#METABOLOMICS WORKBENCH cendalfr_20230605_114108 DATATRACK_ID:4065 STUDY_ID:ST002727 ANALYSIS_ID:AN004420 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        	Negative 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                     	95% water/5% acetonitrile; 1 mM ammonium acetate
CH:SOLVENT_B                     	95% acetonitrile/5% water; 1 mM ammonium acetate
CH:FLOW_GRADIENT                 	0-0.5 min 0% B, 0.5-1.1 min 0-100% B, 1.1-2.75 min hold at 100% B, 2.75-3 min
CH:FLOW_GRADIENT                 	100-0% B, 3-5 min hold at 0% 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                      	NEGATIVE
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-aspartate	1280237	147711.2	206913.4	143916.2	130934.2	516059.3	147554.4	207933.1	183043.6	168872.4	274293.5	212594.7	471213	552020.2	144089	471617.6	241060.5	732706.8	139656	274229.8	437848	269817.6	2472474	652159.6	292093.3	229460.6	345229.9	1360148	1178795	299608	711992.3	1082827
ATP	5914.357	7947.061	67580.62	69968.2	6224.875	7555.184	13264.26	28002.27	99575.91	43456.82	1770.325	14392.69	2512.02	8577.173	1159.033	3053.713	3754.386	2066.554	0	0	0	5525.522	2282.28	9585.789	6536.36	0	0	5089.862	5132.081	37283.51	2387.067	0
ADP	64709.03	0	22178.66	0	0	194785.1	0	10518.49	13052.33	22215.4	11242.46	10253	23808.61	23292.19	9774.098	38857.28	0	43971.22	0	0	6670.852	0	11547.3	81079.76	8288.003	36518.65	17930.47	26476.27	132698.1	37287.09	37905.7	112300.6
CTP	10661.18	8395.362	19061.39	7494.006	9200.95	20858.82	11607.8	5376.941	15392.39	8986.946	7109.117	12135.49	15898.84	10001.27	9594.763	6558.208	7077.367	19084.13	10901.99	40146.38	29509.24	6075.816	18512.37	8542.198	22479.05	13567.79	3664.322	11570.04	11081.82	10503.5	21791.46	18794.85
Thymidine	38130.36	18159.49	58512.52	17010.06	29115.79	0	51686.97	36119.26	17627.24	46261.21	17410.13	60234.04	61837.93	14423.61	41133.08	18252.77	52569.58	687.8574	27646.77	32258.79	15392.95	35015.54	17968.58	41219.83	18769.58	6707.658	19770.16	22677.17	0	26424.62	21065.28	21803.44
UDP	1251603	735800	767760.5	627272	720785	1609504	669192.4	653153.9	552937.1	933655.5	679254.7	605471.6	1127684	745375.8	746275.9	1160583	687907.9	878608.8	950915.3	1342176	894332.7	839129.9	856557.8	949989.8	1211798	733909.3	671120.4	1361480	849865.7	698118.9	971426.6	1198027
IDP	437000.5	430847.5	453879.4	427857.2	297350.2	359519.3	379863	366328.9	342350.2	509264.8	283272.9	460964.4	428579.4	383143.2	463748.8	372148.8	419826.1	334509.6	329362.5	0	13411.86	304071.1	320876.7	437221.3	386123.6	348474.6	385303.1	298679.2	335330.8	341097.8	402887.3	329473.5
Inosine	95931.48	13016.64	1160.374	0	4469.571	604.0537	1654.822	1111.078	1130.291	7259.939	0	666.0211	583.8054	1037.534	600.9387	894.0154	970.6406	891.5024	1027.549	1704.639	2417.479	1792.527	0	510.1975	0	654.8076	6041.604	48769.66	2495.949	0	14098.55	3360.469
5-Hydroxyisourate	28543.4	10162.93	14000.62	10740.61	18099.49	12371.85	9733.197	11343.87	10707.65	29003.65	11861.34	14464.58	28325.83	14273.77	17540.61	13940.21	19955.32	24328.59	39275	27960.33	25630.24	29128.23	8684.827	14865.94	28181.92	10555.03	5000.993	37995	18627.47	14363.12	25030.89	24090.06
