonic Acid RAW_FILE_NAME(File Name )=20240529_JV131_INVIVO_001_MalonicAcid_15 SUBJECT_SAMPLE_FACTORS - Liver 16 Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:Combo HKJS001 and Malonic Acid RAW_FILE_NAME(File Name )=20240529_JV131_INVIVO_001_MalonicAcid_16 SUBJECT_SAMPLE_FACTORS - Standard 2 Sample source:Succinate and Fumarate Reagent Mixture | Standards for Standard Curve and Treatment:Measurable Standard Curve RAW_FILE_NAME(File Name )=20240529_JV131_INVIVO_001_MalonicAcid_17 SUBJECT_SAMPLE_FACTORS - Standard 3 Sample source:Succinate and Fumarate Reagent Mixture | Standards for Standard Curve and Treatment:Measurable Standard Curve RAW_FILE_NAME(File Name )=20240529_JV131_INVIVO_001_MalonicAcid_18 SUBJECT_SAMPLE_FACTORS - Standard 4 Sample source:Succinate and Fumarate Reagent Mixture | Standards for Standard Curve and Treatment:Measurable Standard Curve RAW_FILE_NAME(File Name )=20240529_JV131_INVIVO_001_MalonicAcid_19 SUBJECT_SAMPLE_FACTORS - Standard 5 Sample source:Succinate and Fumarate Reagent Mixture | Standards for Standard Curve and Treatment:Measurable Standard Curve RAW_FILE_NAME(File Name )=20240529_JV131_INVIVO_001_MalonicAcid_20 SUBJECT_SAMPLE_FACTORS - Standard 6 Sample source:Succinate and Fumarate Reagent Mixture | Standards for Standard Curve and Treatment:Measurable Standard Curve RAW_FILE_NAME(File Name )=20240529_JV131_INVIVO_001_MalonicAcid_21 SUBJECT_SAMPLE_FACTORS - Standard 7 Sample source:Succinate and Fumarate Reagent Mixture | Standards for Standard Curve and Treatment:Measurable Standard Curve RAW_FILE_NAME(File Name )=20240529_JV131_INVIVO_001_MalonicAcid_22 SUBJECT_SAMPLE_FACTORS - Standard 8 Sample source:Succinate and Fumarate Reagent Mixture | Standards for Standard Curve and Treatment:Measurable Standard Curve RAW_FILE_NAME(File Name )=20240529_JV131_INVIVO_001_MalonicAcid_23 #COLLECTION CO:COLLECTION_SUMMARY Mice were sacrificed and liver tissue was collected. Tissues were flash frozen CO:COLLECTION_SUMMARY in liquid nitrogen and powderized using a mortar and pestle. Approximately 10 mg CO:COLLECTION_SUMMARY of tissue powder was transferred into an Eppendorf tube. CO:SAMPLE_TYPE Liver #TREATMENT TR:TREATMENT_SUMMARY 13C515N2-glutamine was resuspended to 50 mg/mL in 1X PBS. C57BL/6 mice between TR:TREATMENT_SUMMARY the ages of 10-16 weeks were weighed to allow injection of 0.75 g/kg. Mice that TR:TREATMENT_SUMMARY were injected with HKJS001 was resuspended in DMSO to have a final concentration TR:TREATMENT_SUMMARY of 3.33 mg/mL and 2uL of HKJS001 was injected per gram of mouse. Mice that were TR:TREATMENT_SUMMARY injected with malonic acid was resuspended in water to a molarity of 1M and TR:TREATMENT_SUMMARY 1.6uL of malonic acid was injected per gram of mouse. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY he tissue powder was re-suspended in 800 µL pre-cooled HPLC-grade 60:40 SP:SAMPLEPREP_SUMMARY Methanol:Water and then vortexed for 15 minutes at 4°C. 500 µL of pre-cooled SP:SAMPLEPREP_SUMMARY LC-MS grade chloroform was added to the lysate and again vortexed for 15 minutes SP:SAMPLEPREP_SUMMARY at 4°C. Samples were then centrifuged at 16,000 x g for 10 minutes at 4°C, SP:SAMPLEPREP_SUMMARY creating three layers: the top layer containing polar metabolites, the middle SP:SAMPLEPREP_SUMMARY layer containing protein, and the bottom layer containing non-polar metabolites. SP:SAMPLEPREP_SUMMARY The top layer was transferred into a new Eppendorf tube, dried down in a SP:SAMPLEPREP_SUMMARY Refrigerated CentriVap Benchtop Vacuum Concentrator connected to a CentriVap-105 SP:SAMPLEPREP_SUMMARY Cold Trap, and stored at -80°C until they were re-suspended for LC-MS analysis. SP:SAMPLEPREP_SUMMARY The non-polar layer was discarded, and the remaining protein layer was saved for SP:SAMPLEPREP_SUMMARY protein quantification. