#METABOLOMICS WORKBENCH cwalker_bcm_20200127_072952 DATATRACK_ID:1909 STUDY_ID:ST001309 ANALYSIS_ID:AN002178 PROJECT_ID:PR000890
VERSION             	1
CREATED_ON             	January 29, 2020, 2:04 pm
#PROJECT
PR:PROJECT_TITLE                 	Metabolomic profiling after early-life exposure to an endocrine disrupting
PR:PROJECT_TITLE                 	chemical in the liver.
PR:PROJECT_TYPE                  	Targeted MS analysis
PR:PROJECT_SUMMARY               	Metabolic profiling in the liver (240 days post-natal) after early-life exposure
PR:PROJECT_SUMMARY               	to an endocrine disrupting chemical.
PR:INSTITUTE                     	Baylor College of Medicine
PR:LAST_NAME                     	Walker
PR:FIRST_NAME                    	Cheryl
PR:ADDRESS                       	1 Baylor Plaza, Houston, TX, 77030, USA
PR:EMAIL                         	Cheryl.walker@bcm.edu
PR:PHONE                         	713-798-8219
#STUDY
ST:STUDY_TITLE                   	Metabolite expression in liver after early life exposure to an endocrine
ST:STUDY_TITLE                   	disruptor at 240 days postnatal (part-I)
ST:STUDY_TYPE                    	Metabolite expression after chemical exposure versus control.
ST:STUDY_SUMMARY                 	Our early-life environment has a profound influence on developing organs that
ST:STUDY_SUMMARY                 	impact metabolic function and determines disease susceptibility across the
ST:STUDY_SUMMARY                 	life-course. Using a rat model for exposure to an endocrine disrupting chemical
ST:STUDY_SUMMARY                 	(EDC), we show that early-life exposure causes metabolic dysfunction in
ST:STUDY_SUMMARY                 	adulthood and reprograms histone marks in the developing liver to accelerate
ST:STUDY_SUMMARY                 	acquisition of an adult epigenomic signature. This epigenomic reprogramming
ST:STUDY_SUMMARY                 	persists long after the initial exposure, but many reprogrammed genes remain
ST:STUDY_SUMMARY                 	transcriptionally silent with their impact on metabolism not revealed until a
ST:STUDY_SUMMARY                 	later life exposure to a Western-style diet. Diet-dependent metabolic disruption
ST:STUDY_SUMMARY                 	was largely driven by reprogramming of the Early Growth Response 1 (EGR1)
ST:STUDY_SUMMARY                 	transcriptome and production of metabolites in pathways linked to cholesterol,
ST:STUDY_SUMMARY                 	lipid and one-carbon metabolism. These findings demonstrate the importance of
ST:STUDY_SUMMARY                 	epigenome: environment interactions, which early in life accelerate epigenomic
ST:STUDY_SUMMARY                 	aging, and later in adulthood unlock metabolically restricted epigenetic
ST:STUDY_SUMMARY                 	reprogramming to drive metabolic dysfunction.
ST:INSTITUTE                     	Baylor College of Medicine
ST:DEPARTMENT                    	Molecular and Cellular Biology
ST:LABORATORY                    	Center for Precision Environmental Health
ST:LAST_NAME                     	Walker
ST:FIRST_NAME                    	Cheryl
ST:ADDRESS                       	1 Baylor Plaza, Houston, TX, 77030, USA
ST:EMAIL                         	Cheryl.walker@bcm.edu
ST:PHONE                         	713-798-8219
ST:NUM_GROUPS                    	2
ST:TOTAL_SUBJECTS                	10
ST:NUM_MALES                     	10
#SUBJECT
SU:SUBJECT_TYPE                  	Mammal
SU:SUBJECT_SPECIES               	Rattus norvegicus
SU:TAXONOMY_ID                   	10116
SU:GENOTYPE_STRAIN               	Sprague Dawley
SU:AGE_OR_AGE_RANGE              	240 days
SU:GENDER                        	Male
SU:ANIMAL_ANIMAL_SUPPLIER        	Harlan
SU:ANIMAL_HOUSING                	polycarbonate-free caging
SU:ANIMAL_LIGHT_CYCLE            	14-hr light and 10-hr dark
SU:ANIMAL_FEED                   	Phytoestrogen Reduced II 18-5 (Ziegler Bros, Inc) or D09100301 (Research Diets,
SU:ANIMAL_FEED                   	Inc)
#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           	-	HFD-VEH13	Treatment:vehicle	Day of Liver Harvest Post-Birth=240; BPA_exposure=0 ug/kg post natal days 1, 3, and 5; Diet=phytoestrogen-reduced a diet for first 180 days post birth then diet high in fat (40% kcal), fructose (20% kcal) and cholesterol (2%) (Western-style diet) for 60 days.
