#METABOLOMICS WORKBENCH dmukha_20240309_152107 DATATRACK_ID:4696 STUDY_ID:ST003151 ANALYSIS_ID:AN005169 PROJECT_ID:PR001941 VERSION 1 CREATED_ON March 27, 2024, 10:47 am #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 BT474 breast cancer cell line grown in 20% D2O-containing RPMI-1640 medium ST:STUDY_TITLE treated with Fasnall and GSK2194069 ST:STUDY_TYPE Free fatty acid analysis, D2O tracing ST:STUDY_SUMMARY BT474 breast cancer cell line grown in 20% D2O-containing medium treated with ST:STUDY_SUMMARY Fasnall and GSK2194069. Cells were grown for 24 h 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 7 ST:TOTAL_SUBJECTS 21 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_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 - 01_FFA_Blank Sample source:BT-474 cell lipid extract | Isotopic Tracer:NA | Drug Treatment:NA RAW_FILE_NAME(Raw File Name)=01_FFA_Blank.raw; RAW_FILE_NAME(Raw File Name)=NEG_01_FFA_Blank.mzXML; RAW_FILE_NAME(Raw File Name )=POS_01_FFA_Blank.mzXML SUBJECT_SAMPLE_FACTORS - 02_FFA_Blank Sample source:BT-474 cell lipid extract | Isotopic Tracer:NA | Drug Treatment:NA RAW_FILE_NAME(Raw File Name)=02_FFA_Blank.raw; RAW_FILE_NAME(Raw File Name)=NEG_02_FFA_Blank.mzXML; RAW_FILE_NAME(Raw File Name )=POS_02_FFA_Blank.mzXML SUBJECT_SAMPLE_FACTORS - 03_FFA_Blank Sample source:BT-474 cell lipid extract | Isotopic Tracer:NA | Drug Treatment:NA RAW_FILE_NAME(Raw File Name)=03_FFA_Blank.raw; RAW_FILE_NAME(Raw File Name)=NEG_03_FFA_Blank.mzXML; RAW_FILE_NAME(Raw File Name )=POS_03_FFA_Blank.mzXML SUBJECT_SAMPLE_FACTORS - 04_FFA_D2O_BT474_Control Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:Vehicle RAW_FILE_NAME(Raw File Name)=04_FFA_D2O_BT474_Control.raw; RAW_FILE_NAME(Raw File Name)=NEG_04_FFA_D2O_BT474_Control.mzXML; RAW_FILE_NAME(Raw File Name )=POS_04_FFA_D2O_BT474_Control.mzXML SUBJECT_SAMPLE_FACTORS - 05_FFA_D2O_BT474_GSK0001 Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:1 nM GSK2194069 RAW_FILE_NAME(Raw File Name)=05_FFA_D2O_BT474_GSK0001.raw; RAW_FILE_NAME(Raw File Name)=NEG_05_FFA_D2O_BT474_GSK0001.mzXML; RAW_FILE_NAME(Raw File Name )=POS_05_FFA_D2O_BT474_GSK0001.mzXML SUBJECT_SAMPLE_FACTORS - 06_FFA_D2O_BT474_GSK0040 Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:40 nM GSK2194069 RAW_FILE_NAME(Raw File Name)=06_FFA_D2O_BT474_GSK0040.raw; RAW_FILE_NAME(Raw File Name)=NEG_06_FFA_D2O_BT474_GSK0040.mzXML; RAW_FILE_NAME(Raw File Name )=POS_06_FFA_D2O_BT474_GSK0040.mzXML SUBJECT_SAMPLE_FACTORS - 07_FFA_D2O_BT474_GSK1000 Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:1 uM GSK2194069 RAW_FILE_NAME(Raw File Name)=07_FFA_D2O_BT474_GSK1000.raw; RAW_FILE_NAME(Raw File Name)=NEG_07_FFA_D2O_BT474_GSK1000.mzXML; RAW_FILE_NAME(Raw File Name )=POS_07_FFA_D2O_BT474_GSK1000.mzXML SUBJECT_SAMPLE_FACTORS - 08_FFA_D2O_BT474_Fasnall01 Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:1 uM Fasnall RAW_FILE_NAME(Raw File Name)=08_FFA_D2O_BT474_Fasnall01.raw; RAW_FILE_NAME(Raw File Name)=NEG_08_FFA_D2O_BT474_Fasnall01.mzXML; RAW_FILE_NAME(Raw File Name )=POS_08_FFA_D2O_BT474_Fasnall01.mzXML SUBJECT_SAMPLE_FACTORS - 09_FFA_D2O_BT474_Fasnall05 Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:5 uM Fasnall RAW_FILE_NAME(Raw File Name)=09_FFA_D2O_BT474_Fasnall05.raw; RAW_FILE_NAME(Raw File Name)=NEG_09_FFA_D2O_BT474_Fasnall05.mzXML; RAW_FILE_NAME(Raw File Name )=POS_09_FFA_D2O_BT474_Fasnall05.mzXML SUBJECT_SAMPLE_FACTORS - 10_FFA_D2O_BT474_Fasnall40 Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:40 uM Fasnall RAW_FILE_NAME(Raw