#METABOLOMICS WORKBENCH KeiRodrz30_20220208_122818 DATATRACK_ID:3070 STUDY_ID:ST002080 ANALYSIS_ID:AN003394 PROJECT_ID:PR001320
VERSION             	1
CREATED_ON             	February 8, 2022, 1:51 pm
#PROJECT
PR:PROJECT_TITLE                 	Effects of Ferroptosis on the Metabolome in Cardiac Cells: The Role of
PR:PROJECT_TITLE                 	Glutaminolysis
PR:PROJECT_TYPE                  	GCMS
PR:PROJECT_SUMMARY               	Ferroptosis is a novel iron-dependent regulated cell death mechanism that
PR:PROJECT_SUMMARY               	affects cell metabolism; however, a detailed metabolomic analysis of ferroptotic
PR:PROJECT_SUMMARY               	cells is not yet available. Here, we elucidated the metabolome of H9c2
PR:PROJECT_SUMMARY               	cardioblasts by gas chromatography-mass spectrometry during ferroptosis induced
PR:PROJECT_SUMMARY               	by RSL3, a GPX4 inhibitor, in the presence of ferrostatin-1 (a ferroptosis
PR:PROJECT_SUMMARY               	inhibitor), XJB-5-131 (a mitochondrial-targeted ROS scavenger), or TSM-1005-44
PR:PROJECT_SUMMARY               	(a newly developed cellular ROS scavenger). Results demonstrated that RSL3
PR:PROJECT_SUMMARY               	decreased the levels of amino acids involved in glutathione synthesis more than
PR:PROJECT_SUMMARY               	two-fold. In contrast, saturated fatty acids levels were markedly increased in
PR:PROJECT_SUMMARY               	RSL3-challenged cells, with no effects on unsaturated fatty acids. RSL3
PR:PROJECT_SUMMARY               	significantly altered the levels of mitochondrial tricarboxylic acid cycle
PR:PROJECT_SUMMARY               	intermediates; isocitrate and 2-oxoglutarate were found to increase, whereas
PR:PROJECT_SUMMARY               	succinate was significantly decreased in RSL3-challenged cells. Ferrostatin-1,
PR:PROJECT_SUMMARY               	XJB-5-131, and TSM-1005-44 prevented RSL3-induced cell death and conserved the
PR:PROJECT_SUMMARY               	metabolomic profile of the cells. Since 2-oxoglutarate is involved in the
PR:PROJECT_SUMMARY               	regulation of ferroptosis, particularly through glutamine metabolism, we further
PR:PROJECT_SUMMARY               	assessed the role of glutaminolysis in ferroptosis in H9c2 cardioblasts. Genetic
PR:PROJECT_SUMMARY               	silencing of GLS1, which encodes the K-type mitochondrial glutaminase
PR:PROJECT_SUMMARY               	(glutaminase C), protected against ferroptosis in the early stage. In
PR:PROJECT_SUMMARY               	conclusion, our study demonstrates that RSL3-induced ferroptosis impairs the
PR:PROJECT_SUMMARY               	metabolome of H9c2 cardioblasts.
PR:INSTITUTE                     	University of Puerto Rico, School of Medicine
PR:DEPARTMENT                    	Physiology
PR:LABORATORY                    	Cardiovascular Physiology, DR. Sabzali Javadov's Lab
PR:LAST_NAME                     	Rodriguez-Graciani
PR:FIRST_NAME                    	Keishla M
PR:ADDRESS                       	Medical Sciences Campus, Main Building, 6th Floor, Department of Physiology, San
PR:ADDRESS                       	Juan, Puerto Rico, 00936-5067, USA
PR:EMAIL                         	keishla.rodriguez20@upr.edu
PR:PHONE                         	7877582525x26888
PR:FUNDING_SOURCE                	This research was funded by the National Institutes of Health (Grants
PR:FUNDING_SOURCE                	SC1GM128210, R25GM061838, U54MD007600, P20GM103475, HDTRA1-16-1-0041, and
PR:FUNDING_SOURCE                	U19-AI1068021).
PR:PUBLICATIONS                  	https://www.mdpi.com/1476010, Antioxidants 2022, 11(2), 278;
PR:PUBLICATIONS                  	https://doi.org/10.3390/antiox11020278
PR:CONTRIBUTORS                  	Xavier R. Chapa-Dubocq, Esteban J. Ayala-Arroyo, Ivana Chaves-Negrón, Sehwan
PR:CONTRIBUTORS                  	Jang, Nataliya Chorna, Taber S. Maskrey, Peter Wipf and Sabzali Javadov
#STUDY
ST:STUDY_TITLE                   	Effects of Ferroptosis on the Metabolome in Cardiac Cells: The Role of
ST:STUDY_TITLE                   	Glutaminolysis
ST:STUDY_TYPE                    	GCMS
ST:STUDY_SUMMARY                 	Ferroptosis is a novel iron-dependent regulated cell death mechanism that
ST:STUDY_SUMMARY                 	affects cell metabolism; however, a detailed metabolomic analysis of ferroptotic
ST:STUDY_SUMMARY                 	cells is not yet available. Here, we elucidated the metabolome of H9c2
ST:STUDY_SUMMARY                 	cardioblasts by gas chromatography-mass spectrometry during ferroptosis induced
ST:STUDY_SUMMARY                 	by RSL3, a GPX4 inhibitor, in the presence of ferrostatin-1 (a ferroptosis
ST:STUDY_SUMMARY                 	inhibitor), XJB-5-131 (a mitochondrial-targeted ROS scavenger), or TSM-1005-44
ST:STUDY_SUMMARY                 	(a newly developed cellular ROS scavenger). Results demonstrated that RSL3
ST:STUDY_SUMMARY                 	decreased the levels of amino acids involved in glutathione synthesis more than
ST:STUDY_SUMMARY                 	two-fold. In contrast, saturated fatty acids levels were markedly increased in
ST:STUDY_SUMMARY                 	RSL3-challenged cells, with no effects on unsaturated fatty acids. RSL3
ST:STUDY_SUMMARY                 	significantly altered the levels of mitochondrial tricarboxylic acid cycle
ST:STUDY_SUMMARY                 	intermediates; isocitrate and 2-oxoglutarate were found to increase, whereas
ST:STUDY_SUMMARY                 	succinate was significantly decreased in RSL3-challenged cells. Ferrostatin-1,
ST:STUDY_SUMMARY                 	XJB-5-131, and TSM-1005-44 prevented RSL3-induced cell death and conserved the
ST:STUDY_SUMMARY                 	metabolomic profile of the cells. Since 2-oxoglutarate is involved in the
ST:STUDY_SUMMARY                 	regulation of ferroptosis, particularly through glutamine metabolism, we further
ST:STUDY_SUMMARY                 	assessed the role of glutaminolysis in ferroptosis in H9c2 cardioblasts. Genetic
ST:STUDY_SUMMARY                 	silencing of GLS1, which encodes the K-type mitochondrial glutaminase
ST:STUDY_SUMMARY                 	(glutaminase C), protected against ferroptosis in the early stage. In
ST:STUDY_SUMMARY                 	conclusion, our study demonstrates that RSL3-induced ferroptosis impairs the
ST:STUDY_SUMMARY                 	metabolome of H9c2 cardioblasts.
ST:INSTITUTE                     	University of Puerto Rico, School of Medicine
ST:DEPARTMENT                    	Physiology
ST:LABORATORY                    	Cardiovascular Physiology, DR. Sabzali Javadov's Lab
ST:LAST_NAME                     	Rodriguez-Graciani
ST:FIRST_NAME                    	Keishla M
ST:ADDRESS                       	Medical Sciences Campus, Main Building, 6th Floor, Department of Physiology, San
ST:ADDRESS                       	Juan, Puerto Rico, 00936-5067, USA
ST:EMAIL                         	keishla.rodriguez20@upr.edu
ST:PHONE                         	7877582525x26888
ST:NUM_GROUPS                    	8
ST:PUBLICATIONS                  	https://www.mdpi.com/1476010, Antioxidants 2022, 11(2), 278;
ST:PUBLICATIONS                  	https://doi.org/10.3390/antiox11020278
#SUBJECT
SU:SUBJECT_TYPE                  	Cultured cells
SU:SUBJECT_SPECIES               	Rattus norvegicus
SU:TAXONOMY_ID                   	10116
SU:CELL_BIOSOURCE_OR_SUPPLIER    	American Type Culture Collection (ATCC)
SU:CELL_STRAIN_DETAILS           	H9c2(2-1) is a subclone of the original clonal cell line derived from embryonic
SU:CELL_STRAIN_DETAILS           	BD1X rat heart tissue by B. Kimes and B. Brandt and exhibits many of the
SU:CELL_STRAIN_DETAILS           	properties of skeletal muscle.
