#METABOLOMICS WORKBENCH mdm_20250119_095637 DATATRACK_ID:5542 STUDY_ID:ST003679 ANALYSIS_ID:AN006040 PROJECT_ID:PR002283
VERSION                          	1
CREATED_ON                       	01-23-2025
#PROJECT
PR:PROJECT_TITLE                 	Ketogenic diet suppresses colorectal cancer through the gut microbiome long
PR:PROJECT_TITLE                 	chain fatty acid stearate
PR:PROJECT_SUMMARY               	Our manuscript entitled “Ketogenic diet suppresses colorectal cancer through
PR:PROJECT_SUMMARY               	the gut microbiome long chain fatty acid stearate” describes a reduced colonic
PR:PROJECT_SUMMARY               	tumor burden upon ketogenic diet (KD) consumption in a CRC mouse model with a
PR:PROJECT_SUMMARY               	humanized microbiome. Importantly, we demonstrate a causal relationship through
PR:PROJECT_SUMMARY               	microbiome transplantation into germ-free mice, whereby alterations in the gut
PR:PROJECT_SUMMARY               	microbiota were maintained in the absence of continued selective pressure from
PR:PROJECT_SUMMARY               	the KD. Specifically, we identify a shift toward bacterial species that produce
PR:PROJECT_SUMMARY               	stearic acid in ketogenic conditions, whereas consumers were depleted, resulting
PR:PROJECT_SUMMARY               	in elevated levels of free stearate in the gut lumen. This microbial product
PR:PROJECT_SUMMARY               	demonstrates tumor-suppressing properties by inducing apoptosis in cancer cells
PR:PROJECT_SUMMARY               	and decreasing colonic Th17 immune cell populations. As part of this study, we
PR:PROJECT_SUMMARY               	used different metabolomics workflows to study metabolites in mouse fecal and
PR:PROJECT_SUMMARY               	plasma samples as well as the used rodent diet.
PR:INSTITUTE                     	University of Luxembourg
PR:LAST_NAME                     	Letellier
PR:FIRST_NAME                    	Elisabeth
PR:ADDRESS                       	6, avenue du Swing, Belval, Esch, 4367, Luxembourg
PR:EMAIL                         	madita.brauer@uni.lu
PR:PHONE                         	(+352) 46 66 44 6954
PR:DOI                           	http://dx.doi.org/10.21228/M8HZ6G
#STUDY
ST:STUDY_TITLE                   	Ketogenic diet suppresses colorectal cancer through the gut microbiome long
ST:STUDY_TITLE                   	chain fatty acid stearate - untargeted LCMS data from SPF experiment
ST:STUDY_SUMMARY                 	Mouse fecal pellets from an AOM/DSS CRC mouse experiment performed with specific
ST:STUDY_SUMMARY                 	pathogen-free mice were analyzed by a LC-MS approach to characterize the fecal
ST:STUDY_SUMMARY                 	metabolome of mice from different treatment groups. Metabolites were extracted
ST:STUDY_SUMMARY                 	from mouse fecal samples as described in the method file. Resulting samples were
ST:STUDY_SUMMARY                 	used for a LC-MS untargeted metabolomics screen using a Vanquish UHPLC
ST:STUDY_SUMMARY                 	(ThermoFisher Scientific), coupled to a Q Exactive HF mass spectrometer
ST:STUDY_SUMMARY                 	(ThermoFisher Scientific).
