#METABOLOMICS WORKBENCH cendalfr_20230605_114108 DATATRACK_ID:4065 STUDY_ID:ST002727 ANALYSIS_ID:AN004421 PROJECT_ID:PR001691
VERSION             	1
CREATED_ON             	June 6, 2023, 10:09 am
#PROJECT
PR:PROJECT_TITLE                 	Metabolic and Proteomic Changes in Sickle Cell Disease and B-thalassemia Mouse
PR:PROJECT_TITLE                 	Splenic and Hepatic Macrophages and Peripheral Blood Mononuclear cells
PR:PROJECT_SUMMARY               	Sickle cell disease and Beta-thalassemia represent hemoglobinopathies arising
PR:PROJECT_SUMMARY               	from dysfunctional or under produced beta-globin chains, respectively. In both
PR:PROJECT_SUMMARY               	diseases, red blood cell injury and anemia are the impetus for end organ injury.
PR:PROJECT_SUMMARY               	Because persistent erythrophagocytosis is a hallmark of these genetic maladies
PR:PROJECT_SUMMARY               	it is critical to understand how macrophage phenotype polarizations in tissue
PR:PROJECT_SUMMARY               	compartments can inform on disease progression. Murine models of sickle cell
PR:PROJECT_SUMMARY               	disease and Beta-thalassemia allow for a basic understanding of mechanisms and
PR:PROJECT_SUMMARY               	provide for translation to human disease. A multi-omics approach to
PR:PROJECT_SUMMARY               	understanding macrophage metabolism and protein changes in two murine models of
PR:PROJECT_SUMMARY               	beta-globinopathy was performed on peripheral blood mononuclear cells as well as
PR:PROJECT_SUMMARY               	spleen and liver macrophages isolated from Berkley sickle cell disease (Berk-ss)
PR:PROJECT_SUMMARY               	and heterozygous B1/B2 globin gene deletion (Hbbth3/+) mice. Results from these
PR:PROJECT_SUMMARY               	experiments revealed the metabolome and proteome of macrophages are polarized to
PR:PROJECT_SUMMARY               	a distinct phenotype in Berk-ss and Hbbth3/+ compared each other and their
PR:PROJECT_SUMMARY               	common background mice (C57BL6/J). Further, spleen and liver macrophages
PR:PROJECT_SUMMARY               	revealed distinct disease specific phenotypes, suggesting macrophages become
PR:PROJECT_SUMMARY               	differentially polarized and reprogrammed within tissue compartments. We
PR:PROJECT_SUMMARY               	conclude that tissue recruitment, polarization, metabolic and proteomic
PR:PROJECT_SUMMARY               	reprogramming of macrophages in Berk-ss and Hbbth3/+ mice may be relevant to
PR:PROJECT_SUMMARY               	disease to progression in other tissue.
PR:INSTITUTE                     	University of Colorado School of Medicine
PR:LABORATORY                    	Laboratory of Angelo D'Alessandro in collaboratation with David Irwin
PR:LAST_NAME                     	Cendali
PR:FIRST_NAME                    	Francesca
PR:ADDRESS                       	13199 East Montview Boulevard, Aurora, CO, 80045, USA
PR:EMAIL                         	francesca.cendali@cuanschutz.edu
PR:PHONE                         	3037246131
#STUDY
ST:STUDY_TITLE                   	Metabolic and Proteomic Divergence is Present in Circulating Monocytes and
ST:STUDY_TITLE                   	Tissue Resident Macrophages from Berkeley Sickle Cell Anemia and B-thalassemia
ST:STUDY_TITLE                   	mice (Liver)
ST:STUDY_SUMMARY                 	Sickle cell disease and Beta-thalassemia represent hemoglobinopathies arising
ST:STUDY_SUMMARY                 	from dysfunctional or under produced beta-globin chains, respectively. In both
ST:STUDY_SUMMARY                 	diseases, red blood cell injury and anemia are the impetus for end organ injury.
ST:STUDY_SUMMARY                 	Because persistent erythrophagocytosis is a hallmark of these genetic maladies
ST:STUDY_SUMMARY                 	it is critical to understand how macrophage phenotype polarizations in tissue
ST:STUDY_SUMMARY                 	compartments can inform on disease progression. Murine models of sickle cell
ST:STUDY_SUMMARY                 	disease and Beta-thalassemia allow for a basic understanding of mechanisms and
ST:STUDY_SUMMARY                 	provide for translation to human disease. A multi-omics approach to
ST:STUDY_SUMMARY                 	understanding macrophage metabolism and protein changes in two murine models of
ST:STUDY_SUMMARY                 	beta-globinopathy was performed on peripheral blood mononuclear cells as well as
ST:STUDY_SUMMARY                 	spleen and liver macrophages isolated from Berkley sickle cell disease (Berk-ss)
ST:STUDY_SUMMARY                 	and heterozygous B1/B2 globin gene deletion (Hbbth3/+) mice. Results from these
ST:STUDY_SUMMARY                 	experiments revealed the metabolome and proteome of macrophages are polarized to
ST:STUDY_SUMMARY                 	a distinct phenotype in Berk-ss and Hbbth3/+ compared each other and their
ST:STUDY_SUMMARY                 	common background mice (C57BL6/J). Further, spleen and liver macrophages
ST:STUDY_SUMMARY                 	revealed distinct disease specific phenotypes, suggesting macrophages become
ST:STUDY_SUMMARY                 	differentially polarized and reprogrammed within tissue compartments. We
ST:STUDY_SUMMARY                 	conclude that tissue recruitment, polarization, metabolic and proteomic
ST:STUDY_SUMMARY                 	reprogramming of macrophages in Berk-ss and Hbbth3/+ mice may be relevant to
ST:STUDY_SUMMARY                 	disease to progression in other tissue.