SU:CELL_PASSAGE_NUMBER           	3-10
SU:CELL_COUNTS                   	80-90% confluency
#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           	C	Control_ 1	Treatment:Control	RAW_FILE_NAME=C1_UNK-0010_7132021_10
SUBJECT_SAMPLE_FACTORS           	C	Control_ 2	Treatment:Control	RAW_FILE_NAME=C2_UNK-0030_7132021_30
SUBJECT_SAMPLE_FACTORS           	C	Control_ 3	Treatment:Control	RAW_FILE_NAME=C4_UNK-0021_7132021_21
SUBJECT_SAMPLE_FACTORS           	C	Control_ 4	Treatment:Control	RAW_FILE_NAME=C5_UNK-0017_7132021_17
SUBJECT_SAMPLE_FACTORS           	C	Control_ 5	Treatment:Control	RAW_FILE_NAME=C6_UNK-0041_7132021_41
SUBJECT_SAMPLE_FACTORS           	R	RSL3_1	Treatment:RSL3	RAW_FILE_NAME=R1_UNK-0012_7132021_12
SUBJECT_SAMPLE_FACTORS           	R	RSL3_2	Treatment:RSL3	RAW_FILE_NAME=R2_UNK-0043_7132021_43
SUBJECT_SAMPLE_FACTORS           	R	RSL3_3	Treatment:RSL3	RAW_FILE_NAME=R4_UNK-0037_7132021_37
SUBJECT_SAMPLE_FACTORS           	R	RSL3_4	Treatment:RSL3	RAW_FILE_NAME=R5_UNK-0009_7132021_9
SUBJECT_SAMPLE_FACTORS           	R	RSL3_5	Treatment:RSL3	RAW_FILE_NAME=R6_UNK-0034_7132021_34
SUBJECT_SAMPLE_FACTORS           	R	RSL3_6	Treatment:RSL3	RAW_FILE_NAME=R8_UNK-0040_7132021_40
SUBJECT_SAMPLE_FACTORS           	F	Fer1-1	Treatment:Ferrostatin-1	RAW_FILE_NAME=F1_UNK-0003_7132021_3
SUBJECT_SAMPLE_FACTORS           	F	Fer1-2	Treatment:Ferrostatin-1	RAW_FILE_NAME=F2_UNK-0036_7132021_36
SUBJECT_SAMPLE_FACTORS           	F	Fer1-3	Treatment:Ferrostatin-1	RAW_FILE_NAME=F4_UNK-0046_7132021_46
SUBJECT_SAMPLE_FACTORS           	F	Fer1-4	Treatment:Ferrostatin-1	RAW_FILE_NAME=F5_UNK-0020_7132021_20
SUBJECT_SAMPLE_FACTORS           	F	Fer1-5	Treatment:Ferrostatin-1	RAW_FILE_NAME=F6_UNK-0027_7132021_27
SUBJECT_SAMPLE_FACTORS           	F	Fer1-6	Treatment:Ferrostatin-1	RAW_FILE_NAME=F8_UNK-0011_7132021_11
SUBJECT_SAMPLE_FACTORS           	X	XJB_1	Treatment:XJB-5-131	RAW_FILE_NAME=X1_UNK-0025_7132021_25
SUBJECT_SAMPLE_FACTORS           	X	XJB_2	Treatment:XJB-5-131	RAW_FILE_NAME=X2_UNK-0015_7132021_15
SUBJECT_SAMPLE_FACTORS           	X	XJB_3	Treatment:XJB-5-131	RAW_FILE_NAME=X5_UNK-0019_7132021_19
SUBJECT_SAMPLE_FACTORS           	X	XJB_4	Treatment:XJB-5-131	RAW_FILE_NAME=X6_UNK-0006_7132021_6
SUBJECT_SAMPLE_FACTORS           	X	XJB_5	Treatment:XJB-5-131	RAW_FILE_NAME=X8_UNK-0028_7132021_28
SUBJECT_SAMPLE_FACTORS           	T	TSM_1	Treatment:TSM-1005-44	RAW_FILE_NAME=T1_UNK-0023_7132021_23
SUBJECT_SAMPLE_FACTORS           	T	TSM_3	Treatment:TSM-1005-44	RAW_FILE_NAME=T4_UNK-0032_7132021_32
SUBJECT_SAMPLE_FACTORS           	T	TSM_4	Treatment:TSM-1005-44	RAW_FILE_NAME=T5_UNK-0035_7132021_35
SUBJECT_SAMPLE_FACTORS           	T	TSM_5	Treatment:TSM-1005-44	RAW_FILE_NAME=T6_UNK-0048_7132021_48
SUBJECT_SAMPLE_FACTORS           	T	TSM_6	Treatment:TSM-1005-44	RAW_FILE_NAME=T8_UNK-0008_7132021_8
SUBJECT_SAMPLE_FACTORS           	RF	RF_1	Treatment:RSL3 + Ferrostatin-1	RAW_FILE_NAME=RF1_UNK-0004_7132021_4
SUBJECT_SAMPLE_FACTORS           	RF	RF_2	Treatment:RSL3 + Ferrostatin-1	RAW_FILE_NAME=RF2_UNK-0014_7132021_14
SUBJECT_SAMPLE_FACTORS           	RF	RF_3	Treatment:RSL3 + Ferrostatin-1	RAW_FILE_NAME=RF4_UNK-0024_7132021_24
SUBJECT_SAMPLE_FACTORS           	RF	RF_4	Treatment:RSL3 + Ferrostatin-1	RAW_FILE_NAME=RF5_UNK-0044_7132021_44
SUBJECT_SAMPLE_FACTORS           	RF	RF_5	Treatment:RSL3 + Ferrostatin-1	RAW_FILE_NAME=RF6_UNK-0047_7132021_47
SUBJECT_SAMPLE_FACTORS           	RF	RF_6	Treatment:RSL3 + Ferrostatin-1	RAW_FILE_NAME=RF8_UNK-0029_7132021_29
SUBJECT_SAMPLE_FACTORS           	RX	RX_1	Treatment:RSL3 + XJB-5-131	RAW_FILE_NAME=RX1_UNK-0031_7132021_31
SUBJECT_SAMPLE_FACTORS           	RX	RX_2	Treatment:RSL3 + XJB-5-131	RAW_FILE_NAME=RX2_UNK-0007_7132021_7
SUBJECT_SAMPLE_FACTORS           	RX	RX_3	Treatment:RSL3 + XJB-5-131	RAW_FILE_NAME=RX4_UNK-0016_7132021_16
SUBJECT_SAMPLE_FACTORS           	RX	RX_4	Treatment:RSL3 + XJB-5-131	RAW_FILE_NAME=RX5_UNK-0026_7132021_26
SUBJECT_SAMPLE_FACTORS           	RX	RX_5	Treatment:RSL3 + XJB-5-131	RAW_FILE_NAME=RX6_UNK-0038_7132021_38
SUBJECT_SAMPLE_FACTORS           	RX	RX_6	Treatment:RSL3 + XJB-5-131	RAW_FILE_NAME=RX8_UNK-0013_7132021_13
SUBJECT_SAMPLE_FACTORS           	RT	RT_1	Treatment:RSL3 + TSM-1005-44	RAW_FILE_NAME=RT1_UNK-0042_7132021_42
SUBJECT_SAMPLE_FACTORS           	RT	RT_2	Treatment:RSL3 + TSM-1005-44	RAW_FILE_NAME=RT4_UNK-0022_7132021_22
SUBJECT_SAMPLE_FACTORS           	RT	RT_3	Treatment:RSL3 + TSM-1005-44	RAW_FILE_NAME=RT5_UNK-0039_7132021_39
SUBJECT_SAMPLE_FACTORS           	RT	RT_4	Treatment:RSL3 + TSM-1005-44	RAW_FILE_NAME=RT6_UNK-0001_7152021_1
SUBJECT_SAMPLE_FACTORS           	RT	RT_5	Treatment:RSL3 + TSM-1005-44	RAW_FILE_NAME=RT8_UNK-0045_7132021_45
#COLLECTION
CO:COLLECTION_SUMMARY            	H9c2 cardioblasts were cultured according to the manufacturer’s
CO:COLLECTION_SUMMARY            	recommendations (ATCC, Manassas, VA, USA) with minor modifications. Briefly, the
CO:COLLECTION_SUMMARY            	cells were incubated in DMEM based modified media containing 4 mM L-glutamine,
CO:COLLECTION_SUMMARY            	4.5 g/L glucose, 1 mM sodium pyruvate, and 1.5 g/L sodium bicarbonate, pH 7.4,
CO:COLLECTION_SUMMARY            	and supplemented with 10% fetal bovine serum and 1% antibiotic solution
CO:COLLECTION_SUMMARY            	(HyClone, Logan, UT, USA) in a CO2 incubator containing 95% air and 5% CO2 at 37
CO:COLLECTION_SUMMARY            	◦C. Cells with 80–90% confluence from passages 3–10 were used for
CO:COLLECTION_SUMMARY            	experiments. All chemicals were purchased from Sigma-Aldrich (St. Louis, MO).
