#METABOLOMICS WORKBENCH dmukha_20240309_151143 DATATRACK_ID:4694 STUDY_ID:ST003147 ANALYSIS_ID:AN005163 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                      	NEGATIVE
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	127220.1	1078016	117020.4	228053	306813.9	140956.7	140787.3	322947.2	213801.8
Acetyl-CoA C13-label-1	0	0	0	36115.6	317604	38128.81	80880.13	93488.9	35668.18	47714.04	106812.6	67493.72
Acetyl-CoA C13-label-2	0	0	0	18457.78	193973.3	23306.22	146357.4	155555.4	75718.16	83214.24	132960.5	80586.54
Acetyl-CoA C13-label-3	0	0	0	0	39185.75	3491.678	28131.17	29395.97	15358.74	8169.35	33717.78	10956.36
Acetyl-CoA C13-label-4	0	0	0	0	3945.573	0	4502.181	5850.545	0	4235.344	4401.364	0
Acetyl-CoA C13-label-5	0	0	0	0	0	0	0	0	0	0	0	0
Acetyl-CoA C13-label-6	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	0	0	0	0
Acetyl-CoA C13-label-8	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
Acetyl-CoA C13-label-10	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
Acetyl-CoA C13-label-12	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
Acetyl-CoA C13-label-14	0	0	0	0	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
Acetyl-CoA C13-label-16	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
Acetyl-CoA C13-label-18	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
Acetyl-CoA C13-label-20	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
Acetyl-CoA C13-label-22	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
Citrate C12 PARENT	50749.22	90416.37	68232.96	4.60E+07	5.32E+07	4.72E+07	7318016	8153300	8557449	4.60E+07	5.09E+07	5.87E+07
Citrate C13-label-1	0	8645.016	2483.104	5100949	5734612	5132770	923737.8	1004098	1060814	8780535	8926736	1.02E+07
Citrate C13-label-2	14267.82	6462.361	3033.769	2861848	3419971	3020921	1666841	1705839	1926251	6304330	6139938	6269946
Citrate C13-label-3	0	2347.451	0	236426.2	276655.4	253539.6	245325.8	246538	269890.2	904650.8	831916	714685.8
Citrate C13-label-4	0	0	0	17307.43	18986.1	25381.85	128121.9	118480.1	139395.7	234278.9	217880.4	127077.8
Citrate C13-label-5	0	0	0	0	0	0	21315.98	20300.1	27510.53	24939.67	16404.95	6205.666
Citrate C13-label-6	0	0	0	0	0	0	0	0	0	0	0	0
alpha-Ketoglutaric acid C12 PARENT	2021.681	3060.904	6586.473	3476431	1.16E+07	2314142	139079.5	1454644	953020	2253810	3928362	1241974
alpha-Ketoglutaric acid C13-label-1	0	0	0	310866.9	967246.4	210636.3	8902.117	167419.1	106670.4	306074.4	467557.4	139091.9
alpha-Ketoglutaric acid C13-label-2	2903.609	2288.962	0	135771	385048.8	96160.02	17236.83	203160.5	144882.2	199389.7	251787	66561.59
alpha-Ketoglutaric acid C13-label-3	0	0	0	0	25558.49	0	0	29350.52	6494.592	19202.67	21459.09	0
alpha-Ketoglutaric acid C13-label-4	0	0	0	0	0	0	0	0	0	0	0	0
alpha-Ketoglutaric acid C13-label-5	0	0	0	0	0	0	4574.171	0	0	0	6560.971	0
Succinate C12 PARENT	248731.9	230736.7	300881.9	8354366	7142640	9075381	931865.4	1090647	1353026	1.40E+08	1.65E+08	1.67E+08
Succinate C13-label-1	12947.16	16218.41	13268.65	677721.7	565681.6	748612.2	63877.36	94415.4	115669.1	1.33E+07	1.56E+07	1.41E+07
Succinate C13-label-2	7288.597	8393.331	6492.025	242494.5	212357.2	282946.9	65183.88	86237.12	82598.3	5353816	5886660	4919908
Succinate C13-label-3	0	0	0	16558.2	14545.09	29445.78	4595.92	7030.842	6046.619	505289.2	592762.5	440640.5