ST:INSTITUTE                     	University of Colorado School of Medicine
ST:LABORATORY                    	Laboratory of Angelo D'Alessandro in collaboratation with David Irwin
ST:LAST_NAME                     	Cendali
ST:FIRST_NAME                    	Francesca
ST:ADDRESS                       	13199 East Montview Boulevard, Aurora, CO, 80045, USA
ST:EMAIL                         	francesca.cendali@cuanschutz.edu
ST:PHONE                         	3037246131
#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]Raw file names and additional sample data
SUBJECT_SAMPLE_FACTORS           	-	61	Factor:WT	RAW_FILE_NAME=F2-94-61
SUBJECT_SAMPLE_FACTORS           	-	62	Factor:WT	RAW_FILE_NAME=F2-94-62
SUBJECT_SAMPLE_FACTORS           	-	63	Factor:WT	RAW_FILE_NAME=F2-94-63
SUBJECT_SAMPLE_FACTORS           	-	64	Factor:WT	RAW_FILE_NAME=F2-94-64
SUBJECT_SAMPLE_FACTORS           	-	65	Factor:WT	RAW_FILE_NAME=F2-94-65
SUBJECT_SAMPLE_FACTORS           	-	66	Factor:WT	RAW_FILE_NAME=F2-94-66
SUBJECT_SAMPLE_FACTORS           	-	67	Factor:WT	RAW_FILE_NAME=F2-94-67
SUBJECT_SAMPLE_FACTORS           	-	68	Factor:WT	RAW_FILE_NAME=F2-94-68
SUBJECT_SAMPLE_FACTORS           	-	69	Factor:WT	RAW_FILE_NAME=F2-94-69
SUBJECT_SAMPLE_FACTORS           	-	70	Factor:WT	RAW_FILE_NAME=F2-94-70
SUBJECT_SAMPLE_FACTORS           	-	71	Factor:WT	RAW_FILE_NAME=F2-94-71
SUBJECT_SAMPLE_FACTORS           	-	72	Factor:Berk	RAW_FILE_NAME=F2-94-72
SUBJECT_SAMPLE_FACTORS           	-	73	Factor:Berk	RAW_FILE_NAME=F2-94-73
SUBJECT_SAMPLE_FACTORS           	-	74	Factor:Berk	RAW_FILE_NAME=F2-94-74
SUBJECT_SAMPLE_FACTORS           	-	75	Factor:Berk	RAW_FILE_NAME=F2-94-75
SUBJECT_SAMPLE_FACTORS           	-	76	Factor:Berk	RAW_FILE_NAME=F2-94-76
SUBJECT_SAMPLE_FACTORS           	-	77	Factor:Berk	RAW_FILE_NAME=F2-94-77
SUBJECT_SAMPLE_FACTORS           	-	78	Factor:Berk	RAW_FILE_NAME=F2-94-78
SUBJECT_SAMPLE_FACTORS           	-	79	Factor:Berk	RAW_FILE_NAME=F2-94-79
SUBJECT_SAMPLE_FACTORS           	-	80	Factor:Berk	RAW_FILE_NAME=F2-94-80
SUBJECT_SAMPLE_FACTORS           	-	81	Factor:Berk	RAW_FILE_NAME=F2-94-81
SUBJECT_SAMPLE_FACTORS           	-	82	Factor:Beta Thal	RAW_FILE_NAME=F2-94-82
SUBJECT_SAMPLE_FACTORS           	-	83	Factor:Beta Thal	RAW_FILE_NAME=F2-94-83
SUBJECT_SAMPLE_FACTORS           	-	84	Factor:Beta Thal	RAW_FILE_NAME=F2-94-84
SUBJECT_SAMPLE_FACTORS           	-	85	Factor:Beta Thal	RAW_FILE_NAME=F2-94-85
SUBJECT_SAMPLE_FACTORS           	-	86	Factor:Beta Thal	RAW_FILE_NAME=F2-94-86
SUBJECT_SAMPLE_FACTORS           	-	87	Factor:Beta Thal	RAW_FILE_NAME=F2-94-87
SUBJECT_SAMPLE_FACTORS           	-	88	Factor:Beta Thal	RAW_FILE_NAME=F2-94-88
SUBJECT_SAMPLE_FACTORS           	-	89	Factor:Beta Thal	RAW_FILE_NAME=F2-94-89
SUBJECT_SAMPLE_FACTORS           	-	90	Factor:Beta Thal	RAW_FILE_NAME=F2-94-90
SUBJECT_SAMPLE_FACTORS           	-	91	Factor:Beta Thal	RAW_FILE_NAME=F2-94-91
SUBJECT_SAMPLE_FACTORS           	-	92	Factor:Beta Thal	RAW_FILE_NAME=F2-94-92
#COLLECTION
CO:COLLECTION_SUMMARY            	Whole blood samples (1 mL) were obtained from animals via cardiac puncture,
CO:COLLECTION_SUMMARY            	using a syringe with a 26-gauge needle, and placed in an EDTA treated tube. The
CO:COLLECTION_SUMMARY            	blood was transferred to a 15 mL conical tube and diluted 2:1, sterile PBS:
CO:COLLECTION_SUMMARY            	blood, and gently mixed. Lympholyte® Mammal Cell Separation media (Cedarlane
CO:COLLECTION_SUMMARY            	Labs, product # CL5115) was gently added to the bottom of the blood solution and
CO:COLLECTION_SUMMARY            	spun at 1400 rpm for 30 minutes in a refrigerated centrifuge. After
CO:COLLECTION_SUMMARY            	centrifugation, layers were visualized, the PBMC layer (midlayer) was extracted
CO:COLLECTION_SUMMARY            	and resuspended in a new tube. The isolated PBMC’s were washed using ~14mL
CO:COLLECTION_SUMMARY            	sterile PBS, spun at 1800 rpm for 10 minutes, excess PBS was removed, cells were
CO:COLLECTION_SUMMARY            	resuspended in 1-2mL for counting, and the final pellet was frozen in liquid
CO:COLLECTION_SUMMARY            	nitrogen and stored at -80C.
