#METABOLOMICS WORKBENCH Suzanna_Shirazi04_20250120_171947 DATATRACK_ID:5548 STUDY_ID:ST003704 ANALYSIS_ID:AN006077 PROJECT_ID:PR002299
VERSION             	1
CREATED_ON             	February 4, 2025, 9:00 pm
#PROJECT
PR:PROJECT_TITLE                 	Metabolomic profiling of human bone marrow mesenchymal stromal stem cells in
PR:PROJECT_TITLE                 	high glucose
PR:PROJECT_TYPE                  	Metabolomics
PR:PROJECT_SUMMARY               	A study on the effect of high glucose level on the differentiation and
PR:PROJECT_SUMMARY               	metabolomic changes of human bone marrow mesenchymal stem cells (BMSC). Human
PR:PROJECT_SUMMARY               	BMSC were cultured under normal and high glucose level media with or without the
PR:PROJECT_SUMMARY               	osteogenic or adipogenic conditions. A range of tests were performed on the
PR:PROJECT_SUMMARY               	cells to look at differentiation, stress response and metabolomics changes in
PR:PROJECT_SUMMARY               	the cells.
PR:INSTITUTE                     	University of Adelaide
PR:DEPARTMENT                    	Health and Medical Sciences
PR:LABORATORY                    	MSC Lab
PR:LAST_NAME                     	Shirazi
PR:FIRST_NAME                    	Suzanna
PR:ADDRESS                       	Adelaide University, North Terrace, Adelaide, SA, 5000, Australia
PR:EMAIL                         	suzanna.shirazi@adelaide.edu.au
PR:PHONE                         	0420245950
PR:FUNDING_SOURCE                	NHMRC
#STUDY
ST:STUDY_TITLE                   	Hyperglycemia leads to BMSC(bone marrow mesenchymal stromal cells) impaired
ST:STUDY_TITLE                   	osteogenesis, enhanced adipogenesis, and altered metabolism
ST:STUDY_TYPE                    	metabolomics
ST:STUDY_SUMMARY                 	Diabetes, a major risk factor for osteoporosis, negatively impacts bone health,
ST:STUDY_SUMMARY                 	but the mechanisms underlying the effects of hyperglycemia on bone marrow
ST:STUDY_SUMMARY                 	mesenchymal stromal cells (BMSC) are not fully understood. This study
ST:STUDY_SUMMARY                 	investigates how high glucose conditions influence BMSC differentiation,
ST:STUDY_SUMMARY                 	proliferation, viability, and metabolism. Our results demonstrate that high
ST:STUDY_SUMMARY                 	glucose inhibits osteogenesis, as evidenced by significantly reduced alkaline
ST:STUDY_SUMMARY                 	phosphatase activity, impaired calcium deposition, and downregulation of key
ST:STUDY_SUMMARY                 	osteogenic genes (RUNX2, Osteocalcin, Osteopontin). Additionally, high glucose
ST:STUDY_SUMMARY                 	conditions promote adipogenesis, characterized by increased lipid accumulation,
ST:STUDY_SUMMARY                 	larger fat droplets, and upregulation of adipogenic genes (PPARγ2, CEBPα,
ST:STUDY_SUMMARY                 	Leptin, AdipoQ), suggesting a shift towards fat cell differentiation.
ST:STUDY_SUMMARY                 	Furthermore, BMSCs cultured in high glucose show decreased proliferation,
ST:STUDY_SUMMARY                 	elevated DNA damage, increased oxidative stress, enhanced apoptosis and
ST:STUDY_SUMMARY                 	senescence, particularly in later passages, highlighting the negative impact of
ST:STUDY_SUMMARY                 	hyperglycemia on BMSC viability. Metabolomic profiling of osteogenic and
ST:STUDY_SUMMARY                 	adipogenic differentiation in normal and high glucose conditions revealed key
ST:STUDY_SUMMARY                 	metabolic shifts, with Nicotinamide Adenine Dinucleotide (NAD+) and
ST:STUDY_SUMMARY                 	L-glutamate/α-ketoglutarate (α-KG) identified as critical metabolites driving
ST:STUDY_SUMMARY                 	these processes. Supplementation with NAD+ and α-KG in high glucose conditions
ST:STUDY_SUMMARY                 	significantly enhanced ALP activity and upregulated osteogenic markers,
ST:STUDY_SUMMARY                 	suggesting that these metabolites play a vital role in restoring osteogenic
ST:STUDY_SUMMARY                 	potential under hyperglycemic conditions. Overall, our findings suggest that
ST:STUDY_SUMMARY                 	high glucose promotes adipogenesis at the expense of osteogenesis, exacerbates
ST:STUDY_SUMMARY                 	cellular damage, and accelerates aging in BMSC. The identification of NAD+ and
ST:STUDY_SUMMARY                 	α-KG as key regulators in this process provides new insights into the metabolic
ST:STUDY_SUMMARY                 	mechanisms behind impaired bone health in diabetes and highlights potential
ST:STUDY_SUMMARY                 	therapeutic avenues to counteract these detrimental effects. Further research
ST:STUDY_SUMMARY                 	into the role of NAD+ and α-KG in BMSC differentiation under high glucose may
ST:STUDY_SUMMARY                 	offer novel strategies for managing diabetes-related bone diseases.
