#METABOLOMICS WORKBENCH Renata_Goncalves_20250324_072348 DATATRACK_ID:5773 STUDY_ID:ST003846 ANALYSIS_ID:AN006321 PROJECT_ID:PR002404
VERSION             	1
CREATED_ON             	April 7, 2025, 6:26 pm
#PROJECT
PR:PROJECT_TITLE                 	Hepatic Coenzyme Q (CoQ) Deficiency Drives Reverse Electron Transport and
PR:PROJECT_TITLE                 	Disrupts Hepatic Metabolic Homeostasis in Obesity
PR:PROJECT_SUMMARY               	Mitochondrial reactive oxygen species (mROS) play a crucial role in physiology.
PR:PROJECT_SUMMARY               	While excessive mROS production has been associated with several disease states,
PR:PROJECT_SUMMARY               	its precise sources, regulation, and mechanisms of generation in vivo remain
PR:PROJECT_SUMMARY               	unclear, limiting translational efforts. mROS production is not a single
PR:PROJECT_SUMMARY               	process; at least 11 distinct sites associated with the electron transport chain
PR:PROJECT_SUMMARY               	and matrix substrate oxidation can partially reduce oxygen to superoxide and
PR:PROJECT_SUMMARY               	hydrogen peroxide. These sites exhibit substrate specificity and differ in their
PR:PROJECT_SUMMARY               	capacities to generate mROS. Here, we show that in obesity, hepatic coenzyme Q
PR:PROJECT_SUMMARY               	(CoQ) synthesis in vivo is impaired, leading to decreased CoQ9 levels and an
PR:PROJECT_SUMMARY               	increased CoQH₂/CoQ ratio. These alterations in CoQ metabolism drive excessive
PR:PROJECT_SUMMARY               	mROS production via reverse electron transport (RET) at site IQ of complex I.
PR:PROJECT_SUMMARY               	Using multiple complementary genetic and pharmacological in vivo models, we
PR:PROJECT_SUMMARY               	demonstrate that RET is critical for maintaining metabolic health. Furthermore,
PR:PROJECT_SUMMARY               	we show that in patients with steatosis, hepatic CoQ biosynthesis is also
PR:PROJECT_SUMMARY               	suppressed, and the CoQH₂/CoQ ratio positively correlates with steatosis
PR:PROJECT_SUMMARY               	grade. Our data identify a highly selective mechanism of pathological mROS
PR:PROJECT_SUMMARY               	production in obesity, which can be targeted to preserve metabolic homeostasis.
PR:INSTITUTE                     	Harvard School of Public Health
PR:DEPARTMENT                    	Molecular Metabolism
PR:LABORATORY                    	Hotamisligil Lab
PR:LAST_NAME                     	de Lima Sales Goncalves
PR:FIRST_NAME                    	Renata
PR:ADDRESS                       	665 Huntington Ave bld 1, room 609, Boston, MA, 02115, USA
PR:EMAIL                         	rgoncal@hsph.harvard.edu
PR:PHONE                         	4159402482
#STUDY
ST:STUDY_TITLE                   	Hepatic Coenzyme Q (CoQ) Deficiency Drives Reverse Electron Transport and
ST:STUDY_TITLE                   	Disrupts Hepatic Metabolic Homeostasis in Obesity
ST:STUDY_SUMMARY                 	Mitochondrial reactive oxygen species (mROS) play a crucial role in physiology.
ST:STUDY_SUMMARY                 	While excessive mROS production has been associated with several disease states,
ST:STUDY_SUMMARY                 	its precise sources, regulation, and mechanisms of generation in vivo remain
ST:STUDY_SUMMARY                 	unclear, limiting translational efforts. mROS production is not a single
ST:STUDY_SUMMARY                 	process; at least 11 distinct sites associated with the electron transport chain
ST:STUDY_SUMMARY                 	and matrix substrate oxidation can partially reduce oxygen to superoxide and
ST:STUDY_SUMMARY                 	hydrogen peroxide. These sites exhibit substrate specificity and differ in their
ST:STUDY_SUMMARY                 	capacities to generate mROS. Here, we show that in obesity, hepatic coenzyme Q
ST:STUDY_SUMMARY                 	(CoQ) synthesis in vivo is impaired, leading to decreased CoQ9 levels and an
ST:STUDY_SUMMARY                 	increased CoQH₂/CoQ ratio. These alterations in CoQ metabolism drive excessive
ST:STUDY_SUMMARY                 	mROS production via reverse electron transport (RET) at site IQ of complex I.