3',5'-Cyclic IMP	16011590	10592290	12963520	10812850	8236668	14028880	11331240	10749130	10017150	17946740	9838875	17555910	17691490	15518860	12561450	11879610	12325170	14578640	11860790	8560.511	189804.5	13049860	13181220	15898030	15564760	11936910	12640280	14343080	14342130	11764920	15099690	13075730
Phosphate	7256588	7803654	7798728	8410071	5206138	5961784	8334942	7769982	7996078	7608538	5839602	6001044	5724514	6457512	7621994	6897108	7532302	4876878	6740436	7815210	9793601	5650090	5952278	6415084	6084548	5514398	7267228	5466824	5354460	6904742	5231174	4738452
Diphosphate	73125.47	99641.72	93665.85	121422.8	71949.6	42894.38	99701.12	125285.2	102940.5	110969.9	57038.45	52051.16	48935.48	56471.16	98515.47	70501.27	86173.12	43226.94	79037.2	129121.7	227388.2	49803.53	39366.34	64496.51	42147	60732.42	89802.91	38672.2	37859.5	108909.1	74583.34	30185.2
D-Glucose	14942000	19156430	18122180	19719880	15866690	11614110	24025350	20754620	19991810	20771560	20424720	20067190	14023940	17527010	19022460	16465360	23679820	11449000	21150630	16341850	19647860	15872070	17213020	16986480	18113880	17537850	23500420	11318390	11973200	19362430	13722780	14262400
D-Glucose 6-phosphate	175516.2	11852.26	30471.6	24147.38	0	150994.2	0	19309.58	28112.83	17551.18	23754.67	16250.25	71219.49	53622.88	0	75024.07	0	138488.1	0	54355.59	112951.1	9891.666	0	74149.3	28863.18	20114.85	77222.12	207826.6	153522.5	32697.37	153999.6	194441.2
D-Fructose 1-6-bisphosphate	13090.76	0	81860.43	53580.54	5724.398	74275.16	37166.32	51279.61	149697.1	22125.89	64164.11	15648.41	11767.83	44305.75	4750.27	25877.17	0	4858.896	9265.669	8537.159	22839.49	22687.13	3508.826	100436.9	14908.25	13747.56	105818.4	29419.38	122736	12579.24	63257.95	69647.17
D-Glyceraldehyde 3-phosphate/Glycerone phosphate	182133.7	8618.736	90916.85	25105.41	8085.863	320210.5	0	32344.91	46606.69	14574.2	41257.7	12392.73	175871.6	106556.2	6687.534	100686.9	5732.904	81470.66	5109.118	133104.6	163802.3	29543.2	11761.83	266115.3	51710.05	45305.3	51323.84	189120.1	387405	14094.93	136715.2	278982
1-3-Bisphosphoglycerate	6659.74	6750.912	1579.906	10407.02	1141.217	6852.246	7574.998	4308.788	14595.6	4013.021	2932.421	5751.734	707.6091	6982.536	8196.952	2683.696	1632.559	16537.77	2824.979	8369.253	6542.709	7723.719	9102.21	28913.75	12965.2	4504.878	6279.965	10459.77	10618	2679.746	8359.087	6228.021
2/3-Phospho-D-glycerate	119303.6	35198.36	39546.45	29166.28	0	382547.2	13117.02	25051.49	46842.56	19409.62	31328.81	18505.31	60613.72	77217.67	0	78832.02	0	141374.3	1194.294	15710.5	85050.5	56286.45	48994.81	146989.9	41817.23	45422.08	117424.1	143045.1	352564.5	21642.14	251756.4	178697.5
Pyruvate	402919.1	486965.2	442158.1	434035.5	381697	287729.8	560250.2	493212.4	633222.1	441399	567092.3	478093	343740.5	484205.9	434906	442913.4	581945	300239.1	411477.1	407515.5	431733.1	510757.5	419428.5	495495.1	461013.1	473128.1	571749.1	361099.8	431740.4	457328.9	421017	368442.1
Lactate	6296578	8334104	8756581	9473715	6238778	5848884	10320300	9623167	9487985	9238863	9768227	9198291	6262750	8548546	8783735	7253874	9086943	4883530	8416719	6100610	7654758	7211654	8690459	7705752	7197410	7703618	9640493	5661058	7132016	7602200	7780832	7130916