ST:INSTITUTE                     	University of Adelaide
ST:DEPARTMENT                    	Health and Medical Sciences
ST:LABORATORY                    	MSC Lab
ST:LAST_NAME                     	Shirazi
ST:FIRST_NAME                    	Suzanna
ST:ADDRESS                       	Adelaide University, North Terrace, Adelaide, SA, 5000, Australia
ST:EMAIL                         	suzanna.shirazi@adelaide.edu.au
ST:PHONE                         	0420245950
#SUBJECT
SU:SUBJECT_TYPE                  	Cultured cells
SU:SUBJECT_SPECIES               	Homo sapiens
SU:TAXONOMY_ID                   	9606
#FACTORS
#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           	-	NDO231_Osteo-Low 21D	Sample Class:Osteogenic Media | Timepoint:21D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO231_Osteo-Low 21D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO231_Osteo-Hi 21D	Sample Class:Osteogenic Media | Timepoint:21D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO231_Osteo-Hi 21D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO356_Osteo-Low 21D	Sample Class:Osteogenic Media | Timepoint:21D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO356_Osteo-Low 21D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO356_Osteo-Hi 21D	Sample Class:Osteogenic Media | Timepoint:21D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO356_Osteo-Hi 21D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO362_Osteo-Low 21D	Sample Class:Osteogenic Media | Timepoint:21D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO362_Osteo-Low 21D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO362_Osteo-Hi 21D	Sample Class:Osteogenic Media | Timepoint:21D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO362_Osteo-Hi 21D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO370_Osteo-Low 21D	Sample Class:Osteogenic Media | Timepoint:21D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO370_Osteo-Low 21D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO370_Osteo-Hi 21D	Sample Class:Osteogenic Media | Timepoint:21D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO370_Osteo-Hi 21D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO231_Adipo-Low 21D	Sample Class:Adipogenic Media | Timepoint:21D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO231_Adipo-Low 21D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO231_Adipo-Hi 21D	Sample Class:Adipogenic Media | Timepoint:21D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO231_Adipo-Hi 21D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO356_Adipo-Low 21D	Sample Class:Adipogenic Media | Timepoint:21D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO356_Adipo-Low 21D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO356_Adipo-Hi 21D	Sample Class:Adipogenic Media | Timepoint:21D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO356_Adipo-Hi 21D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO362_Adipo-Low 21D	Sample Class:Adipogenic Media | Timepoint:21D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO362_Adipo-Low 21D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO362_Adipo-Hi 21D	Sample Class:Adipogenic Media | Timepoint:21D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO362_Adipo-Hi 21D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO370_Adipo-Low 21D	Sample Class:Adipogenic Media | Timepoint:21D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO370_Adipo-Low 21D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO370_Adipo-Hi 21D	Sample Class:Adipogenic Media | Timepoint:21D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO370_Adipo-Hi 21D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO231_Cont-Low 21D	Sample Class:Control Media | Timepoint:21D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO231_Cont-Low 21D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO231_Cont-Hi 21D	Sample Class:Control Media | Timepoint:21D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO231_Cont-Hi 21D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO356_Cont-Low 21D	Sample Class:Control Media | Timepoint:21D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO356_Cont-Low 21D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO356_Cont-Hi 21D	Sample Class:Control Media | Timepoint:21D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO356_Cont-Hi 21D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO362_Cont-Low 21D	Sample Class:Control Media | Timepoint:21D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO362_Cont-Low 21D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO362_Cont-Hi 21D	Sample Class:Control Media | Timepoint:21D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO362_Cont-Hi 21D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO370_Cont-Low 21D	Sample Class:Control Media | Timepoint:21D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO370_Cont-Low 21D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO370_Cont-Hi 21D	Sample Class:Control Media | Timepoint:21D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO370_Cont-Hi 21D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO231_Osteo-Low 7D	Sample Class:Osteogenic Media | Timepoint:7D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO231_Osteo-Low 