File Name)=10_FFA_D2O_BT474_Fasnall40.raw; RAW_FILE_NAME(Raw File Name)=NEG_10_FFA_D2O_BT474_Fasnall40.mzXML; RAW_FILE_NAME(Raw File Name )=POS_10_FFA_D2O_BT474_Fasnall40.mzXML SUBJECT_SAMPLE_FACTORS - 11_FFA_D2O_BT474_Control Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:Vehicle RAW_FILE_NAME(Raw File Name)=11_FFA_D2O_BT474_Control.raw; RAW_FILE_NAME(Raw File Name)=NEG_11_FFA_D2O_BT474_Control.mzXML; RAW_FILE_NAME(Raw File Name )=POS_11_FFA_D2O_BT474_Control.mzXML SUBJECT_SAMPLE_FACTORS - 12_FFA_D2O_BT474_GSK0001 Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:1 nM GSK2194069 RAW_FILE_NAME(Raw File Name)=12_FFA_D2O_BT474_GSK0001.raw; RAW_FILE_NAME(Raw File Name)=NEG_12_FFA_D2O_BT474_GSK0001.mzXML; RAW_FILE_NAME(Raw File Name )=POS_12_FFA_D2O_BT474_GSK0001.mzXML SUBJECT_SAMPLE_FACTORS - 13_FFA_D2O_BT474_GSK0040 Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:40 nM GSK2194069 RAW_FILE_NAME(Raw File Name)=13_FFA_D2O_BT474_GSK0040.raw; RAW_FILE_NAME(Raw File Name)=NEG_13_FFA_D2O_BT474_GSK0040.mzXML; RAW_FILE_NAME(Raw File Name )=POS_13_FFA_D2O_BT474_GSK0040.mzXML SUBJECT_SAMPLE_FACTORS - 14_FFA_D2O_BT474_GSK1000 Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:1 uM GSK2194069 RAW_FILE_NAME(Raw File Name)=14_FFA_D2O_BT474_GSK1000.raw; RAW_FILE_NAME(Raw File Name)=NEG_14_FFA_D2O_BT474_GSK1000.mzXML; RAW_FILE_NAME(Raw File Name )=POS_14_FFA_D2O_BT474_GSK1000.mzXML SUBJECT_SAMPLE_FACTORS - 15_FFA_D2O_BT474_Fasnall01 Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:1 uM Fasnall RAW_FILE_NAME(Raw File Name)=15_FFA_D2O_BT474_Fasnall01.raw; RAW_FILE_NAME(Raw File Name)=NEG_15_FFA_D2O_BT474_Fasnall01.mzXML; RAW_FILE_NAME(Raw File Name )=POS_15_FFA_D2O_BT474_Fasnall01.mzXML SUBJECT_SAMPLE_FACTORS - 16_FFA_D2O_BT474_Fasnall05 Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:5 uM Fasnall RAW_FILE_NAME(Raw File Name)=16_FFA_D2O_BT474_Fasnall05.raw; RAW_FILE_NAME(Raw File Name)=NEG_16_FFA_D2O_BT474_Fasnall05.mzXML; RAW_FILE_NAME(Raw File Name )=POS_16_FFA_D2O_BT474_Fasnall05.mzXML SUBJECT_SAMPLE_FACTORS - 17_FFA_D2O_BT474_Fasnall40 Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:40 uM Fasnall RAW_FILE_NAME(Raw File Name)=17_FFA_D2O_BT474_Fasnall40.raw; RAW_FILE_NAME(Raw File Name)=NEG_17_FFA_D2O_BT474_Fasnall40.mzXML; RAW_FILE_NAME(Raw File Name )=POS_17_FFA_D2O_BT474_Fasnall40.mzXML SUBJECT_SAMPLE_FACTORS - 18_FFA_D2O_BT474_Control Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:Vehicle RAW_FILE_NAME(Raw File Name)=18_FFA_D2O_BT474_Control.raw; RAW_FILE_NAME(Raw File Name)=NEG_18_FFA_D2O_BT474_Control.mzXML; RAW_FILE_NAME(Raw File Name )=POS_18_FFA_D2O_BT474_Control.mzXML SUBJECT_SAMPLE_FACTORS - 19_FFA_D2O_BT474_GSK0001 Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:1 nM GSK2194069 RAW_FILE_NAME(Raw File Name)=19_FFA_D2O_BT474_GSK0001.raw; RAW_FILE_NAME(Raw File Name)=NEG_19_FFA_D2O_BT474_GSK0001.mzXML; RAW_FILE_NAME(Raw File Name )=POS_19_FFA_D2O_BT474_GSK0001.mzXML SUBJECT_SAMPLE_FACTORS - 20_FFA_D2O_BT474_GSK0040 Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:40 nM GSK2194069 RAW_FILE_NAME(Raw File Name)=20_FFA_D2O_BT474_GSK0040.raw; RAW_FILE_NAME(Raw File Name)=NEG_20_FFA_D2O_BT474_GSK0040.mzXML; RAW_FILE_NAME(Raw File Name )=POS_20_FFA_D2O_BT474_GSK0040.mzXML SUBJECT_SAMPLE_FACTORS - 21_FFA_D2O_BT474_GSK1000 Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:1 uM GSK2194069 RAW_FILE_NAME(Raw File Name)=21_FFA_D2O_BT474_GSK1000.raw; RAW_FILE_NAME(Raw File Name)=NEG_21_FFA_D2O_BT474_GSK1000.mzXML; RAW_FILE_NAME(Raw