ST:STUDY_SUMMARY                 	Using multiple complementary genetic and pharmacological in vivo models, we
ST:STUDY_SUMMARY                 	demonstrate that RET is critical for maintaining metabolic health. Furthermore,
ST:STUDY_SUMMARY                 	we show that in patients with steatosis, hepatic CoQ biosynthesis is also
ST:STUDY_SUMMARY                 	suppressed, and the CoQH₂/CoQ ratio positively correlates with steatosis
ST:STUDY_SUMMARY                 	grade. Our data identify a highly selective mechanism of pathological mROS
ST:STUDY_SUMMARY                 	production in obesity, which can be targeted to preserve metabolic homeostasis.
ST:INSTITUTE                     	Harvard School of Public Health
ST:DEPARTMENT                    	Molecular Metabolism
ST:LABORATORY                    	Hotamisligil Lab
ST:LAST_NAME                     	de Lima Sales Goncalves
ST:FIRST_NAME                    	Renata
ST:ADDRESS                       	665 Huntington Ave bld 1, room 609
ST:EMAIL                         	rgoncal@hsph.harvard.edu
ST:PHONE                         	4159402482
ST:NUM_GROUPS                    	4
ST:TOTAL_SUBJECTS                	27
ST:NUM_MALES                     	27
#SUBJECT
SU:SUBJECT_TYPE                  	Mammal
SU:SUBJECT_SPECIES               	Mus musculus
SU:TAXONOMY_ID                   	10090
SU:GENOTYPE_STRAIN               	C57BL-6J
SU:AGE_OR_AGE_RANGE              	10 weeks
SU:GENDER                        	Male
SU:ANIMAL_ANIMAL_SUPPLIER        	The Jackson Lab
SU:ANIMAL_HOUSING                	4 mice per cage HSPH Animal facility
SU:ANIMAL_LIGHT_CYCLE            	12/12 dark/light
SU:ANIMAL_FEED                   	ad libtum
SU:ANIMAL_WATER                  	ad libitum
SU:ANIMAL_INCLUSION_CRITERIA     	wildtype lean and lepOB (ob/ob) obese or LepOB (ob/ob) obese mice expressing
SU:ANIMAL_INCLUSION_CRITERIA     	Ciona intestinalis Alternative oxidase (Aox) or green fluorescent protein (GFP)
SU:ANIMAL_INCLUSION_CRITERIA     	as control.