7D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO231_Osteo-Hi 7D	Sample Class:Osteogenic Media | Timepoint:7D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO231_Osteo-Hi 7D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO356_Osteo-Low 7D	Sample Class:Osteogenic Media | Timepoint:7D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO356_Osteo-Low 7D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO356_Osteo-Hi 7D	Sample Class:Osteogenic Media | Timepoint:7D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO356_Osteo-Hi 7D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO362_Osteo-Low 7D	Sample Class:Osteogenic Media | Timepoint:7D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO362_Osteo-Low 7D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO362_Osteo-Hi 7D	Sample Class:Osteogenic Media | Timepoint:7D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO362_Osteo-Hi 7D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO370_Osteo-Low 7D	Sample Class:Osteogenic Media | Timepoint:7D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO370_Osteo-Low 7D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO370_Osteo-Hi 7D	Sample Class:Osteogenic Media | Timepoint:7D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO370_Osteo-Hi 7D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO231_Adipo-Low 7D	Sample Class:Adipogenic Media | Timepoint:7D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO231_Adipo-Low 7D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO231_Adipo-Hi 7D	Sample Class:Adipogenic Media | Timepoint:7D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO231_Adipo-Hi 7D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO356_Adipo-Low 7D	Sample Class:Adipogenic Media | Timepoint:7D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO356_Adipo-Low 7D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO356_Adipo-Hi 7D	Sample Class:Adipogenic Media | Timepoint:7D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO356_Adipo-Hi 7D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO362_Adipo-Low 7D	Sample Class:Adipogenic Media | Timepoint:7D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO362_Adipo-Low 7D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO362_Adipo-Hi 7D	Sample Class:Adipogenic Media | Timepoint:7D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO362_Adipo-Hi 7D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO362_Adipo-Hi 7D DUP	Sample Class:Adipogenic Media | Timepoint:7D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO362_Adipo-Hi 7D DUP-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO370_Adipo-Low 7D	Sample Class:Adipogenic Media | Timepoint:7D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO370_Adipo-Low 7D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO370_Adipo-Hi 7D	Sample Class:Adipogenic Media | Timepoint:7D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO370_Adipo-Hi 7D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO231_Cont-Low 7D	Sample Class:Control Media | Timepoint:7D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO231_Cont-Low 7D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO231_Cont-Hi 7D	Sample Class:Control Media | Timepoint:7D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO231_Cont-Hi 7D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO356_Cont-Low 7D	Sample Class:Control Media | Timepoint:7D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO356_Cont-Low 7D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO356_Cont-Hi 7D	Sample Class:Control Media | Timepoint:7D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO356_Cont-Hi 7D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO362_Cont-Low 7D	Sample Class:Control Media | Timepoint:7D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO362_Cont-Low 7D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO362_Cont-Hi 7D	Sample Class:Control Media | Timepoint:7D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO362_Cont-Hi 7D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO370_Cont-Low 7D	Sample Class:Control Media | Timepoint:7D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO370_Cont-Low 7D-mzML
SUBJECT_SAMPLE_FACTORS           	-	NDO370_Cont-Hi 7D	Sample Class:Control Media | Timepoint:7D | Sample source:Human BMSC	RAW_FILE_NAME(Raw file name)=NDO370_Cont-Hi 7D-mzML
#COLLECTION
CO:COLLECTION_SUMMARY            	BMSC were isolated from the posterior iliac crests of adult volunteers with
CO:COLLECTION_SUMMARY            	informed consent and ethical approval from the Royal Adelaide Hospital. BM
CO:COLLECTION_SUMMARY            	mononuclear cells were cultured in α-MEM with 10% FCS, 2 mM L-glutamine, and
CO:COLLECTION_SUMMARY            	100 μM L-ascorbate-2-phosphate. BMSCs were cultured in media with normal (1
CO:COLLECTION_SUMMARY            	g/L) or high (4.5 g/L) glucose and incubated at 37°C with 5% CO2
CO:SAMPLE_TYPE                   	Cultured cells
#TREATMENT
TR:TREATMENT_SUMMARY             	BMSC from 3 human donors were cultured for 7 and 21 days in normal or high
TR:TREATMENT_SUMMARY             	glucose DMEM media under control (DMEM cat num # 11885-084 500ml and #11995-065