Mannitol	364915.2	358801.2	509141.4	368542.6	131391.2	283611.9	634527.7	514762.9	459180.2	454388	409946.6	523719.8	336654.5	487878.7	426161.8	315044.6	267768.6	177130.8	250308.3	46327.3	56333.14	363587	359374.4	372171.4	404511.9	393860.4	491461.9	268227.2	342056.3	211940.5	392019.2	331950.1
D-Ribose	448611.4	554526.6	576644.2	630686.4	370220	366492.8	572970.8	559072.6	485904.8	567616.5	518302.8	598055.6	466781.5	471834.8	516148.7	455733.3	618632.2	263925.5	607897.1	383206.1	530354.1	371076.5	474287	463298.7	490559.7	510642	732532.8	286993	362286.1	545731.8	400349.5	353272.2
D-Arabitol	8991.994	17923.64	14965.72	22991.7	0	24229.36	33676.77	15766.27	29384.38	56854.96	82163.87	176581.8	17465.47	18543.05	28324.94	19385.88	14819.96	7095.482	4037.274	8115.516	3748.516	5465.831	30457.75	44543.05	5492.287	6559.584	19130.91	23561.38	9509.265	46149.32	45747.39	7118.746
Citrate	4722092	4290648	2718198	3480297	2941980	4456328	5153000	3422163	62063210	51471960	33501950	62646090	2137725	3614834	4649217	2653001	8734631	2380363	2496106	889536.5	1237372	2586740	7635626	4257396	2453926	3684457	6947912	3050791	6475624	2993288	3406408	20729980
2-Oxoglutarate	182523.8	323863.3	1040.237	0	976.3161	5007.502	8795.079	11866.95	0	11167.3	12271.61	1121.737	57388.1	60007.5	6208.895	2428.712	1962.322	7622.292	0	12102.33	1205.321	1140894	1321565	1211688	1170555	867167.5	1134790	166834.7	104764.1	6544.742	0	4918.75
Succinate	243981	390009.1	305031.1	540532.2	226756.9	277035.2	807541.3	520143.5	486282.1	186658.9	341268.4	378173.1	185635.4	269995.3	401541.5	308953.8	141282.6	70374.25	278785.2	71080.95	28962.64	278296.5	284321.1	263539.9	238554.6	274388.4	350246.2	144398.4	205046.7	92976.52	226727.8	152252.2
Fumarate	145567	51911.65	50922.5	35521.93	35015.53	98321.35	57871.11	69091.91	49380.53	90705.34	65323.36	24244.02	80701.26	87348.16	56846.64	108312.9	51862.64	105101.7	15605.33	25056.22	37106.73	59775.72	125658.1	107803.7	73657.26	113041.9	80808.65	128028.8	205818.2	78100.61	116255.3	95779.12
Oxaloacetate	5977.123	7010.869	1331.449	17772.81	3004.74	8266.687	3074.993	4716.98	13296.83	0	0	0	6664.407	710.4986	987.6143	7174.531	2845.127	7857.957	7200.898	17544.75	8409.4	9400.327	4682.604	11291.38	2875.695	2375.331	3199.198	14893.99	21357.81	7088.852	19437.37	12891.72
Malate	1828942	514395.8	898689	668119.4	298307.5	1028689	566251.6	796262	631354.1	973731.2	801404.2	701721.7	956127.2	935928.1	533302.4	1158533	894483.9	1163372	319965.3	350942.8	517126.4	766534.8	1488697	1171968	988347.8	1112188	1184399	1697205	2295809	1214375	1403615	1388260
Itaconate	136144.8	79629.57	53491.67	56046.41	41928	128996.6	79467.99	82708.95	587898.6	1131738	581350.6	776406	79810.34	25538.03	71478.94	67309.31	89678.38	38380.62	31913.56	20413.59	23385.03	79966.3	108601.8	93784.59	76434.79	61935.42	77259.12	138617.5	122076.8	54528.79	110299.6	509497.7
2-Hydroxyglutarate/Citramalate	199856.3	99662.52	80390.54	90706.04	42166.08	161566	104482.4	116644.1	183068.7	154854.5	166005.7	197988.9	74992.05	103539.7	71701.32	85346.77	61561.93	64064.38	52056.46	23766.59	29510.09	113178	133067.4	166204.4	101131.6	91125.91	167582.7	158354.4	155776.4	100207	144001.9	166145.7
5-Phospho-alpha-D-ribose 1-diphosphate	81296.91	63354.19	54076.78	61422.14	19450.53	85261.02	61671.68	30860.05	134073.9	33795.86	67929.65	15264.26	68558.06	37722.25	31658.92	72105.02	33060.17	61894.47	65151.46	71834.19	50483.17	47745.59	52725.43	48266.22	64445	47893.52	44765.54	76896.27	45016.94	55836.79	45382.96	69453.76