#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	Blank	Sample source:Blank | Genotype:Blank	RAW_FILE_NAME(Raw_File_Name)=Blank.mzXML
SUBJECT_SAMPLE_FACTORS           	20191108_1	20191108_aox	Sample source:liver | Genotype:Aox	RAW_FILE_NAME(Raw_File_Name)=20191108_aox.mzXML
SUBJECT_SAMPLE_FACTORS           	20191108_2	20191108_gfp	Sample source:liver | Genotype:GFP	RAW_FILE_NAME(Raw_File_Name)=20191108_gfp.mzXML
SUBJECT_SAMPLE_FACTORS           	20191110_1	20191110_aox	Sample source:liver | Genotype:Aox	RAW_FILE_NAME(Raw_File_Name)=20191110_aox.mzXML
SUBJECT_SAMPLE_FACTORS           	20191110_2	20191110_gfp	Sample source:liver | Genotype:GFP	RAW_FILE_NAME(Raw_File_Name)=20191110_gfp.mzXML
SUBJECT_SAMPLE_FACTORS           	20191111_1	20191111_aox	Sample source:liver | Genotype:Aox	RAW_FILE_NAME(Raw_File_Name)=20191111_aox.mzXML
SUBJECT_SAMPLE_FACTORS           	20191111_2	20191111_gfp	Sample source:liver | Genotype:GFP	RAW_FILE_NAME(Raw_File_Name)=20191111_gfp.mzXML
SUBJECT_SAMPLE_FACTORS           	20191203_1	20191203_aox	Sample source:liver | Genotype:Aox	RAW_FILE_NAME(Raw_File_Name)=20191203_aox.mzXML
SUBJECT_SAMPLE_FACTORS           	20191203_2	20191203_gfp	Sample source:liver | Genotype:GFP	RAW_FILE_NAME(Raw_File_Name)=20191203_gfp.mzXML
SUBJECT_SAMPLE_FACTORS           	20191204_1	20191204_aox	Sample source:liver | Genotype:Aox	RAW_FILE_NAME(Raw_File_Name)=20191204_aox.mzXML
SUBJECT_SAMPLE_FACTORS           	20191204_2	20191204_gfp	Sample source:liver | Genotype:GFP	RAW_FILE_NAME(Raw_File_Name)=20191204_gfp.mzXML
SUBJECT_SAMPLE_FACTORS           	20200304_1LR	20200304_1LR_aox	Sample source:liver | Genotype:Aox	RAW_FILE_NAME(Raw_File_Name)=20200304_1LR_aox.mzXML
SUBJECT_SAMPLE_FACTORS           	20200304_1L	20200304_1L_gfp	Sample source:liver | Genotype:GFP	RAW_FILE_NAME(Raw_File_Name)=20200304_1L_gfp.mzXML
SUBJECT_SAMPLE_FACTORS           	20200304_1N	20200304_1N_aox	Sample source:liver | Genotype:Aox	RAW_FILE_NAME(Raw_File_Name)=20200304_1N_aox.mzXML
SUBJECT_SAMPLE_FACTORS           	20200304_1R	20200304_1R_gfp	Sample source:liver | Genotype:GFP	RAW_FILE_NAME(Raw_File_Name)=20200304_1R_gfp.mzXML.
SUBJECT_SAMPLE_FACTORS           	20200304_3N	20200304_3N_aox	Sample source:liver | Genotype:Aox	RAW_FILE_NAME(Raw_File_Name)=20200304_3N_aox.mzXML
SUBJECT_SAMPLE_FACTORS           	20200304_5LR	20200304_5LR_aox	Sample source:liver | Genotype:Aox	RAW_FILE_NAME(Raw_File_Name)=20200304_5LR_aox.mzXML
SUBJECT_SAMPLE_FACTORS           	20200304_5R	20200304_5R_aox	Sample source:liver | Genotype:Aox	RAW_FILE_NAME(Raw_File_Name)=20200304_5R_aox.mzXML
SUBJECT_SAMPLE_FACTORS           	20200304_L	20200304_L_gfp	Sample source:liver | Genotype:GFP	RAW_FILE_NAME(Raw_File_Name)=20200304_L_gfp.mzXML
SUBJECT_SAMPLE_FACTORS           	20200304_R	20200304_R_gfp	Sample source:liver | Genotype:GFP	RAW_FILE_NAME(Raw_File_Name)=20200304_R_gfp.mzXML
SUBJECT_SAMPLE_FACTORS           	Lean 01	lean_01	Sample source:liver | Genotype:Lean wildtype	RAW_FILE_NAME(Raw_File_Name)=lean_01.mzXML
SUBJECT_SAMPLE_FACTORS           	Lean 01	lean_01_scanevent2	Sample source:liver | Genotype:Lean wildtype	RAW_FILE_NAME(Raw_File_Name)=lean_01_scanevent2.mzXML
SUBJECT_SAMPLE_FACTORS           	Lean 02	lean_02	Sample source:liver | Genotype:Lean wildtype	RAW_FILE_NAME(Raw_File_Name)=lean_02.mzXML
SUBJECT_SAMPLE_FACTORS           	Lean 02	lean_02_scanevent2	Sample source:liver | Genotype:Lean wildtype	RAW_FILE_NAME(Raw_File_Name)=lean_02_scanevent2.mzXML
