#METABOLOMICS WORKBENCH amroilaiwy_20160528_113623_mwtab.txt DATATRACK_ID:640 STUDY_ID:ST000431 ANALYSIS_ID:AN000681 PROJECT_ID:PR000334
VERSION             	1
CREATED_ON             	July 29, 2016, 11:14 am
#PROJECT
PR:PROJECT_TITLE                 	C2C12 stretch cessation models muscle atrophy and anaplerotic changes in
PR:PROJECT_TITLE                 	metabolism
PR:PROJECT_TYPE                  	GC-MS non targeted qualitative analysis
PR:PROJECT_SUMMARY               	Studies of skeletal muscle disuse either in patients on bed rest or
PR:PROJECT_SUMMARY               	experimentally in animals(immobilization) have demonstrated that decreased
PR:PROJECT_SUMMARY               	protein synthesis is common, with transient parallel increases in protein
PR:PROJECT_SUMMARY               	degradation. Muscle disuse atrophy involves a process of transition from slow to
PR:PROJECT_SUMMARY               	fast myosin fiber types 6 . A shift toward glycolysis, decreased capacity for
PR:PROJECT_SUMMARY               	fat oxidation, and substrate accumulation in atrophied muscles have been
PR:PROJECT_SUMMARY               	reported as has accommodation of the liver with an increased gluconeogenic
PR:PROJECT_SUMMARY               	capacity. Recent studies have modeled skeletal muscle disuse by using cyclic
PR:PROJECT_SUMMARY               	stretch of differentiated myotubes (C2C12), which mimics the loading pattern of
PR:PROJECT_SUMMARY               	mature skeletal muscle, followed by cessation of stretch.We utilized this model
PR:PROJECT_SUMMARY               	to determine the metabolic changes using non-targeted metabolomics analysis of
PR:PROJECT_SUMMARY               	the media. We identified increases in amino acids resulting from protein
PR:PROJECT_SUMMARY               	degradation (largely sarcomere) that occurs with muscle atrophy that are
PR:PROJECT_SUMMARY               	involved in feeding the Kreb’s cycle through anaplerosis. Specifically, we
PR:PROJECT_SUMMARY               	identified increased alanine/proline metabolism (significantly elevated proline,
PR:PROJECT_SUMMARY               	alanine, glutamine, and asparagine) and increased -ketoglutaric acid, the
PR:PROJECT_SUMMARY               	proposed Kreb’s cycle intermediate being fed by the alanine/proline metabolic
PR:PROJECT_SUMMARY               	anaplerotic mechanism. Additionally, several unique pathways not clearly
PR:PROJECT_SUMMARY               	delineated in previous studies of muscle unloading were seen, including: 1)
PR:PROJECT_SUMMARY               	elevated ethanolamine and elevated keto-acids (e.g. 2-ketoleucine and
PR:PROJECT_SUMMARY               	2-keovaline) represent intermediates in the Ehlrich amino acid degradation
PR:PROJECT_SUMMARY               	pathway, which feeds into a metabolic pathway supplying acetyl-CoA and
PR:PROJECT_SUMMARY               	2-hydroxybutyrate (also significantly increased); and 2) elevated guanine, an
PR:PROJECT_SUMMARY               	intermediate of purine metabolism, was seen at 12 hours unloading. Given the
PR:PROJECT_SUMMARY               	interest in targeting different aspects of the ubiquitin proteasome system to
PR:PROJECT_SUMMARY               	inhibit protein degradation, this C2C12 system may allow the identification of
PR:PROJECT_SUMMARY               	direct and indirect alterations in metabolism due to anaplerosis or through
PR:PROJECT_SUMMARY               	other yet to be identified mechanisms using a non-targeted metabolomics
PR:PROJECT_SUMMARY               	approach.
