#METABOLOMICS WORKBENCH juliehaines_20211122_122645 DATATRACK_ID:2940 STUDY_ID:ST001999 ANALYSIS_ID:AN003262 PROJECT_ID:000000
VERSION             	1
CREATED_ON             	November 22, 2021, 12:33 pm
#PROJECT
PR:PROJECT_TITLE                 	Polyamine import and accumulation causes immunomodulation in macrophages
PR:PROJECT_TITLE                 	engulfing apoptotic cells
PR:PROJECT_SUMMARY               	Phagocytosis of apoptotic cells, termed efferocytosis, is critical for tissue
PR:PROJECT_SUMMARY               	homeostasis and drives anti-inflammatory programming in engulfing macrophages.
PR:PROJECT_SUMMARY               	Here, we assess metabolites in naïve and inflammatory macrophages following
PR:PROJECT_SUMMARY               	engulfment of multiple cellular and non-cellular targets. Efferocytosis leads to
PR:PROJECT_SUMMARY               	unique increases in the arginine-derived polyamines, spermidine and spermine, in
PR:PROJECT_SUMMARY               	vitro and in vivo. Surprisingly, polyamine accumulation after efferocytosis does
PR:PROJECT_SUMMARY               	not arise from retention of apoptotic cell metabolites or de novo synthesis, but
PR:PROJECT_SUMMARY               	from enhanced polyamine import that is dependent on Rac1, actin, and PI3 kinase.
PR:PROJECT_SUMMARY               	Blocking polyamine import prevents efferocytosis from suppressing macrophage
PR:PROJECT_SUMMARY               	IL-1beta or IL-6. This identifies efferocytosis as a trigger for polyamine
PR:PROJECT_SUMMARY               	import and accumulation, and imported polyamines as mediators of
PR:PROJECT_SUMMARY               	efferocytosis-induced immune reprogramming.
PR:INSTITUTE                     	University of Colorado Denver
PR:LAST_NAME                     	Haines
PR:FIRST_NAME                    	Julie
PR:ADDRESS                       	12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
PR:EMAIL                         	julie.haines@cuanschutz.edu
PR:PHONE                         	3037243339
#STUDY
ST:STUDY_TITLE                   	Polyamine import and accumulation causes immunomodulation in macrophages
ST:STUDY_TITLE                   	engulfing apoptotic cells (Part 4)
ST:STUDY_SUMMARY                 	Phagocytosis of apoptotic cells, termed efferocytosis, is critical for tissue
ST:STUDY_SUMMARY                 	homeostasis and drives anti-inflammatory programming in engulfing macrophages.
ST:STUDY_SUMMARY                 	Here, we assess metabolites in naïve and inflammatory macrophages following
ST:STUDY_SUMMARY                 	engulfment of multiple cellular and non-cellular targets. Efferocytosis leads to
ST:STUDY_SUMMARY                 	unique increases in the arginine-derived polyamines, spermidine and spermine, in
ST:STUDY_SUMMARY                 	vitro and in vivo. Surprisingly, polyamine accumulation after efferocytosis does
ST:STUDY_SUMMARY                 	not arise from retention of apoptotic cell metabolites or de novo synthesis, but
ST:STUDY_SUMMARY                 	from enhanced polyamine import that is dependent on Rac1, actin, and PI3 kinase.
ST:STUDY_SUMMARY                 	Blocking polyamine import prevents efferocytosis from suppressing macrophage
ST:STUDY_SUMMARY                 	IL-1beta or IL-6. This identifies efferocytosis as a trigger for polyamine
ST:STUDY_SUMMARY                 	import and accumulation, and imported polyamines as mediators of
ST:STUDY_SUMMARY                 	efferocytosis-induced immune reprogramming.