CO:SAMPLE_TYPE                   	Macrophages
#TREATMENT
TR:TREATMENT_SUMMARY             	Eight- to ten-week-old female C57Bl/6 WT, Berk-ss, or Hbbth3/+ mice were either
TR:TREATMENT_SUMMARY             	obtained from Jackson Laboratories (Bar Harbor, ME, USA) or our in-house Berk
TR:TREATMENT_SUMMARY             	SCD mouse colony. Mice were housed and bred in an AAALAC accredited animal
TR:TREATMENT_SUMMARY             	facility at the University of Colorado, Denver, Anschutz Medical campus and were
TR:TREATMENT_SUMMARY             	maintained on a 12:12 light-dark cycle with food and water available ad libitum.
TR:TREATMENT_SUMMARY             	Female heterozygous Berk-ss mice were bred with male homozygous Berkss mice to
TR:TREATMENT_SUMMARY             	generate homozygous offspring. Specifically, Berk-ss mice with genotype
TR:TREATMENT_SUMMARY             	Tg(HuminiLCR α1 Gγ Aγ δ βs ) Hba0/0 Hbb0/0 and the hemizygous with genotype
TR:TREATMENT_SUMMARY             	Tg(Hu-miniLCR α1 Gγ Aγ δ βs ) Hba0/0 Hbb0 Hbb+ were littermates. Genotyping
TR:TREATMENT_SUMMARY             	of mice used for breeding and experiments was performed by TransnetYX (Cordova,
TR:TREATMENT_SUMMARY             	TN, USA). A total of 21 mice (C57Bl/6: n=10, Berk-ss mice: n=11, Hbbth3/+ =10)
TR:TREATMENT_SUMMARY             	were used in the present investigation and levels of discomfort and distress
TR:TREATMENT_SUMMARY             	were monitored daily by the in-house animal care staff, with a veterinarian
TR:TREATMENT_SUMMARY             	available as needed. All experimental procedures were conducted under the
TR:TREATMENT_SUMMARY             	guidelines recommended by The Journal of Physiology (11), the National
TR:TREATMENT_SUMMARY             	Institutes of Health and were approved by the Institutional Animal Care and Use
TR:TREATMENT_SUMMARY             	Committee at the University of Colorado, Denver, Anschutz Medical Campus.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Liver macrophages and peripheral blood mononuclear cells (PBMCs) were extracted
SP:SAMPLEPREP_SUMMARY            	in methanol:acetonitrile:water (5:3:2 v/v/v – at a 1x10^6 cells/ml) ). The
SP:SAMPLEPREP_SUMMARY            	samples were vortexed for 30mins at 4°C. The samples were then spun down at
SP:SAMPLEPREP_SUMMARY            	18,213 g for 10 minutes, 4°C and 50uL of supernatant was transferred to
SP:SAMPLEPREP_SUMMARY            	autosampler vial.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	Positive Ion Mode
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Thermo Vanquish
CH:COLUMN_NAME                   	Phenomenex Kinetex C18 (150 x 2.1mm,1.7um)
CH:SOLVENT_A                     	100% water; 0.1% formic acid
CH:SOLVENT_B                     	100% acetonitrile; 0.1% formic acid
CH:FLOW_GRADIENT                 	0-0.5 min 5% B, 0.5-1.1 min 5-95% B, 1.1-2.75 min hold at 95% B, 2.75-3 min
CH:FLOW_GRADIENT                 	95-5% B, 3-5 min hold at 5% B.
CH:FLOW_RATE                     	0.450ml/min
CH:COLUMN_TEMPERATURE            	45
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Thermo Q Exactive Orbitrap
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	POSITIVE
MS:MS_COMMENTS                   	Resolution 70,000, scan range 65-900 m/z, maximum injection time 200 ms,
MS:MS_COMMENTS                   	microscans 2, automatic gain control (AGC) 3 x 10^6 ions, source voltage 4.0 kV,
MS:MS_COMMENTS                   	capillary temperature 320 C, and sheath gas 45, auxiliary gas 15, and sweep gas
MS:MS_COMMENTS                   	0 (all nitrogen). Data converted to mzXML using RawConverter. Metabolites were
MS:MS_COMMENTS                   	annotated and integrated using Maven in conjunction with the KEGG database.