SUBJECT_SAMPLE_FACTORS           	Lean 03	lean_03	Sample source:liver | Genotype:Lean wildtype	RAW_FILE_NAME(Raw_File_Name)=lean_03.mzXML
SUBJECT_SAMPLE_FACTORS           	Lean 03	lean_03_scanevent2	Sample source:liver | Genotype:Lean wildtype	RAW_FILE_NAME(Raw_File_Name)=lean_03_scanevent2.mzXML
SUBJECT_SAMPLE_FACTORS           	Lean 04	lean_04	Sample source:liver | Genotype:Lean wildtype	RAW_FILE_NAME(Raw_File_Name)=lean_04.mzXML
SUBJECT_SAMPLE_FACTORS           	Lean 04	lean_04_scanevent2	Sample source:liver | Genotype:Lean wildtype	RAW_FILE_NAME(Raw_File_Name)=lean_04_scanevent2.mzXML
SUBJECT_SAMPLE_FACTORS           	Obese 01	obese_01	Sample source:liver | Genotype:LepOB	RAW_FILE_NAME(Raw_File_Name)=obese_01.mzXML
SUBJECT_SAMPLE_FACTORS           	Obese 01	obese_01_scanevent2	Sample source:liver | Genotype:LepOB	RAW_FILE_NAME(Raw_File_Name)=obese_01_scanevent2.mzXML
SUBJECT_SAMPLE_FACTORS           	Obese 02	obese_02	Sample source:liver | Genotype:LepOB	RAW_FILE_NAME(Raw_File_Name)=obese_02.mzXML
SUBJECT_SAMPLE_FACTORS           	Obese 02	obese_02_scanevent2	Sample source:liver | Genotype:LepOB	RAW_FILE_NAME(Raw_File_Name)=obese_02_scanevent2.mzXML
SUBJECT_SAMPLE_FACTORS           	Obese 03	obese_03	Sample source:liver | Genotype:LepOB	RAW_FILE_NAME(Raw_File_Name)=obese_03.mzXML
SUBJECT_SAMPLE_FACTORS           	Obese 03	obese_03_scanevent2	Sample source:liver | Genotype:LepOB	RAW_FILE_NAME(Raw_File_Name)=obese_03_scanevent2.mzXML
SUBJECT_SAMPLE_FACTORS           	Obese 04	obese_04	Sample source:liver | Genotype:LepOB	RAW_FILE_NAME(Raw_File_Name)=obese_04.mzXML
SUBJECT_SAMPLE_FACTORS           	Obese 04	obese_04_scanevent2	Sample source:liver | Genotype:LepOB	RAW_FILE_NAME(Raw_File_Name)=obese_04_scanevent2.mzXML
#COLLECTION
CO:COLLECTION_SUMMARY            	Livers were extracted under freeze-clamp conditions using a Wollenberger clamp
CO:COLLECTION_SUMMARY            	and immediately frozen.
CO:SAMPLE_TYPE                   	Liver
CO:STORAGE_CONDITIONS            	-80℃
#TREATMENT
TR:TREATMENT_SUMMARY             	Livers from wild-type lean and LepOB (ob/ob) mice were homogenized in liquid
TR:TREATMENT_SUMMARY             	nitrogen using a cold tissuelyzer. To the pulverized tissues, 40% methanol:40%
TR:TREATMENT_SUMMARY             	acetonitrile (25 mg/mL) was added to each tube. The same protocol was used for
TR:TREATMENT_SUMMARY             	liver samples from mice ectopically expressing Ciona intestinalis alternative
TR:TREATMENT_SUMMARY             	oxidase (AOX) or green fluorescent protein (GFP) as a control.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	20-30mg of tissue were weighted in a previously chilled tube at -20oC and
SP:SAMPLEPREP_SUMMARY            	tissuelyser homogenizer beads. Tissues were homogenized in liquid nitrogen using
SP:SAMPLEPREP_SUMMARY            	a cold tissuelyzer. To the pulverized tissues, 40% methanol:40% acetonitrile (25
SP:SAMPLEPREP_SUMMARY            	mg/mL) was added to each tube. Each tube was vortexed for the 10 sec., incubate
SP:SAMPLEPREP_SUMMARY            	for 10 min on ice and centrifuged 10 min at 3000rpm to pellet debris. 600uL of
SP:SAMPLEPREP_SUMMARY            	the supernatant was transferred to a new tube and centrifuged at max speed for
SP:SAMPLEPREP_SUMMARY            	10’. 80uL of the clean extract was added to a fresh vial and analyzed via mass
SP:SAMPLEPREP_SUMMARY            	spec.