TR:TREATMENT_SUMMARY             	Thermofisher), osteogenic ( Fetal calf serum 5%, Additive’s 1X, Ascorbate-2P
TR:TREATMENT_SUMMARY             	100µM, Dexamethasone (Na/PO4) 0.1µM, KH2PO4 2.6mM), or adipogenic ( Fetal calf
TR:TREATMENT_SUMMARY             	serum 10%, Additive’s 1X, Ascorbate-2P 100µM, Dexamethasone (Na/PO4) 0.1µM,
TR:TREATMENT_SUMMARY             	Indomethacin 60µM) conditions. Media was changed every 3 days. A total of 48
TR:TREATMENT_SUMMARY             	samples were processed by Australian Wine Research Institute, South Australia,
TR:TREATMENT_SUMMARY             	where 110 compounds were detected using HILIC LC-MS. Compound identification was
TR:TREATMENT_SUMMARY             	based on internal and external spectral databases
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	1. 900 uL of a mixture of Methanol and Chloroform (75% methanol, 25% chloroform)
SP:SAMPLEPREP_SUMMARY            	were added to the Eppendorf tube containing the cell media (sample). 2. The
SP:SAMPLEPREP_SUMMARY            	Eppendorf containing the sample and the solvents was vortexed for 1 minutes and
SP:SAMPLEPREP_SUMMARY            	then extracted for 15 minutes using an orbital shaker. 3. After extraction, the
SP:SAMPLEPREP_SUMMARY            	Eppendorf tubes were centrifuged at 14,500 rpm for 15 minutes. The supernatant
SP:SAMPLEPREP_SUMMARY            	was collected and transferred to a new 2mL Eppendorf tube. 4. The remaining
SP:SAMPLEPREP_SUMMARY            	media was submitted to a second extraction using 300 uL of the Methanol
SP:SAMPLEPREP_SUMMARY            	Chloroform mixture (described in point 1) 5. Step 2 and 3 were repeated and the
SP:SAMPLEPREP_SUMMARY            	supernatant obtained from the second extraction was combined to the one obtained
SP:SAMPLEPREP_SUMMARY            	from the first extraction. 6. 500 uL of ice cold MilliQ water was added to the
SP:SAMPLEPREP_SUMMARY            	Eppendorf containing the extract. The Eppendorf tube was vortexed for 1 minute
SP:SAMPLEPREP_SUMMARY            	and centrifuged at 14,500 rpm for 15 minutes. 7. The supernatant (top layer) was
SP:SAMPLEPREP_SUMMARY            	collected, transferred to a glass test tube and dried down using a nitrogen
SP:SAMPLEPREP_SUMMARY            	evaporator (using a nitrogen flow of 3.5 ml/min at 25 degrees.) 8. The dried
SP:SAMPLEPREP_SUMMARY            	samples were resuspended using 75 uL of Mobile phase A (20 mM Ammonim Carbonate
SP:SAMPLEPREP_SUMMARY            	Buffer in MilliQ water) and 25 uL of Mobile Phase B (Acetonitrile) 9. After
SP:SAMPLEPREP_SUMMARY            	resuspension the samples (100uL) were transferred in HPLC vial inserts for LC-MS
SP:SAMPLEPREP_SUMMARY            	analysis
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	Solvent A: 20 mM Ammonim Carbonate Buffer, pH=9; chrom. flow gradient linearly
CH:CHROMATOGRAPHY_SUMMARY        	decreased from 75% B to 50% B from 0.5 to 36min
CH:CHROMATOGRAPHY_TYPE           	HILIC
CH:INSTRUMENT_NAME               	Thermo Vanquish Horizon
CH:COLUMN_NAME                   	Waters Xbridge Amide (150 x 2.1mm, 3.5um)
CH:SOLVENT_A                     	100% water; 20 mM Ammonim Carbonate
CH:SOLVENT_B                     	100% Acetonitrile
CH:FLOW_GRADIENT                 	0-0.5min: 75% B, 0.5 to 36min: linearly decreased from 75% B to 50% B
CH:FLOW_RATE                     	0.4 mL/min
CH:COLUMN_TEMPERATURE            	30°C
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Thermo Orbitrap ID-X Tribrid
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	NEGATIVE
MS:MS_COMMENTS                   	Thermo Compound Discoverer Software (version 3.3) was used to perform molecular
MS:MS_COMMENTS                   	feature extraction and feature grouping into compounds. The term molecular
MS:MS_COMMENTS                   	feature describes a two-dimensional bounded signal: a chromatographic peak
MS:MS_COMMENTS                   	(retention time) and a mass spectral peak (m/z). MS1 signals were acquired for
MS:MS_COMMENTS                   	the sample replicates and the pooled mix replicates (“PBQC”). The PBQC is a
MS:MS_COMMENTS                   	pooled mix of the samples that is used to monitor the instrument performance
MS:MS_COMMENTS                   	along the sequence. In addition, 3 replicates of the pooled mix were analysed in
MS:MS_COMMENTS                   	MS/MS mode, using HCD fragmentation. These MS/MS analyses were used to provide
MS:MS_COMMENTS                   	more reliable putative identifications and structural information of the
MS:MS_COMMENTS                   	detected compounds. The raw data matrix was median normalised to correct for any
MS:MS_COMMENTS                   	possible inter-run instrument variability. Following this, the CV% among the
MS:MS_COMMENTS                   	Pooled Mix replicates was calculated and compounds showing a CV% value above 25
MS:MS_COMMENTS                   	were removed from the data matrix because inconsistent through the analysis
MS:MS_COMMENTS                   	batch. The first 3 PBQC replicates were not included in the CV calculation
MS:MS_COMMENTS                   	because they were used to condition the instrument. The total number of detected
MS:MS_COMMENTS                   	compounds was 109.