ST:INSTITUTE                     	University of Luxembourg
ST:LAST_NAME                     	Letellier
ST:FIRST_NAME                    	Elisabeth
ST:ADDRESS                       	6, avenue du Swing
ST:EMAIL                         	elisabeth.letellier@uni.lu
ST:PHONE                         	(+352) 46 66 44 6954
ST:SUBMIT_DATE                   	2025-01-19
#SUBJECT
SU:SUBJECT_TYPE                  	Mammal
SU:SUBJECT_SPECIES               	Mus musculus
SU:TAXONOMY_ID                   	10090
#SUBJECT_SAMPLE_FACTORS:         	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data
SUBJECT_SAMPLE_FACTORS           	11	KD_11	Sample source:mouse feces | Condition:KD	RAW_FILE_NAME(raw_file_name)=MT003_LC_GlasgowMA_Mouse_feces_KD_11_1.mzML
SUBJECT_SAMPLE_FACTORS           	12	KD_12	Sample source:mouse feces | Condition:KD	RAW_FILE_NAME(raw_file_name)=MT003_LC_GlasgowMA_Mouse_feces_KD_12_1.mzML
SUBJECT_SAMPLE_FACTORS           	13	KD_13	Sample source:mouse feces | Condition:KD	RAW_FILE_NAME(raw_file_name)=MT003_LC_GlasgowMA_Mouse_feces_KD_13_1.mzML
SUBJECT_SAMPLE_FACTORS           	21	KD_21	Sample source:mouse feces | Condition:KD	RAW_FILE_NAME(raw_file_name)=MT003_LC_GlasgowMA_Mouse_feces_KD_21_1.mzML
SUBJECT_SAMPLE_FACTORS           	22	KD_22	Sample source:mouse feces | Condition:KD	RAW_FILE_NAME(raw_file_name)=MT003_LC_GlasgowMA_Mouse_feces_KD_22_1.mzML
SUBJECT_SAMPLE_FACTORS           	23	KD_23	Sample source:mouse feces | Condition:KD	RAW_FILE_NAME(raw_file_name)=MT003_LC_GlasgowMA_Mouse_feces_KD_23_1.mzML
SUBJECT_SAMPLE_FACTORS           	24	KD_24	Sample source:mouse feces | Condition:KD	RAW_FILE_NAME(raw_file_name)=MT003_LC_GlasgowMA_Mouse_feces_KD_24_1.mzML
SUBJECT_SAMPLE_FACTORS           	25	KD_25	Sample source:mouse feces | Condition:KD	RAW_FILE_NAME(raw_file_name)=MT003_LC_GlasgowMA_Mouse_feces_KD_25_1.mzML
SUBJECT_SAMPLE_FACTORS           	1	SD_1	Sample source:mouse feces | Condition:SD	RAW_FILE_NAME(raw_file_name)=MT003_LC_GlasgowMA_Mouse_feces_SD_1_1.mzML
SUBJECT_SAMPLE_FACTORS           	16	SD_16	Sample source:mouse feces | Condition:SD	RAW_FILE_NAME(raw_file_name)=MT003_LC_GlasgowMA_Mouse_feces_SD_16_1.mzML
SUBJECT_SAMPLE_FACTORS           	17	SD_17	Sample source:mouse feces | Condition:SD	RAW_FILE_NAME(raw_file_name)=MT003_LC_GlasgowMA_Mouse_feces_SD_17_1.mzML
SUBJECT_SAMPLE_FACTORS           	18	SD_18	Sample source:mouse feces | Condition:SD	RAW_FILE_NAME(raw_file_name)=MT003_LC_GlasgowMA_Mouse_feces_SD_18_1.mzML
SUBJECT_SAMPLE_FACTORS           	19	SD_19	Sample source:mouse feces | Condition:SD	RAW_FILE_NAME(raw_file_name)=MT003_LC_GlasgowMA_Mouse_feces_SD_19_1.mzML
SUBJECT_SAMPLE_FACTORS           	2	SD_2	Sample source:mouse feces | Condition:SD	RAW_FILE_NAME(raw_file_name)=MT003_LC_GlasgowMA_Mouse_feces_SD_2_1.mzML
SUBJECT_SAMPLE_FACTORS           	20	SD_20	Sample source:mouse feces | Condition:SD	RAW_FILE_NAME(raw_file_name)=MT003_LC_GlasgowMA_Mouse_feces_SD_20_1.mzML
SUBJECT_SAMPLE_FACTORS           	3	SD_3	Sample source:mouse feces | Condition:SD	RAW_FILE_NAME(raw_file_name)=MT003_LC_GlasgowMA_Mouse_feces_SD_3_1.mzML
SUBJECT_SAMPLE_FACTORS           	4	SD_4	Sample source:mouse feces | Condition:SD	RAW_FILE_NAME(raw_file_name)=MT003_LC_GlasgowMA_Mouse_feces_SD_4_1.mzML
#COLLECTION
CO:COLLECTION_SUMMARY            	Mice were subjected to AOM/DSS treatment and received either a ketogenic diet
CO:COLLECTION_SUMMARY            	(KD) or a standard diet (SD) as described in Tsenkova et al. (2025). Throughout
CO:COLLECTION_SUMMARY            	the experiment, feces samples were collected and flesh frozen in liquid nitrogen
CO:COLLECTION_SUMMARY            	and kept at -80℃ until metabolite extraction.