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	Peak Area
MS_METABOLITE_DATA_START
Samples	61	62	63	64	65	66	67	68	69	70	71	72	73	74	75	76	77	78	79	80	81	82	83	84	85	86	87	88	89	90	91	92
Factors	Factor:WT	Factor:WT	Factor:WT	Factor:WT	Factor:WT	Factor:WT	Factor:WT	Factor:WT	Factor:WT	Factor:WT	Factor:WT	Factor:Berk	Factor:Berk	Factor:Berk	Factor:Berk	Factor:Berk	Factor:Berk	Factor:Berk	Factor:Berk	Factor:Berk	Factor:Berk	Factor:Beta Thal	Factor:Beta Thal	Factor:Beta Thal	Factor:Beta Thal	Factor:Beta Thal	Factor:Beta Thal	Factor:Beta Thal	Factor:Beta Thal	Factor:Beta Thal	Factor:Beta Thal	Factor:Beta Thal
L-alanine	1091259	655274.6	655629.4	683801.2	585708.5	1121539	655087.6	713238	1373440	458410.3	694756.8	612201.6	863184.8	730808.4	681358.8	876497.5	654603.8	1324884	452393.8	640745.1	2089544	680022.2	2710308	613184.5	629734.6	495487	1174573	1815907	2201715	1231010	1212856	1511019
L-arginine	1638536	46705.98	65714.88	54975.62	42518.12	708419.9	66414.15	62115.5	159541.8	52414.55	92122.12	94650.31	110440.6	200366.1	100857.8	84808.75	63692.85	142421.2	104753.3	110774.5	132139.8	63236.32	1156382	66473.92	89380.69	96458.62	238967.9	315490.4	229956.2	55067.91	236862.3	308496.7
L-asparagine	72413.98	25296.87	27559.44	26367.53	30207.28	49747.34	30740.29	28888.48	10744.64	21046.57	31203.75	10549.65	36605.7	38837.61	24913.71	10397.93	18471.47	58144.47	23136.9	13095.49	9741.782	29092.03	100765.4	41430.27	24120.08	20123.63	28059.88	87188.07	121061.8	11876.11	75129.4	96735.88
L-glutamate	928008.7	245632	271822.5	149586.6	70133.07	725044.6	167827.6	298274.8	281787.7	160756.2	246382	164542.6	482421	477267.5	132681.4	370306.8	81147.84	488061.8	123118.5	109480.3	275353.3	177838.2	462835	482110.1	310627.4	231682.6	448168.9	830030.2	1060335	160378.3	555061.6	1087345
L-glutamine	179806.7	29181.53	121540.2	89738.55	44006.11	123797.2	121224.3	95150.12	119826.4	83820.7	119265.4	148151	109209.6	125174.4	103218.4	160443.9	43631.39	168956.7	43913.03	28682.12	123773.3	38322.03	120331.6	137017.2	71432.41	46240.89	126395.9	252741.9	159498.9	60976.94	257660.4	230536.5
L-histidine	342794.1	75121.47	109469.2	98383.19	109488.4	185612.2	140599.3	115580.9	194038.3	95670.04	170784.7	133675.6	183795.8	278397.2	141098.7	206437.4	164450.6	282600.5	93948.31	118567.6	219619.5	132447.8	3314721	165920.7	137579.6	121101.6	165271.9	392450.7	719635.7	174192.2	316605.3	401785.2
L-leucine	3258409	4332670	2865440	2889256	952675.2	2998158	3732557	1990468	5919848	1829681	3172127	4007255	1737832	2195863	1081056	2030082	1422460	3237891	1546704	933939.3	2684341	817518	6635440	1702722	1123106	1646266	4619976	5477724	6783904	1914357	3845913	5528248
L-lysine	348826.7	55041.97	107134.2	61900.18	38650.12	187691.5	64930.84	63422.53	79103.05	49329.92	155148.5	67690.38	252930.6	205816.1	44879.31	223000.3	59004.56	336426.8	63832.89	159640.6	277361.8	61628.04	317052.2	138127.8	122236.2	107856.2	188119	602917.9	449567.9	81102.95	382970.7	535679.6
L-methionine	246156.2	1775.062	24114.04	10891.31	0	245194.1	5497.945	6085.787	35918.78	4457.7	17120.38	8822.54	83087.13	88049.82	1774.981	85574.24	1597.596	236536.6	0	46684.2	125306	8107.199	51183.78	59688.8	20404.06	21871.51	78041.71	538130.2	463010	31057.91	328686.5	585969.6
L-phenylalanine	776421.3	1521699	1036859	1431079	451033.4	703749.3	2201434	1029775	2294075	1031507	1501477	2025619	371133.6	688250.9	599947.4	681863.8	606421.8	638798.1	652243.7	297415.5	909881.8	264665	3020311	576316.6	367834	583457.1	1862087	885929.8	1275073	668011.7	1128826	942798
L-proline	3274651	722180.6	1069223	654596.6	498881.4	2360454	767666	740021.7	1221241	594669	1334321	1094545	1484465	1653060	710868.7	1549661	705321	1823371	542062	862835.8	1346282	772760.1	5012143	1553810	1114900	757454.3	2116108	3738182	4396856	686370.4	1772930	3092329
L-serine	157874.8	49727.4	52713.27	40875.04	37637.66	149457.8	37950.68	50002.54	55530.68	46171.65	74021.12	52044.91	94823.4	105737.9	35473.32	80838.03	85230.51	162129.9	40867.68	59226.48	99180.5	45786.36	1544890	83312.25	82051.94	39114.77	82300.65	261075.5	476617.9	94503.12	142194.9	224632.7
L-threonine	250882.4	114851.1	80890.52	58906.57	91131.4	202712.3	71184.34	85296.25	78042.99	56157.47	111079.2	95271.34	139016.5	153667.7	69761.35	128164.8	65145.99	210184.5	52859.58	94200.16	189781.4	86350.45	1012846	94012.05	87968.56	71021.91	123309.6	401683.4	403217.5	86179.92	212414.7	377515.9