File Name )=POS_21_FFA_D2O_BT474_GSK1000.mzXML SUBJECT_SAMPLE_FACTORS - 22_FFA_D2O_BT474_Fasnall01 Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:1 uM Fasnall RAW_FILE_NAME(Raw File Name)=22_FFA_D2O_BT474_Fasnall01.raw; RAW_FILE_NAME(Raw File Name)=NEG_22_FFA_D2O_BT474_Fasnall01.mzXML; RAW_FILE_NAME(Raw File Name )=POS_22_FFA_D2O_BT474_Fasnall01.mzXML SUBJECT_SAMPLE_FACTORS - 23_FFA_D2O_BT474_Fasnall05 Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:5 uM Fasnall RAW_FILE_NAME(Raw File Name)=23_FFA_D2O_BT474_Fasnall05.raw; RAW_FILE_NAME(Raw File Name)=NEG_23_FFA_D2O_BT474_Fasnall05.mzXML; RAW_FILE_NAME(Raw File Name )=POS_23_FFA_D2O_BT474_Fasnall05.mzXML SUBJECT_SAMPLE_FACTORS - 24_FFA_D2O_BT474_Fasnall40 Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:40 uM Fasnall RAW_FILE_NAME(Raw File Name)=24_FFA_D2O_BT474_Fasnall40.raw; RAW_FILE_NAME(Raw File Name)=NEG_24_FFA_D2O_BT474_Fasnall40.mzXML; RAW_FILE_NAME(Raw File Name )=POS_24_FFA_D2O_BT474_Fasnall40.mzXML SUBJECT_SAMPLE_FACTORS - 25_FFA_Blank Sample source:BT-474 cell lipid extract | Isotopic Tracer:NA | Drug Treatment:NA RAW_FILE_NAME(Raw File Name)=25_FFA_Blank.raw; RAW_FILE_NAME(Raw File Name)=NEG_25_FFA_Blank.mzXML; RAW_FILE_NAME(Raw File Name )=POS_25_FFA_Blank.mzXML SUBJECT_SAMPLE_FACTORS - 26_FFA_Blank Sample source:BT-474 cell lipid extract | Isotopic Tracer:NA | Drug Treatment:NA RAW_FILE_NAME(Raw File Name)=26_FFA_Blank.raw; RAW_FILE_NAME(Raw File Name)=NEG_26_FFA_Blank.mzXML; RAW_FILE_NAME(Raw File Name )=POS_26_FFA_Blank.mzXML SUBJECT_SAMPLE_FACTORS - 27_FFA_Blank Sample source:BT-474 cell lipid extract | Isotopic Tracer:NA | Drug Treatment:NA RAW_FILE_NAME(Raw File Name)=27_FFA_Blank.raw; RAW_FILE_NAME(Raw File Name)=NEG_27_FFA_Blank.mzXML; RAW_FILE_NAME(Raw File Name )=POS_27_FFA_Blank.mzXML #COLLECTION CO:COLLECTION_SUMMARY Cells were seeded in 10 cm Petri dishes. At collection, cells were washed with CO:COLLECTION_SUMMARY PBS three times. Then, 1 ml of methanol was added and cells were scraped from CO:COLLECTION_SUMMARY the surface. All content of the plate was transferred into 13 x 100 mm Pyrex CO:COLLECTION_SUMMARY glass tubes. Lipids were extracted by the Folch method. Dried lipids were CO:COLLECTION_SUMMARY redissolved in 1 ml of 0.3 M KOH solution in 90% methanol and incubated at 85 CO:COLLECTION_SUMMARY °C for 1 h. Then, 100 µl of formic acid were added, followed by 800 µl of CO:COLLECTION_SUMMARY hexane for extraction. The hexane phase was transferred to glass LC-MS vials and CO:COLLECTION_SUMMARY dried under the stream of nitrogen. Samples were redissolved in 1 ml of 1:1 CO:COLLECTION_SUMMARY methanol:isopropanol. CO:COLLECTION_PROTOCOL_FILENAME DM_free_fatty_acid_analysis_samples.txt CO:SAMPLE_TYPE Cultured cells CO:COLLECTION_METHOD 100% methanol extraction CO:VOLUMEORAMOUNT_COLLECTED 1 ml 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 20% D2O-containing RPMI-1640 medium and treated with Fasnall TR:TREATMENT_SUMMARY or GSK2194069 at various concentrations. TR:TREATMENT_COMPOUND Fasnall, GSK2194069 TR:TREATMENT_DOSE 1 nM - 40 uM TR:TREATMENT_DOSEDURATION 24 h TR:TREATMENT_VEHICLE DMSO TR:CELL_GROWTH_CONTAINER 6-cm Petri dishes TR:CELL_MEDIA RPMI-1640 TR:CELL_ENVIR_COND 37C, 5% CO2 TR:CELL_PCT_CONFLUENCE ~70% #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Sample preparation for LC-MS free fatty acid analysis Cells were seeded in 10 cm SP:SAMPLEPREP_SUMMARY Petri dishes. At collection, cells were washed with PBS three times. Then, 1 ml SP:SAMPLEPREP_SUMMARY of