Succinate C13-label-4	0	0	0	0	0	0	0	0	0	0	0	0
Malate C12 PARENT	36573.07	46285.29	41278.44	4.00E+07	6.03E+07	4.90E+07	3.72E+07	4.40E+07	3.55E+07	2.89E+07	3.49E+07	4.35E+07
Malate C13-label-1	0	0	0	2969290	4571568	3649595	2607120	3257736	2679131	3609928	4147794	4624518
Malate C13-label-2	0	0	0	727603.2	1147657	923327.3	1574373	1858362	1744982	1411030	1477044	1378425
Malate C13-label-3	0	0	0	23170.33	62751.5	30333.73	306914.8	368230.7	291428.5	162474.6	181065.1	163374.8
Malate C13-label-4	0	0	2126.076	0	0	0	0	24986.53	0	0	0	0
L-Aspartic acid C12 PARENT	25925.12	66489.62	36476.63	9.47E+07	1.09E+08	1.12E+08	4.73E+07	5.31E+07	5.47E+07	7.44E+07	9.83E+07	8.47E+07
L-Aspartic acid C13-label-1	0	0	0	6230810	7073890	7212934	2422576	2684304	2719133	6079556	6854748	6254994
L-Aspartic acid C13-label-2	0	0	0	1126983	1179623	1180277	294887.1	351109.8	347145.5	1569162	1320416	1250205
L-Aspartic acid C13-label-3	0	0	0	75268.63	80419.34	79759.74	72488.16	95122.04	70875.8	216779.1	170736.5	150116
L-Aspartic acid C13-label-4	0	0	0	0	0	0	6561.094	0	6696.408	0	0	0
Palmitic acid C12 PARENT	1.04E+08	1.04E+08	1.05E+08	5.31E+07	6.52E+07	5.83E+07	6.26E+07	5.56E+07	5.77E+07	5.74E+07	6.20E+07	5.21E+07
Palmitic acid C13-label-1	1.83E+07	1.83E+07	1.84E+07	9427677	1.16E+07	1.04E+07	1.11E+07	9871776	1.02E+07	1.01E+07	1.09E+07	9240125
Palmitic acid C13-label-2	1724111	1615721	1673288	2430614	2806947	2666657	1196023	1206827	1128827	1036420	1140792	947820.8
Palmitic acid C13-label-3	141367.7	84580.38	105521.5	376185.4	428367.7	399672.9	123450.2	118687.8	107404.9	77112.01	64880.39	80103.29
Palmitic acid C13-label-4	0	0	0	1490166	1624415	1630882	348671.5	430492.7	356011.3	0	0	0
Palmitic acid C13-label-5	0	0	0	274497.6	297685	308955	66949.24	82110.18	85385.97	0	0	0
Palmitic acid C13-label-6	0	0	0	1105018	1224752	1233814	424114.9	558992.7	505654.9	0	0	0
Palmitic acid C13-label-7	0	0	0	176341.3	208937.6	205238.1	81222.91	103529.2	88767.91	0	0	0
Palmitic acid C13-label-8	0	0	0	748477.9	911732.7	857948.8	472896	558097.2	544537.2	0	0	0
Palmitic acid C13-label-9	0	0	0	120555.1	141719.6	130778.9	94939.66	99141.04	105565.2	0	0	0
Palmitic acid C13-label-10	0	0	0	501419.6	615540.1	555324.4	527312.3	531036.8	560597.4	0	0	0
Palmitic acid C13-label-11	0	0	0	59162.52	79052.54	78661.5	94261.05	93284.06	92246.56	0	0	0
Palmitic acid C13-label-12	0	0	0	276071.5	345315.8	340668.2	492157.8	468898.9	544618.8	4151.292	0	0
Palmitic acid C13-label-13	0	0	0	39139.87	41148.02	42433.81	82236.41	85064.44	94218.68	0	0	0
Palmitic acid C13-label-14	0	0	0	130625	163740.7	160444.1	512657.6	453436.6	497416.5	0	14614.27	8051.78
Palmitic acid C13-label-15	0	0	0	6908.92	0	0	50623.56	47439.67	64053.03	0	0	0
Palmitic acid C13-label-16	0	0	0	40964.19	58666.88	57448.89	309432.2	299572.7	332684.6	0	0	0
Myristic acid C12 PARENT	1714290	1725192	1673096	2817416	3080480	2945368	2910090	2491702	2709714	2204770	2464407	2510177
Myristic acid C13-label-1	253730.8	265883.5	272715.8	429615.6	454263.2	475299.4	418412.8	389446.2	409454.7	332977.9	378217.8	385761.1
Myristic acid C13-label-2	0	0	0	421054.2	450153.7	468869	148661.4	144022.7	136961.1	28588.55	39906.16	36657.48
Myristic acid C13-label-3	0	0	0	84676.56	74196.79	74788.38	21311.14	27352.37	20187.97	0	0	0
