#METABOLOMICS WORKBENCH dmukha_20240309_151143 DATATRACK_ID:4694 STUDY_ID:ST003147 ANALYSIS_ID:AN005164 PROJECT_ID:PR001941 VERSION 1 CREATED_ON March 25, 2024, 3:24 pm #PROJECT PR:PROJECT_TITLE The shutdown of NADH oxidation via Respiratory Complex I mimics fatty acid PR:PROJECT_TITLE biosynthesis inhibition PR:PROJECT_TYPE LC-MS Quantitative Analysis PR:PROJECT_SUMMARY Proliferating cancer cells actively utilize anabolic processes for biomass PR:PROJECT_SUMMARY production, including de novo biosynthesis of amino acids, nucleotides, and PR:PROJECT_SUMMARY fatty acids. The key enzyme of the fatty acid biosynthesis pathway, fatty acid PR:PROJECT_SUMMARY synthase (FASN), is widely recognized as a promising therapeutic target in PR:PROJECT_SUMMARY cancer and other health conditions. Here, we establish a metabolic signature of PR:PROJECT_SUMMARY FASN inhibition using a panel of pharmacological inhibitors (GSK2194069, PR:PROJECT_SUMMARY TVB-2640, TVB-3166, C75, cerulenin, and Fasnall). We find that the activity of PR:PROJECT_SUMMARY some commonly used FASN inhibitors is inconsistent with the metabolic signature PR:PROJECT_SUMMARY of FASN inhibition (accumulation of malonate, succinate, malonyl coenzyme A, PR:PROJECT_SUMMARY succinyl coenzyme A, and other metabolic perturbations). Moreover, we show that PR:PROJECT_SUMMARY one of these putative FASN inhibitors, Fasnall, is a respiratory Complex I PR:PROJECT_SUMMARY inhibitor that mimics FASN inhibition through NADH accumulation and consequent PR:PROJECT_SUMMARY depletion of the tricarboxylic acid cycle metabolites. We demonstrate that PR:PROJECT_SUMMARY Fasnall impairs tumor growth in several oxidative phosphorylation-dependent PR:PROJECT_SUMMARY cancer models, including combination therapy-resistant melanoma patient-derived PR:PROJECT_SUMMARY xenografts. Fasnall administration does not reproduce neurological side effects PR:PROJECT_SUMMARY in mice reported for other Complex I inhibitors. Our results have significant PR:PROJECT_SUMMARY implications for understanding the FASN role in human health and disease and PR:PROJECT_SUMMARY provide evidence of therapeutic potential for Complex I inhibitors with fast PR:PROJECT_SUMMARY systemic clearance. Our findings also highlight the continuing need for PR:PROJECT_SUMMARY validation of small molecule inhibitors to distinguish high-quality chemical PR:PROJECT_SUMMARY probes and to expand the understanding of their application. PR:INSTITUTE Wistar Institute PR:DEPARTMENT Molecular and Cellular Oncogenesis Program, Ellen and Ronald Caplan Cancer PR:DEPARTMENT Center PR:LABORATORY Schug's Lab PR:LAST_NAME Mukha PR:FIRST_NAME Dzmitry PR:ADDRESS 3601 Spruce St., Philadelphia, Pennsylvania 19104, USA PR:EMAIL dmukha@wistar.org PR:PHONE +12154956903 PR:FUNDING_SOURCE This work was supported by grants from the National Institutes of Health (NIH) PR:FUNDING_SOURCE National Cancer Institute (NCI) DP2 CA249950-01 (Z.T.S.), NIH NCI P01 CA114046 PR:FUNDING_SOURCE (Z.T.S.), Melanoma Research Foundation 717173 (Z.T.S.), and Susan G. Komen PR:FUNDING_SOURCE CCR19608782 (Z.T.S.). PR:PUBLICATIONS Submission Pending PR:CONTRIBUTORS Dzmitry Mukha, Jena Dessain, Seamus O’Connor, Katherine Pniewski, Fabrizio PR:CONTRIBUTORS Bertolazzi, Jeet Patel, Mary Mullins, Zachary T. Schug #STUDY ST:STUDY_TITLE BT-474 cells fed with [13C2] acetate and treated with 1 uM Fasnall or 1 uM ST:STUDY_TITLE GSK2194069 for 24 h ST:STUDY_TYPE Intracellular metabolomics, [13C2] acetate ST:STUDY_SUMMARY BT-474 breast cancer cells fed with [13C2] acetate and treated with 1 uM Fasnall ST:STUDY_SUMMARY or 1 uM GSK2194069 for 24 h. Cells were grown in RPMI-1640 with 10% dialyzed ST:STUDY_SUMMARY FBS. ST:INSTITUTE Wistar Institute ST:DEPARTMENT Molecular and Cellular Oncogenesis Program, Ellen and Ronald Caplan Cancer ST:DEPARTMENT Center ST:LABORATORY Schug's Lab ST:LAST_NAME Mukha ST:FIRST_NAME Dzmitry ST:ADDRESS 3601 Spruce St, Philadelphia, PA 19104, USA ST:EMAIL dmukha@wistar.org ST:PHONE 2154956903 ST:NUM_GROUPS 3 ST:TOTAL_SUBJECTS 9 ST:PUBLICATIONS Submission