CO:SAMPLE_TYPE                   	Feces
CO:STORAGE_CONDITIONS            	-80℃
#TREATMENT
TR:TREATMENT_SUMMARY             	Mice were subjected to AOM/DSS treatment and received either a ketogenic diet
TR:TREATMENT_SUMMARY             	(KD) or a standard diet (SD) as described in Tsenkova et al. (2025). Briely,
TR:TREATMENT_SUMMARY             	mice were administered a single intraperitoneal injection of sterile-filtered
TR:TREATMENT_SUMMARY             	AOM (10mg/kg mouse body weight). SPF mice were administered a sterile-filtered
TR:TREATMENT_SUMMARY             	antibiotic mix (vancomycin hydrochloride (Sigma-Aldrich, 500mg/L), ampicillin
TR:TREATMENT_SUMMARY             	(Sigma-Aldrich, 1g/L), neomycin (Sigma-Aldrich, 1g/L) and metronidazole
TR:TREATMENT_SUMMARY             	(Sigma-Aldrich, 1g/L)) via their drinking water for one week prior to AOM
TR:TREATMENT_SUMMARY             	injection. Microbiomes were replaced with donor microbiomes via oral gavage.
TR:TREATMENT_SUMMARY             	Mice were administered a total of three cycles of DSS, with a two-week-long
TR:TREATMENT_SUMMARY             	recovery period between each cycle. DSS was refreshed mid-cycle. Half of the
TR:TREATMENT_SUMMARY             	mice were randomly allocated to each dietary condition at the end of the last
TR:TREATMENT_SUMMARY             	cycle of DSS.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Mouse fecal pellets were placed in 0.5mL Precellys® tubes (VWR) containing five
SP:SAMPLEPREP_SUMMARY            	ceramic beads each and MilliQ® water was added to each sample at a 1:16 dry
SP:SAMPLEPREP_SUMMARY            	weight to water ratio. The samples were homogenized at 6000 rpm, for two
SP:SAMPLEPREP_SUMMARY            	30-second-long cycles at 4°C, in a Precellys®24 Homogenizer (Bertin Corp.).
SP:SAMPLEPREP_SUMMARY            	Samples were then incubated at 4°C for ten minutes, then centrifuged at maximum
SP:SAMPLEPREP_SUMMARY            	speed for 10 minutes at 4°C. Samples were maintained on ice and in the dark.
SP:SAMPLEPREP_SUMMARY            	The supernatant was used for further downstream processing. An ISM was prepared
SP:SAMPLEPREP_SUMMARY            	(2μg/mL of ribitol, pentanedioic-d6 acid and d-mannose and 10μg/mL
SP:SAMPLEPREP_SUMMARY            	tridecanoic-d25 acid, 6-chloropurine riboside, 4-chloro-DL-phenylalanine,
SP:SAMPLEPREP_SUMMARY            	Nε-trifluoroacetyl-L-lysine and thionicotinamide adenine dinucleotide
SP:SAMPLEPREP_SUMMARY            	(Sigma-Aldrich) in MilliQ® water). 40 μL of the ISM was added to 100μL of the
SP:SAMPLEPREP_SUMMARY            	fecal supernatant fluid. 80 μL of this mixture was added to 320μL of methanol,
SP:SAMPLEPREP_SUMMARY            	vortexed thoroughly, incubated for five minutes at 4°C at maximum speed in an
SP:SAMPLEPREP_SUMMARY            	Eppendorf ThermoMixer, and then centrifugated for five minutes at 4°C at
SP:SAMPLEPREP_SUMMARY            	maximum speed. 350 μL of supernatant were added to 280μL of chloroform. 180μL
SP:SAMPLEPREP_SUMMARY            	of MilliQ® water were added, the samples were vortexed thoroughly, incubated
SP:SAMPLEPREP_SUMMARY            	for ten minutes at 4°C at maximum speed in an ThermoMixer (Eppendorf), then
SP:SAMPLEPREP_SUMMARY            	centrifugated for five minutes at 4°C at maximum speed. The extract was then
SP:SAMPLEPREP_SUMMARY            	split – 200 μL of the upper (polar) phase were aliquoted into a GC vial with
SP:SAMPLEPREP_SUMMARY            	a micro-insert and the rest of the upper (polar) phase was filtered through a
SP:SAMPLEPREP_SUMMARY            	PHENEX-RC syringe filter (Phenomenex), and 200 μL were retained in Eppendorf
SP:SAMPLEPREP_SUMMARY            	tubes for further processing. Then, the temperature of the SpeedVac® was
SP:SAMPLEPREP_SUMMARY            	increased to 25°C for 25 minutes to avoid water condensation on the surface of
SP:SAMPLEPREP_SUMMARY            	the glass vial. Samples were protected from light exposure and stored at -80°C.