5-Oxoproline	522831.6	130348.7	196695.4	86096.06	142539.2	496659.1	143591.7	102350.4	132681.6	126305.1	494130.9	457471.5	304314	508658.3	215396.3	320227.4	379086.6	322549.5	114544.3	266326.6	320446.4	363846.7	9202760	394352.5	374959	332075.7	279574.5	676551.8	2133578	487923.2	363992.6	532885.8
Ascorbate	23281.78	19291.83	21124.35	37746.85	29729.22	1667608	9559.128	9708.114	6250.521	9126.157	9822.192	7371.422	28781.66	14304.62	19643.88	48419.95	7483.441	24116.57	10667.91	67854.28	17033.22	26900.73	19127.04	24348.19	23689.77	13687.12	123609.2	35713.09	75324.49	41231.66	13826.86	19479
Dehydroascorbate	144475.2	75621.75	86547.79	82505.31	58274.5	144512.6	77113.48	66963.21	1166772	2386299	1168496	1569501	64199.41	76873.84	63027.26	73920.12	115873.1	57377.3	45757.93	27430.76	50000.96	44516.05	127215.8	102776.8	67007.27	72766.78	112347.5	86277.23	138376.8	69131.09	69378.29	808521.2
3-Phosphonooxypyruvate	56060.74	45041.16	56094.3	46108.89	47595.17	52739	46992	30591.89	55558.86	43584.29	50183.19	57092.48	41671.69	66752.07	46853.43	41337.99	69425.91	55652.76	46580.17	51519.8	54257.01	40060.78	32379.45	55896.85	72628.59	62239.95	58583.62	76479.35	52974.83	46616.64	66182.48	54479.41
5-10-Methenyltetrahydrofolate	42000.97	87938.45	36404.01	120327.1	37322.29	10242.99	116959.7	53584.73	262468.5	159918	69259.93	124146.1	8097.784	33189.1	47338.39	49183.22	103868.9	15146.42	105159.1	164318.8	200580.1	9206.419	173234	9488.084	25907.95	15299.71	51784.52	12347.75	15261.69	113899.9	19324.76	8866.77
Dihydrofolate	7726.6	20124.87	12409.02	24421.57	3740.023	1394.878	24793.36	11970.5	54520.18	20715.47	19470.92	25044.77	0	6255.871	9515.772	8428.169	26426.87	1830.266	22336.14	28411.6	42346.44	1716.157	5538.472	1470.526	1538.472	3551.98	11351	3410.458	1119.78	27851.65	4344.369	0
Ornithine	60929.57	58873.89	77338.61	43564.43	30491.27	54466.56	49567.04	49332.81	91428.74	33221.53	98593.95	90865.97	74126.43	93882.74	68619.29	82525.73	87527.73	90217.43	37074.86	85571.27	138896.3	44845.09	391991.1	100368.1	76982.73	61963.36	83950.06	185433.2	249903.5	71943.56	141885	174473
1-4-beta-D-Xylan	537551.6	738557.8	682012.3	726679.8	487187.4	405491.2	869499.7	684981.5	709842.8	752110.8	748302.1	645855.2	519946.2	632550.7	716238.1	512555	733847.8	446863.6	626588	511632.6	619962.1	612255.8	518009.5	577095.3	518309	585830.4	855162.1	426862.4	429769.2	670127.8	492826.8	471894
Taurine	2914528	153784.1	509169.9	307972.5	106559.4	3033946	147705.9	352200.9	872200.8	401121.5	512546	256763.4	1047305	563610.4	9818.045	761793	42706.34	1418770	90850.95	172339.3	553685.3	138228.1	68773.34	912779.7	358356.3	250641.9	941482.7	2462144	5285080	686460.8	2420434	2716559
L-Selenomethionine	63051.96	89384.91	100984.3	46968.89	71418.27	102512.9	92948.95	115924.7	70442.45	61288.69	116269.4	100686	62000.61	116766	80003.16	101427.5	119638.1	75452.2	73561.55	30339.99	98760.66	123626.6	51825.72	86556.65	62109.38	91711.59	123975.2	84796.06	108882.6	95005.45	123384.4	58129.32