L-tryptophan	235636.9	536799.4	439696.7	557794.4	191761.2	144817.4	947256.3	396046.5	896539	421467.2	508473	614983.1	139262.8	315181.5	187577.4	254874	284971.4	129831.2	326755	189984	332823	88936.27	2240532	173835.5	120322.4	199558.8	513722.4	223632.7	398474.3	273703.3	233262.4	264785.6
L-tyrosine	253157.2	301255.2	170534.2	208191.7	69903.9	293966.2	96178.76	152601.1	319690	137633.9	198974.5	200836.2	123968.4	176663.9	144041.3	147937.5	144073	319652.4	107615.5	91225.38	153000.5	98174.63	699717.7	155842.6	57334.86	125138.7	296360.3	322656.4	642119	157928.9	324304.8	423194.1
L-valine	8014340	4710802	4092325	2745619	2018415	10500000	3229308	4097593	4514348	3301123	10900000	8829963	5139232	4617774	2647524	4163219	0	8994681	1773498	2399983	4472512	3589564	6471854	6709582	6353156	5937518	7705194	10700000	8719952	4587376	9988109	11900000
AMP	80446.88	1618.951	4009.722	2211.152	1576.88	56721.89	2226.787	2232.052	4740.762	6186.924	3089.647	2800.458	19282.41	9088.55	2197.25	18111.74	2162.792	72891.36	0	15901.46	39082.53	1791.2	3671.712	14420.34	11075.89	5283.265	24791.07	57990.86	122439.2	7742.71	45794.81	89691.6
Cytidine	62702.13	108642.8	93134.77	86997.15	124386.4	41803.72	84090.13	24124.79	270359.6	100389	83168.12	153763.3	31196.68	117307.7	79998.8	52782.7	131532.8	19346.84	79688.77	53060.22	45066.06	35861.35	188155.4	41924.66	34347.44	57632.34	48358.78	42580.72	78144.9	85324.1	35652.34	29315.89
Allantoate	15064.7	13729.4	24940.33	13354.92	6361.303	52885.49	127488.6	8583.142	26827.13	12305.89	37190.79	23746.77	30212.96	33181.97	15307.07	38775.66	37135.88	59373.99	12097.86	39520.42	36213.55	14327.55	65931.18	36783.86	32791.98	18804.13	33892.31	108672.1	181831.9	59777.39	73945.2	108883.1
Nicotinamide	2166482	964062.8	1828738	1443926	679523.1	2960663	1028162	1947253	3126484	4266086	2381152	2139008	1158281	1633553	1068644	1609170	822423.2	4026981	539169.1	1104652	1900070	677700.2	637663.5	1741508	952669.8	790941.6	1668481	2325006	3209207	2557800	3418750	3261531
Nicotinate ribonucleotide	7932406	8741152	8072948	11200000	8423871	6090946	10400000	9846657	8842739	7928894	9661837	9319583	7646998	9948648	7917306	8590767	11000000	6074504	10500000	22050.48	37656.53	8084588	7056406	7972282	8150794	8409291	9694759	6864954	6414094	10700000	7590772	6187704
Glutathione	8608.165	0	70797.01	24478.43	0	269229.1	0	45698.79	59071.52	40425.55	36649.8	29110.87	59779.34	16019.73	0	41267.2	0	103437	3755.348	45820.32	45616	0	10784.96	10714	21781.01	9573.994	54382.67	8891.692	302926.5	74688.43	86168.56	80021.01
Spermidine	1586080	1643447	2402771	1703928	1573493	1393047	2100949	2268625	1509126	1476764	2048665	1509134	1713384	1564094	1926289	1881871	2504388	1711488	1717056	1637039	2314360	1117801	1997637	2118496	1323958	2515626	2158585	1244692	883996.2	1774258	1413741	1090494
Spermine	180817.4	160143.8	278569.6	253055.8	160079.7	131094.7	167548.6	250119.6	283726.4	308196.2	213318.2	175089.7	210150.9	165519.2	199153.7	164609.8	291767.8	216896.8	398637.5	204190.8	195849.3	51425.8	400092.7	177976.9	137944.2	217393.6	251881.8	41082.32	22791.27	285907.5	70794.2	85215.04
Creatine	1518814	811691.6	1058349	1102718	412874.7	2015433	1133544	1079776	1194244	984505.1	1359493	1227418	934865.7	958756	954753.3	1146127	380090.3	864699	462839.8	280373	562882.6	707539.1	729765.1	1108234	829775.7	648862.5	1090538	1695897	1555454	768594.8	1422307	1586748
Creatinine	721444.4	987869.3	683401.4	794351	1306371	1440398	2723743	1591190	1353496	1311365	1434320	1266431	806196.3	948100.2	2500544	925032.4	894963.8	1315352	527926.2	436947.4	1555349	2160746	989537.5	770117.2	786953	847445	851445.2	651454.9	0	562893.6	799663.2	578427.9
Guanidinoacetate	111074.6	5258.266	15480.64	4369.436	6782.398	48937.03	4210.634	5817.626	4529.091	4094.313	55792.17	44938.64	42692.84	46666.63	13055.84	63632.89	4653.699	115874.9	6276.533	22084.12	62243.77	4282.718	1982.879	12349.06	11890.08	7658.571	8047.291	81083.62	63391.58	6642.483	44660.23	144022.9
Pantothenate	75524.16	501015.8	279488.8	447213	83256.23	91821.43	877103.3	446573.4	516566.6	360645.3	267239.9	472644.1	69586.31	132725.6	102297.2	93276.76	86933.09	30589.42	154602.4	15277.74	20063.09	73013.18	154920.7	124937.2	91005.27	144201.6	288664.3	56166.11	47712.44	63805.4	105969.5	73275.16
Pantetheine	161865.5	362157.3	284522.6	175143.7	366591.9	74324.59	332170.2	72364.78	549405	221432.9	361262.1	407958.2	51263.34	226253.8	239734.6	98389.22	558622.1	25667.91	230472	226000.6	100309	50224.7	145558	47520.05	54882.42	146856.9	176313.4	75261.17	332217.8	135646	70801.55	40995.15