SP:SAMPLEPREP_SUMMARY            	Samples for LC-MS analysis (polar) were reconstituted in 80 μL 50% ACN in
SP:SAMPLEPREP_SUMMARY            	water, transferred into LC vials for LC-MS analysis and analyzed on a Vanquish
SP:SAMPLEPREP_SUMMARY            	UHPLC (ThermoFisher Scientific), coupled to a Q Exactive HF mass spectrometer
SP:SAMPLEPREP_SUMMARY            	(ThermoFisher Scientific).
#CHROMATOGRAPHY
CH:INSTRUMENT_NAME               	Thermo Vanquish
CH:COLUMN_NAME                   	SeQuant ZIC- pHILIC (150 x 2.1 mm, 5 μm)
CH:COLUMN_TEMPERATURE            	50°C
CH:FLOW_GRADIENT                 	90 % B for 1.5 min, followed by a decrease to 20% B within 15 min, then 20% B
CH:FLOW_GRADIENT                 	for 2 min, increase to 90% B within 2 min, then 90% B for 13 min
CH:FLOW_RATE                     	200 µL/mL
CH:SOLVENT_A                     	100% Water; 20mM Ammonium acetate
CH:SOLVENT_B                     	90% Acetonitrile/10% Water; 0.1% Formic acid; 20 mM Ammonium acetate
CH:CHROMATOGRAPHY_TYPE           	HILIC
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Thermo Q Exactive HF-X Orbitrap
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	ESI
MS:MS_COMMENTS                   	Resolution - 120,000, m/z range - 60-900, AGC target - 1e6, maximum injection
MS:MS_COMMENTS                   	time – 70 ms. ddMS2 was applied using the following settings: Resolution -
MS:MS_COMMENTS                   	30,000, AGC target - 5e5, maximum injection time – 70, topN - 5, Normalized
MS:MS_COMMENTS                   	collision energy – 20. Samples were acquired in positive and negative
MS:MS_COMMENTS                   	ionization mode simultaneously (polarity switching).
MS:ION_MODE                      	UNSPECIFIED
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS         	normalized area
MS_METABOLITE_DATA_START
Samples	KD_11	KD_12	KD_13	KD_21	KD_22	KD_23	KD_24	KD_25	SD_1	SD_16	SD_17	SD_18	SD_19	SD_2	SD_20	SD_3	SD_4
Factors	Sample source:mouse feces | Condition:KD	Sample source:mouse feces | Condition:KD	Sample source:mouse feces | Condition:KD	Sample source:mouse feces | Condition:KD	Sample source:mouse feces | Condition:KD	Sample source:mouse feces | Condition:KD	Sample source:mouse feces | Condition:KD	Sample source:mouse feces | Condition:KD	Sample source:mouse feces | Condition:SD	Sample source:mouse feces | Condition:SD	Sample source:mouse feces | Condition:SD	Sample source:mouse feces | Condition:SD	Sample source:mouse feces | Condition:SD	Sample source:mouse feces | Condition:SD	Sample source:mouse feces | Condition:SD	Sample source:mouse feces | Condition:SD	Sample source:mouse feces | Condition:SD	
2-Hydroxybutyric acid	0.0018	0.0020	0.0038	0.0033	0.0016	0.0016	0.0026	0.0010	0.0056	0.0054	0.0026	0.0237	0.0062	0.0147	0.0023	0.0040	0.0021
2-Hydroxyglutarate	0.0397	0.0194	0.0687	0.0505	0.0742	0.0719	0.0613	0.0486	0.2018	0.1643	0.1680	0.1429	0.4121	0.0611	0.1172	0.0490	0.0351
