#METABOLOMICS WORKBENCH amuirchicago_20250225_104023 DATATRACK_ID:5676 STUDY_ID:ST003759 ANALYSIS_ID:AN006173 PROJECT_ID:PR002342
VERSION             	1
CREATED_ON             	February 28, 2025, 12:51 pm
#PROJECT
PR:PROJECT_TITLE                 	Analysis of metabolite levels in the circulation of pancreatic tumor bearing
PR:PROJECT_TITLE                 	mice with or without depletion of myeloid cells
PR:PROJECT_SUMMARY               	In this study, we assessed the impact of myeloid cells on the levels of
PR:PROJECT_SUMMARY               	circulating metabolites in the plasma of mice bearing tumors. To do so, murine
PR:PROJECT_SUMMARY               	pancreatic ductal adenocarcinoma (PDAC) cells were implanted into C57Bl6J
PR:PROJECT_SUMMARY               	CD11b-DTR host animals. Myeloid cells were then ablated with 25 ng/g diptheria
PR:PROJECT_SUMMARY               	toxin (DT). Serum from DT treated mice was collected for analysis. Additionally,
PR:PROJECT_SUMMARY               	serum was collected from PDAC bearing mice that were vehicle treated. This data
PR:PROJECT_SUMMARY               	allows for identification of how myeloid cells in tumor bearing mice contribute
PR:PROJECT_SUMMARY               	to levels of circulating metabolites.
PR:INSTITUTE                     	University of Chicago
PR:DEPARTMENT                    	Ben May Department for Cancer Research
PR:LAST_NAME                     	Muir
PR:FIRST_NAME                    	Alexander
PR:ADDRESS                       	929 E 57th St
PR:EMAIL                         	amuir@uchicago.edu
PR:PHONE                         	7738346506
#STUDY
ST:STUDY_TITLE                   	Analysis of metabolite levels in the circulation of pancreatic tumor bearing
ST:STUDY_TITLE                   	mice with or without depletion of myeloid cells
ST:STUDY_SUMMARY                 	In this study, we assessed the impact of myeloid cells on the levels of
ST:STUDY_SUMMARY                 	circulating metabolites in the plasma of mice bearing tumors. To do so, murine
ST:STUDY_SUMMARY                 	pancreatic ductal adenocarcinoma (PDAC) cells were implanted into C57Bl6J
ST:STUDY_SUMMARY                 	CD11b-DTR host animals. Myeloid cells were then ablated with 25 ng/kg diptheria
ST:STUDY_SUMMARY                 	toxin (DT). Serum from DT treated mice was collected for analysis. Additionally,
ST:STUDY_SUMMARY                 	serum was collected from PDAC bearing mice that were vehicle treated. This data
ST:STUDY_SUMMARY                 	allows for identification of how myeloid cells in tumor bearing mice contribute
ST:STUDY_SUMMARY                 	to levels of circulating metabolites.