PR:INSTITUTE                     	University of North Carolina
PR:DEPARTMENT                    	McAllister heart Institute, Department of Internal medicine
PR:LABORATORY                    	Multiple Centers
PR:LAST_NAME                     	Ilaiwy, Willis
PR:FIRST_NAME                    	Amro, Monte
PR:ADDRESS                       	111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA
PR:EMAIL                         	amroilaiwy@gmail.com, monte_willis@med.unc.edu
PR:PHONE                         	210-596-0171
PR:FUNDING_SOURCE                	NIH, Fondation Leducq
#STUDY
ST:STUDY_TITLE                   	C2C12 stretch cessation models muscle atrophy and anaplerotic changes in
ST:STUDY_TITLE                   	metabolism
ST:STUDY_TYPE                    	GC-MS non-targeted metabolomic profiling
ST:STUDY_SUMMARY                 	Studies of skeletal muscle disuse either in patients on bed rest or
ST:STUDY_SUMMARY                 	experimentally in animals(immobilization) have demonstrated that decreased
ST:STUDY_SUMMARY                 	protein synthesis is common, with transient parallel increases in protein
ST:STUDY_SUMMARY                 	degradation. Muscle disuse atrophy involves a process of transition from slow to
ST:STUDY_SUMMARY                 	fast myosin fiber types 6 . A shift toward glycolysis, decreased capacity for
ST:STUDY_SUMMARY                 	fat oxidation, and substrate accumulation in atrophied muscles have been
ST:STUDY_SUMMARY                 	reported as has accommodation of the liver with an increased gluconeogenic
ST:STUDY_SUMMARY                 	capacity. Recent studies have modeled skeletal muscle disuse by using cyclic
ST:STUDY_SUMMARY                 	stretch of differentiated myotubes (C2C12), which mimics the loading pattern of
ST:STUDY_SUMMARY                 	mature skeletal muscle, followed by cessation of stretch.We utilized this model
ST:STUDY_SUMMARY                 	to determine the metabolic changes using non-targeted metabolomics analysis of
ST:STUDY_SUMMARY                 	the media. We identified increases in amino acids resulting from protein
ST:STUDY_SUMMARY                 	degradation (largely sarcomere) that occurs with muscle atrophy that are
ST:STUDY_SUMMARY                 	involved in feeding the Kreb’s cycle through anaplerosis. Specifically, we
ST:STUDY_SUMMARY                 	identified increased alanine/proline metabolism (significantly elevated proline,
ST:STUDY_SUMMARY                 	alanine, glutamine, and asparagine) and increased -ketoglutaric acid, the
ST:STUDY_SUMMARY                 	proposed Kreb’s cycle intermediate being fed by the alanine/proline metabolic
ST:STUDY_SUMMARY                 	anaplerotic mechanism. Additionally, several unique pathways not clearly
ST:STUDY_SUMMARY                 	delineated in previous studies of muscle unloading were seen, including: 1)
ST:STUDY_SUMMARY                 	elevated ethanolamine and elevated keto-acids (e.g. 2-ketoleucine and
ST:STUDY_SUMMARY                 	2-keovaline) represent intermediates in the Ehlrich amino acid degradation
ST:STUDY_SUMMARY                 	pathway, which feeds into a metabolic pathway supplying acetyl-CoA and
ST:STUDY_SUMMARY                 	2-hydroxybutyrate (also significantly increased); and 2) elevated guanine, an
ST:STUDY_SUMMARY                 	intermediate of purine metabolism, was seen at 12 hours unloading. Given the
ST:STUDY_SUMMARY                 	interest in targeting different aspects of the ubiquitin proteasome system to
ST:STUDY_SUMMARY                 	inhibit protein degradation, this C2C12 system may allow the identification of
ST:STUDY_SUMMARY                 	direct and indirect alterations in metabolism due to anaplerosis or through
ST:STUDY_SUMMARY                 	other yet to be identified mechanisms using a non-targeted metabolomics
ST:STUDY_SUMMARY                 	approach.
ST:INSTITUTE                     	University of North Carolina
ST:DEPARTMENT                    	McAllister Heart Institute
ST:LABORATORY                    	Mutliple Centers
ST:LAST_NAME                     	Ilaiwy, WIllis
ST:FIRST_NAME                    	Amro, Monte
ST:ADDRESS                       	111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA
ST:EMAIL                         	amroilaiwy@gmail.com, monte_willis@med.unc.edu
ST:PHONE                         	919-3607599
#SUBJECT
SU:SUBJECT_TYPE                  	Animal
SU:SUBJECT_SPECIES               	Mus musculus
SU:TAXONOMY_ID                   	10090
#SUBJECT_SAMPLE_FACTORS:         	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data
SUBJECT_SAMPLE_FACTORS           	-	p0 #1	Group:6 hour stretch	
SUBJECT_SAMPLE_FACTORS           	-	p0 #2	Group:6 hour stretch	
SUBJECT_SAMPLE_FACTORS           	-	p0 #3	Group:6 hour stretch	
SUBJECT_SAMPLE_FACTORS           	-	p1 #1	Group:1 hour reversal	
SUBJECT_SAMPLE_FACTORS           	-	p1 #2	Group:1 hour reversal	
SUBJECT_SAMPLE_FACTORS           	-	p1 #3	Group:1 hour reversal	
SUBJECT_SAMPLE_FACTORS           	-	p3 #1	Group:3 hour reversal	
SUBJECT_SAMPLE_FACTORS           	-	p3 #2	Group:3 hour reversal	
SUBJECT_SAMPLE_FACTORS           	-	p3 #3	Group:3 hour reversal	
SUBJECT_SAMPLE_FACTORS           	-	p6 #1	Group:6 hour reversal	
SUBJECT_SAMPLE_FACTORS           	-	p6 #2	Group:6 hour reversal	
SUBJECT_SAMPLE_FACTORS           	-	p6 #3	Group:6 hour reversal	
SUBJECT_SAMPLE_FACTORS           	-	p12 #1	Group:12 hour reversal	
SUBJECT_SAMPLE_FACTORS           	-	p12 #2	Group:12 hour reversal	
SUBJECT_SAMPLE_FACTORS           	-	p12 #3	Group:12 hour reversal	
SUBJECT_SAMPLE_FACTORS           	-	p0 no stretch #1	Group:no stretch-serum free	
SUBJECT_SAMPLE_FACTORS           	-	p1 no stretch #1	Group:no stretch-serum free	
SUBJECT_SAMPLE_FACTORS           	-	p3 no stretch #1	Group:no stretch-serum free	
SUBJECT_SAMPLE_FACTORS           	-	p6 no stretch #1	Group:no stretch-serum free	
SUBJECT_SAMPLE_FACTORS           	-	serum 6hr no stretch #1	Group:serum-no stretch	
SUBJECT_SAMPLE_FACTORS           	-	serum 6hr no stretch #2	Group:serum-no stretch	
SUBJECT_SAMPLE_FACTORS           	-	serum 6hr no stretch #3	Group:serum-no stretch	
#COLLECTION
CO:COLLECTION_SUMMARY            	Media was collected from all different 6 wells, and stored in -80 C
#TREATMENT
TR:TREATMENT_SUMMARY             	N/A
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	N/A
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	GC/MS methods follow previous studies using a 6890 N GC connected to a 5975
CH:CHROMATOGRAPHY_SUMMARY        	Inert single quadrupole MS (Agilent Technologies, Santa Clara, CA) (Bonikos et
CH:CHROMATOGRAPHY_SUMMARY        	al. 1975; Fiehn 2008; Kind et al. 2009). The two wall-coated, open-tubular GC
CH:CHROMATOGRAPHY_SUMMARY        	columns connected in series are both from J&W/Agilent (part 122–5512), DB5-MS,
CH:CHROMATOGRAPHY_SUMMARY        	15 meters in length, 0.25 mm in diameter, with an 0.25-l m luminal film.