methanol was added and cells were scraped from the surface. All content of SP:SAMPLEPREP_SUMMARY the plate was transferred into 13 x 100 mm Pyrex glass tubes. Lipids were SP:SAMPLEPREP_SUMMARY extracted by the Folch method. Dried lipids were redissolved in 1 ml of 0.3 M SP:SAMPLEPREP_SUMMARY KOH solution in 90% methanol and incubated at 85 °C for 1 h. Then, 100 µl of SP:SAMPLEPREP_SUMMARY formic acid were added, followed by 800 µl of hexane for extraction. The hexane SP:SAMPLEPREP_SUMMARY phase was transferred to glass LC-MS vials and dried under the stream of SP:SAMPLEPREP_SUMMARY nitrogen. Samples were redissolved in 1 ml of 1:1 methanol:isopropanol. SP:SAMPLEPREP_PROTOCOL_FILENAME DM_free_fatty_acid_analysis_samples.txt SP:PROCESSING_METHOD Lipid saponification in 0.3 M KOH SP:EXTRACTION_METHOD 100% 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 Cellular lipids #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Thermo Dionex Ultimate 3000 RS CH:COLUMN_NAME Phenomenex Kinetex XB-C18 (150 x 3 mm, 2.6 um) CH:SOLVENT_A 60:40 Acetonitrile/water; 10 mM ammonium formate CH:SOLVENT_B 90:8:2 Isopropanol/acetonitrile/water; 10 mM ammonium formate CH:FLOW_GRADIENT 0-4.5 min, 15-60% B; 4.5-12 min, 60-82% B; 12-12.75 min, 82-95% B; 12.75-16.5 CH:FLOW_GRADIENT min, 95-100% B; 16.5-22.5 min, 100% B; 22.5-22.6 min, 100-15% B; 22.6-25 min, CH:FLOW_GRADIENT 15% B CH:FLOW_RATE 0-22.5 min, 0.333 ml/min; 22.5-22.6 min, 0.333-0.5 ml/min; 22.6-24.9 min, 0.5 CH:FLOW_RATE ml/min; 24.9-25 min, 0.5-0.333 ml/min CH:COLUMN_TEMPERATURE 65 CH:INJECTION_TEMPERATURE 20 #ANALYSIS AN:ANALYSIS_TYPE MS AN:LABORATORY_NAME Wistar Institute Proteomics & Metabolomic Core AN:OPERATOR_NAME Dzmitry Mukha 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 NEGATIVE MS:MS_COMMENTS The HESI ion source voltage was set to the following parameters: +3,300/-3,500 MS:MS_COMMENTS V, sheath gas 50, auxiliary gas 20, spare gas 0, probe heater 350 °C, capillary MS:MS_COMMENTS temperature 350 °C, S-Lens RF level 80. The mass spectrometer was set to MS:MS_COMMENTS acquire data in polarity-switching mode, one microscan, 60,000 resolution, AGC MS:MS_COMMENTS target 5e6, scan range 130-1950 m/z, IT 200 ms. Data were converted into the MS:MS_COMMENTS mzXML format by ProteoWizard and analyzed in MAVEN. Free cholesterol was MS:MS_COMMENTS measured as a water-loss in-source fragment [M-H2O+H]+. MS:CAPILLARY_TEMPERATURE 350 MS:ION_SOURCE_TEMPERATURE 350 MS:ION_SPRAY_VOLTAGE 3500 MS:IONIZATION Both MS:SOURCE_TEMPERATURE 350 MS:AUTOMATIC_GAIN_CONTROL 5e6 MS:DATAFORMAT RAW #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS AU MS_METABOLITE_DATA_START Samples 01_FFA_Blank 02_FFA_Blank 03_FFA_Blank 04_FFA_D2O_BT474_Control 05_FFA_D2O_BT474_GSK0001 06_FFA_D2O_BT474_GSK0040 07_FFA_D2O_BT474_GSK1000 08_FFA_D2O_BT474_Fasnall01 09_FFA_D2O_BT474_Fasnall05 10_FFA_D2O_BT474_Fasnall40 11_FFA_D2O_BT474_Control 12_FFA_D2O_BT474_GSK0001 13_FFA_D2O_BT474_GSK0040 14_FFA_D2O_BT474_GSK1000 15_FFA_D2O_BT474_Fasnall01 16_FFA_D2O_BT474_Fasnall05 17_FFA_D2O_BT474_Fasnall40 18_FFA_D2O_BT474_Control 19_FFA_D2O_BT474_GSK0001 20_FFA_D2O_BT474_GSK0040 21_FFA_D2O_BT474_GSK1000 22_FFA_D2O_BT474_Fasnall01 23_FFA_D2O_BT474_Fasnall05 24_FFA_D2O_BT474_Fasnall40 25_FFA_Blank 26_FFA_Blank 27_FFA_Blank Factors Sample source:BT-474 cell lipid extract | Isotopic Tracer:NA | Drug Treatment:NA Sample source:BT-474 cell lipid extract | Isotopic Tracer:NA | Drug Treatment:NA Sample source:BT-474 cell lipid extract | Isotopic Tracer:NA | Drug