SUBJECT_SAMPLE_FACTORS           	-	HFD-VEH15	Treatment:vehicle	Day of Liver Harvest Post-Birth=240; BPA_exposure=0 ug/kg post natal days 1, 3, and 5; Diet=phytoestrogen-reduced a diet for first 180 days post birth then diet high in fat (40% kcal), fructose (20% kcal) and cholesterol (2%) (Western-style diet) for 60 days.
SUBJECT_SAMPLE_FACTORS           	-	HFD-VEH18	Treatment:vehicle	Day of Liver Harvest Post-Birth=240; BPA_exposure=0 ug/kg post natal days 1, 3, and 5; Diet=phytoestrogen-reduced a diet for first 180 days post birth then diet high in fat (40% kcal), fructose (20% kcal) and cholesterol (2%) (Western-style diet) for 60 days.
SUBJECT_SAMPLE_FACTORS           	-	HFD-VEH12	Treatment:vehicle	Day of Liver Harvest Post-Birth=240; BPA_exposure=0 ug/kg post natal days 1, 3, and 5; Diet=phytoestrogen-reduced a diet for first 180 days post birth then diet high in fat (40% kcal), fructose (20% kcal) and cholesterol (2%) (Western-style diet) for 60 days.
SUBJECT_SAMPLE_FACTORS           	-	HFD-VEH14	Treatment:vehicle	Day of Liver Harvest Post-Birth=240; BPA_exposure=0 ug/kg post natal days 1, 3, and 5; Diet=phytoestrogen-reduced a diet for first 180 days post birth then diet high in fat (40% kcal), fructose (20% kcal) and cholesterol (2%) (Western-style diet) for 60 days.
SUBJECT_SAMPLE_FACTORS           	-	HFD-BPA12	Treatment:BPA	Day of Liver Harvest Post-Birth=240; BPA_exposure=50 ug/kg post natal days 1, 3, and 5; Diet=phytoestrogen-reduced a diet for first 180 days post birth then diet high in fat (40% kcal), fructose (20% kcal) and cholesterol (2%) (Western-style diet) for 60 days.
SUBJECT_SAMPLE_FACTORS           	-	HFD-BPA13	Treatment:BPA	Day of Liver Harvest Post-Birth=240; BPA_exposure=50 ug/kg post natal days 1, 3, and 5; Diet=phytoestrogen-reduced a diet for first 180 days post birth then diet high in fat (40% kcal), fructose (20% kcal) and cholesterol (2%) (Western-style diet) for 60 days.
SUBJECT_SAMPLE_FACTORS           	-	HFD-BPA14	Treatment:BPA	Day of Liver Harvest Post-Birth=240; BPA_exposure=50 ug/kg post natal days 1, 3, and 5; Diet=phytoestrogen-reduced a diet for first 180 days post birth then diet high in fat (40% kcal), fructose (20% kcal) and cholesterol (2%) (Western-style diet) for 60 days.
SUBJECT_SAMPLE_FACTORS           	-	HFD-BPA16	Treatment:BPA	Day of Liver Harvest Post-Birth=240; BPA_exposure=50 ug/kg post natal days 1, 3, and 5; Diet=phytoestrogen-reduced a diet for first 180 days post birth then diet high in fat (40% kcal), fructose (20% kcal) and cholesterol (2%) (Western-style diet) for 60 days.
SUBJECT_SAMPLE_FACTORS           	-	HFD-BPA18	Treatment:BPA	Day of Liver Harvest Post-Birth=240; BPA_exposure=50 ug/kg post natal days 1, 3, and 5; Diet=phytoestrogen-reduced a diet for first 180 days post birth then diet high in fat (40% kcal), fructose (20% kcal) and cholesterol (2%) (Western-style diet) for 60 days.
#COLLECTION
CO:COLLECTION_SUMMARY            	Liver tissue was harvested on post-natal day 240 after challenge with
CO:COLLECTION_SUMMARY            	Western-style diet.Tissue was snap-frozen in liquid nitrogen.
CO:SAMPLE_TYPE                   	Liver
#TREATMENT
TR:TREATMENT_SUMMARY             	Neonatal rats were treated with vehicle (sesame oil) or bisphenol A (BPA; 50
TR:TREATMENT_SUMMARY             	µg/kg dissolved in sesame oil) orally via pipette tip on post-natal days 1, 3,
TR:TREATMENT_SUMMARY             	and 5. Littermates were randomly assigned to the treatment groups. BPA was
TR:TREATMENT_SUMMARY             	obtained from the National Institute of Environmental Health Sciences (NIEHS).