ST:INSTITUTE                     	University of Colorado Denver
ST:LAST_NAME                     	Haines
ST:FIRST_NAME                    	Julie
ST:ADDRESS                       	12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
ST:EMAIL                         	julie.haines@cuanschutz.edu
ST:PHONE                         	3037243339
#SUBJECT
SU:SUBJECT_TYPE                  	Cultured cells
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           	-	ctrl 1	Category:unexposed macrophages	RAW_FILE_NAME=7+
SUBJECT_SAMPLE_FACTORS           	-	ctrl 2	Category:unexposed macrophages	RAW_FILE_NAME=8+
SUBJECT_SAMPLE_FACTORS           	-	ctrl 3	Category:unexposed macrophages	RAW_FILE_NAME=9+
SUBJECT_SAMPLE_FACTORS           	-	1 -AC 8h	Category:non-engulfing	RAW_FILE_NAME=10+
SUBJECT_SAMPLE_FACTORS           	-	2 -AC 8h	Category:non-engulfing	RAW_FILE_NAME=11+
SUBJECT_SAMPLE_FACTORS           	-	3 -AC 8h	Category:non-engulfing	RAW_FILE_NAME=12+
SUBJECT_SAMPLE_FACTORS           	-	1 +AC 8h	Category:engulfing	RAW_FILE_NAME=13+
SUBJECT_SAMPLE_FACTORS           	-	2 +AC 8h	Category:engulfing	RAW_FILE_NAME=14+
SUBJECT_SAMPLE_FACTORS           	-	3 +AC 8h	Category:engulfing	RAW_FILE_NAME=15+
#COLLECTION
CO:COLLECTION_SUMMARY            	LPS-primed murine peritoneal macrophages were fed fluorescently-tagged apoptotic
CO:COLLECTION_SUMMARY            	Jurkat cells for 1h. Unengulfed targets were washed off and macrophages were
CO:COLLECTION_SUMMARY            	left to degrade targets for a further 7 hours. Macrophages were then collected
CO:COLLECTION_SUMMARY            	and sorted into non-engulfing or engulfing populations, alongside control
CO:COLLECTION_SUMMARY            	unexposed macrophages. Sorted cells were pelleted at 400xg and dry pellets were
CO:COLLECTION_SUMMARY            	snap frozen and stored at -80C until metabolite extraction.
CO:SAMPLE_TYPE                   	Macrophages
#TREATMENT
TR:TREATMENT_SUMMARY             	LPS-primed murine peritoneal macrophages were fed fluorescently-tagged apoptotic
TR:TREATMENT_SUMMARY             	Jurkat cells for 1h. Unengulfed targets were washed off and macrophages were
TR:TREATMENT_SUMMARY             	left to degrade targets for a further 7 hours. Macrophages were then collected
TR:TREATMENT_SUMMARY             	and sorted into non-engulfing or engulfing populations, alongside control
TR:TREATMENT_SUMMARY             	unexposed macrophages. Sorted cells were pelleted at 400xg and dry pellets were
TR:TREATMENT_SUMMARY             	snap frozen and stored at -80C until metabolite extraction.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	To process cells for assessment of intracellular metabolites, cells were
SP:SAMPLEPREP_SUMMARY            	pelleted at 400xg in tubes coated with 0.06% BSA. Supernatant was aspirated and
SP:SAMPLEPREP_SUMMARY            	discarded; residual liquid was carefully wicked away from the pellet with a
SP:SAMPLEPREP_SUMMARY            	kimwipe. Dry pellets were immediately snap frozen and stored at -80C until
SP:SAMPLEPREP_SUMMARY            	processing. To process culture supernatants for assessment of metabolites,
SP:SAMPLEPREP_SUMMARY            	supernatant was centrifuged at 400xg to pellet any cells. Cell-free supernatant
SP:SAMPLEPREP_SUMMARY            	was then transferred to a fresh tube, snap frozen, and stored at -80C until
SP:SAMPLEPREP_SUMMARY            	processing. Ultra-high pressure liquid chromatography-mass spectrometry
SP:SAMPLEPREP_SUMMARY            	(UHPLC-MS) was performed by the University of Colorado School of Medicine
SP:SAMPLEPREP_SUMMARY            	Metabolomics Core. Metabolites from frozen cell pellets were extracted at 2e6
SP:SAMPLEPREP_SUMMARY            	cells/mL in ice cold 5:3:2 MeOH:acetonitrile:water (v/v/v). Media was thawed on
SP:SAMPLEPREP_SUMMARY            	ice and a 10 L aliquot treated with 240 L of the same extraction solution.
SP:SAMPLEPREP_SUMMARY            	Extractions were carried out using vigorous vortexing for 30 min at 4C.