Pending #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 SU:AGE_OR_AGE_RANGE 60 SU:GENDER Female SU:CELL_BIOSOURCE_OR_SUPPLIER ATCC SU:CELL_STRAIN_DETAILS BT-474, breast cancer cell line #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 - Blank-80MeOH Sample source:BT-474 intracellular metabolites | [13C2] Acetate:NA | Drug Treatment:NA RAW_FILE_NAME(Raw File Name)=Blank-80MeOH.raw; RAW_FILE_NAME(Raw File Name)=NEG_Blank-80MeOH.mzXML; RAW_FILE_NAME(Raw File Name )=POS_Blank-80MeOH.mzXML SUBJECT_SAMPLE_FACTORS - Blank-80MeOH_20220211171023 Sample source:BT-474 intracellular metabolites | [13C2] Acetate:NA | Drug Treatment:NA RAW_FILE_NAME(Raw File Name)=Blank-80MeOH_20220211171023.raw; RAW_FILE_NAME(Raw File Name)=NEG_Blank-80MeOH_20220211171023.mzXML; RAW_FILE_NAME(Raw File Name )=POS_Blank-80MeOH_20220211171023.mzXML SUBJECT_SAMPLE_FACTORS - Blank-80MeOH_20220211180427 Sample source:BT-474 intracellular metabolites | [13C2] Acetate:NA | Drug Treatment:NA RAW_FILE_NAME(Raw File Name)=Blank-80MeOH_20220211180427.raw; RAW_FILE_NAME(Raw File Name)=NEG_Blank-80MeOH_20220211180427.mzXML; RAW_FILE_NAME(Raw File Name )=POS_Blank-80MeOH_20220211180427.mzXML SUBJECT_SAMPLE_FACTORS - Blank_NoFlow Sample source:BT-474 intracellular metabolites | [13C2] Acetate:NA | Drug Treatment:NA RAW_FILE_NAME(Raw File Name)=Blank_NoFlow.raw; RAW_FILE_NAME(Raw File Name)=NEG_Blank_NoFlow.mzXML; RAW_FILE_NAME(Raw File Name )=POS_Blank_NoFlow.mzXML SUBJECT_SAMPLE_FACTORS - m22-L025_SO_01 Sample source:BT-474 intracellular metabolites | [13C2] Acetate:NA | Drug Treatment:NA RAW_FILE_NAME(Raw File Name)=m22-L025_SO_01.raw; RAW_FILE_NAME(Raw File Name)=NEG_m22-L025_SO_01.mzXML; RAW_FILE_NAME(Raw File Name )=POS_m22-L025_SO_01.mzXML SUBJECT_SAMPLE_FACTORS - m22-L025_SO_02 Sample source:BT-474 intracellular metabolites | [13C2] Acetate:200 uM | Drug Treatment:NA RAW_FILE_NAME(Raw File Name)=m22-L025_SO_02.raw; RAW_FILE_NAME(Raw File Name)=NEG_m22-L025_SO_02.mzXML; RAW_FILE_NAME(Raw File Name )=POS_m22-L025_SO_02.mzXML SUBJECT_SAMPLE_FACTORS - m22-L025_SO_03 Sample source:BT-474 intracellular metabolites | [13C2] Acetate:200 uM | Drug Treatment:NA RAW_FILE_NAME(Raw File Name)=m22-L025_SO_03.raw; RAW_FILE_NAME(Raw File Name)=NEG_m22-L025_SO_03.mzXML; RAW_FILE_NAME(Raw File Name )=POS_m22-L025_SO_03.mzXML SUBJECT_SAMPLE_FACTORS - m22-L025_SO_04 Sample source:BT-474 intracellular metabolites | [13C2] Acetate:200 uM | Drug Treatment:1 uM Fasnall RAW_FILE_NAME(Raw File Name)=m22-L025_SO_04.raw; RAW_FILE_NAME(Raw File Name)=NEG_m22-L025_SO_04.mzXML; RAW_FILE_NAME(Raw File Name )=POS_m22-L025_SO_04.mzXML SUBJECT_SAMPLE_FACTORS - m22-L025_SO_05 Sample source:BT-474 intracellular metabolites | [13C2] Acetate:200 uM | Drug Treatment:1 uM Fasnall RAW_FILE_NAME(Raw File Name)=m22-L025_SO_05.raw; RAW_FILE_NAME(Raw File Name)=NEG_m22-L025_SO_05.mzXML; RAW_FILE_NAME(Raw File Name )=POS_m22-L025_SO_05.mzXML SUBJECT_SAMPLE_FACTORS - m22-L025_SO_06 Sample source:BT-474 intracellular metabolites | [13C2] Acetate:200 uM | Drug Treatment:1 uM Fasnall RAW_FILE_NAME(Raw File Name)=m22-L025_SO_06.raw; RAW_FILE_NAME(Raw File Name)=NEG_m22-L025_SO_06.mzXML; RAW_FILE_NAME(Raw File Name )=POS_m22-L025_SO_06.mzXML SUBJECT_SAMPLE_FACTORS - m22-L025_SO_07 Sample source:BT-474 intracellular metabolites | [13C2] Acetate:200 uM | Drug Treatment:1 uM GSK2194069 RAW_FILE_NAME(Raw File Name)=m22-L025_SO_07.raw; RAW_FILE_NAME(Raw File Name)=NEG_m22-L025_SO_07.mzXML; RAW_FILE_NAME(Raw File Name )=POS_m22-L025_SO_07.mzXML SUBJECT_SAMPLE_FACTORS - m22-L025_SO_08 Sample source:BT-474 intracellular metabolites | [13C2] Acetate:200 uM | Drug Treatment:1 uM GSK2194069 RAW_FILE_NAME(Raw File Name)=m22-L025_SO_08.raw; RAW_FILE_NAME(Raw File Name)=NEG_m22-L025_SO_08.mzXML; RAW_FILE_NAME(Raw File Name )=POS_m22-L025_SO_08.mzXML SUBJECT_SAMPLE_FACTORS - m22-L025_SO_09 Sample source:BT-474 intracellular metabolites | [13C2] Acetate:200 uM | Drug Treatment:1 uM GSK2194069 RAW_FILE_NAME(Raw File Name)=m22-L025_SO_09.raw; RAW_FILE_NAME(Raw File Name)=NEG_m22-L025_SO_09.mzXML; RAW_FILE_NAME(Raw