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	chromatographic areas with no units
MS_METABOLITE_DATA_START
Samples	NDO231_Adipo-Hi 7D	NDO231_Adipo-Hi 21D	NDO356_Adipo-Hi 7D	NDO356_Adipo-Hi 21D	NDO362_Adipo-Hi 7D	NDO362_Adipo-Hi 7D DUP	NDO362_Adipo-Hi 21D	NDO370_Adipo-Hi 7D	NDO370_Adipo-Hi 21D	NDO231_Adipo-Low 7D	NDO231_Adipo-Low 21D	NDO356_Adipo-Low 7D	NDO356_Adipo-Low 21D	NDO362_Adipo-Low 21D	NDO370_Adipo-Low 7D	NDO370_Adipo-Low 21D	NDO231_Cont-Hi 7D	NDO231_Cont-Hi 21D	NDO356_Cont-Hi 7D	NDO356_Cont-Hi 21D	NDO362_Cont-Hi 7D	NDO362_Cont-Hi 21D	NDO370_Cont-Hi 7D	NDO370_Cont-Hi 21D	NDO231_Cont-Low 7D	NDO231_Cont-Low 21D	NDO356_Cont-Low 7D	NDO356_Cont-Low 21D	NDO362_Cont-Low 7D	NDO362_Cont-Low 21D	NDO370_Cont-Low 7D	NDO370_Cont-Low 21D	NDO231_Osteo-Hi 7D	NDO231_Osteo-Hi 21D	NDO356_Osteo-Hi 7D	NDO356_Osteo-Hi 21D	NDO362_Osteo-Hi 7D	NDO362_Osteo-Hi 21D	NDO370_Osteo-Hi 7D	NDO370_Osteo-Hi 21D	NDO231_Osteo-Low 7D	NDO231_Osteo-Low 21D	NDO356_Osteo-Low 7D	NDO356_Osteo-Low 21D	NDO362_Osteo-Low 21D	NDO370_Osteo-Low 7D	NDO370_Osteo-Low 21D
Factors	Sample Class:Adipogenic Media | Timepoint:7D | Sample source:Human BMSC	Sample Class:Adipogenic Media | Timepoint:21D | Sample source:Human BMSC	Sample Class:Adipogenic Media | Timepoint:7D | Sample source:Human BMSC	Sample Class:Adipogenic Media | Timepoint:21D | Sample source:Human BMSC	Sample Class:Adipogenic Media | Timepoint:7D | Sample source:Human BMSC	Sample Class:Adipogenic Media | Timepoint:7D | Sample source:Human BMSC	Sample Class:Adipogenic Media | Timepoint:21D | Sample source:Human BMSC	Sample Class:Adipogenic Media | Timepoint:7D | Sample source:Human BMSC	Sample Class:Adipogenic Media | Timepoint:21D | Sample source:Human BMSC	Sample Class:Adipogenic Media | Timepoint:7D | Sample source:Human BMSC	Sample Class:Adipogenic Media | Timepoint:21D | Sample source:Human BMSC	Sample Class:Adipogenic Media | Timepoint:7D | Sample source:Human BMSC	Sample Class:Adipogenic Media | Timepoint:21D | Sample source:Human BMSC	Sample Class:Adipogenic Media | Timepoint:21D | Sample source:Human BMSC	Sample Class:Adipogenic Media | Timepoint:7D | Sample source:Human BMSC	Sample Class:Adipogenic Media | Timepoint:21D | Sample source:Human BMSC	Sample Class:Control Media | Timepoint:7D | Sample source:Human BMSC	Sample Class:Control Media | Timepoint:21D | Sample source:Human BMSC	Sample Class:Control Media | Timepoint:7D | Sample source:Human BMSC	Sample Class:Control Media | Timepoint:21D | Sample source:Human BMSC	Sample Class:Control Media | Timepoint:7D | Sample source:Human BMSC	Sample Class:Control Media | Timepoint:21D | Sample source:Human BMSC	Sample Class:Control Media | Timepoint:7D | Sample source:Human BMSC	Sample Class:Control Media | Timepoint:21D | Sample source:Human BMSC	Sample Class:Control Media | Timepoint:7D | Sample source:Human BMSC	Sample Class:Control Media | Timepoint:21D | Sample source:Human BMSC	Sample Class:Control Media | Timepoint:7D | Sample source:Human BMSC	Sample Class:Control Media | Timepoint:21D | Sample source:Human BMSC	Sample Class:Control Media | Timepoint:7D | Sample source:Human BMSC	Sample Class:Control Media | Timepoint:21D | Sample source:Human BMSC	Sample Class:Control Media | Timepoint:7D | Sample source:Human BMSC	Sample Class:Control Media | Timepoint:21D | Sample source:Human BMSC	Sample Class:Osteogenic Media | Timepoint:7D | Sample source:Human BMSC	Sample Class:Osteogenic Media | Timepoint:21D | Sample source:Human BMSC	Sample Class:Osteogenic