2-Picolinic acid	0.3573	0.3156	0.3391	0.4381	0.8253	0.6508	0.5634	0.4472	1.4792	0.7304	2.1599	0.5982	1.1460	0.5307	2.8602	1.1241	1.1525
3-(4-Hydroxyphenyl)propionic acid		0.0019	0.0051	0.0166	0.0029	0.0020	0.0018	0.0053	0.1131	0.4516	0.0271	0.4489	0.2566	0.3414	0.0142	0.3273	0.1262
4-Hydroxy-2-oxoglutaric acid	0.0002	0.0002	0.0013	0.0004	0.0008	0.0004	0.0007	0.0001	0.0004	0.0017	0.0005	0.0008	0.0008	0.0003	0.0019	0.0007	0.0002
4-Hydroxyproline	0.0018	0.0020	0.0021	0.0039	0.0021	0.0023	0.0090		0.0818	0.1722	0.0783	0.0941	0.0612	0.0621	0.0633	0.0835	0.0454
5-Hydroxyindole-3-acetic acid	0.0009	0.0012	0.0019	0.0008	0.0011	0.0008	0.0018	0.0005	0.0030	0.0028	0.0020	0.0020	0.0044	0.0022	0.0017	0.0025	0.0022
Aconitate	0.0009	0.0004	0.0009	0.0021	0.0005	0.0008	0.0063		0.0811	0.0008	0.0005	0.0005	0.0840	0.0006	0.0048	0.0009	0.0005
Adenine (A)	2.2239	0.9072	1.0872	2.1050	0.6653	0.5623	0.7478	1.0155	0.7187	2.1157	1.2528	1.2676	1.9386	0.7616	1.2665	0.9752	1.2669
Adenosine	0.0891	0.1913	0.0919	0.2446	0.0964	0.0877	0.1737	0.3100	0.1311	0.1361	0.1586	0.1293	0.1981	0.1221	0.0979	0.1429	0.1777
a-Ketobutyrate	0.0138	0.0058	0.0085	0.0079	0.0391	0.0368	0.0306	0.0175	0.0376	0.0086	0.0277	0.0134	0.0147	0.0024	0.0437	0.0049	0.0052
a-Ketoglutarate	0.0380	0.0166	0.0230	0.0217	0.1103	0.0984	0.0861	0.0465	0.0929	0.0196	0.0693	0.0311	0.0296	0.0042	0.1119	0.0072	0.0138
Alanine	0.2604	0.1313	0.2094	0.2048	0.0671	0.1733	0.1655	0.0938	0.2529	0.1423	0.1782	0.2211	0.2376	0.2190	0.2574	0.1905	0.1315
AMP							0.0109	0.0007	0.0314	0.0218	0.0067	0.0100	0.0281	0.0036	0.0015	0.0053	0.0033
Arginine	0.0774	0.0360	0.0450	0.1076	0.0202	0.0693	0.0574	0.0550	0.1470	0.0400	0.1215	0.1616	0.3805	0.2520	0.0856	0.0246	0.0209
Asparagine	0.0272	0.0015	0.0205	0.0201	0.0054	0.0256	0.0183	0.0137	0.0012		0.0011	0.0007	0.0013	0.0003	0.0011	0.0009	0.0006
Aspartate	0.2148	0.0696	0.1462	0.2214	0.1204	0.1664	0.1940	0.1366	0.2201	0.2239	0.1732	0.0717	0.3161	0.0782	0.3112	0.2195	0.1974
Betaine	0.1483	0.1224	0.1739	0.4004	0.0879	0.1996	0.8233	0.0739	1.6177	1.7410	0.3441	2.9933	3.0565	1.4881	0.7374	1.3942	0.5529
Citramalic acid	0.0025	0.0022		0.0041	0.0034	0.0038	0.0083	0.0029	0.0309	0.0105	0.0094	0.0075	0.0527	0.0135	0.0117	0.0575	0.0073
Citrate	0.1409	0.0536	0.0177	0.0649	0.0415	0.0429	0.0959	0.0281	2.0503	0.0272	0.0770	0.0467	1.5143	0.2743	0.3017	0.2803	0.1581
Creatine	0.0603	0.0374	0.0220	0.4226	0.0270	0.4778	0.4272	0.0242	3.4799	0.0884	0.0739	0.1591	5.1552	0.2066	0.7005	0.0576	0.0733
Creatinine	0.0051	0.0043	0.0181	0.6689	0.0627	0.0692	2.5218	0.0100	3.3104	0.0274	0.0143	0.0717	4.8406	0.0533	0.7164	0.0163	0.0321
Cystine	0.0031	0.0026	0.0014	0.0140	0.0231	0.0693	0.0487	0.0010								0.0002	
Cytidine	0.1258	0.1128	0.1107	0.1124	0.0848	0.1073	0.0953	0.0883	0.0996	0.0402	0.0804	0.0358	0.0459	0.0804	0.0460	0.0552	0.0805
Cytosine (C)	0.1073	0.0581	0.0801	0.0384	0.0737	0.0766	0.0473	0.0457	0.0215	0.0582	0.0553	0.0502	0.0675	0.0279	0.0961	0.0072	0.0698
Fumarate	0.0127	0.0287	0.0113	0.0100	0.0078	0.0081	0.0051	0.0052	0.0273	0.0488	0.0489	0.0183	0.0262	0.0202	0.0676	0.0458	0.0496