File Name )=POS_m22-L025_SO_09.mzXML #COLLECTION CO:COLLECTION_SUMMARY One PBS washing, 80% methanol extraction. CO:COLLECTION_PROTOCOL_FILENAME DM_metabolomics_samples.txt CO:SAMPLE_TYPE Cultured cells CO:COLLECTION_METHOD 80% methanol extraction CO:VOLUMEORAMOUNT_COLLECTED 500 ul CO:STORAGE_CONDITIONS -80℃ CO:COLLECTION_VIALS 1.5 ml plastic centrifuge tubes CO:STORAGE_VIALS 1.5 ml plastic centrifuge tubes #TREATMENT TR:TREATMENT_SUMMARY Cells were grown in RPMI-1640 supplemented with 200 uM [13C2] acetate and TR:TREATMENT_SUMMARY treated with 1 uM Fasnall or 1 uM GSK2194069. TR:TREATMENT_COMPOUND Fasnall, GSK2194069 TR:TREATMENT_DOSE 0 - 1 uM TR:TREATMENT_DOSEDURATION 24 h TR:TREATMENT_VEHICLE DMSO TR:CELL_GROWTH_CONTAINER 6-well plates TR:CELL_MEDIA RPMI-1640 TR:CELL_ENVIR_COND 37C, 5% CO2 TR:CELL_PCT_CONFLUENCE ~70% #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Metabolites were extracted with ice-cold 80% methanol. SP:SAMPLEPREP_PROTOCOL_FILENAME DM_metabolomics_samples.txt SP:EXTRACTION_METHOD 80% methanol SP:EXTRACT_ENRICHMENT None SP:EXTRACT_CLEANUP None SP:EXTRACT_STORAGE -80℃ SP:SAMPLE_RESUSPENSION None SP:SAMPLE_DERIVATIZATION None SP:SAMPLE_SPIKING None SP:SUBCELLULAR_LOCATION Intracellular metabolites #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Thermo Dionex Ultimate 3000 RS CH:COLUMN_NAME Merck SeQuant ZIC-pHILIC (150 x 2.1mm,5um) CH:SOLVENT_A Water with 0.01% ammonium hydroxide and 20 mM ammonium bicarbonate CH:SOLVENT_B Acetonitrile CH:FLOW_GRADIENT A linear solvent gradient of a total duration 22.5 min was starting with 0.2 CH:FLOW_GRADIENT ml/min flow rate of 80% solvent B, 12.5 min – 30%, 15 min – 30%, 15.2 min CH:FLOW_GRADIENT – 80%, 20 min – 80%, 21 min – flow rate 0.3 ml/min, 22 min – flow rate CH:FLOW_GRADIENT 0.3 ml/min, 22.1 min – flow rate 0.2 ml/min. CH:FLOW_RATE 0.2 ml/min CH:COLUMN_TEMPERATURE 40 #ANALYSIS AN:ANALYSIS_TYPE MS AN:LABORATORY_NAME Wistar Institute Proteomics & Metabolomic Core AN:OPERATOR_NAME Nicole Gorman AN:DETECTOR_TYPE Orbitrap #MS MS:INSTRUMENT_NAME Thermo Q Exactive HF-X Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS The HESI ion source voltage was set to the following parameters: 3,600 V in both MS:MS_COMMENTS polarity modes, sheath gas 30, auxiliary gas 5, spare gas 0, probe heater 200 MS:MS_COMMENTS °C, capillary temperature 325 °C, S-Lens RF level 65. The mass spectrometer MS:MS_COMMENTS was set to acquire data in the polarity-switching mode averaging two microscans MS:MS_COMMENTS with 60,000 resolution, automatic gain control (AGC) target 5e6, scan range MS:MS_COMMENTS 72-1080 m/z, and maximum orbitrap injection time (IT) 200 ms. Data were MS:MS_COMMENTS converted into the mzXML format by ProteoWizard and analyzed in MAVEN. MS:ION_SOURCE_TEMPERATURE 200 MS:ION_SPRAY_VOLTAGE 3600 MS:IONIZATION Both MS:SOURCE_TEMPERATURE 200 MS:AUTOMATIC_GAIN_CONTROL 5e6 MS:DATAFORMAT RAW #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS AU MS_METABOLITE_DATA_START Samples Blank-80MeOH Blank-80MeOH_20220211171023 Blank-80MeOH_20220211180427 Blank_NoFlow m22-L025_SO_01 m22-L025_SO_02 m22-L025_SO_03 m22-L025_SO_04 m22-L025_SO_05 m22-L025_SO_06 m22-L025_SO_07 m22-L025_SO_08 Factors Sample source:BT-474 intracellular metabolites | [13C2] Acetate:NA | Drug Treatment:NA Sample source:BT-474 intracellular metabolites | [13C2] Acetate:NA | Drug Treatment:NA Sample source:BT-474 intracellular metabolites | [13C2] Acetate:NA | Drug Treatment:NA Sample source:BT-474 intracellular metabolites | [13C2] Acetate:NA | Drug Treatment:NA Sample source:BT-474 intracellular metabolites | [13C2] Acetate:NA | Drug Treatment:NA Sample source:BT-474 intracellular metabolites | [13C2] Acetate:200 uM | Drug Treatment:NA Sample source:BT-474 intracellular metabolites | [13C2] Acetate:200 uM | Drug Treatment:NA Sample source:BT-474 intracellular metabolites | [13C2] Acetate:200 uM | Drug Treatment:1 uM Fasnall Sample source:BT-474 intracellular metabolites | [13C2] Acetate:200 uM | Drug Treatment:1 uM Fasnall Sample source:BT-474 intracellular