Media | Timepoint:7D | Sample source:Human BMSC	Sample Class:Osteogenic Media | Timepoint:21D | Sample source:Human BMSC	Sample Class:Osteogenic Media | Timepoint:7D | Sample source:Human BMSC	Sample Class:Osteogenic Media | Timepoint:21D | Sample source:Human BMSC	Sample Class:Osteogenic Media | Timepoint:7D | Sample source:Human BMSC	Sample Class:Osteogenic Media | Timepoint:21D | Sample source:Human BMSC	Sample Class:Osteogenic Media | Timepoint:7D | Sample source:Human BMSC	Sample Class:Osteogenic Media | Timepoint:21D | Sample source:Human BMSC	Sample Class:Osteogenic Media | Timepoint:7D | Sample source:Human BMSC	Sample Class:Osteogenic Media | Timepoint:21D | Sample source:Human BMSC	Sample Class:Osteogenic Media | Timepoint:21D | Sample source:Human BMSC	Sample Class:Osteogenic Media | Timepoint:7D | Sample source:Human BMSC	Sample Class:Osteogenic Media | Timepoint:21D | Sample source:Human BMSC
Cuminaldehyde	2775375	84196	2341598	122862	1295856	1311347	124596	3891625	37488	2796751	138600	2269977	139205	114817	3567230	105327	26330	121656	122171	82207	47339	56640	31397	121698	1272793	93383	110556	62445	44401	20694	2002758	217488	19136	818301	270719	6001025	129317	4757436	173888	3074712	47900	272909	261159	92358	101713	156460	414034
Sphingosine (d18:1)	1836363	344762	1864981	1534223	643441	4796238	334234	2707090	596666	1287863	325092	1749136	3664762	345946	3058171	737568	431785	1717373	1253237	1347511	148218	128101	1095728	3068377	811846	215839	1006235	335105	635002	222675	572630	2478803	1238649	8773	3942227	186644	1794770	462091	1754901	20031	1650330	17275	1162163	7356	37778	1403128	7976
Urocanic acid	1926109	1428180	1681996	2071495	1606532	997770	2414108	3367421	1571456	1977689	2856756	2068284	999295	1853975	1693050	6790146	1684359	1713479	1772169	650742	874917	513792	2149434	2173017	1279871	1187656	1146015	1449550	440189	823608	1762335	1350627	3595640	10568056	4431633	7970753	3487356	17785262	5413598	7541263	5996932	19260220	4903983	13896411	95517285	26662936	14654396
DL-Lysine	2812114	13087369	2422247	13657528	5008467	2643042	16288633	6943400	12826636	3535077	9593107	1700191	10311660	17489357	8380264	11831313	1226159	4322164	2448927	2511688	2136088	3315770	4378569	4992866	2549180	4995739	2645218	3765922	3662986	3511043	3396633	5788238	3308460	6910872	9197064	8699231	15828019	8540067	7843214	6456592	17485200	3373726	4138668	7051709	2721577	4416933	5170834
N-Benzylformamide	14202324	10149193	16152073	13418673	25371071	28017960	7631027	15737098	14619042	12851066	9537310	18397885	16498733	19503329	12672784	19799624	17554096	10541809	19676374	17280659	24622490	18792163	22899968	12859982	23552184	13709400	23129051	26707701	28343885	22840595	19478355	15207947	8716155	2176365	7495871	1991514	33300109	2265748	9104614	4432126	9792785	3588622	7583848	964317	6968307	8680408	2963851
Acetyl-L-carnitine	10705638	4558153	5949140	5822031	9867402	19629277	3970545	4690517	5462818	6372761	6379196	5244643	8284569	7479041	1695894	4777603	1847666	1026344	1979331	1591637	3774609	3293280	1932235	2281619	4109864	2053192	3222650	3484302	8326354	2814531	2397924	1726004	210950	272226	228854	784362	2145070	367450	929692	581463	861207	380629	208203	1591094	129376	384109	318128
Pyrogallol	553743	744444	652846	1720246	639136	764635	758907	396561	4939928	355328	632466	675741	1691898	1083293	403180	704256	497093	1348929	329015	863297	807331	1250541	557049	894730	377376	2336826	1063059	348348	753728	1622606	535143	922621	1791030	1421805	3160393	2545550	2228031	2957871	366968	2248041	4490761	830971	3153288	880685	892887	2545296	823507