TR:TREATMENT_SUMMARY             	The dose and route of administration recapitulates human exposure to BPA. At day
TR:TREATMENT_SUMMARY             	180, adult rats in both treatment groups were fed a diet high in fat (40% kcal),
TR:TREATMENT_SUMMARY             	fructose (20% kcal) and cholesterol (2%) (Western-style diet) for 60 days
TR:TREATMENT_SUMMARY             	(D09100301, Research Diets, Inc). Rats were fasted overnight prior to tissue
TR:TREATMENT_SUMMARY             	collection.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Metabolites were extracted from crushed liver samples and a mouse liver pool was
SP:SAMPLEPREP_SUMMARY            	used for quality control. Twenty-five mg of crushed liver was used for the
SP:SAMPLEPREP_SUMMARY            	metabolic extraction. The extraction step started with the addition of 750 µL
SP:SAMPLEPREP_SUMMARY            	ice-cold methanol:water (4:1) containing 20 µL spiked internal standards to
SP:SAMPLEPREP_SUMMARY            	each tissue sample. Ice-cold chloroform and water were added in a 3:1 ratio for
SP:SAMPLEPREP_SUMMARY            	a final proportion of 1:4:3:1 water:methanol:chloroform:water. The organic
SP:SAMPLEPREP_SUMMARY            	(methanol and chloroform) and aqueous layers were mixed, dried and resuspended
SP:SAMPLEPREP_SUMMARY            	with 50:50 methanol: water. The extract was deproteinized using a 3kDa molecular
SP:SAMPLEPREP_SUMMARY            	filter (Amicon ultracel-3K Membrane; Millipore Corporation, Billerica, MA) and
SP:SAMPLEPREP_SUMMARY            	the filtrate was dried under vacuum (Genevac EZ-2plus; Gardiner, Stone Ridge,
SP:SAMPLEPREP_SUMMARY            	NY). Prior to mass spectrometry, the dried extracts were re-suspended in
SP:SAMPLEPREP_SUMMARY            	identical volumes of injection solvent composed of 1:1 water: methanol and were
SP:SAMPLEPREP_SUMMARY            	subjected to liquid chromatography-mass spectrometry. Fifty µl of sample was
SP:SAMPLEPREP_SUMMARY            	used for preparation. Internal standards were spiked into the samples. Then it
SP:SAMPLEPREP_SUMMARY            	was processed through a 3 kDa filter. After that, 50 µl of sample was diluted
SP:SAMPLEPREP_SUMMARY            	with 450 µl solvent (methanol: water = 50:50 v/v) and subjected to LC/MS
SP:SAMPLEPREP_SUMMARY            	analysis. The injection volume was 10 µl. For internal standards,
SP:SAMPLEPREP_SUMMARY            	high-performance liquid chromatography (HPLC)-grade acetonitrile, methanol, and
SP:SAMPLEPREP_SUMMARY            	water were procured from Burdick & Jackson (Morristown, NJ). Mass
SP:SAMPLEPREP_SUMMARY            	spectrometry-grade formic acid was purchased from Sigma-Aldrich (St Louis, MO).
SP:SAMPLEPREP_SUMMARY            	Calibration solution containing multiple calibrants in a solution of
SP:SAMPLEPREP_SUMMARY            	acetonitrile, trifluroacetic acid, and water was purchased from Agilent
SP:SAMPLEPREP_SUMMARY            	Technologies (Santa Clara, CA). Metabolites and internal standards, including
SP:SAMPLEPREP_SUMMARY            	N-acetyl Aspartic acid-d3, Tryptophan-15N2, Sarcosine-d3, Glutamic acid-d5,
SP:SAMPLEPREP_SUMMARY            	Thymine-d4, Gibberellic acid, Trans-Zeatine, Jasmonic acid, 15N Anthranilic
SP:SAMPLEPREP_SUMMARY            	acid, and Testosterone-d3, were purchased from Sigma-Aldrich (St. Louis, MO).