Myristic acid C13-label-4	0	0	0	323265.4	346751.7	336559.4	153101	176431.5	161893.4	0	0	0
Myristic acid C13-label-5	0	0	0	40961.02	55188.42	61263.29	9379.796	24181.1	15793.77	0	0	0
Myristic acid C13-label-6	0	0	0	230399.9	235669	223795	177915.3	190342.2	191771.6	0	0	0
Myristic acid C13-label-7	0	0	0	21646.89	36953.67	39876.98	26917.96	26266.69	31805.65	0	0	0
Myristic acid C13-label-8	0	0	0	140745.2	149672.5	154085.6	176834.4	187735.5	183469	0	0	0
Myristic acid C13-label-9	0	0	0	12161.4	21181.24	12941.07	31886.87	26617.01	24911.96	0	0	0
Myristic acid C13-label-10	0	0	0	74920.27	96202.96	87331.26	166897.6	167786.9	189636.7	0	0	0
Myristic acid C13-label-11	0	0	0	6955.331	0	6930.664	20386.14	16651.29	9777.247	0	0	0
Myristic acid C13-label-12	0	0	0	31041.35	36591.1	44829.64	151865.5	122017.6	146643.6	0	0	0
Myristic acid C13-label-13	0	0	0	0	0	0	7236.1	6930.512	9003.157	0	0	0
Myristic acid C13-label-14	0	0	0	0	5622.352	8410.413	95269.81	88742.28	79820.75	0	0	0
Palmitoleic acid C12 PARENT	226183.9	125190.7	205576.3	3548621	5456931	4541878	4898222	3941409	4060036	4507328	4922750	5301756
Palmitoleic acid C13-label-1	0	0	0	618302.3	996361	806733.3	755169.6	677032.6	691822.7	794211	791278.8	890294
Palmitoleic acid C13-label-2	0	0	0	0	0	0	0	0	0	0	0	0
Palmitoleic acid C13-label-3	0	0	0	0	0	0	0	0	0	0	0	0
Palmitoleic acid C13-label-4	0	0	0	175194.9	217579.2	214567.5	0	0	0	0	0	0
Palmitoleic acid C13-label-5	0	0	0	18584.53	45033.5	27202.64	0	4867.952	0	0	0	0
Palmitoleic acid C13-label-6	0	0	0	163927.7	260907.2	197059.6	84359.28	69179.3	57242.43	0	0	0
Palmitoleic acid C13-label-7	0	0	0	24051.2	24349.15	31637.5	8027.95	12635.71	6194.175	0	0	0
Palmitoleic acid C13-label-8	0	0	0	126769.5	206652.8	154774.6	66089.53	73486.59	63923.86	0	0	0
Palmitoleic acid C13-label-9	0	0	0	14577.21	26895.42	31544.24	0	0	6750.232	0	0	0
Palmitoleic acid C13-label-10	0	0	0	82307.66	134124	100464	83270.52	70778.57	68129.01	0	0	0
Palmitoleic acid C13-label-11	0	0	0	14004.1	10600.49	17140.93	7537.411	8164.6	5552.021	0	0	0
Palmitoleic acid C13-label-12	0	0	0	53815.04	83377.29	72668.64	87950.23	67515.11	81519.65	0	0	0
Palmitoleic acid C13-label-13	0	0	0	0	0	0	0	0	8246.184	0	0	0
Palmitoleic acid C13-label-14	0	0	0	24543.74	36290.84	26635.88	72282.66	77625.48	51586.29	0	0	0
Palmitoleic acid C13-label-15	0	0	0	0	0	0	0	4921.7	0	0	0	0
Palmitoleic acid C13-label-16	0	0	0	0	8370.884	6721.286	48575.03	47401.07	25199.44	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	8.228556	808.1192	808.1191	444493	C00024	50043
Acetyl-CoA C13-label-1	8.253796	809.1213	808.1191	444493	C00024	50043
Acetyl-CoA C13-label-2	8.244634	810.1243	808.1191	444493	C00024	50043
Acetyl-CoA C13-label-3	8.264252	811.1272	808.1191	444493	C00024	50043
Acetyl-CoA C13-label-4	8.252136	812.1301	808.1191	444493	C00024	50043
Acetyl-CoA C13-label-5	NA	NA	808.1191	444493	C00024	50043	Peak was not detected
Acetyl-CoA C13-label-6	NA	NA	808.1191	444493	C00024	50043	Peak was not detected
Acetyl-CoA C13-label-7	NA	NA	808.1191	444493	C00024	50043	Peak was not detected
Acetyl-CoA C13-label-8	NA	NA	808.1191	444493	C00024	50043	Peak was not detected
Acetyl-CoA C13-label-9	NA	NA	808.1191	444493	C00024	50043	Peak was not detected
Acetyl-CoA C13-label-10	NA	NA	808.1191	444493	C00024	50043	Peak was not detected