GABA	0.0641	0.0350	0.0695	0.0529	0.0649	0.0785	0.0667	0.0463	0.0623	0.0483	0.1047	0.0278	0.0489	0.0228	0.1746	0.0384	0.0425
Glutamate	1.2481	0.6659	1.0477	0.7499	1.1266	1.3128	1.1052	0.8227	1.0621	0.7551	1.4860	0.3897	0.7566	0.3951	2.5369	0.7508	0.8046
Glutamine	0.4277	0.1721	0.3191	0.2212	0.1000	0.1610	0.2227	0.1233	0.2664	0.2085	0.2418	0.2233	0.3611	0.1789	0.2731	0.2213	0.1091
GMP							0.0017		0.0086	0.0012	0.0013	0.0004	0.0042	0.0001	0.0004	0.0007	0.0005
Guanine (G)	0.0389	0.0213	0.0502	0.0570	0.0348	0.0363	0.0484	0.0321	0.3367	0.1107	0.1360	0.0566	0.1792	0.0636	0.0539	0.1159	0.1109
Guanosine	0.0154	0.0186	0.0208	0.0354	0.0110	0.0148	0.0224	0.0113	0.0755	0.0070	0.0274	0.0086	0.0241	0.0159	0.0148	0.0207	0.0169
Hexose-monophosphate_RT13.01			0.0002		0.0012	0.0050	0.0032	0.0001	0.0029	0.0003	0.0019	0.0029	0.0022	0.0003	0.0033	0.0004	0.0005
Hexose-monophosphate_RT13.69		0.0001	0.0006		0.0016	0.0091	0.0058	0.0003	0.0027	0.0005	0.0032	0.0094	0.0046	0.0011	0.0015	0.0006	0.0005
Hexose_RT5.6	0.0075	0.0074	0.0051	0.0176	0.0038	0.0111	0.0204	0.0033	0.1136	1.0665	0.1135	0.5467	0.9320	0.1640	0.0393	0.4624	0.0333
Hexose_RT7.4	0.0020	0.0018	0.0022	0.0033	0.0043	0.0073	0.0036	0.0045	0.0257	0.1868	0.0108	0.3118	0.1615	0.0359	0.0056	0.0235	0.0024
Hexose_RT8.3	0.0020	0.0019	0.0043	0.0029	0.0071	0.0095	0.0080	0.0052	0.0243	0.0045	0.0020	0.0152	0.0167	0.0444	0.0040	0.0312	0.0022
Hexose_RT8.7	0.0353	0.0277	0.0361	0.0273	0.0613	0.0693	0.0456	0.0436	0.0262	0.0294	0.0312	0.0100	0.0182	0.0198	0.0745	0.0248	0.0395
Histidine	0.3496	0.1529	0.1811	0.2133	0.0601	0.1258	0.1445	0.1106	0.1815	0.1971	0.2397	0.3649	0.2252	0.2432	0.2541	0.1479	0.1711
Indole									0.0026	0.0045	0.0022	0.0033	0.0037	0.0023	0.0033	0.0021	0.0021
Indole-3-acetic acid	0.0004	0.0025	0.0012	0.0015	0.0011	0.0009	0.0008	0.0006	0.0008	0.0036	0.0020	0.0021	0.0024	0.0015	0.0019	0.0009	0.0005
Indole-3-carboxaldehyde									0.0031	0.0046	0.0019	0.0027		0.0018		0.0013	0.0024
Indole-3-carboxylic acid				0.0006			0.0021		0.0015	0.0003	0.0004	0.0001	0.0053	0.0012	0.0003	0.0013	0.0013
Indole-3-ethanol									0.0029				0.0073				
Indole-3-glyoxylic acid			0.0008	0.0004		0.0004	0.0007		0.0491	0.0812	0.0469	0.0480	0.0546	0.0417	0.0713	0.0451	0.0480
Indole-3-lactic acid				0.0040			0.0018	0.0006	0.0013	0.0252	0.0005	0.0907	0.0115	0.0221	0.0004	0.0023	0.0004
Indole-3-propionic acid				0.0018	0.0012				0.0033	0.0096	0.0052	0.0056	0.0085	0.0046	0.0018	0.0014	0.0010
Isoleucine	0.3809	0.1881	0.4082	0.4038	0.0787	0.2540	0.2897	0.1528	0.2258	0.0820	0.2184	0.1058	0.3480	0.1351	0.3292	0.2379	0.1365
Kynurenic acid	0.0067	0.0123	0.0204	0.0095	0.0121	0.0098	0.0195	0.0064	0.0154	0.0305	0.0106	0.0171	0.0380	0.0104	0.0083	0.0081	0.0150
Lactate	0.0467	0.0243	0.1204	0.2436	0.0458	0.0476	0.0475	0.0333	0.1230	0.2642	0.0419	4.6899	0.0747	2.0035	0.1226	0.0816	0.0565
Leucine	0.6536	0.2232	0.5428	0.5523	0.1151	0.2933	0.3766	0.2107	0.3054	0.7231	0.2915	1.8840	0.3883	0.3723	0.4087	0.2567	0.1575