SP:SAMPLEPREP_SUMMARY            	Three LC- MS methods were used to separate metabolites. Method A: In ESI
SP:SAMPLEPREP_SUMMARY            	positive mode the HPLC column was waters X-bridge amide 3.5 µm, 4.6 x 100 mm
SP:SAMPLEPREP_SUMMARY            	(Waters, Milford, MA). Mobile phase A and B were 0.1% formic acid in water and
SP:SAMPLEPREP_SUMMARY            	acetonitrile, respectively. Gradient flow: 0-3 min 85% B; 3-12 min 30% B, 12-15
SP:SAMPLEPREP_SUMMARY            	min 2% B, 16 min 95%B, followed by re-equilibration till the end of the gradient
SP:SAMPLEPREP_SUMMARY            	23 min to the initial starting condition of 85% B. Flow rate of the solvents
SP:SAMPLEPREP_SUMMARY            	used for the analysis is 0.3 ml/min. Injection volume was 10 µL. Method B: In
SP:SAMPLEPREP_SUMMARY            	ESI negative mode the HPLC column was waters X-bridge amide 3.5 µm, 4.6 x 100
SP:SAMPLEPREP_SUMMARY            	mm (Waters, Milford, MA). Mobile phase A and B were 20 mM ammonium acetate in
SP:SAMPLEPREP_SUMMARY            	water with pH 9.0 and 100% acetonitrile, respectively. Gradient flow: 0-3 min
SP:SAMPLEPREP_SUMMARY            	85% B, 3-12 min 30% B, 12-15 min 2% B, 15-16 min 85% B followed by
SP:SAMPLEPREP_SUMMARY            	re-equilibration till the end of the gradient 23 min to the initial starting
SP:SAMPLEPREP_SUMMARY            	condition of 85% B. Flow rate of the solvents used for analysis is 0.3 ml/min.
SP:SAMPLEPREP_SUMMARY            	Injection volume was 10 µL. Method C: In ESI positive mode the HPLC column was
SP:SAMPLEPREP_SUMMARY            	Luna 3 µM NH2 100 A0 Chromatography column (Phenomenex, Torrance, CA). Mobile
SP:SAMPLEPREP_SUMMARY            	phase A and B were 20 mM ammonium acetate in water with pH 9.0 and 100%
SP:SAMPLEPREP_SUMMARY            	acetonitrile, respectively. Gradient flow: 0-3 min 85% B, 3-12 min 30% B, 12-15
SP:SAMPLEPREP_SUMMARY            	min 2% B, 15-16 min 85% B followed by re-equilibration till the end of the
SP:SAMPLEPREP_SUMMARY            	gradient 23 min to the initial starting condition of 85% B. Flow rate of the
SP:SAMPLEPREP_SUMMARY            	solvents used for analysis is 0.3 ml/min. Injection volume was 10 µL. For data
SP:SAMPLEPREP_SUMMARY            	acquisition through LC/MS analysis, 10 µL of suspended samples were injected
SP:SAMPLEPREP_SUMMARY            	and analyzed using a 6495 triple quadrupole mass spectrometer (Agilent
SP:SAMPLEPREP_SUMMARY            	Technologies, Santa Clara, CA) coupled to a HPLC system (Agilent Technologies,
SP:SAMPLEPREP_SUMMARY            	Santa Clara, CA) via Multiple reaction monitoring (MRM). Source parameters were
SP:SAMPLEPREP_SUMMARY            	as follows: Gas temperature- 250°C; Gas flow- 14 l/min; Nebulizer - 20psi;
SP:SAMPLEPREP_SUMMARY            	Sheath gas temperature - 350°C; Sheath gas flow- 12 l/min; Capillary - 3000 V
SP:SAMPLEPREP_SUMMARY            	positive and 3000 V negative; Nozzle voltage- 1500 V positive and 1500 V
SP:SAMPLEPREP_SUMMARY            	negative. Approximately 8–11 data points were acquired per detected
SP:SAMPLEPREP_SUMMARY            	metabolite. The data acquired using Agilent mass hunter software and data was
SP:SAMPLEPREP_SUMMARY            	analyzed using mass hunter quantitative analysis software.
SP:SAMPLEPREP_PROTOCOL_FILENAME  	Targeted.MS.method.pdf;unbiased.liver.MS.method.pdf
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_TYPE           	HILIC
CH:INSTRUMENT_NAME               	Agilent 6495 QQQ
CH:COLUMN_NAME                   	Waters XBridge Amide (100 x 4.6mm, 3.5um)
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Agilent 6495 QQQ
MS:INSTRUMENT_TYPE               	Triple quadrupole
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	POSITIVE
MS:MS_COMMENTS                   	For data acquisition through LC/MS analysis, 10 µL of suspended samples were
MS:MS_COMMENTS                   	injected and analyzed using a 6495 triple quadrupole mass spectrometer (Agilent
MS:MS_COMMENTS                   	Technologies, Santa Clara, CA) coupled to a HPLC system (Agilent Technologies,
MS:MS_COMMENTS                   	Santa Clara, CA) via Multiple reaction monitoring (MRM). Source parameters were
MS:MS_COMMENTS                   	as follows: Gas temperature- 250°C; Gas flow- 14 l/min; Nebulizer - 20psi;
MS:MS_COMMENTS                   	Sheath gas temperature - 350°C; Sheath gas flow- 12 l/min; Capillary - 3000 V
MS:MS_COMMENTS                   	positive and 3000 V negative; Nozzle voltage- 1500 V positive and 1500 V
MS:MS_COMMENTS                   	negative. Approximately 8–11 data points were acquired per detected
MS:MS_COMMENTS                   	metabolite. The data acquired using Agilent mass hunter software and data was
MS:MS_COMMENTS                   	analyzed using mass hunter quantitative analysis software.