SP:SAMPLEPREP_SUMMARY            	Supernatants were clarified by centrifugation (10 min, 18,000 g, 4C) and
SP:SAMPLEPREP_SUMMARY            	analyzed using a Thermo Vanquish UHPLC coupled to a Thermo Q Exactive mass
SP:SAMPLEPREP_SUMMARY            	spectrometer.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	Isocratic flow at 250 uL/min using 100% phase A (95% water, 5% acetonitrile, 10
CH:CHROMATOGRAPHY_SUMMARY        	mM ammonium acetate).
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Thermo Vanquish
CH:COLUMN_NAME                   	Phenomenex Kinetex C18 (150 x 2.1mm, 2.6 um)
CH:FLOW_GRADIENT                 	isocratic
CH:FLOW_RATE                     	250 ul/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                   	Global metabolomics analyses were performed using a 3 min isocratic run in
MS:MS_COMMENTS                   	positive and negative ion modes (separate runs) as described previously (Nemkov
MS:MS_COMMENTS                   	et al., 2015, Nemkov et al., 2017); stable isotope tracing samples were analyzed
MS:MS_COMMENTS                   	using a 5 min C18 gradient in positive and negative ion modes (separate runs) as
MS:MS_COMMENTS                   	described (Nemkov et al., 2019, Gehrke et al., 2019). For all analyses, the MS
MS:MS_COMMENTS                   	scanned in MS1 mode across the m/z range of 65 to 950. Peaks were annotated (in
MS:MS_COMMENTS                   	conjunction with the KEGG database), integrated, and quality control performed
MS:MS_COMMENTS                   	using Maven (Princeton University) as described. Stable isotope tracing results
MS:MS_COMMENTS                   	were isotopically corrected for the natural abundance of 13C1, 13C2, 15N1, and
MS:MS_COMMENTS                   	15N2 (Nemkov et al., 2017).
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	peak area
MS_METABOLITE_DATA_START
Samples	ctrl 1	ctrl 2	ctrl 3	1 -AC 8h	2 -AC 8h	3 -AC 8h	1 +AC 8h	2 +AC 8h	3 +AC 8h
Factors	Category:unexposed macrophages	Category:unexposed macrophages	Category:unexposed macrophages	Category:non-engulfing	Category:non-engulfing	Category:non-engulfing	Category:engulfing	Category:engulfing	Category:engulfing
ATP	2.87E+04	3.19E+04	2.75E+04	1.61E+03	2.19E+04	6.80E+03	9.91E+03	2.52E+04	6.89E+03
ADP	9.80E+04	6.13E+04	8.52E+04	4.22E+03	2.03E+04	2.61E+03	9.36E+03	7.41E+03	3.88E+04