Lysine	0.0731	0.0281	0.0664	0.0563	0.0430	0.0696	0.0509	0.0519	0.0396	0.0354	0.0831	0.0446	0.0753	0.0497	0.1514	0.0323	0.0351
Malate	0.0968	0.2475	0.0717	0.1368	0.1068	0.1057	0.0750	0.0686	0.2893	0.5734	0.5304	0.2322	0.2805	0.2476	0.7197	0.5320	0.5311
Methionine	0.5488	0.1986	0.3842	0.3697	0.1258	0.3421	0.3013	0.2041	0.3113	0.1404	0.3277	0.2122	0.4115	0.1780	0.4708	0.2308	0.1994
Methylcitrate	0.0006	0.0009		0.0013	0.0018	0.0018	0.0026	0.0011	0.0035	0.0018	0.0033	0.0070	0.0047	0.0030	0.0068	0.0033	0.0019
N-Acetyl-5-hydroxytryptamine									0.0028	0.0017	0.0023	0.0008	0.0018	0.0009	0.0020	0.0020	0.0026
N-acetylaspartate	0.0233	0.0051	0.0119	0.0905	0.0830	0.0879	0.0976	0.0661	0.0878	0.1542	0.0768	0.1662	0.0937	0.1063	0.0946	0.0965	0.0706
Ornithine	0.0162	0.0084	0.0149	0.0376	0.0170	0.0233	0.0212	0.0130	0.0095	0.0375	0.0147	0.0386	0.0163	0.0117	0.0393	0.0042	0.0081
Pentose-monophosphate_RT12.27			0.0003		0.0026	0.0074	0.0114	0.0003	0.0096		0.0004		0.0005	0.0003	0.0046	0.0001	0.0005
Pentose-monophosphate_RT12.65					0.0048	0.0056	0.0065	0.0008	0.0163	0.0020	0.0031	0.0013	0.0023	0.0005	0.0094	0.0026	0.0048
Pentose-monophosphate_RT12.84		0.0003	0.0010		0.0078	0.0048	0.0069	0.0011	0.0102	0.0011	0.0031	0.0012	0.0019	0.0005	0.0102	0.0020	0.0047
Pentose_RT3.9				0.0019			0.0043		0.0048	0.0115	0.0019	0.0053	0.0188	0.0015	0.0020	0.0039	
Pentose_RT5.9				0.0021			0.0034			0.0126		0.0185	0.0312		0.0049		
Pentose_RT7.7										0.0019			0.0072				
Phenylalanine	0.5150	0.2016	0.3796	0.3668	0.0669	0.2254	0.2392	0.1543	0.2069	0.1215	0.2720	0.2975	0.3751	0.3243	0.2799	0.1895	0.1427
Proline	1.2567	1.1507	1.3154	1.9422	0.6808	1.2657	0.9342	0.5155	2.2487	2.3427	2.2295	1.2101	1.7743	2.8631	2.1030	2.0797	1.9222
Pyruvate	0.0022	0.0007	0.0023	0.0029	0.0049	0.0059	0.0051	0.0025	0.0027	0.0066	0.0031	0.0594	0.0045	0.0010	0.0069	0.0010	0.0011
Quinaldic acid		0.0006	0.0009		0.0009	0.0009			0.0046	0.0049	0.0058	0.0033	0.0057	0.0038	0.0025	0.0033	0.0045
Sarcosine	0.0164	0.0060	0.0102	0.1080	0.0117	0.0387	0.0524	0.0144	0.1143	0.0815	0.0301	0.1914	0.1764	0.1284	0.0413	0.0134	0.0324
Serine	0.0421	0.0226	0.0349	0.0242	0.0084	0.0159	0.0150	0.0105	0.0329	0.0222	0.0203	0.0183	0.0322	0.0196	0.0237	0.0294	0.0153
Succinate	0.0058	0.0081	0.0039	0.1061	0.0067	0.0074	0.0066	0.0055	0.1255	0.1955	0.0827	0.6135	0.1242	0.3251	0.0772	0.1178	0.0753
Threonine	0.1711	0.0906	0.1513	0.0912	0.0401	0.1054	0.0951	0.0585	0.1714	0.0764	0.1243	0.0960	0.1242	0.0873	0.1665	0.1123	0.0863
Thymine (T)	0.0119	0.0079	0.0201	0.0203	0.0122	0.0150	0.0086	0.0088	0.0246	0.0169	0.0305	0.0134	0.0207	0.0153	0.0444	0.0229	0.0208
TMP	0.0059	0.0049	0.0036	0.0028	0.0008	0.0033	0.0221	0.0043	0.0392	0.0105	0.0131	0.0093	0.0121	0.0042	0.0116	0.0224	0.0229
Tryptophan	0.0846	0.0348	0.0608	0.1419	0.0114	0.0371	0.0397	0.0235	0.0414	0.0541	0.0493	0.0613	0.0639	0.0629	0.0422	0.0462	0.0356
Tyrosine	0.4206	0.2052	0.3405	0.3568	0.0806	0.2234	0.2250	0.1433	0.2055	0.1156	0.2301	0.2209	0.3226	0.3286	0.2820	0.1628	0.1560