SP:PROCESSING_STORAGE_CONDITIONS 	-80℃
SP:EXTRACT_STORAGE               	-20℃
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	For small polar metabolite separation and data acquisition in extracted tissues,
CH:CHROMATOGRAPHY_SUMMARY        	a Vanquish Horizon (Thermo Fisher Scientific, Waltham, MA) ultra-high liquid
CH:CHROMATOGRAPHY_SUMMARY        	chromatography (LC) system coupled to a Thermo Q Exactive HF Orbitrap mass
CH:CHROMATOGRAPHY_SUMMARY        	spectrometer (MS) was used. For separation, a Waters (Milford, MA) XBridge BEH
CH:CHROMATOGRAPHY_SUMMARY        	Amide (2.5 μm, 2.1x150 mm) column fitted with a VanGuard (2.5 μm, 2.1x5 mm)
CH:CHROMATOGRAPHY_SUMMARY        	guard column was used. The mobile phases were as follows: Phase A: 95% water/5%
CH:CHROMATOGRAPHY_SUMMARY        	acetonitrile and Phase B: 20% water/80% acetonitrile with 10 mM ammonium acetate
CH:CHROMATOGRAPHY_SUMMARY        	and 10 mM ammonium hydroxide in both phases. The flow rate was held constant at
CH:CHROMATOGRAPHY_SUMMARY        	0.3 ml/min and the following gradient conditions were used: 0 min, 100% B; 3
CH:CHROMATOGRAPHY_SUMMARY        	min, 100% B; 3.2 min, 90% B; 6.2 min, 90% B; 6.5 min, 80% B; 10.5 min, 80% B;
CH:CHROMATOGRAPHY_SUMMARY        	10.7 min, 70% B; 13.5 min, 70% B; 13.7 min, 45% B; 16 min, 45% B, 16.5 min, 100%
CH:CHROMATOGRAPHY_SUMMARY        	B; and 22 min, 100% B. The samples were kept at 4°C, the injection volume was 5
CH:CHROMATOGRAPHY_SUMMARY        	μl and the column was maintained at 25°C. The separated metabolites were
CH:CHROMATOGRAPHY_SUMMARY        	analyzed in both positive and negative ionization modes in the same run
CH:CHROMATOGRAPHY_SUMMARY        	(switching mode). The mass spectra were acquired using a resolution of 120,000
CH:CHROMATOGRAPHY_SUMMARY        	in the 70 – 1,000 m/z range. The ElectroSpray Ionization source parameters for
CH:CHROMATOGRAPHY_SUMMARY        	both modes were as follows: capillary temperature 300°C, spray voltage 3.5kV,
CH:CHROMATOGRAPHY_SUMMARY        	sheath gas 40 (arbitrary units), auxiliary gas 10 (arbitrary units), probe
CH:CHROMATOGRAPHY_SUMMARY        	heater temperature 30°C and S-Lens RF level 45 v.