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Thermo Vanquish CH:COLUMN_NAME Merck SeQuant ZIC-HILIC (150 x 2.1mm,5um) CH:SOLVENT_A 100% water; 0.1% ammonium hydroxide; 20mM ammonium carbonate CH:SOLVENT_B 100% acetonitrile CH:FLOW_GRADIENT 20 min, 80% - 20% B; 0.5 min, 20% - 80% B; 7.5min, 80% B CH:FLOW_RATE 0.15ml/min CH:COLUMN_TEMPERATURE 25 #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive Plus Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS The mass spectrometer had the spray voltage set to 4.0 kV, heated capillary to MS:MS_COMMENTS 350°C, and the HESI probe at 30 °C. The sheath gas flow was set at 10 units, MS:MS_COMMENTS auxiliary gas at 1 units, and sweep gas flow at 1 unit. An additional scan MS:MS_COMMENTS between 220-700 m/z was used to enhance nucleotide detection in the negative MS:MS_COMMENTS mode as well with the maximum injection time set to 80 msec. Data acquired by MS:MS_COMMENTS Thermo Fisher's Xcalibur software and analyzed by their Tracefinder software. MS:MS_COMMENTS The raw files provided contain data from both positive and negative ion mode. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS peak area MS_METABOLITE_DATA_START Samples Liver 1 Liver 2 Liver 3 Liver 4 Liver 5 Liver 6 Liver 7 Liver 8 Liver 9 Liver 10 Liver 11 Liver 12 Liver 13 Liver 14 Liver 15 Liver 16 Standard 2 Standard 3 Standard 4 Standard 5 Standard 6 Standard 7 Standard 8 Factors Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:Vehicle Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:Vehicle Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:Vehicle Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:Vehicle Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:HKJS001 Injected Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:HKJS001 Injected Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:HKJS001 Injected Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:HKJS001 Injected Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:Malonic Acid Injected Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:Malonic Acid Injected Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:Malonic Acid Injected Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:Malonic Acid Injected Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:Combo HKJS001 and Malonic Acid Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:Combo HKJS001 and Malonic Acid Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:Combo HKJS001 and Malonic Acid Sample source:C57/BL6 mouse tissue | Standards for Standard Curve and Treatment:Combo HKJS001 and Malonic Acid Sample source:Succinate and Fumarate Reagent Mixture | Standards for Standard Curve and Treatment:Measurable Standard Curve Sample source:Succinate and Fumarate Reagent Mixture | Standards for Standard Curve and Treatment:Measurable Standard Curve Sample source:Succinate and Fumarate Reagent Mixture | Standards for Standard Curve and Treatment:Measurable Standard Curve Sample source:Succinate and Fumarate Reagent Mixture | Standards for Standard Curve and Treatment:Measurable Standard Curve Sample source:Succinate and Fumarate Reagent Mixture | Standards for Standard Curve and Treatment:Measurable Standard Curve Sample source:Succinate and Fumarate