Treatment:NA Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:Vehicle Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:1 nM GSK2194069 Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:40 nM GSK2194069 Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:1 uM GSK2194069 Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:1 uM Fasnall Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:5 uM Fasnall Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:40 uM Fasnall Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:Vehicle Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:1 nM GSK2194069 Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:40 nM GSK2194069 Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:1 uM GSK2194069 Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:1 uM Fasnall Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:5 uM Fasnall Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:40 uM Fasnall Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:Vehicle Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:1 nM GSK2194069 Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:40 nM GSK2194069 Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:1 uM GSK2194069 Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:1 uM Fasnall Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:5 uM Fasnall Sample source:BT-474 cell lipid extract | Isotopic Tracer:20% D2O | Drug Treatment:40 uM Fasnall Sample source:BT-474 cell lipid extract | Isotopic Tracer:NA | Drug Treatment:NA Sample source:BT-474 cell lipid extract | Isotopic Tracer:NA | Drug Treatment:NA Sample source:BT-474 cell lipid extract | Isotopic Tracer:NA | Drug Treatment:NA Myristic acid C12 PARENT 68226.38 75699.69 41886.86 6005832 5937274 4780492 4437471 5680706 6500344 6163700 5258308 4953594 4991766 4028547 4842136 7109804 6818918 6256852 6226898 6242648 4216980 5891522 5545954 7663040 50534.59 39720.07 53458.52 Myristic acid D-label-1 0 0 0 1009054 1089526 214786.7 0 1235442 107805.6 0 850032.9 879169.5 430957.9 0 1183732 0 0 990215.6 990688.1 308945.1 0 1159644 232372.7 0 0 0 0 Myristic acid D-label-2 0 0 0 1441695 1273000 905916.7 0 1698774 887148.9 106376 1294580 1373070 919646.3 0 1520076 873912.7 107395.8 1454791 1654082 1192598 0 1696987 773728 119706.4 0 0 0 Myristic acid D-label-3 0 0 0 1403610 1161517 863860 0 1318982 733357.2 120957.4 1204736 1195070 879294.7 12302.45 1212291 782668.6 107428.9 1342297 1348791 1079259 0 1240767 585106.4 97514.09 0 0 0 Myristic acid D-label-4 0 0 0 921910.5 779053.7 666245.8 0 758994.6 391690.1 21855.57 558678.7 677099.3 553961.8 0 706011.8 474934.1 56521.06 935888.2 972298.8 652753.6 0 764144.2 377153.9 59204.14 0 0 0 Myristic acid D-label-5 0 0 0 447965 283716.2 105249.4 0 363575.5 158111.9 0 268902.9 257773.4 299023.3 0 248047.5 112742.3 0 508139.8 472849.7 354411.1 0 219119.6 147164.1 27968.87 0 0 0 Myristic acid D-label-6 0 0 0 180990.6 109371.6 129429.7 0 66411.08 37222.46 0 168049.3 124522.5 126464.2 0 75833.13 73970.53 0 202339.4 140692.7 120219.2 0 126084 49923.17 0 0 0 0 Myristic acid D-label-7 0 0 0 31275.05 20001.32 12742.67 0 38070.9 0 0 20346.09 19946.18 16737.5 0 28983.34 0 0 49096.48 19167.76 21325.86 0 31662.46 0 0 0 0 0 Myristic acid D-label-8 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristic acid D-label-9 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristic acid D-label-10 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristic acid D-label-11 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristic acid D-label-12 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristic acid D-label-13 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristic acid D-label-14 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristic acid D-label-15 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristic acid D-label-16 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristic acid D-label-17 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristic acid D-label-18 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristic acid D-label-19 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristic acid D-label-20 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristic acid D-label-21 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristic acid D-label-22 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristic acid D-label-23 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristic acid D-label-24 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristic acid D-label-25 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristic acid D-label-26 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristic acid D-label-27 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Myristic acid D-label-28 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid C12 PARENT 110108.4 0 0 1.08E+07 1.18E+07 9825734 9081803 1.01E+07 7917760 8311804 1.04E+07 1.13E+07 1.02E+07 8721287 9583669 8782116 9296173 1.01E+07 1.07E+07 1.07E+07 8784939 1.06E+07 9077252 8540915 0 0 0 Ximenic acid D-label-1 0 0 0 5342034 5473318 5901752 5233058 4124513 841882 525796.7 4876956 5236834 5666478 5122638 3760117 676573.8 392375.3 4850080 5219994 5572858 5065562 4169905 1709352 0 0 0 0 Ximenic acid D-label-2 0 0 0 2286827 2875168 2566913 3124207 1280004 0 0 2370047 2508792 2960369 3063619 1200408 0 0 2182654 2385207 3162549 2984817 1324810 0 0 0 0 0 Ximenic acid D-label-3 0 0 0 1496278 1568268 1635026 2033536 705400 0 0 1441361 1452409 1744264 2119388 638538.4 34113.57 0 1505161 1605867 1816466 2018599 691911.1 0 0 0 0 0 Ximenic acid D-label-4 0 0 0 702178.8 858175.4 907061.9 1021461 345816.7 0 0 904573.4 868445.6 916828.8 1037852 279273.6 0 0 814575.3 794300.5 921865.7 991354.9 299470.2 0 0 0 0 0 Ximenic acid D-label-5 0 0 0 397795.8 582504 471662.9 434713.2 96313.66 0 0 344520.2 397253.9 529064.1 420173.1 143146.3 0 0 408877 423095.9 378600.1 438980 49738.02 0 0 0 0 0 Ximenic acid D-label-6 0 0 0 204139.5 198256.8 266554.2 97992.1 0 0 0 136640.7 224854.2 262114.9 92803.87 0 0 0 218086.6 200152.8 314927 125027 0 0 0 0 0 0 Ximenic acid D-label-7 0 0 0 48885.4 49669.28 135966 0 0 0 0 154724.3 137229.4 197684.2 0 0 0 0 119752.2 97955 150318.1 24692.22 0 0 0 0 0 0 Ximenic acid D-label-8 0 0 0 69894.62 32057.13 76261.79 0 0 0 0 0 40953.44 37426.78 0 0 0 0 30083.8 36149.76 89217.55 0 0 0 0 0 0 0 Ximenic acid D-label-9 0 0 0 0 0 0 0 0 0 0 0 0 38156.38 0 0 0 0 0 35202.65 0 0 0 0 0 0 0 0 Ximenic acid D-label-10 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-11 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-12 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-13 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-14 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-15 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-16 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-17 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-18 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-19 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-20 