Adenine	1.80E+05	8.48E+04	1.71E+05	1.10E+05	9.33E+04	8.06E+04	1.24E+05	8.44E+04	1.02E+05
GTP	5.56E+03	7.70E+03	6.64E+03	0.00E+00	8.25E+03	7.21E+03	0.00E+00	7.31E+03	0.00E+00
GDP	4.29E+04	7.67E+04	4.07E+04	1.87E+04	1.76E+04	2.28E+04	7.56E+04	3.93E+04	3.73E+04
Cytidine	0.00E+00	0.00E+00	0.00E+00	0.00E+00	3.19E+03	0.00E+00	1.66E+05	1.15E+05	1.16E+05
UTP	1.67E+04	2.36E+04	2.39E+04	6.39E+03	5.95E+03	1.61E+04	1.48E+04	8.86E+03	5.25E+03
UDP	5.29E+04	6.15E+04	4.66E+04	3.15E+04	2.86E+04	3.01E+04	1.10E+05	5.12E+04	4.54E+04
IDP	3.25E+06	3.03E+06	3.36E+06	4.27E+06	3.37E+06	3.89E+06	3.78E+06	3.64E+06	3.48E+06
Inosine	1.08E+04	8.48E+03	5.82E+03	5.35E+03	1.09E+03	2.00E+04	6.90E+04	4.25E+04	3.68E+04
Xanthine	7.51E+04	6.30E+04	4.03E+04	4.35E+04	3.33E+04	1.96E+04	2.74E+05	2.11E+05	2.12E+05
3',5'-Cyclic IMP	7.96E+06	7.61E+06	8.57E+06	1.45E+07	1.12E+07	1.29E+07	9.49E+06	9.41E+06	9.27E+06
UDP-glucose	1.31E+04	2.17E+04	5.00E+03	7.08E+03	5.70E+03	6.45E+03	3.32E+04	2.11E+04	9.16E+03
Diphosphate	2.07E+07	2.42E+07	2.01E+07	1.22E+07	1.27E+07	1.37E+07	2.23E+07	2.30E+07	1.71E+07
D-Glucose	1.22E+07	1.32E+07	1.51E+07	1.43E+07	1.51E+07	1.71E+07	1.46E+07	1.34E+07	1.69E+07
D-Glucose 6-phosphate	9.23E+04	1.20E+05	5.99E+04	9.64E+04	9.79E+04	5.29E+04	3.25E+05	2.42E+05	1.53E+05
D-Fructose 1-6-bisphosphate	7.07E+05	9.32E+05	5.15E+05	5.55E+05	4.26E+05	3.61E+05	1.73E+06	9.75E+05	6.58E+05
D-Glyceraldehyde 3-phosphate/Glycerone phosphate	3.78E+05	4.75E+05	2.41E+05	3.61E+05	3.05E+05	2.57E+05	9.79E+05	6.60E+05	4.44E+05
2/3-Phospho-D-glycerate	3.12E+05	4.73E+05	2.78E+05	3.76E+05	3.92E+05	3.23E+05	1.01E+06	6.07E+05	4.33E+05
Pyruvate	1.25E+06	1.19E+06	1.51E+06	1.40E+06	1.48E+06	1.49E+06	1.39E+06	1.07E+06	1.45E+06
Lactate	4.72E+06	5.27E+06	6.10E+06	5.52E+06	5.47E+06	6.59E+06	5.88E+06	4.82E+06	6.48E+06
Maltose	1.51E+04	4.68E+03	1.95E+04	2.23E+04	0.00E+00	1.99E+04	2.56E+04	6.23E+03	1.27E+04
Mannitol	1.46E+04	2.94E+04	3.75E+05	9.22E+04	3.69E+04	6.06E+04	1.93E+05	2.54E+04	6.76E+04
D-Ribose	3.31E+05	3.37E+05	4.08E+05	3.91E+05	4.18E+05	4.48E+05	4.06E+05	3.35E+05	4.59E+05
Citrate	1.12E+06	1.54E+06	1.12E+06	1.42E+06	9.16E+05	1.06E+06	1.95E+06	1.56E+06	1.05E+06
2-Oxoglutarate	2.49E+04	3.43E+04	2.66E+04	4.45E+04	3.59E+04	3.51E+04	7.59E+04	3.79E+04	4.22E+04
Succinate	3.17E+05	2.81E+05	3.27E+05	4.08E+05	4.07E+05	4.05E+05	6.89E+05	5.08E+05	3.75E+05
Fumarate	2.72E+04	6.09E+04	5.15E+04	8.88E+04	6.18E+04	5.84E+04	1.30E+05	9.16E+04	8.41E+04