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	peak intensity
MS_METABOLITE_DATA_START
Samples	HFD-VEH13	HFD-VEH15	HFD-VEH18	HFD-VEH12	HFD-VEH14	HFD-BPA12	HFD-BPA13	HFD-BPA14	HFD-BPA16	HFD-BPA18
Factors	Treatment:vehicle	Treatment:vehicle	Treatment:vehicle	Treatment:vehicle	Treatment:vehicle	Treatment:BPA	Treatment:BPA	Treatment:BPA	Treatment:BPA	Treatment:BPA
2-Aminooctanoic acid	278549.519	664617.777	340761.8695	299286.0273	301848.8526	170741.0364	234069.0918	189203.2978	133268.9852	366496.1754
Octonyl Carnitine	472930.985	294675.0353	344135.9308	556394.2531	321294.3685	210611.022	379924.3923	358481.9373	413154.3906	259047.6114
1-Stearoyl-sn-glycero-3-phosphocholine	10932955.53	10633554.09	7693075.196	8748290.86	12338814.35	8143491.015	12213536.54	13218814.83	9371530.926	19276929.02
Oleic acid	6604079.717	5062332.038	6743628.44	7195186.73	7575174.317	6764715.643	5605824.425	4783489.752	4712699.616	4896811.127
Acetyl Carnitine	199777374.7	151663536.7	167611336.2	135597588.5	165053765.9	135297866.1	166465986.4	142179576.2	131513503.9	110091970.8
Propeonyl Carnitine	28493905.35	33630241.8	25684275.33	27770720.71	16283386.83	23109113.36	15493295.98	19428314.74	24877452.43	31649932.55
4 Acetamidobutanoic acid	237352.0134	140081.904	168704.9081	211174.2497	102250.3728	148115.2663	122925.945	110682.1135	121667.2705	138838.53
2-Hydroxypyridine	1217558.322	1543523.492	1406751.619	1279497.802	1436948.087	1520877.201	1129003.028	1270017.682	1293388.707	1245434.931
Nicotinamide	184284954	219215778.1	216476372.6	178029224.3	199380459	249971821.6	171891888.5	200002255	208586928.5	183733884.2
1-Methyl tryptophan	117126.1448	121351.4007	101278.1108	108606.2595	117658.777	143210.7124	117157.642	133640.0576	164010.8213	71401.20183
Acetylcholine	2526493.283	2288019.318	1772835.861	1960317.519	1739310.505	1302389.218	1625964.477	1241679.172	1415504.634	1552121.167
Aspartame	180436.7753	170770.4815	164347.1847	122275.4996	189068.7616	196341.2124	144511.8321	116521.2717	163003.1448	146871.3876
Carnitine	34613808.58	36887932.16	37784065.91	32477309.44	41282446.53	27227535.94	32538391	30053293.41	34127772.06	26586000.7
Glutamic acid	104249.4113	139176.1082	6504622.836	83681.28903	82679.03327	77850.19281	70587.57958	83444.81641	82647.74446	88435.82137
Serine	192401.7772	189681.8136	221891.2698	247744.8661	204692.8605	237971.2817	190180.2168	250585.6659	201738.892	235308.7268
Gly-Gly	326243.7576	325199.9896	334162.3193	298338.2189	321833.1274	373439.5573	294773.7462	272276.0933	277527.78	271282.5192
Kynurenic acid	76127.60909	110588.0993	110274.4747	78363.50458	93560.06533	96490.93889	91617.79413	72824.44736	91827.02018	78243.69705
Isoleucine	69818372.22	70670320.8	74165997.88	67054973.3	70635413.83	76914483.46	63643730.31	61242302.45	63729152.01	62002696.46
Phenyl alanine	32705425.04	29298937.19	34765654.96	27894699.42	30847071.94	30076756.37	31753443.58	27209894.69	29333848.04	28020224.49
purine	58299.87087	42005.06711	61228.86505	56515.30028	60472.94397	50650.00602	48944.60827	54715.37169	47377.64024	43503.22651
Urea	15315393.6	19644080.69	13441898.24	11424964.72	11906196.86	14970218.96	22145926.93	14228086.04	11770153.16	14289947.84
4 Hydroxybutyric acid	1515529.061	1356784.906	1554386.156	1701335.818	1356712.113	1662776.677	1286278.972	1713418.018	1449364.446	1632233.276