CO:COLLECTION_SUMMARY            	H9c2 cardioblasts demonstrate similar to the primary cardiomyocytes hypertrophic
CO:COLLECTION_SUMMARY            	response as well as mitochondrial metabolism and morphology.
CO:SAMPLE_TYPE                   	Cultured cells
#TREATMENT
TR:TREATMENT_SUMMARY             	Cultured H9c2 cells reaching 80–90% confluence from passages 3–10 were used
TR:TREATMENT_SUMMARY             	for experiments. Ferroptosis was induced by incubating the cells with 0.5 µM
TR:TREATMENT_SUMMARY             	RSL3 for 3 h. In addition, the cells were exposed to RSL3 in the presence of 1
TR:TREATMENT_SUMMARY             	µM Fer-1, 0.2 µM XJB, and 0.6 µM TSM. Thus, experiments were performed in the
TR:TREATMENT_SUMMARY             	following 8 groups: (i) control (n = 5), (ii) RSL3 (n = 6), (iii) Fer-1 (n = 6),
TR:TREATMENT_SUMMARY             	(iv) XJB (n = 5), (v) TSM (n = 5), (vi) RSL3 + Fer-1 (n = 6), (vii) RSL3 + XJB
TR:TREATMENT_SUMMARY             	(n = 6), and (viii) RSL3 + TSM (n = 5). Fer-1, XJB, or TSM were added to the
TR:TREATMENT_SUMMARY             	culture medium simultaneously with RSL3. At the end of the incubation, the cells
TR:TREATMENT_SUMMARY             	were harvested and processed for analysis by GC-MS.
TR:CELL_PCT_CONFLUENCE           	80-90%
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Metabolites were extracted as previously described using 1 mL of cold MeOH/H2O
SP:SAMPLEPREP_SUMMARY            	(85:15). Samples were sonicated for 15 s (3×) on ice and centrifuged at 1400
SP:SAMPLEPREP_SUMMARY            	rpm × 10 min at 4 ◦C (Rotor model: Eppendorf, FA-45-30-11). For protein
SP:SAMPLEPREP_SUMMARY            	quantification, the pellets were dried for 15 min in a rotary vacuum evaporator,
SP:SAMPLEPREP_SUMMARY            	then resuspended in 60 µL of denaturation buffer and sonicated for 1 min. All
SP:SAMPLEPREP_SUMMARY            	samples were centrifuged for 10 min at the maximum speed and the supernatant was
SP:SAMPLEPREP_SUMMARY            	used for protein quantification using the Bradford method. Supernatants were
SP:SAMPLEPREP_SUMMARY            	collected and evaporated to dryness using a SpeedVac (Savant AS160, Farmingdale,
SP:SAMPLEPREP_SUMMARY            	NY, USA). The metabolite samples were first derivatized through methoxyamination
SP:SAMPLEPREP_SUMMARY            	by adding 30 µL of 20 mg/mL solution of methoxyamine hydrochloride Antioxidants
SP:SAMPLEPREP_SUMMARY            	2022, 11, 278 4 of 16 in pyridine (Sigma-Aldrich, St. Louis, MO, USA) and
SP:SAMPLEPREP_SUMMARY            	incubated at 37 ◦C for 2 h. Afterward, trimethylsilylation was performed by
SP:SAMPLEPREP_SUMMARY            	adding 30 µL of N-tert-butyldimethylsilyl-Nmethyltrifluoroacetamide (MTBSTFA +
SP:SAMPLEPREP_SUMMARY            	1% TBDMSCl; Sigma-Aldrich, St. Louis, MO, USA) and incubated for 1 h at 65 ◦C.
SP:SAMPLEPREP_SUMMARY            	The reaction mixture was centrifuged at 1300 rpm × 10 min at RT. Supernatants
SP:SAMPLEPREP_SUMMARY            	were transferred to analytical vials. Then, 20 µL per sample was added to glass
SP:SAMPLEPREP_SUMMARY            	vials with inserts followed by the addition of 1 mM 2-fluobiphenyl
SP:SAMPLEPREP_SUMMARY            	(Sigma-Aldrich, St. Louis, MO, USA) as an internal standard. Samples were
SP:SAMPLEPREP_SUMMARY            	processed by GC-MS-QP2010 (Shimadzu Scientific Instruments, Inc., Columbia, MD)
SP:SAMPLEPREP_SUMMARY            	using analytical conditions as previously described.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_TYPE           	GC
CH:INSTRUMENT_NAME               	Shimadzu GCMS-QP2010 ultra
CH:COLUMN_NAME                   	Shimadzu SH-RXI (30M x 0.25mm x 0.25um)
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Shimadzu QP2010 Ultra
MS:INSTRUMENT_TYPE               	Single quadrupole
MS:MS_TYPE                       	EI
MS:ION_MODE                      	POSITIVE
MS:MS_COMMENTS                   	Raw chromatography data were obtained and processed in GC-MS Labsolution Postrun
MS:MS_COMMENTS                   	Analysis software (Shimadzu Scientific Instruments, Inc., Columbia, MD) for
MS:MS_COMMENTS                   	identification of metabolites from their electron impact mass spectra by
MS:MS_COMMENTS                   	comparison to the database (NIST14/2014/EPA/NIH) [26]. Peak integration for all
MS:MS_COMMENTS                   	metabolites and multiple searches in the mass spectral library database resulted
MS:MS_COMMENTS                   	in a final database of 52 metabolic features chosen for this analysis. To assess
MS:MS_COMMENTS                   	analytical accuracy and precision, an external quality evaluation was performed
MS:MS_COMMENTS                   	using 2-fluorobiphenyl spiked into derivatization blank samples before running
MS:MS_COMMENTS                   	on the GC-MS. The quantitative analysis corresponding to the metabolite
MS:MS_COMMENTS                   	concentrations in each sample was calculated based on the internal standard in
MS:MS_COMMENTS                   	mM. A table in the format of comma-delimited (* csv) was created and uploaded to
MS:MS_COMMENTS                   	MetaboAnalyst.ca.