metabolites | [13C2] Acetate:200 uM | Drug Treatment:1 uM Fasnall Sample source:BT-474 intracellular metabolites | [13C2] Acetate:200 uM | Drug Treatment:1 uM GSK2194069 Sample source:BT-474 intracellular metabolites | [13C2] Acetate:200 uM | Drug Treatment:1 uM GSK2194069 Acetyl-CoA C12 PARENT 0 0 0 0 134127 1268990 135570.8 272558.7 323059.8 139672.3 172046.9 340255 218387.9 Acetyl-CoA C13-label-1 0 0 0 0 42457.47 367322.8 37888.25 83524.28 96805.95 41254.06 47922.97 103833.5 67788.37 Acetyl-CoA C13-label-2 0 0 0 0 29188.25 231502.5 32888.8 130511.9 174208.8 97970.73 86088.85 165800.1 78632.66 Acetyl-CoA C13-label-3 0 0 0 0 0 19519.54 3690.467 14825.46 45913.85 13946.75 8803.998 39697.46 15517.14 Acetyl-CoA C13-label-4 0 0 0 0 0 0 0 4756.285 0 0 0 6539.866 0 Acetyl-CoA C13-label-5 0 0 0 0 0 0 0 0 3579.619 0 0 0 2280.721 Acetyl-CoA C13-label-6 0 0 0 0 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-7 0 0 0 0 0 0 0 0 3571.288 0 0 0 0 Acetyl-CoA C13-label-8 0 0 0 0 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-9 0 0 0 0 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-10 0 0 0 0 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-11 0 0 0 0 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-12 0 0 0 0 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-13 0 0 0 0 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-14 0 0 1874.099 0 2827.781 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-15 0 0 0 0 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-16 0 0 0 0 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-17 0 0 0 0 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-18 0 0 0 0 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-19 0 0 0 0 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-20 0 0 0 0 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-21 0 0 0 0 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-22 0 0 0 0 0 0 0 0 0 0 0 0 0 Acetyl-CoA C13-label-23 0 0 0 0 0 0 0 0 0 0 0 0 0 Fasnall C12 PARENT 0 0 0 0 0 0 0 5.71E+07 5.77E+07 5.31E+07 0 53236.12 0 Fasnall C13-label-1 0 0 0 0 0 0 0 1.21E+07 1.23E+07 1.12E+07 0 0 0 Fasnall C13-label-2 0 0 0 0 0 0 0 648464.4 0 549699.2 0 0 0 Fasnall C13-label-3 0 0 0 0 0 0 0 11030.35 0 0 0 0 0 Fasnall C13-label-4 0 0 0 0 0 0 0 0 0 0 0 0 0 Fasnall C13-label-5 0 0 0 0 0 0 0 0 0 0 0 0 0 Fasnall C13-label-6 0 0 0 0 0 0 0 0 0 0 0 79835.35 0 Fasnall C13-label-7 0 0 0 0 0 0 0 0 0 0 0 0 8746.782 Fasnall C13-label-8 0 0 0 0 0 0 0 0 0 0 0 0 0 Fasnall C13-label-9 0 0 0 0 0 0 0 0 0 0 0 0 0 Fasnall C13-label-10 0 0 0 0 0 0 0 0 0 0 0 0 0 Fasnall C13-label-11 0 0 0 0 0 0 0 0 0 0 0 0 0 Fasnall C13-label-12 0 0 0 0 0 0 0 0 0 0 0 0 0 Fasnall C13-label-13 0 0 0 0 0 0 0 0 0 0 0 0 0 Fasnall C13-label-14 0 0 0 0 0 0 0 0 0 0 0 0 0 Fasnall C13-label-15 0 0 0 0 0 0 0 0 0 0 0 0 0 Fasnall C13-label-16 0 0 0 0 0 0 0 0 0 0 0 0 0 Fasnall C13-label-17 0 0 0 0 0 0 0 0 0 0 0 0 0 Fasnall C13-label-18 0 0 0 0 0 0 0 0 0 0 0 0 0 Fasnall C13-label-19 0 0 0 0 0 0 0 0 0 0 0 0 0 GSK2194069 C12 PARENT 0 0 0 0 0 0 0 14028.24 0 9365.128 2.41E+07 3.12E+07 2.57E+07 GSK2194069 C13-label-1 0 0 0 0 0 0 0 0 0 0 6685548 8740249 7159852 GSK2194069 C13-label-2 0 0 0 0 0 0 0 0 0 0 968977.8 1245574 1070284 GSK2194069 C13-label-3 0 0 0 0 0 0 0 0 0 0 0 103884.2 104436.8 GSK2194069 C13-label-4 0 0 0 0 0 0 0 0 0 0 0 0 0 GSK2194069 C13-label-5 0 0 0 0 0 0 0 0 0 0 0 0 0 GSK2194069 C13-label-6 0 0 0 0 0 0 0 0 0 0 0 0 0 GSK2194069 C13-label-7 0 0 0 0 0 0 0 0 0 0 0 0 0 GSK2194069 C13-label-8 0 0 0 0 0 0 0 0 0 0 0 0 0 GSK2194069 C13-label-9 0 0 0 0 0 0 0 0 0 0 0 0 0 GSK2194069 C13-label-10 0 0 0 0 0 0 0 0 0 0 0 0 0 GSK2194069 C13-label-11 0 0 0 0 0 0 0 0 0 0 0 0 0 GSK2194069 C13-label-12 0 0 0 0 0 0 0 0 0 0 0 0 0 GSK2194069 C13-label-13 0 0 0 0 0 0 0 0 0 0 0 0 0 GSK2194069 C13-label-14 0 0 0 0 0 0 0 0 0 0 0 0 0 GSK2194069 C13-label-15 0 0 0 0 0 0 0 0 0 0 0 0 0 GSK2194069 C13-label-16 0 0 0 0 0 0 0 0 0 0 0 0 0 GSK2194069 C13-label-17 0 0 0 0 0 0 0 0 0 0 0 0 0 GSK2194069 C13-label-18 0 0 0 0 0 0 0 0 0 0 0 0 0 GSK2194069 C13-label-19 0 0 0 0 0 0 0 0 0 0 0 0 0 