Guanine 1	5083466	16524766	3858333	14826169	7861566	7684086	32457838	6451103	23913908	6873422	16112093	8629652	15098066	17166445	6035784	32127094	20526698	24856053	7531693	10100496	22707778	19206165	25555780	23124876	14144952	39754555	42752762	22045100	25192896	25861674	22430815	18648032	3005061	916876	1909996	891395	10556150	833882	3252411	1907809	6386336	1001654	6914576	1473671	2571878	1874729	1170259
4-Hydroxybenzaldehyde	2342380	1648910	2746039	2200001	4431563	4909765	2388461	2602968	2427587	2131080	1557778	3198298	2754546	3189742	2010441	3401355	3015686	1633543	3349944	2863218	4244508	3136337	3934620	2134485	3997195	2241734	3956007	4549753	8680142	3791852	3281335	2559292	1447810	299076	1188727	236329	5456296	314355	1558190	731605	1669423	382524	1426388	151128	1119620	1483495	304424
Pipecolic acid	938090	5243950	1314031	5242987	2511731	456648	6039278	3134035	5110195	1947453	4066087	1025649	3544344	6012966	3121905	4788771	471501	2216410	1673351	581568	629909	612671	1989587	2894038	1574057	1312241	1464418	1443487	1145801	883672	1881731	3133227	3033534	1740367	4955617	3584141	6287215	3606044	4558600	2932257	6835799	2611838	1828866	4208140	621470	1566262	1996700
Valine	319757737	170693973	360232994	219402717	482271080	214793861	240211459	239471519	220944129	216601877	159077403	406123554	242527651	347127152	259147600	306008070	240827375	193638501	315938084	322136071	380385709	266632751	400240114	209683088	417825278	222641807	329108576	423911046	777154260	381306283	367427826	248668949	125450374	107489618	134891771	102821917	491889145	99992340	147404147	116671053	148492511	95656634	119758396	44582918	112736393	153498313	101668040
Trigonelline	213957	1456291	203443	2158086	559450	206431	7714461	324954	9917427	161423	1051632	200353	1927961	1674787	444118	1145337	724510	1870820	474633	1576057	1405780	2620822	881541	1360014	487174	3366082	1777882	1782555	507873	2682950	894832	1105511	6563472	2142210	1247891	3761650	3912802	5081818	6613474	4845905	14649331	681017	3052853	542854	1472436	5996258	631170
Creatinine	72131192	45110365	69815067	44080349	119742354	159815289	54812739	67426446	32418963	56422229	50149637	46075246	54142538	99873190	57131913	60202072	46863468	40659462	53268004	54439821	82637885	45076142	61186411	33356150	94379553	32963415	64250841	93423183	143900292	98306128	70791590	35119937	14155210	14686174	20197355	25610833	16750330	26951997	17568019	36243855	32388489	6541625	12442844	8539999	8991064	16540853	8416172
Nicotinamide	35934764	56828880	33864636	56640889	18261026	55777820	53724207	45659805	87453188	40889850	80861050	43823512	53117512	86424107	39369304	104131704	58415830	3660175	57500085	33753578	46765293	25704918	65301945	58557744	19605977	29522876	25471125	32034501	42278862	26376127	48116588	59081239	14474320	37768619	13507042	19222337	26323900	15918997	18030378	37890539	41802899	10484097	22884382	15082186	28921308	19510018	17139619
Taurine	119018993	34259664	88101650	27912460	50579381	71892542	8083619	101749761	10529759	96359554	35564251	46898947	26180557	42519522	72115718	22508516	53358776	47221317	55361655	46424882	50737566	34360948	74218981	63780110	118819444	64439264	50904749	79241354	121993298	57010128	63965578	68561115	2384101	1485215	3663739	623470	38661421	656901	4353420	1741498	7637756	197870	1846642	125371	257471	2093028	244004