CH:CHROMATOGRAPHY_SUMMARY        	Positive ions generated with conventional electron-ionization at 70 eV are
CH:CHROMATOGRAPHY_SUMMARY        	scanned broadly from 600 to 50 m/z in the detector throughout the 45 min cycle
CH:CHROMATOGRAPHY_SUMMARY        	time.
CH:CHROMATOGRAPHY_TYPE           	GC
CH:INSTRUMENT_NAME               	Agilent 6890N
CH:COLUMN_NAME                   	Agilent DB5-MS (15m × 0.25mm, 0.25um)
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:MS_COMMENTS                   	-
MS:INSTRUMENT_NAME               	Agilent 5975
MS:INSTRUMENT_TYPE               	Single quadrupole
MS:MS_TYPE                       	EI
MS:ION_MODE                      	POSITIVE
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS         	Peak values (Log transformed)
MS_METABOLITE_DATA_START
Samples	p0 #1	p0 #2	p0 #3	p1 #1	p1 #2	p1 #3	p3 #1	p3 #2	p3 #3	p6 #1	p6 #2	p6 #3	p12 #1	p12 #2	p12 #3	p0 no stretch #1	p1 no stretch #1	p3 no stretch #1	p6 no stretch #1	serum 6hr no stretch #1	serum 6hr no stretch #2	serum 6hr no stretch #3
Factors	Group:6 hour stretch	Group:6 hour stretch	Group:6 hour stretch	Group:1 hour reversal	Group:1 hour reversal	Group:1 hour reversal	Group:3 hour reversal	Group:3 hour reversal	Group:3 hour reversal	Group:6 hour reversal	Group:6 hour reversal	Group:6 hour reversal	Group:12 hour reversal	Group:12 hour reversal	Group:12 hour reversal	Group:no stretch-serum free	Group:no stretch-serum free	Group:no stretch-serum free	Group:no stretch-serum free	Group:serum-no stretch	Group:serum-no stretch	Group:serum-no stretch
Glyoxylic acid	13.08	12.66	13.58		13.08	12.94		13.08	13.34	12.85										12.8	12.96	13
C10 hydrocarbon	13.29	13.57	14.07	14.87	13.76	14.93	15.02	14.7	14.88	15.27	13.54	14.63	14.66	14.87	14.84	14.65	14.82	14.87	15.29	15.04	14.73	13.38
epsilon-Caprolactam		12.61	13.15	12.46	11.74	14.34	12.04	13.88	13.41	12.73	13.09		14.41	13.84	14.46	14.31	13.46	14.07	11.24	14.7	14.37	13.09
2-Hydroxypyridine	12.14				12.27		11.48			14.04		11.86			14.38				11.37		12.73	
Pyruvic acid	22.39	22.32	22.99	22.5	22.36	22.7	22.44	22.41	22.54	22.28	22.43	22.57	21.9	21.93	21.81	22.35	22.8	22.22	21.68	21.21	21.34	21.46
Lactic acid	26.03	25.81	26.19	25.84	26.15	26.25	26.35	26.26	26.23	26.48	26.87	26.63	26.84	27.11	26.93	25.98	26.24	26.48	26.41	27.24	27.7	27.63
Glycolic acid	14.16			13.99	14.12		14.15	14.18	14.1	13.96	14.04	14.22	15.27	15.28	14.86	13.72		11.92		15.42	15.78	15.33
Alanine	21.96	21.91	22.43	22.16	22.23	22.57	22.46	22.4	22.42	22.71	23.01	23.05	23.13	23.27	23.19	22.29	22.49	22.65	22.68	22.76	23.3	23.11
C11 hydrocarbon	15.31	15.69	15.41	16.22	15.46	15.83	14.93	15.65	13.58	13.84	15.41	15.6	13.73	15.8	13.75	13.93	13.71	13.83	16.02	15.9	15.61	15.19
2-Hydroxy-2-methylbutyric acid		15.12	15.9		15.63	15.1	15.25				15.48		14.75	15.81				15.36	15.3			
2-Ketovaline	16.61	16.27	16.96	16.27	16.19	17.11	16.83	17.35	17.02	17.2	17.52	17.65	18.01	18.31	18.21	16.91	17.28	17.42	17.42	17.88	18.5	18.4
2-Hydroxybutyric acid	23.17	23.19	23.82	23.71	23.61	23.87	23.36	23.33	23.89	23.29	23.69	23.86	23.23	23.3	23.57	23.4	23.6	23.4	22.99	23.18	24.12	23.59
Sulfuric acid					16.57		17.4	12.47	13.85		12.55	14.6		15.39				17	16.18			
Dodecamethylpentasiloxane	17.14	16.68	16.83	16.29	16.84	16.92	17.54	16.64	17.05	17.39	17.71	17.71	17.61	17.66	17.55	15.93	16.35	15.87	15.89	17.43	17.92	18.07