Acetyl-CoA C13-label-11	NA	NA	808.1191	444493	C00024	50043	Peak was not detected
Acetyl-CoA C13-label-12	NA	NA	808.1191	444493	C00024	50043	Peak was not detected
Acetyl-CoA C13-label-13	NA	NA	808.1191	444493	C00024	50043	Peak was not detected
Acetyl-CoA C13-label-14	NA	NA	808.1191	444493	C00024	50043	Peak was not detected
Acetyl-CoA C13-label-15	NA	NA	808.1191	444493	C00024	50043	Peak was not detected
Acetyl-CoA C13-label-16	NA	NA	808.1191	444493	C00024	50043	Peak was not detected
Acetyl-CoA C13-label-17	NA	NA	808.1191	444493	C00024	50043	Peak was not detected
Acetyl-CoA C13-label-18	NA	NA	808.1191	444493	C00024	50043	Peak was not detected
Acetyl-CoA C13-label-19	NA	NA	808.1191	444493	C00024	50043	Peak was not detected
Acetyl-CoA C13-label-20	NA	NA	808.1191	444493	C00024	50043	Peak was not detected
Acetyl-CoA C13-label-21	NA	NA	808.1191	444493	C00024	50043	Peak was not detected
Acetyl-CoA C13-label-22	NA	NA	808.1191	444493	C00024	50043	Peak was not detected
Acetyl-CoA C13-label-23	NA	NA	808.1191	444493	C00024	50043	Peak was not detected
Citrate C12 PARENT	11.00825	191.0197	191.0197	311	C00158	37071
Citrate C13-label-1	11.00965	192.023	191.0197	311	C00158	37071
Citrate C13-label-2	11.00648	193.0263	191.0197	311	C00158	37071
Citrate C13-label-3	11.01685	194.0297	191.0197	311	C00158	37071
Citrate C13-label-4	11.00274	195.0328	191.0197	311	C00158	37071
Citrate C13-label-5	11.00976	196.0364	191.0197	311	C00158	37071
Citrate C13-label-6	NA	NA	191.0197	311	C00158	37071	Peak was not detected
alpha-Ketoglutaric acid C12 PARENT	9.830396	145.0142	145.0142	51	C00026	37135
alpha-Ketoglutaric acid C13-label-1	9.835419	146.0176	145.0142	51	C00026	37135
alpha-Ketoglutaric acid C13-label-2	9.826958	147.0209	145.0142	51	C00026	37135
alpha-Ketoglutaric acid C13-label-3	9.84732	148.0244	145.0142	51	C00026	37135
alpha-Ketoglutaric acid C13-label-4	NA	NA	145.0142	51	C00026	37135	Peak was not detected
alpha-Ketoglutaric acid C13-label-5	9.851441	150.0316	145.0142	51	C00026	37135
Succinate C12 PARENT	9.777125	117.0193	117.0193	1110	C00042	1966
Succinate C13-label-1	9.769395	118.0227	117.0193	1110	C00042	1966
Succinate C13-label-2	9.78619	119.0259	117.0193	1110	C00042	1966
Succinate C13-label-3	9.728387	120.0293	117.0193	1110	C00042	1966
Succinate C13-label-4	NA	NA	117.0193	1110	C00042	1966	Peak was not detected
Malate C12 PARENT	10.12774	133.0142	133.0142	222656	C00149	37105
Malate C13-label-1	10.13109	134.0175	133.0142	222656	C00149	37105
Malate C13-label-2	10.12268	135.0211	133.0142	222656	C00149	37105
Malate C13-label-3	10.11847	136.0243	133.0142	222656	C00149	37105
Malate C13-label-4	10.09459	137.0279	133.0142	222656	C00149	37105
L-Aspartic acid C12 PARENT	9.575432	132.0302	132.0302	5960	C0004	37126
L-Aspartic acid C13-label-1	9.564569	133.0335	132.0302	5960	C0004	37126
L-Aspartic acid C13-label-2	9.562537	134.0371	132.0302	5960	C0004	37126
L-Aspartic acid C13-label-3	9.572783	135.0405	132.0302	5960	C0004	37126
L-Aspartic acid C13-label-4	9.578674	136.0442	132.0302	5960	C0004	37126
Palmitic acid C12 PARENT	1.339877	255.2329	255.2329	985	C00249	23
Palmitic acid C13-label-1	1.339877	256.2363	255.2329	985	C00249	23
Palmitic acid C13-label-2	1.341462	257.2393	255.2329	985	C00249	23