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	mM
MS_METABOLITE_DATA_START
Samples	Control_ 1	Control_ 2	Control_ 3	Control_ 4	Control_ 5	RSL3_1	RSL3_2	RSL3_3	RSL3_4	RSL3_5	RSL3_6	Fer1-1	Fer1-2	Fer1-3	Fer1-4	Fer1-5	Fer1-6	XJB_1	XJB_2	XJB_3	XJB_4	XJB_5	TSM_1	TSM_3	TSM_4	TSM_5	TSM_6	RF_1	RF_2	RF_3	RF_4	RF_5	RF_6	RX_1	RX_2	RX_3	RX_4	RX_5	RX_6	RT_1	RT_2	RT_3	RT_4	RT_5
Factors	Treatment:Control	Treatment:Control	Treatment:Control	Treatment:Control	Treatment:Control	Treatment:RSL3	Treatment:RSL3	Treatment:RSL3	Treatment:RSL3	Treatment:RSL3	Treatment:RSL3	Treatment:Ferrostatin-1	Treatment:Ferrostatin-1	Treatment:Ferrostatin-1	Treatment:Ferrostatin-1	Treatment:Ferrostatin-1	Treatment:Ferrostatin-1	Treatment:XJB-5-131	Treatment:XJB-5-131	Treatment:XJB-5-131	Treatment:XJB-5-131	Treatment:XJB-5-131	Treatment:TSM-1005-44	Treatment:TSM-1005-44	Treatment:TSM-1005-44	Treatment:TSM-1005-44	Treatment:TSM-1005-44	Treatment:RSL3 + Ferrostatin-1	Treatment:RSL3 + Ferrostatin-1	Treatment:RSL3 + Ferrostatin-1	Treatment:RSL3 + Ferrostatin-1	Treatment:RSL3 + Ferrostatin-1	Treatment:RSL3 + Ferrostatin-1	Treatment:RSL3 + XJB-5-131	Treatment:RSL3 + XJB-5-131	Treatment:RSL3 + XJB-5-131	Treatment:RSL3 + XJB-5-131	Treatment:RSL3 + XJB-5-131	Treatment:RSL3 + XJB-5-131	Treatment:RSL3 + TSM-1005-44	Treatment:RSL3 + TSM-1005-44	Treatment:RSL3 + TSM-1005-44	Treatment:RSL3 + TSM-1005-44	Treatment:RSL3 + TSM-1005-44
L-Isoleucine	0.023855059	0.015279335	0.014545088	0.022637229	0.021945321	0.050670915	0.014616325	0.003900552	0.01927543	0.032504553	0.016565365	0.016415414	0.014712377	0.017001869	0.016562404	0.013118972	0.01568373	0.029007589	0.014969662	0.017246555	0.012363484	0.025104264	0.029031745	0.017385558	0.02136262	0.018028206	0.014678176	0.016185173	0.028696025	0.039483825	0.011957117	0.027405923	0.014321944	0.030243222	0.011542392	0.007411324	0.009537635	0.023201998	0.00568887	0.018364857	0.017640258	0.0151836	0.008833551	0.01138563
Lactic acid	0.125939804	0.09337406	0.129137651	0.107762424	0.126047214	0.290160595	0.219909143	0.047871635	0.229090977	0.213727032	0.171058054	0.099142715	0.092978155	0.137681627	0.122138602	0.095156206	0.112740426	0.20080092	0.119458102	0.134235546	0.104478705	0.147034117	0.124025908	0.171012849	0.148407457	0.101565986	0.123562074	0.111703872	0.173516058	0.172300966	0.085654391	0.146647882	0.121644252	0.182441848	0.082091554	0.130021092	0.073764815	0.130506573	0.106830095	0.133670191	0.11004941	0.1309109	0.075495975	0.11368298
Glycolic acid	0.003298776	0.002750013	0.00279402	0.00225361	0.002337595	0.008359305	0.00928996	0.005528053	0.008023406	0.004075016	0.007062717	0.002854614	0.002689416	0.002704701	0.002724213	0.002101356	0.003206289	0.005256121	0.003379105	0.002769912	0.00278366	0.003064505	0.003892295	0.004619428	0.003239358	0.002078777	0.003094245	0.003272378	0.004418248	0.00407853	0.001797896	0.002541607	0.003040179	0.004709193	0.002570095	0.007665224	0.001704929	0.002923804	0.00465892	0.003328813	0.00298039	0.002979955	0.001582257	0.002760576
Alanine	0.019916353	0.0124674	0.023820018	0.007436345	0.015180484	0.005457954	0.007740356	0.001858652	0.012240356	0.012090271	0.003304846	0.025523637	0.017383314	0.013572095	0.024339889	0.016856794	0.015847044	0.041730264	0.040934403	0.034221722	0.057073815	0.013994412	0.004819665	0.032199667	0.022640662	0.016491615	0.02545502	0.009007378	0.038142154	0.023005063	0.014487864	0.020140526	0.024724845	0.033358202	0.014606169	0.007120641	0.013942522	0.017292926	0.011129884	0.031020603	0.024222223	0.016493338	0.022994569	0.018048993
Glycine	0.063840525	0.055057513	0.072067617	0.024409483	0.052730001	0.012455841	0.019791921	0.004031489	0.033243655	0.047786735	0.011219615	0.067215485	0.07845006	0.05836014	0.076499691	0.064205831	0.065271092	0.12748852	0.111181359	0.088308854	0.090294642	0.060506428	0.01950663	0.100818504	0.082785148	0.060985084	0.088640106	0.034444617	0.119104367	0.076702189	0.053142508	0.060760672	0.085520447	0.096927682	0.044709419	0.018230028	0.047627833	0.053244145	0.030408211	0.087822205	0.08649512	0.070001044	0.051205763	0.069497966
Uracil	0.003162802	0.001429961	0.002009151	0.002401971	0.001621994	0.003262433	0.003176915	0.001582265	0.002487188	0.001544836	0.00000291	0.002658518	0.001721513	0.001493722	0.00123582	0.001013025	0.001103685	0.002510905	0.002192024	0.001785973	0.001606453	0.000801079	0.00173	0.001141859	0.0012855	0.001030812	0.001241423	0.002421561	0.001955378	0.002287316	0.001186378	0.001141896	0.000996756	0.0024939	0.001715902	0.002234229	0.001307734	0.00073331	0.001432043	0.001668969	0.001238628	0.000994586	0.001063623	0.001031085
beta-Alanine	0.011396068	0.01234891	0.013041611	0.001847765	0.008002482	0.00000215	0.001886448	0.00000304	0.004131464	0.007797701	0.00000291	0.013948524	0.01822585	0.009334721	0.017109656	0.016923626	0.01206034	0.0278	0.02855452	0.0211	0.021619516	0.009560429	0.000996	0.019132842	0.018813057	0.012603783	0.01843297	0.003974625	0.025698802	0.012172658	0.0113064	0.009325586	0.016088631	0.019742929	0.009140778	0.002897013	0.011472803	0.006917111	0.004890424	0.018848649	0.019229682	0.012208137	0.012511541	0.012592513
Urea	0.025203151	0.021177962	0.02105714	0.004486013	0.011758218	0.013045691	0.022690503	0.013805086	0.032874656	0.011604238	0.017193939	0.025987716	0.016250056	0.009802435	0.017986778	0.01552988	0.015578684	0.045296618	0.027834518	0.025615686	0.025055257	0.02017146	0.006068766	0.016815185	0.017004105	0.013560557	0.035363828	0.00661209	0.048122711	0.027918167	0.011511029	0.024877044	0.016415371	0.034104469	0.009460894	0.012440713	0.009558572	0.009997798	0.013172927	0.027555483	0.016481395	0.012539299	0.008635967	0.011491087
Valine	0.020787539	0.014269447	0.022485845	0.004873203	0.015756248	0.012161129	0.016355478	0.005846182	0.024094407	0.018629614	0.014179758	0.024051179	0.015893344	0.011651557	0.023011971	0.020650298	0.014564074	0.03309837	0.030163173	0.027814001	0.037959146	0.023039821	0.003626005	0.024327372	0.020962109	0.018524942	0.021619451	0.007433427	0.038921444	0.031898968	0.013817861	0.02869375	0.018854831	0.033690348	0.012627753	0.008780852	0.013498171	0.020783212	0.011965586	0.027968478	0.020281415	0.014792831	0.015394921	0.013783072
Capric acid	0.004974783	0.003301786	0.00424601	0.003879527	0.004294504	0.010000442	0.014341568	0.009766327	0.01286882	0.00000153	0.011966246	0.004389389	0.004132422	0.000454279	0.004146455	0.003677007	0.004969304	0.006964006	0.005569322	0.004595295	0.004616695	0.00542	0.004479175	0.006026406	0.004506783	0.003411426	0.005339432	0.005319985	0.006811212	0.006178835	0.002975503	0.004434405	0.004891581	0.006900755	0.004111617	0.012046049	0.002766703	0.002961368	0.007084671	0.004586982	0.004960107	0.004863502	0.002669682	0.004365559
Leucine	0.016711224	0.010679271	0.02005207	0.002923072	0.012816949	0.006743363	0.012452284	0.003228068	0.017757923	0.013559507	0.009061017	0.019799853	0.012984735	0.009176078	0.01929537	0.017083137	0.011621984	0.028694287	0.02731802	0.024318887	0.036607297	0.017865028	0.001881491	0.021018692	0.017069881	0.01561812	0.018663946	0.005097496	0.032113724	0.024267202	0.011505864	0.023792654	0.017341417	0.028298857	0.010063972	0.005914626	0.011147035	0.016873182	0.009926102	0.025326094	0.016561839	0.012356541	0.013399795	0.012074539