Malonic acid	17.24	17.75	17.17	16.45	17.88	16.97	17.85	16.92	17.27	15.99	17.1	17.11	16.5	17.47	16.94	16.45	16.52	17.27	17.07	17.44	17.23	16.58
2-Ketoleucine	19.5	19.23	20.09	19.51	18.94	20.05	19.95	19.84	19.99	20.11	20.36	20.49	20.69	20.94	20.77	19.76	20.03	20.04	20.14	20.28	21	21
Valine	23.48	23.26	24.58	24.05	23.53	24.71	23.28	23.96	24.43	24.08	24.06	24.47	24.56	23.94	24.82	24.42	24.77	24.06	24.3	24.21	24.84	24.75
C12 hydrocarbon	13.57	14.66	13.98	14.57	13.82	14.82	14.93	14.2	14.51	14.42	14.24	14.44	14.18	14.36	13.85	14.36	14.36	14.32	14.58	14.49	15.16	14.64
Benzoic acid	16.69	16.67	16.76	16.84	16.66	16.8	16.86	16.83	17.05	17.03	16.68	16.76	16.83	16.84	16.84	16.64	16.67	16.69	16.33	17.06	17.23	16.91
Ethanolamine	16.38	16.37	16.97	16.69	16.63	16.82	17.05	16.77	17.03	16.9	16.87	17.1	17.7	17.65	17.57	16.97	17.37	17.22	17.5	17.36	17.84	17.68
Phosphoric acid	17.65	17.45		17.46	16.87	16.85	18.11	17.35	17.34	17.2	18.41	17.11	17.68	17.34	17.02	16.97	17.12	16.53	17.99	17.97	17.67	17.59
1,3-Dihydroxyacetone	23.24	23.35			23.62		23.72	23.42	23.9	23.75	23.63		24	23.82				23.71				
Leucine	24.08	24.25	24.84	24.13	24.42	25	24.53	24.04	24.69	24.5	24.49	24.69	24.88	24.64	25.03	24.59	25	24.31	24.54	24.46	24.85	24.91
Glycerol	23.24	23.35	21.41	19.76	23.62	25	21.28	23.42	23.9	23.75	20.72	20.6	21.5	22.04	22.78	24.59	20.63	23.71	20.44	20.82	20.37	22.56
Ethylmalonic acid	23.24	23.35			23.62	22.48	23.72	23.42	23.9	23.75	23.63		24	23.82	22.78			23.71				22.56
C13 hydrocarbon	16.42	16.85	16.77	17.18	16.57	17	17.08	16.78	16.97	17.16	16.65	16.67	17.06	16.89	16.78	16.59	16.75	16.83	17.09	17.03	17.02	16.28
Isoleucine	24.28	24.64	24.97	24.35	24.55	25.05	24.86	24.47	24.76	24.35	24.75	24.78	24.96	24.92	24.97	24.63	25.09	24.75	24.68	24.41	24.91	24.98
Proline	16.21	16.91	17.72	17.12	17.22	18.01	17.81	17.55	17.95	18	18.45	18.62	19.29	19.29	19.54	17.83	18.2	18.6	18.36	19.61	20.37	20.34
Glycine	23.6	23.7	24.03	23.72	23.75	24.31	23.98	23.87	24.08	23.7	24.14	24.24	24.39	24.29	24.53	24.03	24.4	24.14	23.95	23.98	24.66	24.65
Succinic acid	16.71	17.08			17.48		17.92	17.11		17.47	17.57		17.59	17.87	17.37	17.39	17.74	16.79		17.64	18.37	18.24
Glyceric acid																				13.71	16.25	15.27
MSTFA artifact	13.04	14.54	15.91		15.17	15.86	14.2	15.31	16.15	15.58	15.36	15.42	14.45	15.05	17.39	15.93	15.97	15.58	15.45	12.87	15.92	15.93
Fumaric acid	11.28	11.84	12.48	12.02	12.11	12.35	13.1	12.33	12.5	13.18	13.22	11.41	13.53	13.92	13.84	12.01	13.85	12.57	12.51	14.2	15.32	14.88
Serine	23.26	23.43	23.76	23.22	23.54	23.73	23.64	23.42	23.63	22.76	23.64	23.54	23.72	23.64	23.65	23.55	24.01	23.59	23.31	22.14	22.88	22.78
Threonine	24.83	24.9	25.34	24.83	25.06	25.2	25.14	24.96	25.2	24.78	25.14	25.28	25.34	25.36	25.43	25.04	25.4	25.18	25.08	24.67	25.25	25.26
Hydroquinone	14.78	14.88		15.05	14.98	15.84	14.95	15.03	15.44	15.7	15.16	15.48	15.14	15.41	15.92	15.48	15.6				16.69	16
C14 Hydrocarbon		13.33		14.11			14.12	13.61		13.49	13.18		14.21			13.14	13.52				14.11	
beta-Alanine			12.53			12.52	12.19		11.83		9.32		12.65	12.47	12.85		12.57		12.3			12.43
Threose/erythrose	14.21	12.9			12.94	13.14	13.1	13.13	11.07		13.53			12.95	12.98		12.91		12.57	11.64	14.15	