ST:INSTITUTE                     	University of Chicago
ST:LAST_NAME                     	Muir
ST:FIRST_NAME                    	Alexander
ST:ADDRESS                       	929 E 57th St
ST:EMAIL                         	amuir@uchicago.edu
ST:PHONE                         	7738346506
#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           	-	serum_DT_1	Sample source:Blood serum | Treatment:Diptheria_toxin	RAW_FILE_NAME(Raw_data_file_name)=190723_QE2_AlexanderMuir_Job1623_35.raw
SUBJECT_SAMPLE_FACTORS           	-	serum_DT_2	Sample source:Blood serum | Treatment:Diptheria_toxin	RAW_FILE_NAME(Raw_data_file_name)=190723_QE2_AlexanderMuir_Job1623_36.raw
SUBJECT_SAMPLE_FACTORS           	-	serum_DT_3	Sample source:Blood serum | Treatment:Diptheria_toxin	RAW_FILE_NAME(Raw_data_file_name)=190723_QE2_AlexanderMuir_Job1623_37.raw
SUBJECT_SAMPLE_FACTORS           	-	serum_DT_4	Sample source:Blood serum | Treatment:Diptheria_toxin	RAW_FILE_NAME(Raw_data_file_name)=190723_QE2_AlexanderMuir_Job1623_38.raw
SUBJECT_SAMPLE_FACTORS           	-	serum_DT_5	Sample source:Blood serum | Treatment:Diptheria_toxin	RAW_FILE_NAME(Raw_data_file_name)=190723_QE2_AlexanderMuir_Job1623_39.raw
SUBJECT_SAMPLE_FACTORS           	-	serum_DT_6	Sample source:Blood serum | Treatment:Diptheria_toxin	RAW_FILE_NAME(Raw_data_file_name)=190723_QE2_AlexanderMuir_Job1623_40.raw
SUBJECT_SAMPLE_FACTORS           	-	serum_noDT_1	Sample source:Blood serum | Treatment:vehicle	RAW_FILE_NAME(Raw_data_file_name)=190723_QE2_AlexanderMuir_Job1623_47.raw
SUBJECT_SAMPLE_FACTORS           	-	serum_noDT_2	Sample source:Blood serum | Treatment:vehicle	RAW_FILE_NAME(Raw_data_file_name)=190723_QE2_AlexanderMuir_Job1623_48.raw
SUBJECT_SAMPLE_FACTORS           	-	serum_noDT_3	Sample source:Blood serum | Treatment:vehicle	RAW_FILE_NAME(Raw_data_file_name)=190723_QE2_AlexanderMuir_Job1623_49.raw
SUBJECT_SAMPLE_FACTORS           	-	serum_noDT_4	Sample source:Blood serum | Treatment:vehicle	RAW_FILE_NAME(Raw_data_file_name)=190723_QE2_AlexanderMuir_Job1623_50.raw
SUBJECT_SAMPLE_FACTORS           	-	serum_noDT_5	Sample source:Blood serum | Treatment:vehicle	RAW_FILE_NAME(Raw_data_file_name)=190723_QE2_AlexanderMuir_Job1623_51.raw
SUBJECT_SAMPLE_FACTORS           	-	serum_noDT_6	Sample source:Blood serum | Treatment:vehicle	RAW_FILE_NAME(Raw_data_file_name)=190723_QE2_AlexanderMuir_Job1623_52.raw
#COLLECTION
CO:COLLECTION_SUMMARY            	Mice were euthanised 24 hours after treatment by CO2 asphyxiation and blood
CO:COLLECTION_SUMMARY            	collected through cardiac puncture. Serum was isolated using BD microtainer
CO:COLLECTION_SUMMARY            	serum separator tubes as per manufactures’ instructions and stored at -80
CO:COLLECTION_SUMMARY            	until analysis.
CO:SAMPLE_TYPE                   	Blood (serum)
#TREATMENT
TR:TREATMENT_SUMMARY             	CD11b-DTR mice were produced as an F1 cross between
TR:TREATMENT_SUMMARY             	B6.FVB-Tg(ITGAM-DTR/EGFP)34Lan/J mice and C57BL/6J mice (Jackson Laboratory).