Reagent Mixture | Standards for Standard Curve and Treatment:Measurable Standard Curve Sample source:Succinate and Fumarate Reagent Mixture | Standards for Standard Curve and Treatment:Measurable Standard Curve Fumaric acid 211671367.5 370460351.7 262491574.9 288489904.2 356535392.1 254598450.1 604334551.5 454599031 480914759.5 385998674.3 478320429 381808058.3 321395980.4 332499116.9 254636400.4 435963238.2 193926237.8 95755799.98 61718942.11 24661696.88 10506760.37 1495476.431 804715.6607 Fumaric acid_13C1 33426013.3 54336747.86 53442472.2 51776418.07 114572693.9 43343595.11 120758544.7 133610716.7 93161960.17 30362731.65 72825579.08 95085869.85 65912362.45 90464512.89 70307028.06 125401842.3 8833187.777 4336841.849 2676160.912 993452.1348 470198.6059 40163.8984 28733.47309 Fumaric acid_13C2 14253002.19 24418476.86 21045335.47 23172567.16 62093209.85 19799080.56 59065845.01 68640554.04 46006858.52 12084468.63 29850988.93 55028382.61 33676603.46 50167335.4 35223410.57 62847823.65 1651258.402 732157.434 476215.4079 186875.7323 54693.11152 4534.085122 13406.24698 Fumaric acid_13C3 12283304.63 22996465.91 16629506.85 20849617.52 63306614.57 11084410.32 63676709.14 68774225.12 42938369.05 18856891.41 29780357.28 77613395.78 32676410 47341072.11 33599080.54 56433477.48 51227.3505 0 0 0 10908.50174 5265.894488 14808.66257 Fumaric acid_13C4 14550504.99 30214403.44 14023507.32 27938672.89 52053297.42 11875982.4 83051820.29 56730024.86 55329315.83 29869117.12 40063604.04 94146165.2 30682534.69 46478817.75 34401723.08 44481390.65 0 6768.652588 0 0 5647.519419 0 5707.406848 Succinic acid 1206007551 1159221144 865998413 1213490724 1513477846 1274916392 1743275973 1926680024 1964909665 2495488986 1612873432 1077875440 1619922906 1342295314 1119260326 1699685270 108000911.6 50476754.91 30987109.9 12934625.91 5317147.001 919546.3328 823269.1367 Succinic acid_13C1 196911665.7 186539868.5 185540097.7 240166036.5 475300925.3 246534008.5 347936925.8 527444264.1 394419130.8 207923431.6 256031835.5 248461095.9 322413206.1 348271684 289219031.4 460557081.3 5241943.223 2479691.276 1482968.309 770908.9567 211352.0412 137738.5453 54230.53859 Succinic acid_13C2 97619556.28 97689426.56 94164939.89 128998918.2 343389231.5 136932338.2 228306900.4 344107892 233316160.1 97725642.53 129241069.4 191501650.6 208763741.3 221518063.1 195427968.7 286107532.6 892228.4761 0 0 0 0 0 0 Succinic acid_13C3 70687106.64 74909675.76 63288902.2 106391321.6 261579045.6 72094561.21 186075086.2 267747409.8 156495789.9 129048419 101289931.5 225965842 148521851.9 159629420 131378015.7 186504228.5 162719.9622 97138.43633 0 79159.06413 97568.21995 51735.7694 29728.17243 Succinic acid_13C4 113564866 134429429.8 84901797.23 181537935.9 355180108.8 102971930.5 330541090.8 368827194 316229831.4 254718672.7 173473744.6 411223717.8 223631136.3 259963234.2 225978549.5 283110652.4 424342.2354 166108.1371 0 0 0 306605.7409 0 Citric acid 521012984.3 931990645.3 540201865.3 7.44E+08 1003707540 774407037.3 1228766722 851141510.9 1664154886 1310178860 1.12E+09 9.98E+08 8.07E+08 7.56E+08 8.31E+08 9.52E+08 2.48E+08 80941608 38708700 10629422 4005764 884728.4 172036.9 Citric acid_13C5 22544518.93 42327548.6 25346758.18 44436752 194830556.5 55342203.73 198746166.8 94685133.24 345536782.7 52197684.74 1.27E+08 2.17E+08 93888781 1.60E+08 85974095 1.30E+08 12957.12 0 0 0 8585.372 26205.25 5196.77 Aspartic acid 232317630.8 198128442.7 202003992.9 2.42E+08 302387347.3 195770348.4 357994130.9 304949156.2 272031897 440427127.6 3.67E+08 5.45E+08 2.57E+08 