Malate	5.18E+05	6.42E+05	5.19E+05	5.43E+05	3.96E+05	4.27E+05	1.59E+06	1.01E+06	8.23E+05
Itaconate	1.07E+06	1.77E+06	7.65E+05	8.12E+05	6.82E+05	7.66E+05	4.24E+06	3.23E+06	1.97E+06
5-Oxoproline	2.68E+05	2.27E+05	2.35E+05	1.40E+05	1.45E+05	1.50E+05	1.97E+05	1.21E+05	1.92E+05
Ascorbate	1.86E+05	2.03E+05	2.11E+05	4.19E+05	3.04E+05	3.65E+05	2.92E+05	2.57E+05	2.66E+05
UDP-N-acetyl-D-glucosamine	2.83E+04	3.01E+04	9.78E+03	1.31E+04	1.95E+04	1.53E+04	3.49E+04	3.71E+04	2.35E+04
Phosphocreatine	2.72E+04	2.28E+04	2.11E+04	9.18E+03	2.21E+04	2.28E+04	2.59E+04	2.72E+04	2.02E+04
3D-(3-5/4)-Trihydroxycyclohexane-1-2-dione	3.41E+05	3.64E+05	4.42E+05	6.06E+05	5.13E+05	5.48E+05	4.86E+05	4.42E+05	4.90E+05
Glycerol 3-phosphate	3.88E+04	5.49E+04	2.92E+04	5.04E+04	4.42E+04	3.86E+04	9.72E+04	5.40E+04	3.46E+04
Ethanolamine phosphate	5.92E+05	4.48E+05	3.21E+05	1.47E+05	1.88E+05	9.38E+04	3.94E+05	2.61E+05	2.86E+05
Hexanoic acid (caproate)	7.32E+05	6.19E+05	7.41E+05	7.74E+05	7.49E+05	6.39E+05	6.21E+05	6.46E+05	6.74E+05
Heptanoic acid	6.85E+05	4.25E+05	7.00E+05	8.09E+05	7.11E+05	6.99E+05	6.91E+05	5.40E+05	7.24E+05
Octanoic acid (caprylic acid)	1.62E+07	1.12E+07	1.96E+07	6.28E+07	3.16E+07	2.93E+07	3.30E+07	2.10E+07	2.78E+07
Nonanoic acid (pelargonic acid)	9.50E+05	4.18E+05	1.08E+06	2.33E+06	1.01E+06	8.72E+05	1.01E+06	6.53E+05	1.20E+06
Decanoic acid (capric acid)	1.04E+05	4.91E+04	8.84E+04	1.44E+05	9.93E+04	9.37E+04	1.05E+05	7.13E+04	9.48E+04
Dodecanoic acid (lauric acid)	8.20E+04	5.76E+04	9.92E+04	1.31E+05	9.21E+04	8.86E+04	8.96E+04	9.22E+04	7.72E+04
Tetradecanoic acid (myristic acid)	2.23E+05	1.29E+05	2.42E+05	3.31E+05	1.95E+05	3.42E+05	1.97E+05	2.19E+05	1.93E+05
Hexadecanoic acid (palmitic acid)	5.96E+06	2.65E+06	9.34E+06	8.66E+06	6.55E+06	9.34E+06	3.27E+06	6.65E+06	5.85E+06
Octadecanoic acid (stearic acid)	3.95E+06	1.89E+06	6.52E+06	6.18E+06	4.85E+06	6.42E+06	2.69E+06	4.85E+06	4.87E+06
Tetradecenoic acid (myristoleic acid)	1.01E+04	4.19E+03	7.40E+03	2.10E+04	9.89E+03	2.20E+04	9.92E+03	1.48E+04	9.06E+03
Hexadecenoic acid (Palmitoleic acid)	7.64E+04	2.95E+04	5.05E+04	1.07E+05	4.15E+04	1.94E+05	7.44E+04	1.08E+05	5.04E+04
Octadecenoic acid (Oleic acid)	4.23E+05	2.83E+05	6.61E+05	8.76E+05	5.53E+05	8.09E+05	4.45E+05	6.55E+05	4.91E+05
Linoleic acid ((9Z,12Z)-Octadecadienoic acid)	2.14E+05	2.05E+05	2.42E+05	7.33E+05	4.10E+05	3.76E+05	2.94E+05	3.91E+05	2.86E+05