Aminomalonic acid	20.71	19.23	20.55	20.24	20.88	20.6	20.63	19.77	19.96	19.74	21.26	20.4	20.19	20.76	20.37	20	20.75	19.8	20.1	19.86	22.15	21.05
Aspartic acid	17.29	15.5	17.25	16.78	17.64	16.92	17.28	15.68	16.05	17.46	18.13	17.05	16.79	18.39	17.74	15.97	17.12	16.47	17.72	16.33	19.97	18.45
Malic acid	15.68	15.52	15.95	15.65	16.26	16.5	16.71	16.2	16.36	16.35	17.12	17.05	17.28	17.56	17.94	16	16.2	16.65	16.28	17.12	18.51	18.41
Nicotinamide	16.1	16.66	16.9	15.54	16.46	17.07	16.92	16.59	16.7		17.33	16.95	17.14	17.58		17.08	17.29	17.02	16.8	13.98	17.51	17.51
C15 hydrocarbon								17.01	17.05			16.98	17.19		17.04			17.4		17.33	14.87	
Hydroxyprolines			13.72		11.84	12.99	11.97	11.85	13.14		12.81	13.34		12.68	13.31		12.65	12.98	12.95			13.34
Threitol/erythritol							15				14.85	15.41	15.64	15.97	15.97	15.51		15.79	15.59	15.06	16.23	16.66
Methionine	21.5	21.66	22.42	21.56	21.74	22.18	21.77	21.83	22.23	21.5	22.11	22.21	22.31	22.3	22.35	22.07	22.55	22.07	22.13	21.33	22.18	22.26
Glutamic acid	25.32	25.22	25.61	24.87	25.51	25.6	25.56	25.32	25.56	25.17	25.81	25.67	25.71	25.95	25.99	25.48	25.71	25.55	25.5	25.05	25.87	25.88
Erythronic acid/threonic acid	14.67		15.24		15.05	15.08	15.35	14.95	14.58	14.64	15.6	15.31	15.68	15.69	15.97	14.76	15.39	15.43	15.65	15.33	16.39	16.31
Cysteine	16.39	16.98	17.76	16.81	16.89	17.84	17.27	17.34	18.02	17.31	17.77	18.36	17.89	18.03	18.45	17.88	18.12	18.14	18	17.76	18.18	18.67
alpha Ketoglutaric acid		16.1	16.88	15.84	16.07	16.96	16.45	16.63	17.23	16.39	17	17.52	17.46	17.89	18.21	16.01	16.56	16.72	16.94	17.03	18.31	18.22
2-Isopropylmalic acid	21.24	18.8	20.2	19.72	20.94	19.35	20.21	17.88	17.7	19.76	20.7	18.5	18.69	20.28	18.82	18.7	20.02	18.54	20.02	18.24	22.22	20.17
Phenylalanine	23.9	23.77	24.38	23.59	23.95	24.35	24.09	23.87	24.18	23.69	24.26	24.39	24.38	24.43	24.56	24.22	24.45	24.22	24.15	23.54	24.44	24.4
Succinylacetone	13.18	13.46	13.85	13.3	13.65	13.68	13.45	13.98		12.43		13.81	13.19	14.05		13.69	13.88	13.94	13.49			
Asparagine	16.54	17.17	17.87	17.04	17.57	17.32	17.76	17.69	17.54	17.6	18.11	18.1	18.77	18.74	18.9	17.52	17.76	18.04	17.81	18.2	19.06	18.97
Cysteinesulfinic acid	14.94	13.02			13.84	13.54	13.34	13.11							13.21	13.22	13.61	12.64		14.12	11.59	12.53
Pentitols	13.81	14.01	14.12		13.73	13.46	15.2	14.02	14.31	14.58	14.61	15.13	16.41	16.83	16.89	14.61	14.36	15.99	15.77	16.49	17.71	17.55
Putrescine	13.63	13.38	14.28	13.11		14.3	13.87	13.9	13.97	13.87	14.29	14.18			13.61		13.24	13.62	13.61	14.87	16.1	15.73
Glutamine	26.35	26.44	26.73	26.31	26.48	26.61	26.51	26.46	26.62	26.1	26.59	26.72	26.6	26.59	26.66	26.57	26.85	26.56	26.55	26.14	26.47	26.74
Gentisic acid			14.83	14.52				13.6	14.93	15.75	13.97	15.32		14.94	15.61				15.38			
Unoximated aldohexose																				12.28	14.78	13.26
Hypoxanthine						14.69		14.03	13.56	14.29	14.07	14.86		15.13	15.42	12.46		14.85	14.51			
Citric acid/isocitric acid	19.35	19.53	18.89	19.5	20.2	20.08	20.48	20.05	19.47	19.71	21.16	20.9	21.65	21.69	21.69	19.84	19.97	20.38	20.28	22.22	23.04	23.08
Ornithine	20.57	18.71	19.42	18.61	19.12	19.38	19.4	19.1	19.53	19.15	19.63	19.79	19.58	20.01	19.93	19.06	19.46	19.7	19.51	19.6	20.25	20.49
Arginine/Citrulline	18.05	18.06	18.83	17.74	18.42	19.12	18.51	18.16	18.97	18.54	18.83	19.03	18.63	19.01	19.82	18.65	19.3	18.87	18.95	16.27	18.53	18.67