Nicotinamide N-oxide	0.00592835	0.003754863	0.00467984	0.00024683	0.003576068	0.002911867	0.003612366	0.00079789	0.004745878	0.005106227	0.001406985	0.00539224	0.004611871	0.004170515	0.005393097	0.003703522	0.004126976	0.007739983	0.005581857	0.006159536	0.005158855	0.004553699	0.00000133	0.005818934	0.00598125	0.003415925	0.005289264	0.001221427	0.007635139	0.006748857	0.003062027	0.004283403	0.004952085	0.00649	0.003308456	0.003041291	0.003017948	0.004120671	0.002452072	0.005032907	0.00476337	0.004335304	0.002809818	0.003740921
Isoleucine	0.016457639	0.011137714	0.019215012	0.002642118	0.013302931	0.00620341	0.010198287	0.002179869	0.016964999	0.014577685	0.007854586	0.019302386	0.012644601	0.009529272	0.019312243	0.0171309	0.011644083	0.028588968	0.025856808	0.024346699	0.036555308	0.018807974	0.002241494	0.02068118	0.01776113	0.016215847	0.017956655	0.004821652	0.032815108	0.026307814	0.01147224	0.024670974	0.015800568	0.027361851	0.010102372	0.004273026	0.011715826	0.018021415	0.008109857	0.0247067	0.017139275	0.011897637	0.012915785	0.011135413
5-oxoprolinate	0.045893503	0.034936198	0.031363909	0.098743207	0.051221782	0.11116158	0.014674322	0.00247599	0.023131999	0.07531309	0.020008806	0.017199658	0.028393791	0.053001295	0.037748695	0.026448178	0.029852092	0.052716846	0.014271896	0.033711698	0.015504803	0.053299401	0.140169477	0.040128297	0.060896067	0.043523893	0.021107561	0.057264596	0.058329373	0.085288127	0.024628653	0.058349468	0.03089371	0.05616077	0.026006916	0.011245135	0.025024594	0.081115667	0.005180568	0.031497623	0.040509975	0.035130337	0.023193764	0.020795909
Proline	0.013602531	0.011306159	0.015639253	0.006642107	0.009390786	0.00000215	0.00000308	0.00000304	0.00000346	0.00000153	0.00000291	0.0145	0.012400678	0.011959959	0.016349986	0.016050642	0.010803889	0.0219	0.0231	0.0213	0.028	0.011	0.00000133	0.018106809	0.015420932	0.012047229	0.015881187	0.00000101	0.022798308	0.0152	0.010149154	0.0128	0.015309812	0.0172	0.010690827	0.00000258	0.010430888	0.00000141	0.007222242	0.01542592	0.014463693	0.004649499	0.011562772	0.007276708
DL-Phenylalanine	0.012581463	0.008255818	0.007390117	0.016155214	0.011860658	0.028195791	0.006250167	0.00000304	0.008064901	0.01683354	0.007331471	0.007695679	0.007026093	0.009703644	0.009125882	0.006823166	0.00772665	0.013687601	0.0063041	0.00914	0.007229454	0.013363015	0.017991573	0.008863763	0.013175776	0.009506744	0.007271802	0.01051546	0.014781919	0.023101169	0.005531548	0.013876977	0.007590943	0.014079225	0.006370904	0.003311994	0.00525744	0.015110433	0.002907351	0.007809166	0.009027144	0.008163136	0.004901185	0.005538638
Sarcosine	0.007072394	0.004541216	0.00000021	0.005803044	0.005762509	0.00000215	0.00000308	0.00000304	0.00000346	0.00000153	0.00000291	0.00577	0.00593621	0.00000103	0.000000986	0.000000867	0.00000135	0.00000147	0.00753	0.00000104	0.000000884	0.00000147	0.00000133	0.008979589	0.00000123	0.00000093	0.007153757	0.00000101	0.00934	0.0092	0.00392	0.00601	0.00701	0.00999	0.000000936	0.00000258	0.000000704	0.00000141	0.00000188	0.00657	0.007135597	0.00622	0.00000071	0.00652
Glycerol	0.02232121	0.020662053	0.02616469	0.022147071	0.029562954	0.027941609	0.071093649	0.040483878	0.029116543	0.049421423	0.065764857	0.009679918	0.02973163	0.034791653	0.027441633	0.01905677	0.017262208	0.030163193	0.022498852	0.024270017	0.017111327	0.036431692	0.035889968	0.04572767	0.037226097	0.032503159	0.014912235	0.017420233	0.036650573	0.039296022	0.021388097	0.035957547	0.027860714	0.032300144	0.017354618	0.051010733	0.018245126	0.038746269	0.023293096	0.026968546	0.038584557	0.023403707	0.019327622	0.024017758
Oxoglutaric Acid	0.002882513	0.00162	0.00173	0.00101	0.0014	0.003084288	0.003747063	0.00234334	0.003497851	0.002140029	0.003109714	0.002173809	0.002095069	0.001493222	0.001701986	0.00156301	0.001860643	0.003642472	0.003284981	0.002279353	0.001733062	0.001870925	0.001401388	0.00284829	0.00187116	0.00130578	0.002283662	0.001881103	0.002672374	0.002207466	0.001465366	0.001435732	0.002308715	0.00281496	0.001343381	0.003476132	0.001180338	0.000474301	0.00233908	0.002938834	0.002214279	0.001722588	0.001242434	0.000240042
Taurine	0.000760029	0.00046333	0.001412973	0.000000167	0.001661404	0.00000215	0.00000308	0.00000304	0.00000346	0.00000153	0.00000291	0.000000866	0.000000919	0.00000103	0.000000986	0.000000867	0.00000135	0.00000147	0.000877	0.00000104	0.000000884	0.00000147	0.00000133	0.00000137	0.00232	0.000644	0.00000108	0.000492	0.00000156	0.00000122	0.000000796	0.00000116	0.00000124	0.00000125	0.000000936	0.00000258	0.000000704	0.00000141	0.00000188	0.00000127	0.00000139	0.0000011	0.00000071	0.00000107
Phosphoric acid	0.120988609	0.110822402	0.121381593	0.110787999	0.115535169	0.19792258	0.280560144	0.110703578	0.257993372	0.1859379	0.092859362	0.095160039	0.126614777	0.138561211	0.115005526	0.100777983	0.117673988	0.199742474	0.144806778	0.126529069	0.099288257	0.121315814	0.126247951	0.179946135	0.147225375	0.097193405	0.134143783	0.127705151	0.174239464	0.151632008	0.083712175	0.117005709	0.145816785	0.17062203	0.093839295	0.26055228	0.075031509	0.134659461	0.15264418	0.128652968	0.145008784	0.14364549	0.08073705	0.133929356
5-oxoproline	0.128676005	0.114496532	0.132799642	0.097051473	0.134340909	0.120354168	0.123912674	0.020974438	0.166135972	0.165946419	0.10049738	0.101865802	0.131449744	0.135904954	0.134623135	0.109818909	0.123829456	0.213141601	0.159874379	0.14776077	0.12477698	0.148172892	0.018731651	0.189112675	0.15919145	0.120097393	0.14175552	0.098402005	0.204106908	0.166226946	0.09405281	0.159648884	0.151423709	0.192686025	0.094565007	0.073494433	0.08520344	0.17913479	0.083633707	0.149727645	0.159185346	0.14982088	0.09164245	0.13984058
Methionine	0.006343864	0.004494439	0.007259292	0.00196191	0.007066836	0.003697826	0.003519369	0.002403511	0.005073187	0.004911741	0.002977017	0.009511137	0.006143692	0.004045949	0.006328642	0.007014562	0.005393159	0.010925642	0.008867865	0.009962322	0.013540752	0.006866014	0.001330042	0.009834899	0.007262064	0.006540129	0.008992496	0.002940643	0.011900896	0.010295324	0.005144874	0.011299559	0.00830638	0.012635213	0.0034444	0.001538108	0.004326041	0.007106848	0.002896473	0.010913529	0.006585876	0.006406498	0.004919554	0.006282225
N-Acetyl-aspartic acid	0.00538637	0.004490372	0.005218436	0.004511335	0.007060524	0.014375862	0.010196006	0.002212768	0.010488599	0.005632394	0.004856534	0.006369072	0.005444553	0.004296781	0.004454831	0.004499706	0.00599905	0.008540732	0.00556286	0.006489715	0.007142175	0.007580771	0.00282326	0.008425582	0.006319152	0.005183141	0.007183433	0.005082644	0.010551086	0.01042511	0.003882215	0.008908972	0.006820775	0.010485747	0.00406169	0.005785717	0.003507314	0.006852683	0.006381183	0.007680689	0.007282493	0.006682544	0.003707909	0.00578137