TR:TREATMENT_SUMMARY             	Male mice between 8-12 weeks of age were used experimentally. Orthotopic PDAC
TR:TREATMENT_SUMMARY             	tumours were established from 5x104 KPC-7940B PDAC cells injected into the
TR:TREATMENT_SUMMARY             	pancreas were established as tumors for 14 days. Mice were then treated with
TR:TREATMENT_SUMMARY             	diphtheria toxin (Enzo Life Science) at a concentration of 25 ng/kg or vehicle
TR:TREATMENT_SUMMARY             	by intraperitoneal injection. Mice were euthanised 24 hours after treatment by
TR:TREATMENT_SUMMARY             	CO2 asphyxiation and blood collected through cardiac puncture. Serum was
TR:TREATMENT_SUMMARY             	isolated using BD microtainer serum separator tubes as per manufactures’
TR:TREATMENT_SUMMARY             	instructions and stored at -80 until analysis.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Metabolites were quantified as described in Sullivan et al eLife 2019 (PMID:
SP:SAMPLEPREP_SUMMARY            	30990168). Briefly, in order to quantitate metabolites in serum samples, we
SP:SAMPLEPREP_SUMMARY            	first constructed a library of 149 chemical standards of plasma polar
SP:SAMPLEPREP_SUMMARY            	metabolites. These compounds were selected to encompass a number of metabolic
SP:SAMPLEPREP_SUMMARY            	processes and have previously been included in efforts to profile plasma polar
SP:SAMPLEPREP_SUMMARY            	metabolites by LC/MS (Cantor et al., 2017; Evans et al., 2009; Lawton et al.,
SP:SAMPLEPREP_SUMMARY            	2008; Mazzone et al., 2016). We pooled these metabolites into 7 separate
SP:SAMPLEPREP_SUMMARY            	chemical standard pools. To do this, each metabolite in a given pool was weighed
SP:SAMPLEPREP_SUMMARY            	and then mixed (6 cycles of 1 min. mixing at 25 Hz followed by 3 min. resting)
SP:SAMPLEPREP_SUMMARY            	using a Mixer Mill MM301 (Retsch, Düsseldorf, Germany), and mixed metabolite
SP:SAMPLEPREP_SUMMARY            	powder stocks were stored at -20°C prior to resuspension and analysis. Stock
SP:SAMPLEPREP_SUMMARY            	solutions of the mixed standards pools containing ~5mM, ~1mM, ~300µM, ~100µM,
SP:SAMPLEPREP_SUMMARY            	~30µM, ~10µM, ~3µM and ~1µM of each metabolite were made in HPLC grade water
SP:SAMPLEPREP_SUMMARY            	and were stored at -80°C. We refer to these stock solutions as “external
SP:SAMPLEPREP_SUMMARY            	standard pools” throughout. External standard pools were used to confirm the
SP:SAMPLEPREP_SUMMARY            	retention time and m/z for each analyte and provide standards to quantitate
SP:SAMPLEPREP_SUMMARY            	concentrations of stable isotope labeled internal standards used in downstream
SP:SAMPLEPREP_SUMMARY            	analysis, as well as to quantitate metabolite concentrations in TIF and plasma
SP:SAMPLEPREP_SUMMARY            	samples directly where internal standards were not available (see below for
SP:SAMPLEPREP_SUMMARY            	details). To extract polar metabolites from plasma, TIF or the external standard
SP:SAMPLEPREP_SUMMARY            	pools, 5µL of serum or external sample pools was mixed with 45uL of
SP:SAMPLEPREP_SUMMARY            	acetonitrile:methanol:formic acid (75:25:0.1) extraction buffer including the
SP:SAMPLEPREP_SUMMARY            	following isotopically labeled internal standards: 13C labeled yeast extract
SP:SAMPLEPREP_SUMMARY            	(Cambridge Isotope Laboratory, Andover, MA, ISO1), 13C3 lactate (Sigma Aldrich,
SP:SAMPLEPREP_SUMMARY            	Darmstadt, Germany, 485926), 13C3 glycerol (Cambridge Isotope Laboratory,
SP:SAMPLEPREP_SUMMARY            	Andover, MA, CLM-1510), 13C6 15N2 cystine (Cambridge Isotope Laboratory,
SP:SAMPLEPREP_SUMMARY            	Andover, MA, CNLM-4244), 2H9 choline (Cambridge Isotope Laboratory, Andover, MA,
SP:SAMPLEPREP_SUMMARY            	DLM-549), 13C4 3-hydroxybutyrate (Cambridge Isotope Laboratory, Andover, MA,
SP:SAMPLEPREP_SUMMARY            	CLM-3853), 13C6 glucose (Cambridge Isotope Laboratory, Andover, MA, CLM-1396),
SP:SAMPLEPREP_SUMMARY            	13C2 15N taurine (Cambridge Isotope Laboratory, Andover, MA, CNLM-10253), 2H3
SP:SAMPLEPREP_SUMMARY            	creatinine (Cambridge Isotope Laboratory, Andover, MA, DLM-3653), 8-13C adenine
SP:SAMPLEPREP_SUMMARY            	(Cambridge Isotope Laboratory, Andover, MA, CLM-1654), 13C5 hypoxanthine