deuterated Myristic Acid (RTL std)	22.05	22.23	22.39	22.04	22.27	22.42	21.99	22.37	22.39	22.26	22.24	22.38	22.34	22.14	22.49	22.38	22.17	22.38	22.29	22.1	22.35	22.55
Myristic acid or Pentadecanol	14.96		16.33	14.75	16.25	13.31	14.22	16.52	14.98	14.93	16.07	16.24	14.19	14.66	15.07	15.01	14.38	14.67		15.12	14.38	16.08
Methionine sulfoxide	15.68	15.84	13.52	14.41	17.22	15	16.71	16.14	14.72	14.82	14.63	14.36	15	14.56	16.28	15.16	14.75	14.9		15.28	15.36	15.25
Fructose or similar ketohexose	22.71	22.52	22.13	19.55	22.15	21.92	21.95	21.43	21.75	21.59	21.91	21.23	22	21.9	22.09	21.06	21.87	21.51	22.08	20.84	21.3	21.89
Gluconic acid or similar sugar acid	22.93	22.58	23.24	22.38	22.97	23.29	23.14	23.08	23.06	22.87	23.3	23.11	23	23.28	23.53	22.78	23.33	23.09	23.15	20.84	21.3	22.74
Pyridoxine	18.68	18.96	20.53	18.56	18.86	20.74	18.78	19.51	16.82	19.09	19.02	22.15	19.69	19.23	20.31	19.71	19.56	19.88	18.83	21.42	18.97	19.35
Glucose and other aldohexoses 2	27.68	27.72	27.97	27.58	27.57	27.93	27.16	27.84	28.18	27.74	27.62	27.95	27.9	27.7	27.87	27.98	27.94	27.98	27.96	27.78	26.93	27.79
Glucose and other aldohexoses 1	27.68	27.72	27.97	27.58	27.57	27.93	26.96	27.84	28.18	27.74	27.62	27.95	27.9	27.7	27.87	27.98	27.94	27.98	27.96	27.78	27.58	27.79
Dehydroascorbic acid�	25.56	23.69	25.41	23.55	25.21	24.88	24.67	25.35	25.31	25.86	25.5	25.59	25.6	24.82	25.43	26.94	25.91	24.87	25.4	23.1	22.7	27
Hexitols	26.83	26.87	27.02	26.65	27.07	27.34	26.96	26.96	27.19	26.95	27.05	27.15	27.07	27.02	27.17	26.94	27.17	27.01	27.16	26.97	26.93	27.34
Histidine	20.4	20.78	21.02	20.69	20.78	21.04	20.86	20.72	20.84	20.6	21	20.96	21.08	21.08	21.28	21.05	21.11	20.97	20.63	20.58	21.18	21.23
Lysine	25.16	25.19	25.55	25.2	25.38	25.51	25.39	25.25	25.42	25.13	25.39	25.49	25.52	25.68	25.75	25.41	25.66	25.51	25.52	25.01	25.49	25.52
Tyrosine	24.36	24.4	24.78	24.28	24.54	24.67	24.6	24.39	24.56	24.19	24.75	24.74	24.78	24.88	25.03	24.6	24.89	24.63	24.66	24.14	24.71	24.87
Sedoheptulose or similar sugar	23.47	23.55	23.9	23.52	23.65	23.77	23.79	23.58	23.65	23.36	23.79	23.78	23.92	23.88	24.01	23.83	24.03	23.74	23.73	23.14	23.57	23.78
Glucose and other aldohexoses 3	26.61	26.39	26.1	26.2	26.84	25.94	26.64	25.46	24.14	25.93	27.2	25.45	25.97	26.8	25.76	25.85	26.51	25.1	26.21	25.99	27.38	26.65
Pantothenic acid	19.83	19.58	19.48	19.34	20.1	19.75	19.88	19.01	19.13	19.38	20.01	19.42	19.44	19.97	19.61	19.91	19.96	19.32	19.9	19.13	19.98	20.09
Palmitic acid	20.66	21.8	20.93	22.07	20.89	21.91	22.01	20.94	21.96	21.86	20.87	20.78	22.12	21.79	22.05	21.6	21.72	21.73	22.06	21.99	21.94	20.65
Myoinositol	22.15	22.12	22.27	21.87	22.23	22.34	22.28	22.04	22.07	21.79	22.49	22.38	22.18	22.27	22.41	22.25	22.47	22.22	22.27	21.84	22.42	22.52
Guanine										12.06	12.91	12.42	14.2	14.65	15.12	13.8	13.47	13.75	14.84			
C21 Hydrocarbon		14.58		14.83			15.88				14.15	14.21	15.01					15.02		15.12		
Methyl stearate		17.48	17.09		18.78	18.68	17.88	16.82	18.35	19	17.09		18.19	17.89		16.74		18.04	16.6			
Tetracosamethylundecasiloxane			14.8		14.72	13.72		12.42					13.5	12.12								
Heptadecanoic acid/Octadecanol	13.64	13.89	15.13					15.24		13.25	15.21											15.04
C22 hydrocarbon		13.92		14.6		14.51	14.48	13.72	14.4	14.07		13.21	14.3	14.12	14.47	13.92	13.93	14.23	14.41	14.07	13.76	