N-Acetyl-glutamic acid	0.164825465	0.150494275	0.157296139	0.159160428	0.162843668	0.121306564	0.068791993	0.007481262	0.144300591	0.249107276	0.026006664	0.109653338	0.15568902	0.192086865	0.158561826	0.128420037	0.148219192	0.246578513	0.158089069	0.167794257	0.116941882	0.171785679	0.147597108	0.209491617	0.217547059	0.138026856	0.15587407	0.168889297	0.226877735	0.215328942	0.111837596	0.153532847	0.1702987	0.211755358	0.118002631	0.125397193	0.104732736	0.188558645	0.061824711	0.153453968	0.183921661	0.175656088	0.1106552	0.145072624
Serine	0.106100208	0.093773153	0.077564976	0.140212347	0.11068827	0.055884191	0.053029393	0.005224395	0.078200931	0.176033219	0.014407168	0.065131363	0.07057596	0.105889777	0.095477212	0.075237315	0.099712294	0.142513248	0.141215068	0.123652075	0.101503522	0.108323602	0.124496107	0.161424493	0.146467124	0.09986049	0.120622941	0.114578619	0.172235675	0.120984531	0.084794751	0.140087266	0.122032985	0.13840832	0.063385045	0.070459847	0.094368488	0.098954348	0.055170577	0.137234973	0.151980058	0.139989932	0.074815483	0.111467555
Fumaric Acid	0.012821704	0.002954312	0.005169022	0.002690813	0.001759206	0.007824747	0.010208502	0.00595156	0.009106159	0.004903642	0.008278559	0.005575336	0.005010674	0.004325836	0.004907991	0.004004585	0.005186278	0.008241042	0.006949508	0.006055347	0.005625736	0.004851302	0.002914683	0.00689217	0.005550162	0.003720844	0.006510021	0.003707353	0.007839345	0.005829301	0.003119936	0.004466819	0.005597978	0.006837304	0.00411267	0.00846178	0.003078843	0.004875647	0.006011514	0.005141262	0.005594584	0.00557942	0.003245768	0.004632054
Threonine	0.037431741	0.025076351	0.035401782	0.009632452	0.045178799	0.010507435	0.019198344	0.00000304	0.019110298	0.022294361	0.00000291	0.033140131	0.043426901	0.028222558	0.040509457	0.03406041	0.02884009	0.071840105	0.055267766	0.0531	0.046240585	0.032122888	0.00517	0.068702061	0.043096471	0.042565889	0.039344576	0.018595731	0.052441585	0.051301546	0.035164956	0.058753946	0.050955354	0.065390293	0.017380012	0.013937495	0.02561982	0.041438779	0.018042876	0.06399282	0.045736707	0.035306096	0.033289193	0.050546546
Azelaic acid	0.005049156	0.005968772	0.007291422	0.005172823	0.00586692	0.013863295	0.017501754	0.010536147	0.016855325	0.007948808	0.014165882	0.00705836	0.006570059	0.006529387	0.007699167	0.005462106	0.0083758	0.012178068	0.009071533	0.007928983	0.007699097	0.007128824	0.005962524	0.009161322	0.008090877	0.00577943	0.0084	0.007558938	0.010738549	0.008546454	0.005034893	0.006598378	0.008104291	0.009342423	0.00596526	0.014431648	0.004415268	0.007383049	0.009952508	0.007538175	0.007271652	0.007563992	0.004160091	0.00000107
(2R)-Pyrrolidine	0.044177701	0.033288555	0.028768675	0.027218273	0.045378638	0.01086137	0.011822166	0.001126125	0.018193279	0.035568506	0.002077666	0.036870916	0.044309376	0.03439649	0.041788223	0.039779912	0.031837044	0.071785475	0.055982437	0.05069964	0.037893094	0.02744187	0.019313915	0.042113077	0.047760461	0.032431146	0.037627917	0.038463614	0.055290289	0.057029973	0.032647787	0.03142564	0.04638505	0.055410051	0.023268143	0.018116977	0.029596978	0.024130753	0.016257286	0.046347682	0.043661715	0.036778712	0.026651896	0.029132456
Myristic acid	0.008282244	0.005556793	0.006582844	0.005376153	0.00570148	0.011921147	0.013983247	0.009248585	0.014712132	0.007889027	0.01172599	0.007585885	0.006393904	0.00616334	0.006901225	0.004929387	0.007055656	0.01230644	0.008361248	0.007237151	0.007089993	0.006358322	0.005770744	0.009877226	0.008623478	0.004671261	0.008033163	0.007807508	0.009793734	0.008610932	0.004457288	0.006987243	0.007969687	0.010436441	0.004896303	0.014807145	0.003872962	0.006823164	0.008127854	0.008107714	0.006589326	0.007305413	0.003692001	0.005891791
Phenylalanine	0.018012637	0.010350195	0.015673607	0.002737505	0.014847184	0.00744277	0.009964326	0.001575289	0.014019983	0.011325276	0.005689059	0.021367879	0.015568074	0.008394259	0.017578495	0.015089254	0.011906697	0.031718192	0.025343563	0.021955349	0.027398634	0.014669159	0.000479472	0.022164724	0.01578767	0.013396246	0.019461407	0.005688488	0.027282557	0.023039823	0.01185404	0.023264364	0.017095096	0.029354071	0.00818918	0.003777387	0.009764319	0.013577647	0.00826486	0.024492847	0.016119043	0.014480512	0.010810139	0.013114481
Malic acid	0.042037953	0.022989728	0.039704651	0.031590398	0.03129084	0.043375153	0.030471572	0.003441549	0.041412073	0.048729406	0.008332921	0.033224098	0.030001279	0.037767816	0.038503798	0.027470293	0.037031193	0.061635161	0.035551108	0.04592742	0.034360791	0.036620102	0.023457658	0.056536858	0.049694661	0.027168737	0.044196099	0.042093558	0.039180865	0.050294473	0.028504826	0.028501988	0.049736645	0.04954884	0.02022185	0.040701906	0.024266464	0.018492559	0.033914487	0.032788851	0.030984314	0.042342274	0.02654173	0.036870741
Aspartic acid	0.053808295	0.039830528	0.046801476	0.006077402	0.03575348	0.000733283	0.006811066	0.000967959	0.017324213	0.016353057	0.00000291	0.052610318	0.056168395	0.028803576	0.051343484	0.040444866	0.037238784	0.097097403	0.092878616	0.069626094	0.062674825	0.030005771	0.000654	0.066544408	0.051288724	0.033346342	0.067125068	0.019589793	0.07276699	0.047206686	0.038443684	0.037675151	0.059598858	0.067103574	0.022781583	0.013777207	0.030810477	0.02182452	0.020204527	0.065395953	0.056011788	0.04648922	0.035878026	0.047654733
Cysteine	0.003687739	0.002232804	0.003954751	0.000219751	0.00375566	0.000797818	0.000738919	0.00000304	0.000918542	0.002497561	0.00102027	0.005370221	0.003653945	0.001953341	0.00365955	0.003746028	0.002687393	0.00798446	0.006767251	0.00505	0.006283105	0.003979657	0.00000133	0.004745373	0.003685005	0.003107129	0.00553648	0.000944735	0.00595745	0.00413	0.003153986	0.005078013	0.003821748	0.00824	0.001430185	0.00000258	0.002449284	0.001448096	0.000758099	0.006658221	0.003030085	0.002641867	0.002575897	0.002892686
Hypoxanthine	0.012603478	0.010041046	0.011666077	0.000332884	0.003218064	0.00000215	0.003403765	0.00093381	0.004730943	0.004529443	0.001169039	0.009600892	0.01301742	0.008450655	0.010650342	0.006624884	0.003793452	0.0209	0.013464818	0.009313022	0.008932992	0.00381422	0.00000133	0.024967696	0.009041399	0.005386574	0.01243633	0.006859686	0.015403523	0.011918083	0.005314014	0.004523014	0.019980455	0.0157	0.006684094	0.001601287	0.003794584	0.00452235	0.00765299	0.011833709	0.013628058	0.016395616	0.004751999	0.009983919
Palmitoleic Acid	0.005400579	0.003195201	0.003937695	0.001604624	0.003947455	0.015013897	0.005600217	0.003030533	0.00554165	0.002533149	0.002844892	0.008759197	0.004629218	0.006370899	0.005002939	0.003769871	0.003107945	0.015986332	0.00692722	0.006021826	0.005494643	0.002360456	0.002735323	0.006781339	0.005054622	0.003133099	0.005872247	0.004287805	0.006091438	0.004839494	0.003454929	0.004228772	0.004915619	0.007105273	0.001959889	0.003261194	0.002639353	0.002624007	0.002300676	0.006270224	0.003890881	0.00416894	0.002956481	0.001532651