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-21 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-22 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-23 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-24 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-25 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-26 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-27 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-28 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-29 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-30 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-31 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-32 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-33 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-34 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-35 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-36 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-37 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-38 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-39 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-40 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-41 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-42 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-43 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-44 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-45 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-46 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-47 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-48 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-49 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ximenic acid D-label-50 0 0 0 0 0 0 0 0 0 0 0 0 0 0 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 Myristic acid C12 PARENT 5.706242 227.2023 227.2023 11005 C06424 36 Myristic acid D-label-1 5.704849 228.208 227.2023 11005 C06424 36 Myristic acid D-label-2 5.696602 229.2148 227.2023 11005 C06424 36 Myristic acid D-label-3 5.694521 230.2211 227.2023 11005 C06424 36 Myristic acid D-label-4 5.688298 231.2276 227.2023 11005 C06424 36 Myristic acid D-label-5 5.685366 232.2337 227.2023 11005 C06424 36 Myristic acid D-label-6 5.682484 233.2395 227.2023 11005 C06424 36 Myristic acid D-label-7 5.679576 234.246 227.2023 11005 C06424 36 Myristic acid D-label-8 NA NA 227.2023 11005 C06424 36 Peak was not detected Myristic acid D-label-9 NA NA 227.2023 11005 C06424 36 Peak was not detected Myristic acid D-label-10 NA NA 227.2023 11005 C06424 36 Peak was not detected Myristic acid D-label-11 NA NA 227.2023 11005 C06424 36 Peak was not detected Myristic acid D-label-12 NA NA 227.2023 11005 C06424 36 Peak was not detected Myristic acid D-label-13 NA NA 227.2023 11005 C06424 36 Peak was not detected Myristic acid D-label-14 NA NA 227.2023 11005 C06424 36 Peak was not detected Myristic acid D-label-15 NA NA 227.2023 11005 C06424 36 Peak was not detected Myristic acid D-label-16 NA NA 227.2023 11005 C06424 36 Peak was not detected Myristic acid D-label-17 NA NA 227.2023 11005 C06424 36 Peak was not detected Myristic acid D-label-18 NA NA 227.2023 11005 C06424 36 Peak was not detected Myristic acid D-label-19 NA NA 227.2023 11005 C06424 36 Peak was not detected Myristic acid D-label-20 NA NA 227.2023 11005 C06424 36 Peak was not detected Myristic acid D-label-21 NA NA 227.2023 11005 C06424 36 Peak was not detected Myristic acid D-label-22 NA NA 227.2023 11005 C06424 36 Peak was not detected Myristic acid D-label-23 NA NA 227.2023 