SP:SAMPLEPREP_SUMMARY            	(Cambridge Isotope Laboratory, Andover, MA, CLM-8042), 8-13C guanine (Cambridge
SP:SAMPLEPREP_SUMMARY            	Isotope Laboratory, Andover, MA, CLM-1019), 13C3 serine (Cambridge Isotope
SP:SAMPLEPREP_SUMMARY            	Laboratory, Andover, MA, CLM-1574) and 13C2 glycine (Cambridge Isotope
SP:SAMPLEPREP_SUMMARY            	Laboratory, Andover, MA, CLM-1017). All solvents used in the extraction buffer
SP:SAMPLEPREP_SUMMARY            	were HPLC grade. Samples were then vortexed for 10 min. at 4°C and insoluble
SP:SAMPLEPREP_SUMMARY            	material was sedimented by centrifugation at 15kg for 10 min. at 4°C. 20µL of
SP:SAMPLEPREP_SUMMARY            	the soluble polar metabolite extract was taken for LC/MS analysis.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_TYPE           	HILIC
CH:INSTRUMENT_NAME               	Thermo Dionex Ultimate 3000
CH:COLUMN_NAME                   	SeQuant ZIC-pHILIC (150 x 2.1mm,5um)
CH:SOLVENT_A                     	100% water; 20 mM ammonium carbonate, 0.1% ammonium hydroxide
CH:SOLVENT_B                     	100% acetonitrile
CH:FLOW_GRADIENT                 	linear gradient from 80% to 20% B; 20-20.5 min: linear gradient from 20% to 80%
CH:FLOW_GRADIENT                 	B; 20.5-28min: hold at 80% B.
CH:FLOW_RATE                     	0.150 mL/min
CH:COLUMN_TEMPERATURE            	25
#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                   	LC/MS analysis was performed using a QExactive orbitrap mass spectrometer using
MS:MS_COMMENTS                   	an Ion Max source and heated electrospray ionization (HESI) probe coupled to a
MS:MS_COMMENTS                   	Dionex Ultimate 3000 UPLC system (Thermo Fisher Scientific, Waltham, MA).
MS:MS_COMMENTS                   	External mass calibration was performed every 7 days. 2μL of each sample was
MS:MS_COMMENTS                   	injected onto a ZIC-pHILIC 2.1 × 150 mm analytical column equipped with a 2.1
MS:MS_COMMENTS                   	× 20 mm guard column (both 5 μm particle size, EMD Millipore). The autosampler
MS:MS_COMMENTS                   	and column oven were held at 4°C and 25°C, respectively. Buffer A was 20 mM
MS:MS_COMMENTS                   	ammonium carbonate, 0.1% ammonium hydroxide; buffer B was acetonitrile. The
MS:MS_COMMENTS                   	chromatographic gradient was run at a flow rate of 0.150 mL/min as follows: 0-20
MS:MS_COMMENTS                   	min: linear gradient from 80% to 20% B; 20-20.5 min: linear gradient from 20% to
MS:MS_COMMENTS                   	80% B; 20.5-28min: hold at 80% B. The mass spectrometer was operated in full
MS:MS_COMMENTS                   	scan, polarity-switching mode with the spray voltage set to 3.0 kV, the heated
MS:MS_COMMENTS                   	capillary held at 275°C, and the HESI probe held at 350°C. The sheath gas flow
MS:MS_COMMENTS                   	rate was set to 40 units, the auxiliary gas flow was set to 15 units, and the
MS:MS_COMMENTS                   	sweep gas flow was set to 1 unit. The MS data acquisition was performed in a
MS:MS_COMMENTS                   	range of 70-1000 m/z, with the resolution set to 70,000, the AGC target at 1e6,
MS:MS_COMMENTS                   	and the maximum injection time at 20 msec. Metabolite identification and
MS:MS_COMMENTS                   	quantification was performed with XCalibur 2.2 software (Thermo Fisher
MS:MS_COMMENTS                   	Scientific, Waltham, MA) using a 5ppm mass accuracy and a 0.5 min. retention
MS:MS_COMMENTS                   	time window. For metabolite identification, external standard pools were used
MS:MS_COMMENTS                   	for assignment of metabolites to peaks at given m/z and retention time, and to
MS:MS_COMMENTS                   	determine the limit of detection for each metabolite. Metabolite quantification
MS:MS_COMMENTS                   	was performed by two separate methods. Where internal standards were available,
MS:MS_COMMENTS                   	first, comparison of the peak areas of the stable isotope labeled internal
MS:MS_COMMENTS                   	standards with the external standard pools allowed for quantification of the
MS:MS_COMMENTS                   	concentration of labeled internal standards in the extraction buffer.