L-Methionine S-oxide	108983.5	63513	66876.33	38016.56	8217.505	62872.33	47262.29	31508.06	88448.32	19327.21	66497.8	39374.85	55509.99	74450.44	5717.756	65377.81	21792.83	74976.95	22083.17	29244.31	79357.32	8297.084	69112.15	44597.12	38006.53	46981.41	118685.3	173829.6	127686.2	38493.63	101778.6	130520.8
Indole	34707.32	0	44167.3	19430.74	0	137685.1	196730.3	128275.4	201754.5	192687.4	163467.3	210314.7	180920.5	197401.8	182314.6	215731.2	141495.4	164331.7	155051.4	139525.2	146510.2	189096.2	195853.3	192664	153724.8	179327	215379.1	148192.1	213639.2	186869.7	141785.9	179191.9
L-Adrenaline	439722	20424.24	47120	75861.8	23013.12	455756.2	100053.4	52869.79	51414.36	44214.29	122728.9	67222.62	170381.5	111008.4	31360.88	167731.6	13290.75	421951	20618.33	40614.21	176771.6	77170.2	21816.44	189018.2	88173.6	69754.4	143087.2	314578.6	544586.2	109221.5	380642.8	383306.1
Ectoine	43390.72	46708.31	59783.57	55339.82	55163.16	56564.56	50535.48	0	43155.31	50188.22	66380.28	84454.27	38381.09	41385.48	71460.23	43358.77	45413.75	54083.61	42944.18	37715.09	79407.45	61492.09	43885.36	55494.86	38170.29	42182.25	44922.31	51280.55	57193.64	47700.93	40044.86	36249.98
Sphingosine	1164458	1081989	2396214	1344551	824342.7	1951989	749031.3	2783623	2353458	2829871	4512242	2479061	2994735	3272359	1659108	5547036	1880294	4186441	1687822	4834433	5416796	1510752	4055681	2391290	3442105	6929218	3949048	5615926	1318196	4841618	4640114	3541454
Sphingosine 1-phosphate	19049.98	17912.54	39319.22	26133.72	21157.11	46580.52	41124.79	55910.8	35738.21	39116.52	69484.01	39446.39	33946.66	56954.86	52063.74	72550.27	65088.88	38988.79	43933.64	37869.77	41982.02	99424.09	39850.92	96170.88	49628.36	50441.77	28986.27	32694.03	47717.82	48136.05	38667.39	32741.5
Acetylcholine	206375.3	66710.66	142166.2	92762.16	24810.96	280558.4	90215.46	94760.05	98997.38	64661.18	118205.6	48671.95	143741	148187	87330.46	432250.2	29100.4	281449.3	27955.91	105794.1	214536.2	89810.72	63980.71	156432.2	107759.9	159108	111719.9	430993	325531.8	125826.9	395311.1	429712.2
Choline	9850035	2600185	3552537	3669435	1408212	5984886	2828157	3568509	3443601	1496721	3930700	2912474	4502618	5015564	3162829	6428838	1680818	2070925	1810390	3326143	3327707	2569158	3222753	5125624	2096834	1947240	3706930	5607534	5428296	1599233	4674684	4991004
L-Carnitine	4061166	275657.8	483420.6	298185.5	133758.5	2420279	337803.2	475139.6	590133.5	408420.1	873754.7	714348.8	866979.4	947893.6	281629.5	1098708	143183.2	2242739	204572.7	618248.1	1021561	436458.8	442843.4	1086516	685011.6	622383.7	941308.8	2188885	2473082	1088438	2634385	3575748
propionyl-carnitine (acyl-C3)	166991.3	85565.95	59122.35	91655.63	13368.14	97237.28	111454.1	78781.53	110253	67331.02	60337.68	74618.8	47538.7	57276.9	38762.12	59309.78	12411.98	78697.72	61913.32	9504.947	35514.63	35958.15	39242.95	54222.41	44701.69	39623.54	56269.26	93775.44	118003.9	18526.59	63987.79	118037
butanoyl-l-carnitine (acyl-C4)	40791.64	58691.51	74794.55	114305.1	20047.24	29173.36	176479.9	75541.32	112804.8	57464.7	62521.95	85033.91	16669.43	35129.91	28271.37	36529.86	25181.13	7986.041	27742.08	14941.16	18125.88	18071.18	23544.85	24155.8	15237.84	30308.8	51454.45	24674.59	23417.65	29656.43	27750.62	25964.99
acyl-C4-OH (Hydroxybutyrylcarnitine)	87135.97	2282.574	48256.8	16372.43	8054.074	122018.4	8451.295	11887.28	54326.22	14464.03	11345.84	17080.82	23332.85	20400.04	5428.111	8279.535	3809.89	36539.49	4582.04	7176.467	24168.66	3225.808	2473.696	22466.67	9239.86	5778.402	31960.28	100259.6	75932.63	25078.53	89433.78	120501.2
hexanoyl-L-carnitine (acyl-C6)	60918.88	53178.9	63314.1	96814.34	36659.64	72368.3	107812.9	66769.59	118679.7	43740.66	130244.5	112206.9	61493.28	77173.38	65164.68	50374.93	36491.11	20444.79	33774.01	58570.29	65989.96	58055.86	78425.11	85006.81	47795.25	123499.8	55113.49	79666.63	69481.13	68197.66	32833	63782.43
O-tetradecanoyl-L-carnitine (acyl-C14)	46375.39	20970.85	76118.96	50466.01	8723.608	142546.6	103449.8	33979.63	181010.9	6655.089	163382.4	98307.35	88597.46	151754.5	156745.2	96068.66	21571.52	26307.88	52265.7	82230.66	351323.5	57948.97	41429.12	62673.37	42672.99	122290	152393.9	135618.2	90363.38	28516.9	40140.26	97474.01
Tetradecenoyl Carnitine (acyl-C14:1)	0	6169.637	15642.63	13407.21	0	13045.27	31377.34	0	58998.84	4425.419	28037.14	0	15416.18	18560.88	14749.67	17377.68	0	0	15880.65	16440.41	48279.12	12436.45	5958.042	0	0	8387.405	19281.21	14763.79	0	0	0	5323.473