GSK2194069 C13-label-20 0 0 0 0 0 0 0 0 0 0 0 0 0 GSK2194069 C13-label-21 0 0 0 0 0 0 0 0 0 0 0 0 0 GSK2194069 C13-label-22 0 0 0 0 0 0 0 0 0 0 0 0 0 GSK2194069 C13-label-23 0 0 0 0 0 0 0 0 0 0 0 0 0 GSK2194069 C13-label-24 0 0 0 0 0 0 0 0 0 0 0 0 0 GSK2194069 C13-label-25 0 0 0 0 0 0 0 0 0 0 0 0 0 O-Acetyl-L-carnitine C12 PARENT 0 43632.04 47079.86 50215.22 2.52E+07 3.52E+07 2.66E+07 3.47E+07 3.53E+07 3.31E+07 3.93E+07 4.93E+07 5.34E+07 O-Acetyl-L-carnitine C13-label-1 0 0 0 0 2483257 3503697 2559625 3383232 3471455 3382514 4219606 5603478 5944914 O-Acetyl-L-carnitine C13-label-2 0 2542.458 0 0 2202923 2820786 2441456 9942639 1.01E+07 1.09E+07 5418600 6133474 5709726 O-Acetyl-L-carnitine C13-label-3 0 0 0 0 185869.9 243344 176133.1 730670.7 773169.2 773708.5 423696.4 0 419481.5 O-Acetyl-L-carnitine C13-label-4 0 0 0 0 0 0 0 0 0 0 0 0 0 O-Acetyl-L-carnitine C13-label-5 0 0 1873.382 0 0 0 0 0 0 0 0 0 0 O-Acetyl-L-carnitine C13-label-6 0 0 0 0 0 0 0 0 0 0 0 0 0 O-Acetyl-L-carnitine C13-label-7 0 0 0 0 0 0 0 0 0 0 0 0 0 O-Acetyl-L-carnitine C13-label-8 0 0 0 0 0 0 0 0 0 0 0 0 0 O-Acetyl-L-carnitine C13-label-9 0 0 0 0 0 0 0 0 0 0 0 0 0 L-Carnitine C12 PARENT 0 14007.05 16159.94 14728.11 2.07E+07 2.22E+07 2.33E+07 2.17E+07 2.86E+07 2.39E+07 2.30E+07 2.70E+07 2.80E+07 L-Carnitine C13-label-1 0 0 0 0 1578573 1685429 1764883 1643358 2201011 1802166 1762505 2068152 2119048 L-Carnitine C13-label-2 0 0 0 0 0 0 0 0 0 0 0 0 0 L-Carnitine C13-label-3 0 0 0 0 0 0 0 0 0 0 0 0 0 L-Carnitine C13-label-4 0 0 0 0 0 0 0 0 0 0 0 0 0 L-Carnitine C13-label-5 0 0 0 0 0 0 0 0 0 0 0 0 0 L-Carnitine C13-label-6 0 0 0 0 0 0 0 0 0 0 0 0 0 L-Carnitine C13-label-7 0 0 0 0 0 0 0 0 0 0 0 0 0 Palmitoyl-L-carnitine C12 PARENT 0 0 0 0 852951.2 2257988 1270304 1192343 1165075 821626.2 70997.4 109263.4 79818.6 Palmitoyl-L-carnitine C13-label-1 0 0 0 0 0 0 0 0 0 0 0 0 0 Palmitoyl-L-carnitine C13-label-2 0 0 0 0 505514.9 1313263 658872.8 91955.09 0 0 0 0 0 Palmitoyl-L-carnitine C13-label-3 0 0 0 0 113119.2 277055 163000.1 47474.14 0 32064.72 8602.604 0 0 Palmitoyl-L-carnitine C13-label-4 0 0 0 0 290999.2 707056.1 441697.9 221887.9 221352.7 154433 0 0 0 Palmitoyl-L-carnitine C13-label-5 0 0 0 0 86259.88 215512.2 112147.3 59090.97 65382 28377.08 0 0 0 Palmitoyl-L-carnitine C13-label-6 0 0 0 0 197679.1 484794 302954.2 205204.1 261999.6 164270.7 0 0 0 Palmitoyl-L-carnitine C13-label-7 0 0 0 0 41233.5 112916.7 66314.51 48503.93 59089.39 46963.67 0 0 0 Palmitoyl-L-carnitine C13-label-8 0 0 0 0 126402.3 304617.4 177961.6 225287.5 218098.2 172799.6 0 0 0 Palmitoyl-L-carnitine C13-label-9 0 0 0 0 12609.78 55362.67 33537.07 57569.48 68571.03 47554.45 0 0 0 Palmitoyl-L-carnitine C13-label-10 0 0 0 0 73555.67 209306.4 117081.4 190076 198254.1 161781.9 0 0 0 Palmitoyl-L-carnitine C13-label-11 0 0 0 0 0 51609.42 28541.86 29657.18 27725.98 36299.62 0 0 0 Palmitoyl-L-carnitine C13-label-12 0 0 0 0 45713.51 0 0 173206.7 142131.5 114572.6 0 0 0 Palmitoyl-L-carnitine C13-label-13 0 0 0 0 0 0 6565.801 0 26574.45 0 0 0 0 Palmitoyl-L-carnitine C13-label-14 0 0 0 0 15914.85 0 33698.67 147240.8 116213.7 82074.15 0 0 0 Palmitoyl-L-carnitine C13-label-15 0 0 0 0 0 0 0 23640.45 10338.05 17326.3 0 0 0 Palmitoyl-L-carnitine C13-label-16 0 0 0 0 0 14673 0 88231.75 91486.88 67847.73 0 0 0 Palmitoyl-L-carnitine C13-label-17 0 0 0 0 0 0 0 0 0 0 0 0 0 Palmitoyl-L-carnitine C13-label-18 0 0 0 0 0 0 0 0 0 0 0 0 0 Palmitoyl-L-carnitine C13-label-19 0 0 0 0 0 0 0 0 0 0 0 0 0 Palmitoyl-L-carnitine C13-label-20 0 0 0 0 0 0 0 0 0 0 0 0 0 Palmitoyl-L-carnitine C13-label-21 0 0 0 0 0 0 0 0 0 0 0 0 0 Palmitoyl-L-carnitine C13-label-22 0 0 0 0 0 0 0 0 0 0 0 0 0 Palmitoyl-L-carnitine C13-label-23 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristoyl-L-carnitine C12 PARENT 0 0 0 0 464930 969884.2 682946.4 441495.2 383072.5 285094.7 0 9027.361 5692.772 Myristoyl-L-carnitine C13-label-1 0 0 0 0 133808 252272.4 176909.8 119475.4 89999.38 68154.26 0 0 0 Myristoyl-L-carnitine C13-label-2 0 0 0 0 322531.5 548276.9 447712.9 180830.9 170748.5 115013.7 0 0 0 Myristoyl-L-carnitine C13-label-3 