MS:MS_COMMENTS                   	Subsequently, we compared the peak area of a given metabolite in the TIF and
MS:MS_COMMENTS                   	plasma samples with the peak area of the internal standard to quantitate the
MS:MS_COMMENTS                   	concentration of that metabolite in the serum sample. For metabolites without
MS:MS_COMMENTS                   	internal standards, the peak area of each analyte was normalized to the peak
MS:MS_COMMENTS                   	area of a labeled amino acid internal standard that eluted at roughly the same
MS:MS_COMMENTS                   	retention time to account for differences in recovery between samples. From the
MS:MS_COMMENTS                   	normalized peak areas of metabolites in the external standard pools, we
MS:MS_COMMENTS                   	generated a standard curve describing the relationship between metabolite
MS:MS_COMMENTS                   	concentration and normalized peak area. The standard curves were linear with
MS:MS_COMMENTS                   	fits typically at or above r2=0.95. Metabolites which did not meet these
MS:MS_COMMENTS                   	criteria were excluded from further analysis. These equations were then used to
MS:MS_COMMENTS                   	convert normalized peak areas of analytes in the serum samples into analyte
MS:MS_COMMENTS                   	concentration in the samples. The relationship between metabolite concentration
MS:MS_COMMENTS                   	and normalized peak area is matrix dependent, and the external standards are
MS:MS_COMMENTS                   	prepared in water, which is a different matrix than serum. Therefore, we
MS:MS_COMMENTS                   	consider metabolite measurements using this external standard method
MS:MS_COMMENTS                   	semi-quantitative.
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	micromole/L
MS_METABOLITE_DATA_START
Samples	serum_DT_1	serum_DT_2	serum_DT_3	serum_DT_4	serum_DT_5	serum_DT_6	serum_noDT_1	serum_noDT_2	serum_noDT_3	serum_noDT_4	serum_noDT_5	serum_noDT_6
Factors	Sample source:Blood serum | Treatment:Diptheria_toxin	Sample source:Blood serum | Treatment:Diptheria_toxin	Sample source:Blood serum | Treatment:Diptheria_toxin	Sample source:Blood serum | Treatment:Diptheria_toxin	Sample source:Blood serum | Treatment:Diptheria_toxin	Sample source:Blood serum | Treatment:Diptheria_toxin	Sample source:Blood serum | Treatment:vehicle	Sample source:Blood serum | Treatment:vehicle	Sample source:Blood serum | Treatment:vehicle	Sample source:Blood serum | Treatment:vehicle	Sample source:Blood serum | Treatment:vehicle	Sample source:Blood serum | Treatment:vehicle
3-methyl-2-oxopentanoic-acid-ketoisoleucine	4.959751978	7.083232525	9.252156987	6.19876889	4.819853816	8.50887137	7.176097659	3.370555346	17.79212529	21.76751671	4.689716292	19.62867907