L-Palmitoylcarnitine (acyl-C16)	414668.8	157093.4	804225.2	366821.8	109628.3	1532889	443921.5	386195.5	1155625	74099.68	1072655	624084.3	395020.5	586202.2	1109325	433963.6	71942.59	215192.4	92497.59	354889.2	1432734	220825.7	152989.5	520451.6	230235.6	687780.6	1070248	812130.3	534530.7	251522.3	427361.4	873474.9
Hexadecenoyl-carnitine (acyl-C16:1)	16370.76	15120.57	29507.17	27195.97	15530.61	46197.74	66884.18	6300.731	90463.02	0	43213.37	23206.31	31084.61	48463.39	49197.94	37127.05	5477.277	5539.333	19527.44	0	48652.86	22069.54	12494.8	16987.01	12991.71	31594.93	41757.53	38897.66	4682.358	9938.187	6044.398	19817.2
O-octadecenoyl-L-carnitine (acyl-C18:1)	131560.3	39482.66	165159.7	112440	25596.37	112883.4	128844.9	60590.96	331559.3	11094.12	155796	111399.6	91389.24	139394.5	254657.6	82979.02	14363.32	26873.38	35931.97	78146.52	255171.7	52246.95	12234.43	80068.74	45561.53	219807.8	210723.3	216959	51349.11	12508.25	28567.41	96596.7
acyl-C18:2 (Linoleoyl-CoA)	15546.35	15030.5	54491.12	62904.62	5926.594	33287.9	30282.38	12676.53	67974.8	5759.309	73109.94	51144.31	33214.08	106124.1	186060.1	75191.25	8216.219	19973.98	23295.48	34043.6	117477.8	21654.4	6478.732	47648.24	19322.74	119456	48661.53	134547.3	7809.436	15408.67	27724.96	39251.69
acyl-C18:2-OH	0	0	7716.551	12640.76	0	13473.95	5528.614	4519.904	5779.124	4218.482	6385.586	0	6606	7360.895	0	0	0	6508.528	0	0	4700.116	0	0	14327.54	0	4114.287	0	9525.99	0	0	8334.43	12339.95
acyl-C20:4	41515.81	31695.56	53092.48	19238.63	4250.458	37665.83	16391.58	20018.46	92278.62	0	82591.69	56456.52	41222.31	99358.45	503557.9	59478.56	13999.95	27368.5	14947.35	52282.16	239430.2	16373.52	11284.36	77166.77	68095.23	293310.2	56988.88	136785.1	0	27263.03	24109.78	62383.03
Acetylcarnitine	2905970	362659.5	1073567	638192.6	102023.9	4357449	464829.5	799184.3	1545359	520470.5	812644.2	551959.7	1547211	1091339	138076.2	924098	149746.3	1645312	158794.7	455102	541753.1	286556.8	189399.8	1180400	672778.3	296761.1	1088450	3247484	2454397	820850.4	2077774	4464908
Isovalerylcarnitine	26901.48	75082.87	79193.13	110642.7	39203.81	18590.71	199812.6	65215.55	141498.8	92726.57	90481.21	108835.7	49555.44	110547.5	50919.18	130026.2	90283.37	93489.52	60958.1	232795.7	117225.6	208006.4	516311.7	359287.9	217260.4	140667.7	42381.48	131052.6	49270.73	179237.7	122278.9	141493.8
Hexenoylcarnitine	9281.122	26549.07	28946.94	29519.46	29211.17	15713.86	11844.24	10638.95	18919.19	21445.76	26732.25	37588.57	34381.84	23575.2	64568.84	7487.879	28324.17	16118.73	20308.89	33360.62	17840.57	15157.65	13304.75	22421.38	16690.65	89798.22	7135.642	18841.79	28498.95	34469.16	19000.12	17692.6
Hexanoylcarnitine	60918.88	53178.9	63314.1	96814.34	36659.64	72368.3	107812.9	66769.59	118679.7	43740.66	130244.5	112206.9	61493.28	77173.38	65164.68	50374.93	36491.11	20444.79	33774.01	58570.29	65989.96	58055.86	78425.11	85006.81	47795.25	123499.8	55113.49	79666.63	69481.13	68197.66	32833	63782.43
Dimethylheptanoyl carnitine	49687.47	65457.51	62747.34	41058.44	51947.3	48148.82	51206.76	53639.53	38930.18	49069.2	69476.37	41286.96	45021.97	82915.15	49014.22	78889.27	54656.46	61553.09	34017.25	67141.87	46688.45	70618.34	60729.83	64496.61	44911.73	41218.67	60706.11	35166.88	45034.75	56566.67	42376.91	47368.66
Pimelylcarnitine	15188.47	43953.99	33608.68	60842.44	34750.34	6425.703	78442.63	41190.12	66658.21	39415.18	34148.32	59355.55	11172.36	25961.01	25282.15	28823.9	44775.16	4259.706	51698.06	31796.71	17859.27	12451.42	18311.36	16221.52	20271.35	21696.68	28016.11	12336.14	10005.43	25983.32	14305.42	7744.549
Hydroxyoctanoyl carnitine	39897.86	64342.02	64548.29	57813.5	66271.74	44929.55	80244.91	31291.91	121250	53974.86	78896.41	110239.4	22690.46	37517.09	71287.45	18686.99	116276.9	20424.18	61100.18	35794.54	39887.16	19132.31	64322.48	39413.5	10572.2	33870	36675.08	34948.24	35141.71	52404.97	31552.53	29628.25
Decanoylcarnitine	8540.272	7899.172	23031.56	6089.807	7531.318	14945.4	8457.546	15734.36	8894.284	7374.75	7503.668	6561.377	6141.385	13871.16	6436.507	6597.852	6985.293	5052.189	10164.05	36700.36	27858.17	15430.2	10799.28	7049.223	8531.067	7069.791	6750.148	16829.24	9560.753	8131.766	7707.025	6830.822
Hydroxynonanoyl carnitine	8818.108	16924.29	24284.23	18833.18	14812.52	13915.23	18269.06	11332.05	54999.72	16479.19	23308.34	28464.55	17449.42	20813.6	22922.48	208258.7	24169.93	7993.904	27010.22	35011.23	10608.07	17597.98	20117.83	10353.11	21750.34	14867.99	14731.79	21016.18	18033.08	0	22140.47	13910.83