a-Linolenic acid (Octadecatrienoic acid)	1.74E+04	1.89E+04	1.89E+04	7.21E+04	3.89E+04	4.37E+04	4.70E+04	3.34E+04	2.72E+04
Arachidonic acid (Eicosatetraenoic acid)	5.94E+04	5.92E+04	5.46E+04	9.92E+04	6.07E+04	6.27E+04	1.06E+05	8.22E+04	5.35E+04
Adrenic acid (7,10,13,16-Docosatetraenoic acid)	3.14E+04	2.99E+04	2.97E+04	4.87E+04	3.13E+04	3.11E+04	3.82E+04	3.05E+04	2.41E+04
Dihomo-gamma-linolenic acid ((8Z-11Z-14Z)-Icosatrienoic acid)	2.23E+04	2.17E+04	2.74E+04	4.93E+04	2.91E+04	3.63E+04	4.34E+04	4.50E+04	2.63E+04
Docosapentaenoic acid ((7Z-10Z-13Z-16Z-19Z)-Docosa-7-10-13-16-19-pentaenoic acid)	1.44E+04	2.30E+04	2.06E+04	3.36E+04	2.63E+04	2.84E+04	4.18E+04	3.06E+04	2.58E+04
Cholate	2.12E+04	3.05E+04	3.52E+04	1.22E+05	7.16E+04	7.32E+04	3.26E+04	5.74E+04	5.03E+04
Glycochenodeoxycholate	5.10E+04	7.02E+04	6.53E+04	1.88E+05	1.86E+05	1.37E+05	1.04E+05	8.51E+04	7.46E+04
Glycocholate	1.34E+04	2.38E+04	2.86E+04	8.13E+04	7.68E+04	7.19E+04	5.97E+04	4.57E+04	3.33E+04
Taurodeoxycholate	2.27E+04	2.63E+04	2.73E+04	9.03E+04	8.06E+04	5.74E+04	3.95E+04	4.03E+04	3.75E+04
Taurocholate	6.94E+03	8.68E+03	8.75E+03	2.86E+04	2.99E+04	2.76E+04	1.52E+04	1.63E+04	9.40E+03
Prostaglandin D2/Thromboxane A2 (isobars)	1.91E+04	1.04E+04	5.21E+04	1.94E+04	6.43E+04	2.34E+04	1.38E+04	7.28E+04	3.03E+04
Prostaglandin D3 (isobars)	4.62E+03	6.30E+03	5.78E+03	4.83E+03	5.96E+03	1.51E+04	3.92E+03	9.39E+03	6.99E+03
Prostaglandin G2	1.30E+04	6.05E+03	2.31E+04	1.41E+04	2.16E+04	1.20E+04	1.35E+04	1.88E+04	1.24E+04
Cholesterol sulfate	1.58E+04	7.96E+03	1.27E+04	1.37E+04	1.00E+04	1.43E+04	1.77E+04	1.07E+04	1.76E+04
Pentobarbital	3.43E+03	4.81E+03	6.18E+04	5.03E+04	2.09E+04	2.76E+04	1.39E+04	1.26E+04	5.86E+03
2-Methyleneglutarate	1.51E+06	1.66E+06	1.95E+06	1.74E+06	1.82E+06	1.98E+06	1.67E+06	1.46E+06	1.94E+06
6-Pyruvoyltetrahydropterin	3.04E+04	2.24E+04	3.80E+04	6.64E+04	3.94E+04	5.29E+04	5.07E+04	2.85E+04	2.91E+04
D-glucono-1,5-lactone	9.08E+05	9.10E+05	1.16E+06	1.34E+06	1.25E+06	1.31E+06	1.20E+06	9.84E+05	1.19E+06
Catechol	5.26E+05	4.24E+05	4.73E+05	4.84E+05	3.82E+05	4.11E+05	5.53E+05	5.63E+05	4.96E+05
Catechin	7.46E+05	5.29E+05	8.27E+05	1.23E+06	1.10E+06	1.13E+06	2.25E+05	1.17E+06	1.06E+06
D-Ribitol 5-phosphate	6.11E+04	4.67E+04	4.40E+04	1.17E+05	7.35E+04	1.07E+05	9.17E+04	8.89E+04	6.80E+04
Glucosinolate	1.51E+05	3.66E+05	1.43E+05	1.13E+05	1.72E+05	2.14E+05	2.05E+05	2.58E+05	1.19E+05
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	KEGG ID	m/z	r.t.