Palmitic acid C13-label-3	1.347659	258.2424	255.2329	985	C00249	23
Palmitic acid C13-label-4	1.336807	259.2462	255.2329	985	C00249	23
Palmitic acid C13-label-5	1.346431	260.2498	255.2329	985	C00249	23
Palmitic acid C13-label-6	1.333683	261.2529	255.2329	985	C00249	23
Palmitic acid C13-label-7	1.352558	262.2566	255.2329	985	C00249	23
Palmitic acid C13-label-8	1.33032	263.2597	255.2329	985	C00249	23
Palmitic acid C13-label-9	1.343081	264.2633	255.2329	985	C00249	23
Palmitic acid C13-label-10	1.339848	265.2664	255.2329	985	C00249	23
Palmitic acid C13-label-11	1.336688	266.2697	255.2329	985	C00249	23
Palmitic acid C13-label-12	1.364719	267.273	255.2329	985	C00249	23
Palmitic acid C13-label-13	1.336975	268.2766	255.2329	985	C00249	23
Palmitic acid C13-label-14	1.345827	269.2797	255.2329	985	C00249	23
Palmitic acid C13-label-15	1.344468	270.283	255.2329	985	C00249	23
Palmitic acid C13-label-16	1.33034	271.2866	255.2329	985	C00249	23
Myristic acid C12 PARENT	1.371463	227.2016	227.2016	11005	C06424	36
Myristic acid C13-label-1	1.386577	228.2048	227.2016	11005	C06424	36
Myristic acid C13-label-2	1.376782	229.2082	227.2016	11005	C06424	36
Myristic acid C13-label-3	1.384542	230.2118	227.2016	11005	C06424	36
Myristic acid C13-label-4	1.371615	231.215	227.2016	11005	C06424	36
Myristic acid C13-label-5	1.397231	232.2185	227.2016	11005	C06424	36
Myristic acid C13-label-6	1.368551	233.2216	227.2016	11005	C06424	36
Myristic acid C13-label-7	1.371328	234.2251	227.2016	11005	C06424	36
Myristic acid C13-label-8	1.365228	235.2283	227.2016	11005	C06424	36
Myristic acid C13-label-9	1.422878	236.2323	227.2016	11005	C06424	36
Myristic acid C13-label-10	1.387419	237.2352	227.2016	11005	C06424	36
Myristic acid C13-label-11	1.376315	238.2383	227.2016	11005	C06424	36
Myristic acid C13-label-12	1.383951	239.2418	227.2016	11005	C06424	36
Myristic acid C13-label-13	1.393341	240.2455	227.2016	11005	C06424	36
Myristic acid C13-label-14	1.428358	241.2484	227.2016	11005	C06424	36
Palmitoleic acid C12 PARENT	1.363024	253.2173	253.2173	445638	C08362	490
Palmitoleic acid C13-label-1	1.344916	254.2207	253.2173	445638	C08362	490
Palmitoleic acid C13-label-2	NA	NA	253.2173	445638	C08362	490	Peak was not detected
Palmitoleic acid C13-label-3	NA	NA	253.2173	445638	C08362	490	Peak was not detected
Palmitoleic acid C13-label-4	1.330609	257.2305	253.2173	445638	C08362	490
Palmitoleic acid C13-label-5	1.394058	258.2333	253.2173	445638	C08362	490
Palmitoleic acid C13-label-6	1.339797	259.2371	253.2173	445638	C08362	490
Palmitoleic acid C13-label-7	1.375131	260.2401	253.2173	445638	C08362	490
Palmitoleic acid C13-label-8	1.346047	261.2438	253.2173	445638	C08362	490
Palmitoleic acid C13-label-9	1.361002	262.2469	253.2173	445638	C08362	490
Palmitoleic acid C13-label-10	1.343238	263.2505	253.2173	445638	C08362	490
Palmitoleic acid C13-label-11	1.368292	264.2533	253.2173	445638	C08362	490
Palmitoleic acid C13-label-12	1.346535	265.257	253.2173	445638	C08362	490
Palmitoleic acid C13-label-13	1.335138	266.2609	253.2173	445638	C08362	490
Palmitoleic acid C13-label-14	1.381363	267.2639	253.2173	445638	C08362	490
Palmitoleic acid C13-label-15	1.469797	268.2673	253.2173	445638	C08362	490
Palmitoleic acid C13-label-16	1.35712	269.2706	253.2173	445638	C08362	490
METABOLITES_END
#END