3-methyl-2-oxobutyrate-keto-isovaleric-acid	3.254135251	6.273914241	11.45472189	37.54668526	9.296835751	4.804580056	9.132572566	3.320854488	9.644646288	9.030450908	3.747580445	6.167704034
pipecolate	4.581089047	4.359180803	5.146100084	7.244539197	5.001384874	4.13190797	6.359276088	6.851752544	7.98320349	11.56053887	7.385009517	5.875464959
orotate	0.713227205	0.95727733	1.291783359	1.494555739	1.381846976	0.776686805	0.288019342	0.597377617	2.685251532	1.754480055	1.408781914	0.911014999
2HG	13.66857674	12.53050155	16.82557578	18.92721015	19.66897539	11.53220866	7.232493584	14.522768	15.71950451	12.24812285	16.25899812	6.153527601
UDP-GlcNAc	0	0	0	0	0	0	63.00400306	106.0210866	79.77367097	74.38800685	108.5791185	15.53774364
pyruvate	388.2881877	381.2080316	371.0257413	520.5543433	285.5860612	454.3066942	69.80326829	77.10380154	141.2392871	45.41097981	292.6489279	53.58547687
succinate	197.3280307	197.2203993	100.1920962	214.8756298	107.255962	461.0136586	71.56244467	165.0948979	122.5502135	146.7505733	204.564441	48.11432608
akg	18.88896785	31.70929059	15.74116987	17.5634183	11.40904508	24.74063341	7.442660729	6.208714072	35.34456503	17.56520401	4.736118722	9.528372289
malate	41.45628418	52.81391931	31.15861756	41.63263131	24.85702104	108.9927748	189.7570892	208.0402709	180.8040001	196.9761607	360.5117032	58.56112641
glucose-6-phosphate	0	0	0	0	0	4.948571604	0	0	100.471338	4529.119614	0	0
citrate	259.2826805	244.3556198	263.3349595	303.5546	215.4415576	280.7343638	319.7259247	205.1366291	331.2566397	364.9041899	325.216945	227.5561971
uridine	20.46351952	19.96788164	11.64789627	27.25335164	13.86945051	76.96155976	9.985389925	10.48308562	0	0	12.74393899	9.844857419
AMP	0	0	0	0	0	0	126.1027493	450.8604861	171.0741779	182.7340036	393.5100474	53.95759751
a-aminobutyrate	16.32547754	13.06517564	18.16942132	0	21.16344241	18.25992451	12.21437574	15.79623175	24.45578687	10.61384172	13.61868712	14.22301812
argininosuccinate	0.214822125	0	0.126751601	0.216798212	0	0.698023024	0.685027454	0.504870161	0	0.887848447	1.24721154	0.172916474
GMP	0	0	0	0	0	0.975183519	27.00031129	91.0480432	52.00480373	42.18140062	77.46568985	10.38070338
lactate	16038.0607	15759.7016	19272.41738	22031.38162	13222.18717	18097.08814	14155.44583	10637.52209	17043.15924	18817.34775	20591.71697	10000.57487
glucose	9751.398657	8170.998618	6154.08856	16294.12026	5318.700009	12797.90067	5120.521596	611.8661412	5103.917478	8678.413049	2697.493813	3410.07749
serine	152.2790851	259.2956415	217.2635009	171.4690183	119.7936627	134.8289427	335.6402038	307.845245	377.2536978	361.913793	555.8812075	185.0250667
glutamate	73.54001111	72.09733157	94.71723656	79.01763356	88.69929094	80.30653462	1638.161436	2331.478426	1879.616879	2185.672799	2497.638738	553.5469469