L-Cysteate	48313.76	45295.8	46136.66	55918.62	43714.67	20742.82	44096.5	54274.02	49589.64	57377.34	59442.67	45768.86	18834.68	57706.95	31288.02	44835.43	65038.94	53605.54	54115.31	64794.97	69300.77	52715.39	43012.95	47184.05	58540.85	46687.91	49869.03	46659.48	52539.12	36336.56	57496.94	42983.77
1D-myo-Inositol 1-4-bisphosphate	13090.76	0	81860.43	53580.54	5724.398	74275.16	37166.32	51279.61	149697.1	22125.89	64164.11	15648.41	11767.83	44305.75	4750.27	25877.17	0	4858.896	9265.669	8537.159	22839.49	22687.13	3508.826	100436.9	14908.25	13747.56	105818.4	29419.38	122736	12579.24	63257.95	69647.17
Ethanolamine phosphate	277579.8	13610.35	11480.84	927.2377	0	101197.9	0	13080.04	16009.24	964.528	10420.49	1187.976	23128.66	14039.62	0	33649.64	0	38303.18	1821.049	13661.01	31608.04	0	65364.17	69806.29	12444.4	9999.857	33958.97	71723.54	87839.03	3653.161	64110.21	85969.87
Butanoic/Butyric acid	42686.3	70858.2	46248.57	53348.11	37695.8	38342.77	45915.15	61338.42	39087.5	83100.47	62134.2	102517.2	34578.02	46582.55	17177.8	56300.33	45086.32	70967.23	35800.79	47474.05	23143.07	39657.3	42550.94	77967.36	60240.91	41344.21	59010.63	27323.09	41305.75	49915.99	45420.34	64216.43
Pentanoate (valerate)	119634.4	76503.73	58448.58	64001.84	38552.39	110327.9	55724.96	38509.79	73026.3	50703.07	67705.71	73600.96	69956.84	54502.14	45847.98	47943.16	63622.91	95766.91	74756.94	76056.6	93302.56	78523.99	82759.18	77015.34	106145	50776.98	73018.97	118197.3	100343.4	74097.62	76750.62	83959.99
Heptanoic acid	219452.2	272303.8	224502.3	241582.1	147600.7	213473	255255	228030.3	237380.4	201073.6	251655.2	253104.2	227865.9	204590	195228.2	219383.6	248539.5	248055.2	232990.3	239420.1	233246.3	228890	236994.9	235756.3	201831.4	190922.2	216785.6	153932.4	137035.1	176056.1	164955.7	212686.4
Octanoic acid (caprylic acid)	3120966	12070920	7150830	7567450	3449967	1536501	13873040	11828240	11193620	16147210	10262360	17199640	3790782	6131504	6075350	4133254	7481210	2043920	6905882	3342564	4856936	4538074	7644830	6189120	2724334	5007192	8002348	1208979	1000303	1841562	1772318	1354612
Nonanoic acid (pelargonic acid)	379024.8	808410.7	392873.5	449033.6	297755.8	483389.1	530937.3	562954.8	612581.6	425493.5	602531.2	596742.5	536431.5	553415.7	570825.6	362954.8	624947.7	545912.2	620284.8	542164	537494.1	520276.7	538049	596726.1	449060.5	472348.6	582267.4	412810.6	474441.8	520422.5	614852.2	539923.1
Decanoic acid (capric acid)	265053.2	426547.7	381105.2	351158.9	98879.66	203141.8	296799.6	257861.4	305154.3	228136	272872.8	289658.1	252774.2	252937.1	253760.5	183861.8	345702.6	269040	453140.5	340098.2	235629.1	206501.5	308039.6	313392.4	207360.5	265982.2	260716.2	260295.5	186109.6	243849.9	300455	222250.5
Dodecanoic acid (lauric acid)	464234.9	1376093	930260.9	401208.7	391915.7	337377.2	519085.6	413489.6	1285919	694559.6	550401.1	772839.6	291349.7	639689.7	549988.8	276045.5	568043.2	259024.8	1709141	570912.3	415725.7	277890.3	1157258	275007.5	122636.5	505806.8	489993.4	326468.2	484059	720941.5	328429	311207.6
Tetradecanoic acid (myristic acid)	1703099	4350155	2386477	1015176	3068824	949441.3	1271482	1138600	7066278	3553321	2579409	1842403	1155455	2902112	1668575	1557370	3833384	1863879	9254246	2363700	3220629	830248.9	5163084	935527	1414822	1502263	3215624	1095001	1881422	3412359	1334685	1992440