0 0 0 0 72461.82 128661.5 97551.01 0 28444.63 24488.73 0 0 0 Myristoyl-L-carnitine C13-label-4 0 0 0 0 216395.8 329241.4 307203.8 222844.8 222893.3 151219.9 0 0 0 Myristoyl-L-carnitine C13-label-5 519.1772 0 0 0 43100.06 97065.12 52387.38 0 60087.43 44784.41 0 0 0 Myristoyl-L-carnitine C13-label-6 0 0 0 0 102739.9 201048.8 159723.2 217009 217361.4 164036.6 0 0 0 Myristoyl-L-carnitine C13-label-7 0 0 0 0 24810.04 56036.23 25791.42 54981.25 46115.98 39875.8 0 0 0 Myristoyl-L-carnitine C13-label-8 0 0 0 0 67177.44 92182.01 104145 208780.1 196311.3 148745.6 0 0 0 Myristoyl-L-carnitine C13-label-9 0 0 0 0 6697.133 0 9647.78 29767.85 25889.45 0 0 0 0 Myristoyl-L-carnitine C13-label-10 0 0 0 0 40902.66 65684.95 52774.74 161540.7 144093 125813.6 0 0 0 Myristoyl-L-carnitine C13-label-11 0 0 0 0 18470.64 0 7646.524 38931.26 34706.79 0 0 0 0 Myristoyl-L-carnitine C13-label-12 0 0 0 0 24134.12 14742.97 25260.24 134905.5 117765.4 84189.67 0 0 0 Myristoyl-L-carnitine C13-label-13 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristoyl-L-carnitine C13-label-14 0 0 0 0 0 0 0 71434.1 56180.67 51452.49 0 0 0 Myristoyl-L-carnitine C13-label-15 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristoyl-L-carnitine C13-label-16 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristoyl-L-carnitine C13-label-17 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristoyl-L-carnitine C13-label-18 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristoyl-L-carnitine C13-label-19 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristoyl-L-carnitine C13-label-20 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristoyl-L-carnitine C13-label-21 0 0 0 0 0 0 0 0 0 0 0 0 0 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name retention time quantified m/z parent pubchem id kegg id regno comments Acetyl-CoA C12 PARENT 8.264252 810.1323 810.1323 444493 C00024 50043 Acetyl-CoA C13-label-1 8.264318 811.1351 810.1323 444493 C00024 50043 Acetyl-CoA C13-label-2 8.282574 812.1376 810.1323 444493 C00024 50043 Acetyl-CoA C13-label-3 8.284739 813.1407 810.1323 444493 C00024 50043 Acetyl-CoA C13-label-4 8.28975 814.1436 810.1323 444493 C00024 50043 Acetyl-CoA C13-label-5 8.273425 815.1458 810.1323 444493 C00024 50043 Acetyl-CoA C13-label-6 NA NA 810.1323 444493 C00024 50043 Peak was not detected Acetyl-CoA C13-label-7 8.3211 817.1536 810.1323 444493 C00024 50043 Acetyl-CoA C13-label-8 NA NA 810.1323 444493 C00024 50043 Peak was not detected Acetyl-CoA C13-label-9 NA NA 810.1323 444493 C00024 50043 Peak was not detected Acetyl-CoA C13-label-10 NA NA 810.1323 444493 C00024 50043 Peak was not detected Acetyl-CoA C13-label-11 NA NA 810.1323 444493 C00024 50043 Peak was not detected Acetyl-CoA C13-label-12 NA NA 810.1323 444493 C00024 50043 Peak was not detected Acetyl-CoA C13-label-13 NA NA 810.1323 444493 C00024 50043 Peak was not detected Acetyl-CoA C13-label-14 8.241442 824.1783 810.1323 444493 C00024 50043 Acetyl-CoA C13-label-15 NA NA 810.1323 444493 C00024 50043 Peak was not detected Acetyl-CoA C13-label-16 NA NA 810.1323 444493 C00024 50043 Peak was not detected Acetyl-CoA C13-label-17 NA NA 810.1323 444493 C00024 50043 Peak was not detected Acetyl-CoA C13-label-18 NA NA 810.1323 444493 C00024 50043 Peak was not detected Acetyl-CoA C13-label-19 NA NA 810.1323 444493 C00024 50043 Peak was not detected Acetyl-CoA C13-label-20 NA NA 810.1323 444493 C00024 50043 Peak was not detected Acetyl-CoA C13-label-21 NA NA 810.1323 444493 C00024 50043 Peak was not detected Acetyl-CoA C13-label-22 NA NA 810.1323 444493 C00024 50043 Peak was not detected Acetyl-CoA C13-label-23 NA NA 810.1323 444493 C00024 50043 Peak was not detected Fasnall C12 PARENT 1.678357 339.1631 339.1631 16230362 NA NA Fasnall C13-label-1 1.701267 340.1664 339.1631 16230362 NA NA Fasnall C13-label-2 1.658297 341.1711 339.1631 16230362 NA NA Fasnall C13-label-3 1.723833 342.1743 339.1631 16230362 NA NA Fasnall C13-label-4 NA NA 339.1631 16230362 NA NA Peak was not detected Fasnall C13-label-5 NA NA 339.1631 16230362 NA NA Peak was not detected Fasnall