CH:CHROMATOGRAPHY_TYPE           	HILIC
CH:INSTRUMENT_NAME               	Thermo Vanquish Horizon
CH:COLUMN_NAME                   	Waters XBridge BEH Amide (150 x 2.1mm, 2.5um) with VanGuard (5 x 2.1mm, 2.5um)
CH:COLUMN_NAME                   	guard column
CH:SOLVENT_A                     	95% water/5% acetonitrile; 10 mM ammonium acetate; 10 mM ammonium hydroxide
CH:SOLVENT_B                     	20% water/80% acetonitrile; 10 mM ammonium acetate; 10 mM ammonium hydroxide
CH:FLOW_GRADIENT                 	0 min, 100% B; 3 min, 100% B; 3.2 min, 90% B; 6.2 min, 90% B; 6.5 min, 80% B;
CH:FLOW_GRADIENT                 	10.5 min, 80% B; 10.7 min, 70% B; 13.5 min, 70% B; 13.7 min, 45% B; 16 min, 45%
CH:FLOW_GRADIENT                 	B, 16.5 min, 100% B; and 22 min, 100% B
CH:FLOW_RATE                     	0.3 mL/min
CH:COLUMN_TEMPERATURE            	25
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
AN:LABORATORY_NAME               	Hui lab
AN:OPERATOR_NAME                 	Clement Rosique and Tony Shen Hui
#MS
MS:INSTRUMENT_NAME               	Thermo Q Exactive HF-X Orbitrap
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	POSITIVE
MS:MS_COMMENTS                   	For small polar metabolite separation and data acquisition in extracted tissues,
MS:MS_COMMENTS                   	a Vanquish Horizon (Thermo Fisher Scientific, Waltham, MA) ultra-high liquid
MS:MS_COMMENTS                   	chromatography (LC) system coupled to a Thermo Q Exactive HF Orbitrap mass
MS:MS_COMMENTS                   	spectrometer (MS) was used. For separation, a Waters (Milford, MA) XBridge BEH
MS:MS_COMMENTS                   	Amide (2.5 μm, 2.1x150 mm) column fitted with a VanGuard (2.5 μm, 2.1x5 mm)
MS:MS_COMMENTS                   	guard column was used. The mobile phases were as follows: Phase A: 95% water/5%
MS:MS_COMMENTS                   	acetonitrile and Phase B: 20% water/80% acetonitrile with 10 mM ammonium acetate
MS:MS_COMMENTS                   	and 10 mM ammonium hydroxide in both phases. The flow rate was held constant at
MS:MS_COMMENTS                   	0.3 ml/min and the following gradient conditions were used: 0 min, 100% B; 3
MS:MS_COMMENTS                   	min, 100% B; 3.2 min, 90% B; 6.2 min, 90% B; 6.5 min, 80% B; 10.5 min, 80% B;
MS:MS_COMMENTS                   	10.7 min, 70% B; 13.5 min, 70% B; 13.7 min, 45% B; 16 min, 45% B, 16.5 min, 100%
MS:MS_COMMENTS                   	B; and 22 min, 100% B. The samples were kept at 4°C, the injection volume was 5
MS:MS_COMMENTS                   	μl and the column was maintained at 25°C. The separated metabolites were
MS:MS_COMMENTS                   	analyzed in both positive and negative ionization modes in the same run
MS:MS_COMMENTS                   	(switching mode). The mass spectra were acquired using a resolution of 120,000
MS:MS_COMMENTS                   	in the 70 – 1,000 m/z range. The ElectroSpray Ionization source parameters for
MS:MS_COMMENTS                   	both modes were as follows: capillary temperature 300°C, spray voltage 3.5kV,
MS:MS_COMMENTS                   	sheath gas 40 (arbitrary units), auxiliary gas 10 (arbitrary units), probe
MS:MS_COMMENTS                   	heater temperature 30°C and S-Lens RF level 45 v. Each single raw data file is
MS:MS_COMMENTS                   	processed into two separate ones each of one polarity mode. In this repository,
MS:MS_COMMENTS                   	the data of each polarity is deposited separately for clarity.