Hexadecanoic acid (palmitic acid)	34112280	76908300	56920330	33354180	47326820	30180010	27647690	40151850	119857800	123773100	64115070	60791840	41019400	62483920	101446100	34504620	103340400	110853100	64828970	140183700	194854800	26566030	59221160	46672990	35128950	95858180	94738860	55120080	35209290	189315100	52882600	97963460
Octadecanoic acid (stearic acid)	13882230	14483990	18736740	9867518	25711020	15349160	6225012	12910660	19874560	28953620	20459320	17338020	16490800	24902970	34511270	13413590	37821830	41122480	15689820	46893680	46990880	14446700	29809490	11587800	18353170	40958420	20581010	20700930	14819000	60667920	20137550	31729060
Tetradecenoic acid (myristoleic acid)	231813.9	613204.9	430602	150284.1	323388.4	140300	219936.2	116910.7	809823.2	516303.7	338561	470147.9	92204.52	325040.6	259920.5	272990.5	399353.5	96286.06	3746832	304871.8	346573.7	111291.3	481704.3	160797	40728.08	285844.4	349508	197317.4	235844.5	269534.3	153920.7	187349.2
Hexadecenoic acid (Palmitoleic acid)	3696263	15963130	10608430	3998696	7777794	3346490	4013391	5825964	29922700	19515580	9370028	9718549	3254973	7203760	4907559	3592663	7671010	6915522	34215320	7745034	13745000	1918964	10496040	6199742	5199888	7319586	24259680	4370072	4787446	14106580	5854750	7857822
Octadecenoic acid (Oleic acid)	11852200	58710690	47946960	29507270	18266810	7243168	16414220	28596100	75653620	57930020	30080610	24249950	20177190	18945730	33346600	10463310	19919740	38368570	38097360	42556380	83861380	5769940	24061610	22665640	25996370	47860400	85401150	30703930	10909540	66649970	24444430	35804920
Linoleic acid ((9Z,12Z)-Octadecadienoic acid)	4798516	47522750	27858750	26354370	6489568	3586103	9663870	12800910	22458960	61192720	21877350	18841420	11251480	13091160	12022900	8824645	16751750	44612060	10914620	27964860	43360890	4403088	9862607	17264300	22355310	45152460	25561080	35289450	4263484	75997740	22862820	36963930
a-Linolenic acid (Octadecatrienoic acid)	176507	3008888	1417272	1930897	492089.7	181777	879691.1	871912.4	2051615	3258552	1206599	1077322	558045.4	728064.2	338297.6	576961.2	868853.2	2121592	1076619	1499679	1361977	230177.8	958559.4	1449900	1624359	1846420	1519951	1339186	162862	3100857	1245063	1727144
Arachidonic acid (Eicosatetraenoic acid)	8036948	20120330	19512500	11312810	5094514	8414217	5748794	12746740	20894050	53068290	21944390	17171650	12200400	11399080	27060140	8000046	12349100	47614180	4686308	22428250	47540540	3018903	8352062	23439530	19166470	48557320	21384350	32456700	2623925	59640630	15883420	27683600
Docosahexaenoic acid	1714447	6018738	6342306	3172938	2341241	901968.5	1478239	3717699	3181523	19153530	5011960	3836233	2303746	2670602	4511038	2179228	6196794	19339110	1464487	9503585	14128190	758303.2	3047143	4267762	4410683	7521734	4977621	4870136	343650.2	20290390	3917948	6382058
Eicosapentaenoic acid	1174320	2608116	788273.2	69300.13	2194626	409840.8	311043.2	0	18446080	899736.2	1755038	2165560	411542.5	2006690	1457689	592969.7	2644877	264745.8	1494380	1961236	1027716	270295.9	655955	194889.3	279086.1	890114.5	1041271	342813.5	1142613	1235820	667171.1	382648.6