5-Hydroxyindoleacetic acid	18985.4084	35516.12481	27304.40018	18487.28437	15718.77198	58678.13536	24439.0998	52021.47566	24747.80837	25751.8214
Aminobutyric acid	13442434	13662317.25	14184593.31	15765298.46	13187913.79	15130211.81	12245285.69	15742738.85	12760988.62	15719077.78
Choline	9076224.518	8730056.569	9442366.635	10755323.72	8639829.471	10277971.09	8168409.064	10771603.19	8236308.723	9436485.812
Kynurenine	215333.1746	240282.0759	221470.1324	215880.5196	177855.4513	311300.7129	152946.9515	225865.7116	178441.7364	178039.9145
Hypoxanthine	77452925.16	74320049.71	79761750.19	65360656.89	71572692.71	84739374.32	64483287.48	77751715.62	72256526.1	66415624.45
Creatinine	6727901.918	6933413.591	5666988.715	5513563.415	6279322.988	6426114.437	8766222.156	5480249.441	6118568.338	5831279.371
Gly-Leu	1925102.984	1330296.701	1699166.298	1557218.623	1747869.693	857422.7661	1118317.289	660513.8343	998062.4359	839235.901
Tryptophan	4592615.421	3699323.41	4006414.956	3189527.993	3797550.323	3013047.57	3136303.516	2796037.906	2973152.001	2565350.293
Valine	20240772.16	19316793.96	22733504.16	21149310.98	20180867.32	24207927.65	18614007.06	18203285.2	16249576.23	19416131.9
2-Methyl glutamic acid	413090.1537	465120.8703	392620.5899	416205.0704	465780.3488	341498.4009	415058.8881	385841.4202	380678.587	436752.7927
Xanthine	5424478.395	5111632.332	5064203.647	5183808.492	4582585.076	5827105.965	4321111.584	4286763.917	4250289.214	5252042.343
Guanidine acetic acid	12279274.3	11079159.95	7737469.586	10994907.7	10311905.61	8662454.195	16797731.77	12326222.53	10764725.2	10522305.08
N-Acetyl Lysine	319433.5085	378025.3918	290874.7747	253800.7318	235091.2527	389818.6613	308203.1012	232691.8444	283127.9826	406307.3178
Tyrosine	1399435.183	1399520.364	1439807.296	1499905.084	1321291.354	1856708.473	1235083.656	1090927.315	959039.971	1280225.078
Homocysteine	367450.7515	367135.1045	283856.7807	337066.0449	268226.1005	364230.4078	368162.6884	233144.1775	218338.4968	272959.8912
Betaine	137408906.2	128662684.9	123722596	118171690.5	113152608.4	132970502.1	136700515.9	133096664.4	138645393.8	126077915.9
Methyl Nicotinamide	2004586.645	1745367.835	2113669.637	1753786.114	2304486.869	1941021.689	1802439.952	2031393.481	1742916.875	1779955.831
Proline	4331215.055	4340633.037	4049893.286	4410807.095	3574375.431	5045184.007	3631487.759	3373840.181	3779756.123	4069363.57
Sarcosine/Alanine	1071077.128	894241.0442	1022489.586	974527.6984	956780.0652	1347980.763	1032811.498	743874.5592	919434.2274	867660.8684
Guanosine	42232.73243	39634.2594	31902.7711	26909.99657	29991.64849	18639.57828	46323.34639	48624.48259	31317.54563	24222.96447
N, N Dimethyl Glycine	3545532.537	3405308.965	4278795.386	4788711.496	3886666.662	3191320.421	4215886.179	4434488.952	3353475.864	3080378.975
Dimethyl Arginine	228114.2541	295697.8286	241182.1822	120962.3343	188622.7053	532426.3817	173022.9883	116017.4527	124584.2259	122444.4913
Glycine	88791.66113	95741.54298	89658.982	93693.48155	75321.91376	86618.15343	93981.58279	117498.2959	86174.75707	104268.788
Ergothioneine	53086.87006	42615.21106	67049.03308	62922.48954	48310.59	71529.1823	75643.38731	94463.71384	71682.69401	49537.38564
1-Methyl-Histidine	629894.3426	837321.6483	688717.1332	569238.5054	566450.9125	615015.6162	856003.6449	720160.4315	560989.378	568539.6967
Spermidine	26436.21861	49286.86427	19278.95208	18136.59591	13408.55842	8809.739113	27720.90881	15709.83979	10391.04528	14355.88144