Succinic Acid	0.011322986	0.007248162	0.008603387	0.007739112	0.011380083	0.004721837	0.004751181	0.000586165	0.004770014	0.007522711	0.002342106	0.016885666	0.013711137	0.011226468	0.016662507	0.012551824	0.014729481	0.030588878	0.019015645	0.018303302	0.017104004	0.016783733	0.017707611	0.01644609	0.01624334	0.012363518	0.016225333	0.0115771	0.021176851	0.026089002	0.012213874	0.016032963	0.015631506	0.027311745	0.009730898	0.01939676	0.009995153	0.010567267	0.019695718	0.021838579	0.014679718	0.012097897	0.010970896	0.013504049
Glutamic acid	0.130366424	0.115397463	0.13554732	0.044106736	0.125406239	0.02145544	0.077446225	0.007401358	0.143112462	0.112458888	0.010874607	0.109154969	0.143207795	0.133127669	0.133984853	0.114879118	0.123096857	0.227565078	0.173558938	0.151856775	0.123799692	0.125099944	0.001144323	0.201401295	0.158820937	0.110543603	0.152567309	0.099701165	0.193373084	0.158491354	0.099201842	0.13249267	0.160878297	0.188151207	0.08718369	0.102521395	0.087642976	0.129357926	0.113200361	0.150252302	0.155168828	0.152593939	0.093998276	0.144562246
Palmitic Acid	0.074509329	0.062894936	0.069335595	0.054702858	0.068666401	0.14010547	0.192415421	0.128173854	0.172064068	0.098658382	0.164565139	0.059308791	0.067941414	0.072968626	0.076270542	0.054877171	0.076115572	0.114865971	0.092675412	0.073181084	0.065311474	0.079432061	0.057614827	0.102814791	0.089125406	0.055271078	0.080056523	0.077772486	0.105385111	0.08998844	0.051292431	0.075168012	0.084615221	0.100012095	0.055517194	0.174147519	0.040875091	0.07827497	0.103800242	0.074170357	0.081161864	0.074227622	0.042058765	0.07398654
Asparagine	0.002478965	0.001737498	0.001787708	0.000000167	0.001854091	0.00000215	0.00000308	0.00000304	0.001790598	0.00000153	0.00000291	0.002981421	0.003538032	0.001315468	0.002688572	0.00238779	0.000720982	0.00558	0.00387	0.00342	0.003632298	0.00000147	0.00000133	0.005312901	0.002108509	0.002167583	0.002263773	0.00062642	0.003812822	0.00124	0.001922525	0.002836507	0.004327502	0.00409	0.000369601	0.00000258	0.001130547	0.001158164	0.001547609	0.004180133	0.002922175	0.003767956	0.001367528	0.00281
Lysine	0.007978151	0.002439664	0.007411387	0.000000167	0.01069009	0.00000215	0.00000308	0.00000304	0.00000346	0.00000153	0.00000291	0.008595977	0.009942097	0.007419826	0.008738474	0.005469809	0.00534368	0.00000147	0.0146	0.00814	0.0105	0.0123	0.00000133	0.008739069	0.007553605	0.004831058	0.009159315	0.003185596	0.014373667	0.008402179	0.008573682	0.008314998	0.008529585	0.0133	0.00651348	0.010289642	0.002517756	0.005260466	0.006575068	0.00947	0.012217942	0.010668734	0.004723595	0.00698705
Glutamine	0.007901031	0.005292178	0.00985931	0.000000167	0.007188397	0.00000215	0.008898593	0.001091752	0.000481363	0.00000153	0.00000291	0.015241304	0.017479088	0.004037214	0.012934806	0.010256724	0.00896105	0.0292	0.038494621	0.020502748	0.024324611	0.00417	0.00000133	0.035285606	0.006850334	0.008689067	0.022594694	0.001935604	0.016676782	0.005583804	0.012614786	0.017511282	0.020351837	0.0222	0.002032055	0.003473447	0.00537707	0.003721572	0.010634556	0.026427324	0.012700942	0.015031139	0.009720459	0.01960162
Oleic Acid	0.027919734	0.016362848	0.019987063	0.014222238	0.028637066	0.062872068	0.054003917	0.021900388	0.03348799	0.020757224	0.023528808	0.027773201	0.01854003	0.01806398	0.019493255	0.015628309	0.018357279	0.052708388	0.025349156	0.017728467	0.017862837	0.00000147	0.015224847	0.02768897	0.023040786	0.017722833	0.019369514	0.017645365	0.029676904	0.040761669	0.015637756	0.042079898	0.020718404	0.051611439	0.010072084	0.042876705	0.008747897	0.017022567	0.022268943	0.040392012	0.016320077	0.0167533	0.008390129	0.0167
Stearic acid	0.095315884	0.083183212	0.089543537	0.074922887	0.089295945	0.196752973	0.304801485	0.188164323	0.235823905	0.138334349	0.248729374	0.075460294	0.086984431	0.094369901	0.092949323	0.075050462	0.095035174	0.146631826	0.115229471	0.09357279	0.081779221	0.109435591	0.064418442	0.122202933	0.106936529	0.075000779	0.101233188	0.098795847	0.140594942	0.117202377	0.062440164	0.10344721	0.102374058	0.12960998	0.074285204	0.232850717	0.053367519	0.104287776	0.138023931	0.09836817	0.101335915	0.099936058	0.055179339	0.0949045
Citric acid	0.056152853	0.042749471	0.043146043	0.014053492	0.047981615	0.03353904	0.086260165	0.023634518	0.083116046	0.070928212	0.046372523	0.042111256	0.04891529	0.039705893	0.043676863	0.034172975	0.039563252	0.072852675	0.062875666	0.050298441	0.043398977	0.053950742	0.00000133	0.0381055	0.046443503	0.037828522	0.044307383	0.02993164	0.075624014	0.078721353	0.03127913	0.052734279	0.04538096	0.082	0.03425967	0.052683728	0.029569012	0.03419704	0.037873783	0.051079591	0.049578104	0.038143718	0.026435348	0.031810435
Tyrosine	0.030935585	0.019017757	0.02626668	0.002961756	0.0273143	0.014084846	0.02808103	0.005343837	0.028396489	0.017310368	0.011427078	0.03381467	0.027916623	0.015010427	0.029140958	0.024988391	0.022606871	0.05622667	0.043825445	0.035035562	0.039859037	0.025485261	0.00000133	0.040673505	0.025714057	0.023789648	0.033094738	0.008580446	0.045161204	0.034060386	0.020590238	0.041370537	0.03146477	0.0531	0.011761257	0.013558875	0.015031716	0.021948778	0.018978979	0.041984389	0.028288502	0.026324742	0.017103657	0.026466748
Pantothenic acid	0.012722606	0.010484868	0.011388821	0.006716299	0.017068893	0.012750239	0.02479739	0.00000304	0.014724217	0.01769584	0.012861225	0.008515419	0.013184065	0.009046916	0.011285844	0.011683973	0.012088358	0.029902221	0.02259979	0.0162	0.014757854	0.018251386	0.009389134	0.024798601	0.02254491	0.016778461	0.015032802	0.004720348	0.025820038	0.02224886	0.010775602	0.022742592	0.018520485	0.029713284	0.008154264	0.015924445	0.009279949	0.013582187	0.017831138	0.025834896	0.020634057	0.013527133	0.011100048	0.018130699
Isocitric Acid	0.041492961	0.028811482	0.029896187	0.011581434	0.081847199	0.174130143	0.165227325	0.021622106	0.048529324	0.059677482	0.066107327	0.035350002	0.038483613	0.018916372	0.023177613	0.024877778	0.033016426	0.100146547	0.041218235	0.02160536	0.025810359	0.069835763	0.039762964	0.017553059	0.031630978	0.049096002	0.021956094	0.104952227	0.149	0.036445687	0.032834064	0.116187313	0.028634014	0.14287616	0.000000936	0.022556725	0.010293779	0.021147711	0.032197112	0.104560319	0.029137414	0.015420366	0.01187768	0.010123469
Erythronic Acid	0.015629851	0.008026883	0.007440381	0.003973736	0.029974036	0.07317196	0.061243365	0.005316342	0.014250463	0.013009311	0.012519271	0.014271136	0.012911884	0.005096956	0.006252021	0.007490274	0.010941801	0.037561761	0.013794576	0.005792807	0.007900039	0.017212007	0.012354806	0.005044517	0.00697867	0.013254354	0.007635921	0.036168603	0.0514	0.010637583	0.01031943	0.044533948	0.008989006	0.052993858	0.000000936	0.005446684	0.002711336	0.004935047	0.011456356	0.039330607	0.007604371	0.004567034	0.002979058	0.003442801
Doconexent	0.005264463	0.002224213	0.00228244	0.002071983	0.003997353	0.005821268	0.00655643	0.001977954	0.004702206	0.00281631	0.001798357	0.005043663	0.003607299	0.002424632	0.002090606	0.001994351	0.003905367	0.007849101	0.003841821	0.003136095	0.003177819	0.002674335	0.00212007	0.003640135	0.002914862	0.002585722	0.00419	0.004071735	0.005784481	0.004402171	0.006568714	0.00490501	0.003674837	0.007279915	0.001561501	0.004764867	0.001367373	0.002714627	0.003119998	0.005257441	0.002150621	0.002641187	0.001266928	0.00000107