UMP	0.0001	0.0003	0.0004	0.0007	0.0000	0.0001	0.0078	0.0003	0.0157	0.0129	0.0044	0.0075	0.0158	0.0008	0.0008	0.0032	0.0014
Uracil (U)	0.0972	0.0484	0.1497	0.0618	0.0510	0.0680	0.0451	0.0304	0.1032	0.0326	0.0928	0.0194	0.0519	0.0297	0.1515	0.0654	0.0677
Uridine	0.0074	0.0087	0.0078	0.0179	0.0071	0.0106	0.0127	0.0093	0.0541	0.0208	0.0329	0.0120	0.0339	0.0274	0.0215	0.0263	0.0193
Valine	0.4774	0.1716	0.4613	0.6308	0.0843	0.2449	0.3143	0.1717	0.2313	0.1653	0.2291	0.2207	0.3414	0.1882	0.3228	0.1760	0.1200
Xanthurenic acid	0.0024	0.0119	0.0051	0.0347	0.0109	0.0137	0.0437	0.0184	0.0566	0.0125	0.0044	0.0059	0.0643	0.0144	0.0133	0.0049	0.0112
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	pubchem_id	inchi_key	kegg_id	other_id	other_id_type	ri	ri_type	moverz_quant	
2-Hydroxybutyric acid									
2-Hydroxyglutarate									
2-Picolinic acid									
3-(4-Hydroxyphenyl)propionic acid									
4-Hydroxy-2-oxoglutaric acid									
4-Hydroxyproline									
5-Hydroxyindole-3-acetic acid									
Aconitate									
Adenine (A)									
Adenosine									
a-Ketobutyrate									
a-Ketoglutarate									
Alanine									
AMP									
Arginine									
Asparagine									
Aspartate									
Betaine									
Citramalic acid									
Citrate									
Creatine									
Creatinine									
Cystine									
Cytidine									
Cytosine (C)									
Fumarate									
GABA									
Glutamate									
Glutamine									
GMP									
Guanine (G)									
Guanosine									
Hexose-monophosphate_RT13.01									
Hexose-monophosphate_RT13.69									
Hexose_RT5.6									
Hexose_RT7.4									
Hexose_RT8.3									
Hexose_RT8.7									
Histidine									
Indole									
Indole-3-acetic acid									
Indole-3-carboxaldehyde									
Indole-3-carboxylic acid									
Indole-3-ethanol									
Indole-3-glyoxylic acid									
Indole-3-lactic acid									
Indole-3-propionic acid									
Isoleucine									
Kynurenic acid									
Lactate									
Leucine									
Lysine									
Malate									
Methionine									
Methylcitrate									
N-Acetyl-5-hydroxytryptamine									
N-acetylaspartate									
Ornithine									
Pentose-monophosphate_RT12.27									
Pentose-monophosphate_RT12.65									
Pentose-monophosphate_RT12.84									
Pentose_RT3.9									
Pentose_RT5.9									
Pentose_RT7.7									
Phenylalanine									
Proline									
Pyruvate									
Quinaldic acid									
Sarcosine									
Serine									
Succinate									
Threonine									
Thymine (T)									
TMP									
Tryptophan									
Tyrosine									
UMP									
Uracil (U)									
Uridine									
Valine									
Xanthurenic acid									
METABOLITES_END
#END

#METABOLOMICS WORKBENCH mdm_20250119_095637 DATATRACK_ID:5542 STUDY_ID:ST003679 ANALYSIS_ID:AN006040 PROJECT_ID:
VERSION                          	1
CREATED_ON                       	01-23-2025
#PROJECT
#STUDY
#SUBJECT
#SUBJECT_SAMPLE_FACTORS:         	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data
#COLLECTION
#TREATMENT
#SAMPLEPREP
#CHROMATOGRAPHY
#ANALYSIS
#END