Dihomo-gamma-linolenic acid ((8Z-11Z-14Z)-Icosatrienoic acid)	841234	2892940	1942784	1464317	649050.8	482824.7	857765.2	1465666	2370421	6788458	1826193	1785416	846060.7	937370.4	2524929	533876.5	1245199	2531590	1275504	1628138	4509744	399531.5	791519.8	1932642	1120559	2486021	3598674	1544467	282747	5778004	1186051	2236182
Docosapentaenoic acid ((7Z-10Z-13Z-16Z-19Z)-Docosa-7-10-13-16-19-pentaenoic acid)	1031298	1574959	1101165	917661.2	405025.4	383324.8	501475.3	944826.7	855767.1	4275120	1347085	1432512	626122.4	763627.3	4057843	635122.6	1071180	4034941	461684.1	1670048	3110199	265147.6	1202354	1320215	1098338	2148736	1539670	2177095	209537.8	4184839	1170865	1643502
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	parent	medRt
L-aspartate	132.0286	0.2227178
ATP	505.985	0.2138207
ADP	426.022	0.2180567
CTP	481.9821	0.2664376
Thymidine	241.0816	0.2690611
UDP	402.9995	0.2548533
IDP	427.007	0.2153879
Inosine	267.0732	0.3145487
5-Hydroxyisourate	183.0127	0.266116
3',5'-Cyclic IMP	329.0298	0.2133768
Phosphate	96.96804	0.2231005
Diphosphate	176.9345	0.2170104
D-Glucose	179.0549	0.2606818
D-Glucose 6-phosphate	259.0221	0.2189058
D-Fructose 1-6-bisphosphate	338.9884	0.2148654
D-Glyceraldehyde 3-phosphate/Glycerone phosphate	168.9883	0.246715
1-3-Bisphosphoglycerate	264.9521	0.2533868
2/3-Phospho-D-glycerate	184.9845	0.2174905
Pyruvate	87.00715	0.2424555
Lactate	89.0228	0.2444777
Mannitol	181.0708	0.2483564
D-Ribose	149.0447	0.2487217
D-Arabitol	151.0599	0.2749905
Citrate	191.0188	0.2201223
2-Oxoglutarate	145.0131	0.2235973
Succinate	117.0177	0.2222333
Fumarate	115.0023	0.2221624
Oxaloacetate	130.9999	0.2429924
Malate	133.0127	0.2219913
Itaconate	129.0177	0.2229531
2-Hydroxyglutarate/Citramalate	147.0285	0.221474
5-Phospho-alpha-D-ribose 1-diphosphate	388.9401	0.2577492
5-Oxoproline	128.034	0.2310195
Ascorbate	175.0259	0.2279899
Dehydroascorbate	173.008	0.2196947
3-Phosphonooxypyruvate	182.967	0.2679472
5-10-Methenyltetrahydrofolate	455.1561	0.6580315
Dihydrofolate	442.1525	0.648059
Ornithine	131.0811	0.2741969
1-4-beta-D-Xylan	131.0334	0.2464157
Taurine	124.0059	0.2385396
L-Selenomethionine	195.9914	0.2561352
L-Cysteate	167.9961	0.2682212
1D-myo-Inositol 1-4-bisphosphate	338.9884	0.2121459
Ethanolamine phosphate	140.0107	0.2335196
Butanoic/Butyric acid	87.04357	0.2538966
Pentanoate (valerate)	101.0593	0.2563013
Heptanoic acid	129.0908	0.560181
Octanoic acid (caprylic acid)	143.1064	0.5674003
Nonanoic acid (pelargonic acid)	157.1222	0.6033562
Decanoic acid (capric acid)	171.1379	0.6261744
Dodecanoic acid (lauric acid)	199.1698	0.6735848
Tetradecanoic acid (myristic acid)	227.2011	0.7278498
Hexadecanoic acid (palmitic acid)	255.2325	0.7951986
Octadecanoic acid (stearic acid)	283.2638	0.8666071
Tetradecenoic acid (myristoleic acid)	225.1855	0.6929401
Hexadecenoic acid (Palmitoleic acid)	253.2168	0.7480394
Octadecenoic acid (Oleic acid)	281.2481	0.8121029
Linoleic acid ((9Z,12Z)-Octadecadienoic acid)	279.2325	0.7616185
a-Linolenic acid (Octadecatrienoic acid)	277.2168	0.7287952
Arachidonic acid (Eicosatetraenoic acid)	303.2326	0.7545494
Docosahexaenoic acid	327.2327	0.7403131
Eicosapentaenoic acid	301.2169	0.7940361
Dihomo-gamma-linolenic acid ((8Z-11Z-14Z)-Icosatrienoic acid)	305.2481	0.7873595
Docosapentaenoic acid ((7Z-10Z-13Z-16Z-19Z)-Docosa-7-10-13-16-19-pentaenoic acid)	329.2484	0.7703723
METABOLITES_END
#END