aspartate	33.51440262	25.90898298	45.24786148	40.50963601	30.66769607	36.16769575	443.540742	500.4307194	504.0436413	560.35853	633.6565077	132.9816818
taurine	1235.394746	1177.081546	1532.89347	1489.679741	907.6748332	1776.335985	8765.142623	8795.987732	7375.19982	8581.662169	13332.62904	1974.836914
aKB	2.02	4.73	0.69	5.54	6.4	4.96	0	0	2.19	0	0	0.5
formyl-methionine	0	0	0	0	0	0	0	0	0	0	0	0
aHB	14.96	26.56	14.72	63.95	32.15	54.51	6.28	9.35	10.07	14.06	12.64	13.29
mannose	0	0	0	0	0	0	0	60.07	0	0	0	0
3-hydroxyisobutyric acid	14.88	26.44	23.7	33.37	0	0	9.48	11	18.63	30.87	0	16.62
FAD	0	0	0	0	0	0	0	0	0	0	0	0
uric acid	0	0	0	0	0	0	0	0	0	0	0	0
cysteic acid	0	0	0	0	0	0	0	0	0	0	0	0
carbamoylaspartate	0	0	0	0	0	0	0	0	25.73	450.15	0	0
uracil	32.11	41.88	34.85	48.7	34.74	67.53	157.19	233.78	212.45	266.56	377.59	111.76
fumarate	0	0	0	0	0	0.49	0	0	12.74	1151.99	0	0
DHAP	0	0	0	0	0	0	0	0	0	0	0	0
3-phosphoglycerate	0	0	0	0	0	0	0	0	0	0	0	0
cis-aconitate	3.1	2.49	2.81	3.58	2.39	2.11	4.49	3.56	2.91	5.65	3.97	2.58
thymidine	3.95	4.81	6.31	9.73	5.91	8.84	3.39	1.74	2.72	3.75	1.93	3.06
acetylglycine	1.52	1.95	2.82	1.5	1.92	4.41	1.26	1.15	1.87	0	0.75	0
ribose-5-phosphate	13.4	10.65	26.53	20.45	39.47	17.89	108.72	74	112.43	182.06	75.43	35.25
ATP	0	0	0	0	0	0	0	0	0	0	0	0
dCTP	0	0	0	0	0	0	0	0	0	0	0	0
UTP	0	0	0	0	0	0	0	0	0	0	0	0
CTP	0	0	0	0	0	0	0	0	0	0	0	0
GTP	0	0	0	0	0	0	0	0	0	0	0	0
p-aminobenzoate	0	0.44	0.03	2.01	0.63	0	0	0.32	0.61	1.07	0.18	1.26
glycerate	0.46	1.83	0.38	0	1.13	1.76	0.52	2.67	3.66	3.39	0	2.17
sorbitol	0	21.3	0	0	0	0	3.86	3.18	6.09	19.47	11.02	1.02
allantoin	8.88	11.28	35.44	58.54	61.28	34.65	14.85	18.3	25.24	48.01	18.14	50.24
myo-inositol	29.88	30.93	45.2	52.68	46.34	49.74	147.18	245.07	158.88	363.5	221.19	56.53
UDP-glucose	0	0	0	0	0	0	0	0	0	0	0	0
2-hydroxybutyrate	9.44	17.36	9.28	42.91	21.17	36.43	3.52	5.61	6.1	8.82	7.85	8.38
thiamine	0	0	0	0	0	0	0	0	0	0	0	0
xanthine	0	0	0	0	31.12	0	125.16	102.01	90.03	116.66	143.62	34.37
fructose	33.01	31.18	30.43	37.6	17.79	18.46	49.35	22.69	0	43.47	26.76	17.15
dTMP	0	0	0	0	0	0	0.2	0.76	0.46	0.22	1.99	0.01
o-phosphoethanolamine	0	0	0	0	0	0	0	0	0	0	0	0
glucose-1-phosphate	0	0	0	0	0	0	0	0	0	0	0	0
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name
3-methyl-2-oxopentanoic-acid-ketoisoleucine
3-methyl-2-oxobutyrate-keto-isovaleric-acid
pipecolate
orotate
2HG
UDP-GlcNAc
pyruvate
succinate
akg
malate
glucose-6-phosphate
citrate
uridine
AMP
a-aminobutyrate
argininosuccinate
GMP
lactate
glucose
serine
glutamate
aspartate
taurine
aKB
formyl-methionine
aHB
mannose
3-hydroxyisobutyric acid
FAD
uric acid
cysteic acid
carbamoylaspartate
uracil
fumarate
DHAP
3-phosphoglycerate
cis-aconitate
thymidine
acetylglycine
ribose-5-phosphate
ATP
dCTP
UTP
CTP
GTP
p-aminobenzoate
glycerate
sorbitol
allantoin
myo-inositol
UDP-glucose
2-hydroxybutyrate
thiamine
xanthine
fructose
dTMP
o-phosphoethanolamine
glucose-1-phosphate
METABOLITES_END
#END