C13-label-6 1.573583 345.1847 339.1631 16230362 NA NA Fasnall C13-label-7 1.701867 346.1877 339.1631 16230362 NA NA Fasnall C13-label-8 NA NA 339.1631 16230362 NA NA Peak was not detected Fasnall C13-label-9 NA NA 339.1631 16230362 NA NA Peak was not detected Fasnall C13-label-10 NA NA 339.1631 16230362 NA NA Peak was not detected Fasnall C13-label-11 NA NA 339.1631 16230362 NA NA Peak was not detected Fasnall C13-label-12 NA NA 339.1631 16230362 NA NA Peak was not detected Fasnall C13-label-13 NA NA 339.1631 16230362 NA NA Peak was not detected Fasnall C13-label-14 NA NA 339.1631 16230362 NA NA Peak was not detected Fasnall C13-label-15 NA NA 339.1631 16230362 NA NA Peak was not detected Fasnall C13-label-16 NA NA 339.1631 16230362 NA NA Peak was not detected Fasnall C13-label-17 NA NA 339.1631 16230362 NA NA Peak was not detected Fasnall C13-label-18 NA NA 339.1631 16230362 NA NA Peak was not detected Fasnall C13-label-19 NA NA 339.1631 16230362 NA NA Peak was not detected GSK2194069 C12 PARENT 1.684213 429.1911 429.1914 67376285 NA NA GSK2194069 C13-label-1 1.65017 430.1946 429.1914 67376285 NA NA GSK2194069 C13-label-2 1.65017 431.1976 429.1914 67376285 NA NA GSK2194069 C13-label-3 1.656594 432.2006 429.1914 67376285 NA NA GSK2194069 C13-label-4 NA NA 429.1914 67376285 NA NA Peak was not detected GSK2194069 C13-label-5 NA NA 429.1914 67376285 NA NA Peak was not detected GSK2194069 C13-label-6 NA NA 429.1914 67376285 NA NA Peak was not detected GSK2194069 C13-label-7 NA NA 429.1914 67376285 NA NA Peak was not detected GSK2194069 C13-label-8 NA NA 429.1914 67376285 NA NA Peak was not detected GSK2194069 C13-label-9 NA NA 429.1914 67376285 NA NA Peak was not detected GSK2194069 C13-label-10 NA NA 429.1914 67376285 NA NA Peak was not detected GSK2194069 C13-label-11 NA NA 429.1914 67376285 NA NA Peak was not detected GSK2194069 C13-label-12 NA NA 429.1914 67376285 NA NA Peak was not detected GSK2194069 C13-label-13 NA NA 429.1914 67376285 NA NA Peak was not detected GSK2194069 C13-label-14 NA NA 429.1914 67376285 NA NA Peak was not detected GSK2194069 C13-label-15 NA NA 429.1914 67376285 NA NA Peak was not detected GSK2194069 C13-label-16 NA NA 429.1914 67376285 NA NA Peak was not detected GSK2194069 C13-label-17 NA NA 429.1914 67376285 NA NA Peak was not detected GSK2194069 C13-label-18 NA NA 429.1914 67376285 NA NA Peak was not detected GSK2194069 C13-label-19 NA NA 429.1914 67376285 NA NA Peak was not detected GSK2194069 C13-label-20 NA NA 429.1914 67376285 NA NA Peak was not detected GSK2194069 C13-label-21 NA NA 429.1914 67376285 NA NA Peak was not detected GSK2194069 C13-label-22 NA NA 429.1914 67376285 NA NA Peak was not detected GSK2194069 C13-label-23 NA NA 429.1914 67376285 NA NA Peak was not detected GSK2194069 C13-label-24 NA NA 429.1914 67376285 NA NA Peak was not detected GSK2194069 C13-label-25 NA NA 429.1914 67376285 NA NA Peak was not detected O-Acetyl-L-carnitine C12 PARENT 6.304233 204.1229 204.1229 7045767 C02571 NA O-Acetyl-L-carnitine C13-label-1 6.308872 205.1259 204.1229 7045767 C02571 NA O-Acetyl-L-carnitine C13-label-2 6.296321 206.1294 204.1229 7045767 C02571 NA O-Acetyl-L-carnitine C13-label-3 6.300106 207.1328 204.1229 7045767 C02571 NA O-Acetyl-L-carnitine C13-label-4 NA NA 204.1229 7045767 C02571 NA Peak was not detected O-Acetyl-L-carnitine C13-label-5 6.25845 209.1394 204.1229 7045767 C02571 NA O-Acetyl-L-carnitine C13-label-6 NA NA 204.1229 7045767 C02571 NA Peak was not detected O-Acetyl-L-carnitine C13-label-7 NA NA 204.1229 7045767 C02571 NA Peak was not detected O-Acetyl-L-carnitine C13-label-8 NA NA 204.1229 7045767 C02571 NA Peak was not detected O-Acetyl-L-carnitine C13-label-9 NA NA 204.1229 7045767 C02571 NA Peak was not detected L-Carnitine C12 PARENT 7.897175 162.1124 162.1124 10917 C00318 42914 L-Carnitine C13-label-1 7.906517 163.1158 162.1124 10917 C00318 42914 L-Carnitine C13-label-2 NA NA 162.1124 10917 C00318 42914 Peak was not detected L-Carnitine C13-label-3 NA NA 162.1124 10917 C00318 42914 Peak was not detected L-Carnitine C13-label-4 NA NA 162.1124 