2.60E+08 2.29E+08 3.05E+08 44756027 14210726 7470141 2436629 1363325 245350 245126.6 Aspartic acid_13C3 4902555.557 2993297.552 2342064.04 11492645 203452.6688 4589518.595 0 9684110.166 11389365.93 8948229.757 0 0 802792.8 485543 0 823055 0 0 0 0 0 0 0 Aspartic acid_13C4 8968513.688 14599393 7653817.527 16186393 24683201.81 5914992.589 29445173.22 22530762.77 18622865.92 17467580.93 17145854 90976334 10405633 19561571 19195600 21157590 189535 138747.6 0 39372.08 44096.97 72722.81 98869.84 Malic acid 1430076957 2538330826 1824133319 1.87E+09 2320023675 1649236487 3725238386 2904325163 3055376193 2405064894 3.09E+09 2.48E+09 2.26E+09 2.29E+09 1.77E+09 2.83E+09 7.36E+08 3.62E+08 2.34E+08 87801763 38136975 6094115 2605206 Malic acid_13C1 222405420.3 372258063.7 361378757.6 3.41E+08 725833693.1 275251244.6 747788922.6 846151168.8 614245012.7 196290574.7 4.77E+08 6.21E+08 4.80E+08 6.25E+08 4.81E+08 8.39E+08 32972677 16159258 10350407 3936721 1858398 420417.7 235081.8 Malic acid_13C2 108933747.5 186735178 163277425.4 1.72E+08 473016736.8 145686024.1 410663626 509130935.3 322552330.7 90556854.04 2.22E+08 3.90E+08 2.61E+08 3.82E+08 2.90E+08 4.54E+08 8026752 3549592 2335995 812223.2 208245.2 44276.38 43465.83 Malic acid_13C3 86409794.06 160099511.3 120316586.8 1.39E+08 416736750.4 83428926.62 385221268 442306566.7 274954405.6 119720225.2 2.00E+08 5.01E+08 2.40E+08 3.40E+08 2.44E+08 3.76E+08 343964.2 160962 90508.61 58253.88 8967.032 35664.95 8303.409 Malic acid_13C4 101845227 205936402 97135599.09 1.75E+08 332712763.8 80734580.09 505000517.1 378654258.3 350491155.3 194053377.9 2.63E+08 5.79E+08 2.23E+08 3.38E+08 2.59E+08 2.97E+08 54901.51 127407.4 78887.25 81069.11 132708.5 108758.7 74600.06 ATP 1866181446 1387861155 2068643764 2930054075 3054845219 3009956249 3178301079 3045291861 2474829896 757559935 1583786806 1314198657 1433255815 1167686011 951623417 1461360111 68331871.6 29545663.5 17622400.4 6080830.38 2570720.12 344044.397 194622.864 ADP 1764682303 1924853582 1669438966 2796443931 2747541856 2693795048 2972722112 3175705405 2918381708 1718743990 2453680132 2194279522 2420810785 1989596710 1754132067 2599100080 192979622 82476851.5 50175392 20020747.6 8726110.28 1500099.57 891770.149 NADH 449793484 342031209 336712973 296722033 842591595 751656243 917881389 646656255 618689052 573710978 503142478 423625544 342808236 700298784 528622902 696099946 0 1280.75485 0 0 1134.35278 0 0 UTP 141622674 66156321.5 159158769 202284842 257789324 224664827 221280240 232687524 288778473 58536390.2 156371184 166271677 198226185 145085080 129173788 240780432 0 0 0 0 0 0 0 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Retention index Quantified m/z PubChem ID KEGG ID Fumaric acid 10.15 115.00368 444972 C00122 Fumaric acid_13C1 10.15 116.00704 Fumaric acid_13C2 10.15 117.01039 Fumaric acid_13C3 10.15 118.01375 Fumaric acid_13C4 10.15 119.0171 Succinic acid 9.62 117.01933 1110 C00042 Succinic acid_13C1 9.62 118.02269 Succinic acid_13C2 9.62 119.02604 Succinic acid_13C3 9.62 120.0294 Succinic acid_13C4 9.62 121.03275 Citric acid 11.35 191.01973 311 C00158 Citric acid_13C5 11.35 196.0365 Aspartic acid 9.45 132.03023 5960 C00049 Aspartic acid_13C3 9.45 135.0403 Aspartic acid_13C4 9.45 136.04365 Malic acid 10.18 133.01425 222656 C00149 Malic acid_13C1 10.18 134.0176 Malic acid_13C2 10.18 135.02096 Malic acid_13C3 10.18 136.02431 Malic acid_13C4 10.18 137.02767 ATP 10.77 505.98847 5957 C00002 ADP 9.59 426.02214 6022 C00008 NADH 8.86 664.11749 439153 C00004 UTP 12.02 482.96125 6133 C00075 METABOLITES_END #END