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	peak area
MS_METABOLITE_DATA_START
Samples	lean_01_scanevent2	lean_02_scanevent2	lean_03_scanevent2	lean_04_scanevent2	obese_01_scanevent2	obese_02_scanevent2	obese_03_scanevent2	obese_04_scanevent2
Factors	Sample source:liver | Genotype:Lean wildtype	Sample source:liver | Genotype:Lean wildtype	Sample source:liver | Genotype:Lean wildtype	Sample source:liver | Genotype:Lean wildtype	Sample source:liver | Genotype:LepOB	Sample source:liver | Genotype:LepOB	Sample source:liver | Genotype:LepOB	Sample source:liver | Genotype:LepOB
Beta-Alanine	2424160	2433213	1705227	1602715	1029024	918828.7	972610.8	804995
2-aminobutyric acid	135857.5	187008.2	152208.4	109146.7	163952.1	175070.6	136585.2	177836
Choline	95857130	96295660	76335210	85748490	45809560	47300250	51968320	46413800
Creatinine	3162414	2619612	2046739	1910662	1391390	1090659	1086043	1503362
Betaine	104197300	96933820	106895100	85416900	60538040	74216710	67088380	76281620
Nicotinamide	1860894	392654.5	1413813	5973880	1421234	2280371	248914.1	991282.8
4-Imidazoleacetate	216351.1	94581.09	70934.58	57812.92	46759.64	36365.59	26043.22	44000.27
Pipecolate	495015.4	485746.7	417088.9	284366.3	395644.5	356078	393753	670783.3
5-Aminolevulinate	1120277	1340471	663696.5	366543.6	457324.8	171097.4	127377.5	97695.88
Creatine	18405910	20560520	21348600	14777000	8484239	9967250	8455845	14013890
Ornithine	2832589	1913710	1875888	2004543	957019.4	657878.9	789071.8	1026837
Methylnicotinamide	1217428	1562320	592921.7	605416.9	3356204	3600687	3827209	4446492
O-Phosphorylethanolamine	1178125	943206.4	1298206	1008739	400355	333680	418124.3	452429.1
Carnitine	80512480	79286330	84734840	67795500	84704670	75397180	67842410	74228010
cis-aconitic acid	41441.74	97070.66	16982.76	29645.74	14771.98	19859.94	38605.37	33937.89
Guanidinosuccinic acid	941540.7	341923.3	337171.6	138620.4	225184.7	183063.5	94898.16	83698.07
Serotonin	129541.8	29456.95	28137.1	47260.76	35131	29832.25	43210.55	54541.93
3-phosphoglycerate	41847.74	56195.18	57159.8	29083.19	29902.92	43979.47	14805.66	16664.84
Acetylglutamine	286213.8	314924.1	340970.6	281923.2	250119.2	172371.3	210830.1	79547.52
N-Acetyl-lysine	499804.8	629337.7	488014.8	331005.8	334860.3	286655.8	264926.2	229905.7
Homoarginine	123985.3	125855.2	118823	107755.9	40803.26	30753.44	8770.954	9280.509
O-Acetyl-L-carnitine	61711560	41512650	54430720	36896750	52946270	47984660	50226590	61377200
Anserine	68964.88	42087.86	38458.25	16316.34	5440.573	7439.043	5633.527	8154.857
Adenosine	11068930	5714202	6994450	6058026	3716263	1462147	4536758	4055679
Glutaryl-carnitine	8549857	7581160	6821796	5643868	2687934	2123291	2017989	2943058
Ophthalmate	472909	427221.8	941423.5	1425840	49686.19	78442.66	75889.44	36628.25
AMP	11592140	7618350	9254059	11278080	3130627	3358099	4497058	3658587
GMP	904700.7	389225.8	419304.9	1874482	277323.4	450633.4	188746.3	271121.5
S-Adenosyl-homocysteine	1475662	3085465	1302041	1154859	439722.6	547157.6	744627.2	907168.2
S-Adenosylmethionine	1351542	0	0	0	678690.8	0	0	233318.6