Cholesterol	0.092273888	0.03418243	0.014223492	0.031637829	0.092256547	0.16050364	0.21052374	0.060500925	0.047496294	0.006620348	0.014389302	0.074066686	0.071873712	0.024545282	0.02738179	0.02543229	0.065836806	0.135726526	0.083119227	0.028395252	0.035570025	0.036442798	0.067201304	0.051734549	0.029812383	0.027831903	0.061405474	0.081617162	0.003579931	0.118985659	0.034873924	0.073137125	0.066745041	0.100143334	0.007435461	0.101531957	0.010927928	0.004092978	0.071332456	0.08211544	0.044083873	0.062954521	0.009023298	0.075302247
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	Characteristic Fragment Ions	Retention time	HMDB	PubChem	KEGG	Formula	Exact mass	Super class	Main class	Sub class
L-Isoleucine	86, 170, 188	8.737	HMDB0000172	6306	C00407	C6H13NO2	131	Organic acids	Amino acids and peptides	Amino acids
Lactic acid	189, 233, 261	10.048	HMDB0000190	61503	C00256	C3H6O3	90.0317	Organic acids	Short-chain acids	Short-chain acids
Glycolic acid	189, 247	10.282	HMDB0000115	757	C03547	C2H4O3	76.016	Fatty Acyls	Fatty acids	Hydroxy FA
Alanine	158, 232, 260	10.754	HMDB0000161	5950	C00041	C3H7NO2	89.0477	Organic acids	Amino acids and peptides	Amino acids
Glycine	218, 246	11.071	HMDB0000123	750	C00037	C2H5NO2	75.032	Organic acids	Amino acids and peptides	Amino acids
Uracil	75,  179,  195	11.632	HMDB0000300	1174	C00106	C4H4N2O2	112.0273	Nucleic acids	Pyrimidines	Pyrimidines
beta-Alanine	144, 218, 260	12.236	HMDB0000056	239	C00099	C3H7NO2	89.0477	Organic acids	Amino acids and peptides	Amino acids
Urea	73, 147, 231	12.371	HMDB0000294	2447	C00086	CH4N2O	60.0324	Organic acids	Carboximidic acids	Isoureas
Valine	186, 260, 288	12.471	HMDB0000883	6287	C00183	C5H11NO2	117.079	Organic acids	Amino acids and peptides	Amino acids
Capric acid	75, 229	12.83	HMDB0000511	2969	C01571	C10H20O2	172.1463	Fatty Acyls	Fatty acids	Saturated FA
Leucine	200, 274, 302	13.04	HMDB0000687	6106	C00123	C6H13NO2	131.0946	Organic acids	Amino acids and peptides	Amino acids
Nicotinamide N-oxide	75,  136,  180	13.209	HMDB0001406	936	C00153	C6H6N2O	122.048	Organoheterocyclic compounds	Pyridinecarboxylic acids	Nicotinamides
Isoleucine	200, 274, 302	13.48	HMDB0000172	6306	C00407	C6H13NO2	131.0946	Organic acids	Amino acids and peptides	Amino acids
5-oxoprolinate	84, 158, 186	13.578	HMDB0000805	499	C02237	C5H7NO3	129.0426	Organoheterocyclic compounds	Pyrroline carboxylic acids	Pyrroline carboxylic acids
Proline	184, 258, 286	13.991	HMDB0000162	145742	C00148	C5H9NO2	115.0633	Organic acids	Amino acids and peptides	Amino acids
DL-Phenylalanine	120, 130, 146	14.32	HMDB0250791	994	C02265	NA	165.079	Organic acids	Amino acids and peptides	Amino acids
Sarcosine	158, 232, 260	14.873	HMDB0000271	1088	C00213	C3H7NO2	89.0477	Organic acids	Amino acids and peptides	Amino acids
Glycerol	245, 289, 377	15.43	HMDB00131	753	C00116	C3H8O3	92.0473	Organic Oxygen	Organooxygen	Sugar alcohol
Oxoglutaric Acid	157, 215, 317	19.22	HMDB0000208	51	C00026	C5H6O5	146.0215	Organic acids	TCA acids	TCA acids
Taurine	88, 158, 296	15.844	HMDB0000251	1123	C00245	C2H7NO3S	125.0147	Organic acids	Sulfonic acids	Sulfonic acids
Phosphoric acid	124, 383, 384	16.013	HMDB0001429	1004	C00009	H3PO4	97.9769	Non-metal compounds	Non-metal oxanionic compounds	Non-metal phosphates
5-oxoproline	198, 272, 300	16.349	HMDB0000267	7405	C01879	C5H7NO3	129.0426	Organoheterocyclic compounds	Pyrroline carboxylic acids	Pyrroline carboxylic acids
Methionine	218, 292, 320	16.551	HMDB0000696	6137	C00073	C5H11NO2S	149.0511	Organic acids	Amino acids and peptides	Amino acids
N-Acetyl-aspartic acid	73, 225, 346	16.601	HMDB0000812	65065	C01042	C6H9NO5	175.0481	Organic acids	Amino acids and peptides	Amino acids
N-Acetyl-glutamic acid	186, 216, 318	16.8	HMDB0001138	185	C00624	C7H11NO5	189.0637	Organic acids	Amino acids and peptides	Amino acids
Serine	288, 362, 390	16.823	HMDB0000187	5951	C00065	C3H7NO3	105.0426	Organic acids	Amino acids and peptides	Amino acids
Fumaric Acid	147, 287, 329	17.02	HMDB00134	444972	C00122	C4H4O4	116.011	Organic acids	TCA acids	TCA acids
Threonine	303, 376, 404	17.212	HMDB0000167	6288	C00188	C4H9NO3	119.0582	Organic acids	Amino acids and peptides	Amino acids
Azelaic acid	129, 359, 401	17.57	HMDB00784	2266	C08261	C9H16O4	188.1049	Fatty Acyls	Fatty Acids	Saturated FA
(2R)-Pyrrolidine	184, 214, 330	17.487	NA	91742919	NA	C18H37NO4Si2	387.7	Organoheterocyclic compounds	Pyrroline carboxylic acids	Pyrroline carboxylic acids
Myristic acid	75, 129, 285	17.892	HMDB0000806	11005	C06424	C14H28O2	228.2089	Fatty Acyls	Fatty acids	Saturated FA
Phenylalanine	234, 302, 336	18.02	HMDB0000159	6140	C00079	C9H11NO2	165.079	Organic acids	Amino acids and peptides	Amino acids
Malic acid	115, 287, 419	18.233	HMDB0000744	525	C03668	C4H6O5	134.0215	Organic acids	TCA acids	TCA acids
Aspartic acid	302, 316, 418	18.933	HMDB0000191	5960	C00049	C4H7NO4	133.0375	Organic acids	Amino acids and peptides	Amino acids
Cysteine	302, 378, 406	19.315	HMDB0000574	5862	C00097	C3H7NO2S	121.0198	Organic acids	Amino acids and peptides	Amino acids
Hypoxanthine	73,  193,  307	19.554	HMDB0000157	790	C00262	C5H4N4O	136.0385	Nucleic acids	Purines	Hypoxanthines
Palmitoleic Acid	75, 311, 353	20.051	HMDB0003229	5312427	C08362	C16H30O2	254.2246	Fatty Acyls	Fatty acids	Saturated FA
Succinic Acid	73,147, 289, 331	14.028	HMDB0000254	1110	C00042	C4H6O4	118.0266	Organic acids	TCA acids	TCA acids
Glutamic acid	272, 330, 432	20.026	HMDB0000148	33032	C00025	C5H9NO4	147.0532	Organic acids	Amino acids and peptides	Amino acids
Palmitic Acid	75, 117, 129, 313	20.12	HMDB0000220	985	C00249	C16H32O2	256.2402	Fatty Acyls	Fatty acids	Saturated FA
Asparagine	302, 400, 417	20.361	HMDB0000168	6267	C00152	C4H8N2O3	132.0535	Organic acids	Amino acids and peptides	Amino acids
Lysine	198, 272, 300	21.144	HMDB0000182	5962	C00047	C6H14N2O2	146.1055	Organic acids	Amino acids and peptides	Amino acids
Glutamine	198, 271, 431	21.596	HMDB0000641	5961	C00064	C5H10N2O3	146.0691	Organic acids	Amino acids and peptides	Amino acids
Oleic Acid	75, 129, 339	21.946	HMDB0000207	445639	C00712	C18H34O2	282.2559	Fatty Acyls	Fatty acids	Unsaturated FA
Stearic acid	75, 341	22.176	HMDB0000827	5281	C01530	C18H36O2	284.2715	Fatty Acyls	Fatty acids	Saturated FA
Citric acid	431, 459, 591	23.494	HMDB0000094	311	C00158	C6H8O7	192.027	Organic acids	TCA acids	TCA acids
Tyrosine	302, 364, 466	23.7	HMDB0000158	6057	C00082	C9H11NO3	181.0739	Organic acids	Amino acids and peptides	Amino acids
Pantothenic acid	75, 87, 504	24.138	HMDB0000210	6613	C00864	C9H17NO5	219.11	Organic Oxygen	Organooxygen	Secondary alcohols
Isocitric Acid	73, 147, 202, 345	24.435	HMDB0000193	1198	C00311	C6H8O7	192.027	Organic acids	TCA acids	TCA acids
Erythronic Acid	261, 344, 476	24.56	HMDB0000613	2781043	NA	C4H8O5	136.0372	Carbohydrates	Monosaccharides	Sugar acids
Doconexent	75, 209, 385, 427	25.482	HMDB0002183	445580	C06429	C22H32O2	328.2402	Fatty Acyls	Fatty acids	Unsaturated FA
Cholesterol	75, 367, 443	35.95	HMDB0000067	5997	C00187	C27H46O	386.3549	Sterol Lipids	Sterols	Cholesterols
METABOLITES_END
#END