Tryptamine/Norepinephrine	13.61	15.14	15.08	14.76	14.94	14.79	14.76	14.7	15.82	14.65	14.25	15.74	15.16	15.15	16.19	14.75	14.53	15.49	15.74	14.09		15.09
Tryptophan	20.43	20.52	21.25	20.15	20.6	21.42	20.67	20.5	21.07	20.15	21.2	21.36	20.92	21.18	21.93	21.01	21.59	21.09	21.15	20.46	21.82	21.87
Stearic acid	21.46	22.16	21.63	22.16	21.77	22.25	22.23	21.94	22.11	22.31	21.68	21.76	22.24	22.15	22.35	21.95	22.06	22.12	22.27	22.32	22.16	21.51
Hexacosemethyldodecasiloxane			13.16		13	12.51		14.23			13.96	13.15					12.33	13.27			13.68	14.73
Cystine	22.62	22.51	22.52	22.19	22.56	22.75	22.77	22.35	22.21	21.72	23.13	22.8	22.81	22.8	22.94	22.67	23.04	22.44	22.49	22.4	22.87	22.97
Hexose-6-phosphate	18.89	19.13		17.13			18.74	18.49	16.49		18.9	18.24	17.13		17.11	19.34				19.79	18.24	19.36
5-Hydroxytryptophan			14.6	14.34		14.73			14.44	15.25		15.87	17.06	17.33	17.46		15.1	15.18	15.74	16.61	18.61	17.95
beta-Monopalmitin	17.26	16.81	15.86	16.63	16.82	16.64	16.76	16.65	16.23	16.39	18.19	16.21	16.75	16.7	16.81	16.77	16.72	16.43	14.04	16.85	18.88	17.19
alpha-Monopalmitin	21.8	21.85	21.55	21.97	21.8	22.11	22.08	21.72	22.06	21.87	22.08	22.05	21.91	22.22	22.38	22.24	22.25	22	21.77	22.1	22	22.07
Sucrose and similar disaccharides	13.67			13.37	13.28			11.93	12.12	12.9	13.76	13.71	12.06			13.57			13.93			13.02
Lactose or similar disaccharide	18.3	18.19	18.88	17.93	18.25	18.55	18.57	18.47	18.83	17.92	18.53	19.19	19.06	18.63	19.31	18.42	18.81	18.96	18.48			
beta-Monostearin	19.56	19.52	19.08	19.96	20.16	19.57	19.79	19.11	19.7	19.81	19.74	19.52	19.65	19.98	20.05	19.62	20.22	19.79	19.77	19.94	19.99	19.93
Maltose or similar disaccharide	16.42	18.39	18.61	17.42	18.27	18.91	17.58	18.64	18.58	18.29	17.61	18.49	17.8	19.28	18.37	18.55	19.43	17.29	18.5	17.29	18.41	18.21
alpha-Monostearin	24.83	24.92	24.67	25.16	25	25.24	25.21	24.79	25.2	25.04	25.05	25.1	25.08	25.28	25.4	25.35	25.39	25.23	24.93	25.14	24.97	25.08
Isomaltose or similar disaccharide	19.67	19.5	19.73	19.12	19.52	19.78	19.86	19.42	19.49	19.02	20.09	19.96	19.86	19.85	20.07	19.72	20.11	19.71	19.73	19.24	20.21	20.34
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	CAS	Retention time	Reverse	SimPeak
Glyoxylic acid	563-96-2	5.916	73.5	7
C10 hydrocarbon	124-18-5	6.185	81.2	8
epsilon-Caprolactam	105-60-2	6.2574	82.5	9
2-Hydroxypyridine	142-08-5	6.4525	85.2	13
Pyruvic acid	127-17-3	6.5922	99	14
Lactic acid	79-33-4	6.7791	99.8	16
Glycolic acid	79-14-1	7.0133	88.1	20
Alanine	56-41-7	7.4056	99	23
C11 hydrocarbon	1120-21-4	7.4866	75.1	24
2-Hydroxy-2-methylbutyric acid	3739-30-8	7.8051	68.9	28
2-Ketovaline	759-05-7	7.8521	94.9	29
2-Hydroxybutyric acid	565-70-8	7.9649	74	32
Sulfuric acid	7664-93-9	8.3068	86.9	35
Dodecamethylpentasiloxane		8.3661	0	36
Malonic acid	141-82-2	8.9088	84.9	46
2-Ketoleucine	816-66-0	9.0128	99.3	49
Valine	72-18-4	9.0754	99.2	50
C12 hydrocarbon	112-40-3	9.2413	74.3	53
Benzoic acid	65-85-0	9.5983	98.4	60
Ethanolamine	141-43-5	9.8107	90.5	64
Phosphoric acid	7664-38-2	9.8485	91.8	65
1,3-Dihydroxyacetone	96-26-4	9.8665	65.6	66
Leucine	61-90-5	9.8713	97.9	67
Glycerol	56-81-5	9.8815	93.4	69
Ethylmalonic acid	601-75-2	9.8799	69.9	70
C13 hydrocarbon	629-50-5	10.0037	77.7	72
Isoleucine	443-79-8	10.1788	100	74
Proline	147-85-3	10.2512	96	75
Glycine	56-40-6	10.3698	99	77
Succinic acid	110-15-6	10.4499	75.6	79