Allo-Threonine	946637.0257	964646.3176	1079572.821	1090240.454	1143276.65	1184924.506	1182415.049	1125083.063	942958.7396	1136376.98
Homoserine	946637.0257	964646.3176	1079572.821	1090240.454	1143276.65	1184924.506	1182415.049	1125083.063	942958.7396	1136376.98
N-Acetylputrescine	105017.3638	83677.09934	115325.8823	81415.0168	87099.67107	103419.4714	78270.77869	89009.84516	73611.10496	72818.5916
citrulline	41650.52585	42554.56067	45926.75052	47264.29635	36132.76131	49571.43707	41548.52321	71924.99791	51865.44213	80717.06143
Lysine	13934602.01	13724201.97	15543811.4	16197041.26	16526530.11	16380193.9	14679282.6	16035784.39	14981738	15752010.26
Methylglutaric acid	532842.178	465030.3626	530054.4528	479709.6929	533848.5146	470201.4135	564188.3997	559368.7153	524864.911	481789.1081
5-oxo-proline	54223270.17	55030576.38	56599334.53	61565002.64	63270918.03	59431643.81	56428693.44	55219922.65	51147315.28	53329841.41
DL-Pipecolic acid	54223270.17	55030576.38	56599334.53	61565002.64	63270918.03	59431643.81	56428693.44	55219922.65	51147315.28	53329841.41
SAH	1389382.221	1543236.076	1744539.82	1517493.077	1509874.228	1646979.937	1319156.259	1638059.585	929834.5941	1851616.798
dCMP	116202.0617	151065.3363	131082.8721	124897.1084	93296.00893	116619.2212	70641.26398	130889.7035	116412.606	140408.6118
Arginine	323911.4205	392149.8403	234268.511	250297.4312	262669.3405	363432.1708	295308.6418	287690.2494	195040.9656	302337.1234
2-Aminobutyraldehyde	111019.0864	110413.7895	123677.7648	145670.4804	137463.3031	116327.0815	131817.9125	109797.7166	97703.18325	153786.4725
Ethylmalonate	122160.4449	160076.4898	83844.59556	122239.9024	97286.73756	113256.3247	159972.4694	109103.1059	134841.8858	156915.5794
Glutamyl alanine	194996.133	105572.8651	166204.3768	146617.5576	166937.0088	171923.479	158139.0351	243345.9481	143190.3205	220488.2103
Phospho Choline	4747335.787	2288498.621	3392731.617	3578291.131	2560301.907	1812204.898	3522761.42	2573573.582	2847246.724	3168470.154
3-Hydroxykynurenine	1046036.812	1275335.928	1075152.087	1099791.305	992652.8363	1092445.166	1009589.451	1224381.405	935701.6376	748600.9278
Deoxy carnitine	11088329.54	11330718.32	6895616.663	8992318.97	6776040.888	5280820.984	6364028.974	5706458.745	5727537.135	6721819.455
3-phospho-serine	374821.0354	211219.1398	273217.34	293989.7815	241919.3346	200541.4655	325199.9465	205460.1354	254688.1245	277524.4985
Glu-Tyr	351520.3704	358009.3262	327618.4733	323286.2387	263151.2322	337492.0923	264667.0053	348326.7584	302770.5192	251952.3321
Asparagine	633676.9846	568317.6997	713734.6295	720992.3567	742730.2478	805917.8008	680779.3554	501997.4513	525022.4457	584017.926
MYRISTATE (14:0)+Na	93993.26973	77478.84256	116575.4893	95414.76666	94712.37323	106824.7954	114656.6757	134043.5277	94511.42493	99007.12187
S-ribosyl-L-homocysteine	50250.43869	55473.58725	46784.84302	43691.18426	41205.72423	53213.5723	66993.03444	50475.21626	54358.44328	51521.85814
Nicotinamide ribotide	3798283.779	3106430.551	6732885.777	6162627.429	6309741.614	9169815.858	5099274.535	5392148.087	4474689.709	6609341.667
AMP	92531424.43	105792766.3	76008605.07	75518668.12	53000371.95	52993455.78	71802851.09	86410481.61	80394793.04	85555594.53
acadesine	175882.0857	146945.3154	137640.7798	137759.9783	75903.20694	171526.5871	168501.822	135563.941	137525.7765	123670.4617