Thiamine pyrophosphate	47406.42	35804.5	37395.49	19748.35	31357.91	20642.76	17855.67	30423.81
Adenosine 3 5-diphosphate	1509753	1411841	1326296	1037642	792625.4	1162849	802213.3	1161797
cytidine 5'-diphosphocholine (CDP-choline)	620932.8	500176.9	563526.2	424999.8	111615.2	53390.44	262278.6	91426.63
ATP	0	121688.2	0	21119.98	196696.7	302233.5	107942.2	185514.7
Adenosine 5'-diphosphoribose (ADP-ribose)	218093.6	62132.95	134054.3	586669.3	169311.5	313771.6	13769.82	128081.1
flavin adenine dinucleotide (FAD)	143656.7	138905.5	108326.6	53251.94	116104.2	180596.4	96628.79	136414.5
Acetyl CoA	143212.6	200855.5	93033.35	34945.84	167812	142939	131022.1	176680.8
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	retention time (min)	m/z	HMDB	ChEBI	PubChem	KEGG	METLIN
Beta-Alanine	6.7	90.055504	HMDB0000056
2-aminobutyric acid	6.2	104.071154	HMDB0000452	35619	80283
Choline	5.2	104.107539	HMDB0000939	16680	439155	C00021	296
Creatinine	2.7	114.066737	HMDB0000562	16737	588	C00791	8
Betaine	4.5	118.086804	HMDB0000056	16958	239	C00099	5119
Nicotinamide	1.8	123.055838	HMDB0001406	17154	936	C00153	1497
4-Imidazoleacetate	5.7	127.050753	HMDB0000050	16335	60961	C00212	86
Pipecolate	5.4	130.086804	HMDB0000070	17964	849	C00408	50
5-Aminolevulinate	6.7	132.066069	HMDB0001149
Creatine	6.2	132.077302	HMDB0000271	15611	1088	C00213	51
Ornithine	12.7	133.097703	HMDB0000214		75606
Methylnicotinamide	5.5	137.071488	HMDB0000064	16919	586	C00300	7
O-Phosphorylethanolamine	9.6	142.026922	HMDB0000224	17553	1015	C00346	5233
Carnitine	6.4	162.113019	HMDB0000062	16347	10917	C00318	52
cis-aconitic acid	9	175.024265	HMDB0000072	32805
Guanidinosuccinic acid	8.3	176.067132	HMDB0003157	17072	439918	C03139	10
Serotonin	3.3	177.102788	HMDB0000259	28790	5202	C00780	74
3-phosphoglycerate	8.6	187.000768	HMDB0000807	17050	724	C00597	150
Acetylglutamine	4.4	189.087533	HMDB0006029	21553
N-Acetyl-lysine	7.5	189.123918	HMDB0001372	9532	1132	C00068	2832
Homoarginine	13	189.135151	HMDB0000670	27747	9085	C01924	5640
O-Acetyl-L-carnitine	5.3	204.123584	HMDB0005765	84058	7018721	C21016
Anserine	8.6	241.130066	HMDB0000026	18261	111	C02642	5097
Adenosine	2.6	268.10458	HMDB0000050
Glutaryl-carnitine	8.3	276.144714	HMDB0013130
Ophthalmate	8.3	290.135212	HMDB0005765
AMP	8.9	348.070913	HMDB0000045	16027	6083	C00020	5111
GMP	9.7	364.065828	HMDB0001397	17345	6804	C00144	6216
S-Adenosyl-homocysteine	7.8	385.129416	HMDB0000099	17482	439258	C02291	39
S-Adenosylmethionine	11.2	399.145066	HMDB0001185	15414	34756	C00019	6064
Thiamine pyrophosphate	11.7	425.044974	HMDB0001078	49728	439198	C00275	5987
Adenosine 3 5-diphosphate	9.9	428.037246	HMDB0000061
cytidine 5'-diphosphocholine (CDP-choline)	9.3	489.115162	HMDB0001983	17319	439182	C05198
ATP	11.5	508.003579	HMDB0000538	15422	5957	C00002	5523
Adenosine 5'-diphosphoribose (ADP-ribose)	8.8	560.079506	HMDB0001206	15351	444493	C00024	6082
flavin adenine dinucleotide (FAD)	8.1	786.164967	HMDB0001248
Acetyl CoA	8.8	810.133609	HMDB0000201		7045767	C02571	5213
METABOLITES_END
#END