11005 C06424 36 Peak was not detected Myristic acid D-label-24 NA NA 227.2023 11005 C06424 36 Peak was not detected Myristic acid D-label-25 NA NA 227.2023 11005 C06424 36 Peak was not detected Myristic acid D-label-26 NA NA 227.2023 11005 C06424 36 Peak was not detected Myristic acid D-label-27 NA NA 227.2023 11005 C06424 36 Peak was not detected Myristic acid D-label-28 NA NA 227.2023 11005 C06424 36 Peak was not detected Ximenic acid C12 PARENT 10.87266 393.3743 393.3744 5282775 C17278   NA Ximenic acid D-label-1 10.86089 394.3791 393.3744 5282775 C17278   NA Ximenic acid D-label-2 10.86457 395.3858 393.3744 5282775 C17278   NA Ximenic acid D-label-3 10.85699 396.3919 393.3744 5282775 C17278   NA Ximenic acid D-label-4 10.85668 397.399 393.3744 5282775 C17278   NA Ximenic acid D-label-5 10.85848 398.405 393.3744 5282775 C17278   NA Ximenic acid D-label-6 10.8503 399.4113 393.3744 5282775 C17278   NA Ximenic acid D-label-7 10.83568 400.4183 393.3744 5282775 C17278   NA Ximenic acid D-label-8 10.84956 401.4235 393.3744 5282775 C17278   NA Ximenic acid D-label-9 10.84198 402.4321 393.3744 5282775 C17278   NA Ximenic acid D-label-10 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-11 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-12 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-13 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-14 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-15 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-16 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-17 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-18 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-19 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-20 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-21 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-22 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-23 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-24 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-25 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-26 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-27 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-28 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-29 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-30 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-31 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-32 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-33 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-34 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-35 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-36 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-37 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-38 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-39 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-40 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-41 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-42 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-43 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-44 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-45 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-46 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-47 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-48 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-49 NA NA 393.3744 5282775 C17278   NA Peak was not detected Ximenic acid D-label-50 NA NA 393.3744 5282775 C17278   NA Peak was not detected METABOLITES_END #END