Dimethylnonanoyl carnitine	23661.22	27497.47	45312.2	16297.39	30824.78	19998.79	38801.76	34665.01	37799.78	28176.98	42689.83	36620.05	29275.62	56219.35	0	57006.17	35573.54	30310.62	18253.59	33041.25	35068.78	28950.12	43317.49	45734.73	47383.8	41720.3	26767.92	37273.24	281614	28896.18	29838.09	39382.78
Hydroxydecanoyl carnitine	21722.78	5043.33	46462.7	14663.08	10144.81	32346.74	7536.272	18832.76	53309.97	9048.671	18948.88	12180.82	22255.8	16746.16	14482.29	35816.42	7866.007	7287.227	12362.49	26420.63	27374.34	8853.825	14916.42	15928.77	14304.39	11199.04	29959.17	29252.47	33855.56	27979.89	14013.23	22027.93
Tetradecanoylcarnitine	46375.39	20970.85	76118.96	50466.01	8723.608	142546.6	103449.8	33979.63	181010.9	6655.089	163382.4	98307.35	88597.46	151754.5	156745.2	96068.66	21571.52	26307.88	52265.7	82230.66	351323.5	57948.97	41429.12	62673.37	42672.99	122290	152393.9	135618.2	90363.38	28516.9	40140.26	97474.01
Pentadecanoylcarnitine	11790.61	7957.163	21343.07	20101.77	12434.83	13639.27	5971.559	4181.446	6162.445	0	12414.35	6835.013	27883.1	10320.22	113963.4	15181.46	3975.267	3255.427	11803.3	3923.646	27325.12	8581.651	12846.4	24009.05	0	185763.6	10209.54	21612.16	18454.35	23702.37	4206.702	5786.418
Hydroxytetradecanoyl carnitine	16753.51	6254.947	66598.13	29642.56	5701.42	66091.51	19455.02	25788.45	99476.12	4794.568	24296.11	21097.26	11345.05	29247.71	6256.297	22999.15	8020.295	0	5373.696	4170.988	11970.2	5760.297	17032.13	13663.98	8954.392	13050.44	40534.74	22028.45	34394.88	21931.84	22873.99	40555.22
Hydroxyhexadecanoyl carnitine	36027.04	0	98133.59	24361.38	10471.7	171514.3	57168.73	37825.86	182303.5	9629.452	75398.02	34460.21	64187.82	66829.85	37516.47	52985.04	0	26139.83	6905.943	17634.7	70387.97	5254.515	15079.97	45479.86	18304.93	51374.52	119981.3	66087.54	70309.2	79885.35	120685.2	135923.5
Biliverdin	0	0	7156.768	11274.91	0	0	6422.701	0	24110.66	0	11773.37	8753.056	19832.67	40538.27	0	35251.86	0	65194.42	24902.04	61904.04	42147.82	7778.793	5247.174	11119.45	12189	71877.71	0	34303.27	0	0	24560.86	4341.983
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	parent	medRt
L-alanine	90.0556	0.219455
L-arginine	175.119	0.1675275
L-asparagine	133.0605	0.1724712
L-glutamate	148.0604	0.1716918
L-glutamine	147.0762	0.1766035
L-histidine	156.0764	0.1806751
L-leucine	132.1018	0.1926388
L-lysine	147.1125	0.1673335
L-methionine	150.0579	0.1897255
L-phenylalanine	166.0859	0.2613297
L-proline	116.0709	0.1708287
L-serine	106.0501	0.1696804
L-threonine	120.0656	0.1827708
L-tryptophan	205.097	0.4112165
L-tyrosine	182.0809	0.1901361
L-valine	118.0865	0.1923036
AMP	348.0695	0.2152835
Cytidine	244.0954	0.6292967
Allantoate	177.0615	0.1533905
Nicotinamide	123.0555	0.1786006
Nicotinate ribonucleotide	337.0609	0.2050757
Glutathione	308.0901	0.1980568
Spermidine	146.165	0.1514117
Spermine	203.2227	0.150648
Creatine	132.0769	0.1705798
Creatinine	114.0664	0.1895912
Guanidinoacetate	118.0614	0.1697755
Pantothenate	220.1175	0.3217335
Pantetheine	279.1414	0.7404448
L-Methionine S-oxide	166.0527	0.2012817
Indole	118.0654	0.812836
L-Adrenaline	184.0949	0.1543104
Ectoine	143.0813	0.2132641
Sphingosine	300.2891	0.7764517
Sphingosine 1-phosphate	380.2552	0.7651757
Acetylcholine	146.1173	0.1636865
Choline	104.1074	0.162063
L-Carnitine	162.1122	0.1686187
propionyl-carnitine (acyl-C3)	218.1385	0.1804759
butanoyl-l-carnitine (acyl-C4)	232.1542	0.3579879
acyl-C4-OH (Hydroxybutyrylcarnitine)	248.1488	0.1956434
hexanoyl-L-carnitine (acyl-C6)	260.1855	0.6265142
O-tetradecanoyl-L-carnitine (acyl-C14)	372.3102	0.7846905
Tetradecenoyl Carnitine (acyl-C14:1)	370.2946	0.761014
L-Palmitoylcarnitine (acyl-C16)	400.3411	0.8316876
Hexadecenoyl-carnitine (acyl-C16:1)	398.3258	0.7990546
O-octadecenoyl-L-carnitine (acyl-C18:1)	426.3567	0.8402839
acyl-C18:2 (Linoleoyl-CoA)	424.341	0.8111843
acyl-C18:2-OH	440.3368	0.7481034
acyl-C20:4	448.341	0.8092763
Acetylcarnitine	204.1229	0.172105
Isovalerylcarnitine	246.1698	0.6123381
Hexenoylcarnitine	258.1697	0.6563342
Hexanoylcarnitine	260.1855	0.6275252
Dimethylheptanoyl carnitine	302.2322	0.8198106
Pimelylcarnitine	304.1752	0.6294311
Hydroxyoctanoyl carnitine	304.211	0.6134121
Decanoylcarnitine	316.2478	0.7397916
Hydroxynonanoyl carnitine	318.2267	0.6402744
Dimethylnonanoyl carnitine	330.2632	0.8124963
Hydroxydecanoyl carnitine	332.2427	0.6854022
Tetradecanoylcarnitine	372.3103	0.7853739
Pentadecanoylcarnitine	386.3256	0.8065909
Hydroxytetradecanoyl carnitine	388.3046	0.734615
Hydroxyhexadecanoyl carnitine	416.3359	0.7778495
Biliverdin	583.2534	0.7262135
METABOLITES_END
#END