Glyceric acid	473-81-4	10.663	71.7	83
MSTFA artifact	24589-78-4	10.7331	86.4	84
Fumaric acid	17013-01-3	10.961	86.3	87
Serine	56-45-1	11.0755	99	88
Threonine	72-19-5	11.4268	99	92
Hydroquinone	123-31-9	11.8527	51	95
C14 Hydrocarbon	629-59-4	11.9049	73.3	96
beta-Alanine	107-95-9	12.0056	75.6	97
Threose/erythrose	95-43-2	12.2856	73.5	98
Aminomalonic acid	1068-84-4	12.5131	65.3	101
Aspartic acid	16504-56-6	12.5295	71.6	102
Malic acid	617-48-1	12.7361	87.3	105
Nicotinamide	98-92-0	12.7709	92.5	106
C15 hydrocarbon	629-62-9	12.8566	82.1	107
Hydroxyprolines	4298-08-2	12.9591	75.8	110
Threitol/erythritol	149-32-6	12.9845	81.5	111
Methionine	63-68-3	13.1669	86.8	112
Glutamic acid	56-86-0	13.199	99.7	113
Erythronic acid/threonic acid	13752-84-6	13.4125	75.4	115
Cysteine	52-90-4	13.5897	93.1	119
alpha Ketoglutaric acid	328-50-7	13.8052	82.2	121
2-Isopropylmalic acid	16048-89-8	14.1449	77	125
Phenylalanine	63-91-2	14.4664	97	128
Succinylacetone	51568-18-4	14.6441	78.8	131
Asparagine	70-47-3	14.9279	93.3	136
Cysteinesulfinic acid	1115-65-7	15.0536	75.6	140
Pentitols	488-82-4	15.5475	81.4	149
Putrescine	110-60-1	15.7483	81	151
Glutamine	56-85-9	16.1276	98	154
Gentisic acid	490-79-9	16.2475	75	157
Unoximated aldohexose	138-59-0	16.4398	55.7	159
Hypoxanthine	68-94-0	16.4736	85.3	160
Citric acid/isocitric acid	5949-29-1	16.5612	98.2	162
Ornithine	70-26-8	16.5768	92.1	163
Arginine/Citrulline	74-79-3	16.6427	96.1	164
deuterated Myristic Acid (RTL std)	60658-41-5	16.7219	99	165
Myristic acid or Pentadecanol	629-76-5	16.9412	94	169
Methionine sulfoxide	3226-65-1	17.001	81.4	171
Fructose or similar ketohexose	87-81-0	17.1235	98.8	173
Gluconic acid or similar sugar acid	1198-69-2	17.2245	83.9	176
Pyridoxine	65-23-6	17.39	93	183
Glucose and other aldohexoses 2	2595-98-4	17.5606	98.7	184
Glucose and other aldohexoses 1	50-99-7	17.5983	99.3	187
Dehydroascorbic acid�	490-83-5	17.5577	73.3	188
Hexitols	87-78-5	17.5814	90.7	189
Histidine	5934-29-2	17.6883	81.1	191
Lysine	56-87-1	17.7219	99	192
Tyrosine	60-18-4	17.8678	99	195
Sedoheptulose or similar sugar	469-90-9	18.0049	75.7	200
Glucose and other aldohexoses 3	50-99-7	18.1978	97.4	206
Pantothenic acid	137-08-6	18.3531	94.6	208
Palmitic acid	57-10-3	18.908	99.9	217
Myoinositol	87-89-8	19.3215	97	221
Guanine	73-40-5	19.6127	85	225
C21 Hydrocarbon	629-94-7	19.6866	66.9	227
Methyl stearate	112-61-8	19.7165	87.8	228
Tetracosamethylundecasiloxane	107-53-9	19.7579	62.7	229
Heptadecanoic acid/Octadecanol	506-12-7	19.8307	81.3	230
C22 hydrocarbon	629-97-0	20.0663	66.5	232
Tryptamine/Norepinephrine	61-54-1	20.235	73	233
Tryptophan	73-22-3	20.4606	100	236
Stearic acid	57-11-4	20.7175	99.1	240
Hexacosemethyldodecasiloxane	2471-08-1	21.0244	64.8	246
Cystine	56-89-3	21.1313	99.4	247
Hexose-6-phosphate	3672-15-9	21.3133	78.7	248
5-Hydroxytryptophan	4350-09-8	22.1245	69.7	255
beta-Monopalmitin	23470-00-0	23.2523	96.1	264
alpha-Monopalmitin	542-44-9	23.5166	99.8	266
Sucrose and similar disaccharides	57-50-1	23.8844	75.2	268
Lactose or similar disaccharide	63-42-3	24.6518	81.6	273
beta-Monostearin	31566-31-1	24.6937	96.6	275
Maltose or similar disaccharide	69-79-4	24.8427	89.5	278
alpha-Monostearin	123-94-4	24.9608	100	279
Isomaltose or similar disaccharide	499-40-1	25.6047	98.2	282
METABOLITES_END
#END