ATP	C00002	505.9875	0.59
ADP	C00008	426.0217	0.58
Adenine	C00147	134.0460	0.67
GTP	C00044	521.9828	0.58
GDP	C00035	442.0164	0.58
Cytidine	C00475	242.0782	0.67
UTP	C00075	482.9605	0.55
UDP	C00015	402.9943	0.56
IDP	C00104	427.0068	0.56
Inosine	C00294	267.0724	0.65
Xanthine	C00385	151.0250	0.68
3',5'-Cyclic IMP	C00943	329.0299	0.55
UDP-glucose	C00029	565.0470	0.55
Diphosphate	C00013	176.9348	0.61
D-Glucose	C00031	179.0552	0.64
D-Glucose 6-phosphate	C02965	259.0217	0.57
D-Fructose 1-6-bisphosphate	C00354	338.9883	0.53
D-Glyceraldehyde 3-phosphate/Glycerone phosphate	C00118	168.9895	0.56
2/3-Phospho-D-glycerate	C00631	184.9846	0.56
Pyruvate	C00022	87.0072	0.65
Lactate	C01432	89.0228	0.64
Maltose	C00208	341.1086	0.65
Mannitol	C00392	181.0708	0.65
D-Ribose	C00121	149.0446	0.64
Citrate	C00158	191.0186	0.59
2-Oxoglutarate	C00026	145.0131	0.58
Succinate	C00042	117.0178	0.62
Fumarate	C00122	115.0032	0.61
Malate	C00149	133.0130	0.58
Itaconate	C00490	129.0180	0.60
5-Oxoproline	C01879	128.0339	0.62
Ascorbate	C00072	175.0240	0.65
UDP-N-acetyl-D-glucosamine	C00043	606.0737	0.56
Phosphocreatine	C02305	210.0277	0.56
3D-(3-5/4)-Trihydroxycyclohexane-1-2-dione	C04287	159.0288	0.65
Glycerol 3-phosphate	C00093	171.0052	0.59
Ethanolamine phosphate	C00346	140.0105	0.62
Hexanoic acid (caproate)	C01585	115.0750	1.73
Heptanoic acid	C17714	129.0907	1.86
Octanoic acid (caprylic acid)	C06423	143.1064	1.96
Nonanoic acid (pelargonic acid)	C01601	157.1221	2.08
Decanoic acid (capric acid)	C01571	171.1379	2.16
Dodecanoic acid (lauric acid)	C02679	199.1693	2.33
Tetradecanoic acid (myristic acid)	C06424	227.2008	2.56
Hexadecanoic acid (palmitic acid)	C00249	255.2324	2.93
Octadecanoic acid (stearic acid)	C01530	283.2638	3.46
Tetradecenoic acid (myristoleic acid)	C08322	225.1852	2.36
Hexadecenoic acid (Palmitoleic acid)	C08362	253.2168	2.62
Octadecenoic acid (Oleic acid)	C00712	281.2481	2.97
Linoleic acid ((9Z,12Z)-Octadecadienoic acid)	C01595	279.2325	2.65
a-Linolenic acid (Octadecatrienoic acid)	C06427	277.2168	2.45
Arachidonic acid (Eicosatetraenoic acid)	C00219	303.2325	2.55
Adrenic acid (7,10,13,16-Docosatetraenoic acid)	C16527	331.2639	2.82
Dihomo-gamma-linolenic acid ((8Z-11Z-14Z)-Icosatrienoic acid)	C03242	305.2482	2.73
Docosapentaenoic acid ((7Z-10Z-13Z-16Z-19Z)-Docosa-7-10-13-16-19-pentaenoic acid)	C16513	329.2482	2.58
Cholate	C00695	407.2815	1.92
Glycochenodeoxycholate	C05466	448.3064	1.77
Glycocholate	C01921	464.3009	1.74
Taurodeoxycholate	C05463	498.2890	1.77
Taurocholate	C05122	514.2836	1.74
Prostaglandin D2/Thromboxane A2 (isobars)	C00696	351.2201	1.96
Prostaglandin D3 (isobars)	C13802	349.2040	1.92
Prostaglandin G2	C05956	367.2140	1.93
Cholesterol sulfate	C18043	465.3054	2.01
Pentobarbital	C07422	225.1236	1.94
2-Methyleneglutarate	C02930	143.0336	0.65
6-Pyruvoyltetrahydropterin	C03684	236.0781	1.74
D-glucono-1,5-lactone	C00198	177.0396	0.65
Catechol	C00090	109.0281	1.86
Catechin	C17590	289.0675	0.56
D-Ribitol 5-phosphate	C01068	231.0273	0.64
Glucosinolate	C17260	316.9843	0.71
METABOLITES_END
#END