Cystathionine	451134.0832	494743.3149	568897.4248	454817.2515	391456.7047	821000.0794	355358.5627	308310.1012	276603.4254	469480.735
aminoimidazole carboxamide ribonucleotide	58719.40468	48008.49159	51937.47646	75330.13214	34900.37569	37290.57936	79654.56943	69270.05131	57718.81445	66896.77521
thiamine	21721.99763	14588.26763	20651.56555	11408.25356	22136.99814	27081.79919	8243.163126	17784.1462	13018.66396	26997.12811
Cytosine	1112103.173	1469541.672	1447006.718	1314588.5	1331404.737	1345178.057	863591.9153	1051794.254	1002470.274	1030745.621
Cytidine 5 -Monophosphate (5-CMP)	3721799.847	3425109.497	2811681.31	3056355.027	2451363.494	2489212.855	2777717.219	3785349.862	2913097.496	4494155.937
Inositol-1P	52710.75123	75430.91048	24179.93625	51684.28793	28051.76743	23138.83926	38621.93355	89522.98424	69278.14898	44901.77881
N-acetyl Glucosamine 6 Phos	49482.33237	58546.66893	61883.0748	63196.67733	47068.2074	99802.35244	64842.80735	50579.26062	41475.12027	77176.02028
Glutamine	25902314.65	25524329.7	29402360.27	29967813.11	32504656.94	24365420.37	26146145.27	26185023.08	25684730.95	29215402.58
Histidine	43982343.31	47997815.2	50619671.67	36107057.03	42032581.15	50851413.15	41974057.42	42585870.78	35275151.18	51706004.62
NAD	4200508.151	1417521.793	4711784.741	1643686.36	1884911.886	1769207.887	2552713.099	3048417.708	2242441.574	2981572.974
GMP	11613724.15	11262154.51	9562788.635	7978575.989	4588533.039	2956871.51	11054372.05	9547622.453	10987427.81	8188013.054
Glu-Glu	136004.9324	120148.7278	130917.9694	101138.3534	140947.9671	125599.1389	115567.6875	125247.0794	111110.6889	147061.6369
xanthosine-5-phosphate	168960.2284	202889.7	85284.26367	173375.9996	102086.6581	141192.4237	124892.6685	166735.6991	120047.5149	158318.4102
UMP	787321.0187	859021.7343	457266.9014	506216.6082	393032.05	248762.1454	534710.4159	813569.0385	634767.0153	461000.4277
L-Creatinine (internal standard)	6157139.722	5728782.935	5592526.099	5072963.773	5264991.607	5716845.615	4677357.851	4968603.857	5411106.852	4248299.067
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name
2-Aminooctanoic acid
Octonyl Carnitine
1-Stearoyl-sn-glycero-3-phosphocholine
Oleic acid
Acetyl Carnitine
Propeonyl Carnitine
4 Acetamidobutanoic acid
2-Hydroxypyridine
Nicotinamide
1-Methyl tryptophan
Acetylcholine
Aspartame
Carnitine
Glutamic acid
Serine
Gly-Gly
Kynurenic acid
Isoleucine
Phenyl alanine
purine
Urea
4 Hydroxybutyric acid
5-Hydroxyindoleacetic acid
Aminobutyric acid
Choline
Kynurenine
Hypoxanthine
Creatinine
Gly-Leu
Tryptophan
Valine
2-Methyl glutamic acid
Xanthine
Guanidine acetic acid
N-Acetyl Lysine
Tyrosine
Homocysteine
Betaine
Methyl Nicotinamide
Proline
Sarcosine/Alanine
Guanosine
N, N Dimethyl Glycine
Dimethyl Arginine
Glycine
Ergothioneine
1-Methyl-Histidine
Spermidine
Allo-Threonine
Homoserine
N-Acetylputrescine
citrulline
Lysine
Methylglutaric acid
5-oxo-proline
DL-Pipecolic acid
SAH
dCMP
Arginine
2-Aminobutyraldehyde
Ethylmalonate
Glutamyl alanine
Phospho Choline
3-Hydroxykynurenine
Deoxy carnitine
3-phospho-serine
Glu-Tyr
Asparagine
MYRISTATE (14:0)+Na
S-ribosyl-L-homocysteine
Nicotinamide ribotide
AMP
acadesine
Cystathionine
aminoimidazole carboxamide ribonucleotide
thiamine
Cytosine
Cytidine 5 -Monophosphate (5-CMP)
Inositol-1P
N-acetyl Glucosamine 6 Phos
Glutamine
Histidine
NAD
GMP
Glu-Glu
xanthosine-5-phosphate
UMP
L-Creatinine (internal standard)
METABOLITES_END
#END