10917 C00318 42914 Peak was not detected L-Carnitine C13-label-5 NA NA 162.1124 10917 C00318 42914 Peak was not detected L-Carnitine C13-label-6 NA NA 162.1124 10917 C00318 42914 Peak was not detected L-Carnitine C13-label-7 NA NA 162.1124 10917 C00318 42914 Peak was not detected Palmitoyl-L-carnitine C12 PARENT 1.718306 400.3422 400.3422 11953816 C02990 NA Palmitoyl-L-carnitine C13-label-1 NA NA 400.3422 11953816 C02990 NA Peak was not detected Palmitoyl-L-carnitine C13-label-2 1.716446 402.3474 400.3422 11953816 C02990 NA Palmitoyl-L-carnitine C13-label-3 1.70899 403.3514 400.3422 11953816 C02990 NA Palmitoyl-L-carnitine C13-label-4 1.720637 404.3551 400.3422 11953816 C02990 NA Palmitoyl-L-carnitine C13-label-5 1.733581 405.3579 400.3422 11953816 C02990 NA Palmitoyl-L-carnitine C13-label-6 1.7332 406.3619 400.3422 11953816 C02990 NA Palmitoyl-L-carnitine C13-label-7 1.701859 407.3648 400.3422 11953816 C02990 NA Palmitoyl-L-carnitine C13-label-8 1.710953 408.3682 400.3422 11953816 C02990 NA Palmitoyl-L-carnitine C13-label-9 1.678883 409.3712 400.3422 11953816 C02990 NA Palmitoyl-L-carnitine C13-label-10 1.742961 410.3751 400.3422 11953816 C02990 NA Palmitoyl-L-carnitine C13-label-11 1.707467 411.3792 400.3422 11953816 C02990 NA Palmitoyl-L-carnitine C13-label-12 1.712608 412.3818 400.3422 11953816 C02990 NA Palmitoyl-L-carnitine C13-label-13 1.775325 413.3849 400.3422 11953816 C02990 NA Palmitoyl-L-carnitine C13-label-14 1.707227 414.3881 400.3422 11953816 C02990 NA Palmitoyl-L-carnitine C13-label-15 1.738667 415.3915 400.3422 11953816 C02990 NA Palmitoyl-L-carnitine C13-label-16 1.713333 416.3953 400.3422 11953816 C02990 NA Palmitoyl-L-carnitine C13-label-17 NA NA 400.3422 11953816 C02990 NA Peak was not detected Palmitoyl-L-carnitine C13-label-18 NA NA 400.3422 11953816 C02990 NA Peak was not detected Palmitoyl-L-carnitine C13-label-19 NA NA 400.3422 11953816 C02990 NA Peak was not detected Palmitoyl-L-carnitine C13-label-20 NA NA 400.3422 11953816 C02990 NA Peak was not detected Palmitoyl-L-carnitine C13-label-21 NA NA 400.3422 11953816 C02990 NA Peak was not detected Palmitoyl-L-carnitine C13-label-22 NA NA 400.3422 11953816 C02990 NA Peak was not detected Palmitoyl-L-carnitine C13-label-23 NA NA 400.3422 11953816 C02990 NA Peak was not detected Myristoyl-L-carnitine C12 PARENT 1.811613 372.3102 372.3102 53477791 NA NA Myristoyl-L-carnitine C13-label-1 1.815706 373.3137 372.3102 53477791 NA NA Myristoyl-L-carnitine C13-label-2 1.815631 374.3171 372.3102 53477791 NA NA Myristoyl-L-carnitine C13-label-3 1.811353 375.3202 372.3102 53477791 NA NA Myristoyl-L-carnitine C13-label-4 1.799856 376.3235 372.3102 53477791 NA NA Myristoyl-L-carnitine C13-label-5 1.820578 377.3264 372.3102 53477791 NA NA Myristoyl-L-carnitine C13-label-6 1.81275 378.3306 372.3102 53477791 NA NA Myristoyl-L-carnitine C13-label-7 1.797006 379.3344 372.3102 53477791 NA NA Myristoyl-L-carnitine C13-label-8 1.822125 380.3367 372.3102 53477791 NA NA Myristoyl-L-carnitine C13-label-9 1.822008 381.3404 372.3102 53477791 NA NA Myristoyl-L-carnitine C13-label-10 1.828425 382.344 372.3102 53477791 NA NA Myristoyl-L-carnitine C13-label-11 1.802525 383.3481 372.3102 53477791 NA NA Myristoyl-L-carnitine C13-label-12 1.806289 384.3501 372.3102 53477791 NA NA Myristoyl-L-carnitine C13-label-13 NA NA 372.3102 53477791 NA NA Peak was not detected Myristoyl-L-carnitine C13-label-14 1.808983 386.3571 372.3102 53477791 NA NA Myristoyl-L-carnitine C13-label-15 NA NA 372.3102 53477791 NA NA Peak was not detected Myristoyl-L-carnitine C13-label-16 NA NA 372.3102 53477791 NA NA Peak was not detected Myristoyl-L-carnitine C13-label-17 NA NA 372.3102 53477791 NA NA Peak was not detected Myristoyl-L-carnitine C13-label-18 NA NA 372.3102 53477791 NA NA Peak was not detected Myristoyl-L-carnitine C13-label-19 NA NA 372.3102 53477791 NA NA Peak was not detected Myristoyl-L-carnitine C13-label-20 NA NA 372.3102 53477791 NA NA Peak was not detected Myristoyl-L-carnitine C13-label-21 NA NA 372.3102 53477791 NA NA Peak was not detected METABOLITES_END #END