Leucylproline 1	1000634	2199479	1222355	3194029	2158494	1722218	2778442	1089921	1683869	1081893	1711057	784775	1791549	2504640	1804574	1875047	5688576	7731008	2652123	8319249	7484786	12685457	4576829	3988366	4014504	10047879	2816395	8766484	7174809	10344459	3352911	2875786	2545690	309209	3836192	637272	4855696	461641	9342566	548108	1919365	1779074	1336920	528291	2510098	6674252	1044672
Guanine 2	2673191	9636810	5537251	6371267	8813230	15280264	17878045	6163117	17927307	5996407	10083312	10076977	8400587	8456773	4080208	16799566	14606554	14067455	8809484	6099702	17910193	12638062	21245919	14576965	10582174	24080284	43014304	16078127	27502404	17535693	17057956	11384101	4273387	2024050	1799389	2065620	13699728	2663862	1888451	4726461	5217080	1588546	5626917	351355	5035453	1527413	1795483
L-2-Aminoadipic acid 1	4727755	1801440	8539203	2698022	16059118	24743895	2080806	3696388	3313813	5922803	1909221	9757607	4341590	3988962	4337852	5197569	9965655	3454219	13135260	3522724	8729251	3428909	9056838	2699469	13486815	1326463	17049229	5592248	14100764	4728481	12949444	4481495	347815	336891	544214	567225	9583585	423863	337397	318978	1677038	680392	1980514	522001	338322	470016	458648
D-(+)-Proline	1019886576	792268231	1303696601	1081584635	1430024932	1932497656	721606491	1334375116	1018056062	1001243019	733945153	1324676244	1159821301	1339363060	1134138380	1128609852	1042791424	1047250670	1228912927	1153255477	1318949889	1081239646	1637122683	1002095451	1407023924	1069943210	1327360146	1418937792	2137632520	1380136066	1347039248	1174309678	454995139	211059177	486361018	202140673	1895180124	213444855	668414770	360765768	749880556	185192878	546057524	68224174	364228388	501369389	219723100
L-Aspartic acid 1	51598768	19897056	56350354	18451956	55350613	95993741	22548185	51506344	20797021	38219105	23589516	68241386	30242434	44107517	17377703	44782876	19606183	13236527	26718159	19952150	31093143	22931380	26637133	20060066	48502229	14081971	38925907	35255166	87338144	32831562	47514444	27541557	245212	220128	471508	203595	17086644	235450	342789	199279	4270354	406099	2870731	1000507	232977	412943	637529
L-2-Aminoadipic acid 2	3939054	1256709	3240752	1946436	27888	6583959	1436450	3141468	1013809	5981828	1377694	4152944	2155885	3486450	4079667	1679306	6524561	4866540	8863023	4500888	620998	4377980	9851944	4081737	7796872	3692499	6512267	4118098	10311359	3704437	5211406	3158026	498186	908269	528886	1079455	2992725	874300	555478	200486	1460380	438028	896525	547921	393096	431908	294298
trans-3-Indoleacrylic acid	19764784	13391718	24558648	16247190	37850834	50878790	17874798	24272013	18216589	17627128	13042231	29132098	20848733	26221071	19171184	25396743	18642177	12135781	27627769	12699063	34419766	29495107	31972470	18089816	35229985	17429676	31124601	42241964	94411850	33144018	28847354	23157932	6312682	1383974	8955062	1704515	54189062	1903879	8246898	3747306	11100415	1536131	8014303	1183239	3191977	8780251	1519568
Methionine	63475329	50573373	67466832	63294219	95538381	114447985	53901994	58665990	69200688	54038521	47695998	71407704	67995399	83259392	49734928	84304424	94157604	82336181	77934664	103391765	135998158	107991979	87076516	55250963	124420209	92909796	95275991	137951017	169207562	123980439	70256429	58098374	45857362	5417637	35667245	4124458	197658746	3586882	52580122	10322504	45654095	27069783	44592602	10319631	27454945	33972830	23250713
2-Hydroxyphenylalanine	62984717	43205377	71016255	58148486	111355489	127599768	57958427	66740621	62130290	56867321	40305714	79729984	70785319	86753982	55596917	85024276	77000061	45166252	86180110	77451183	111000865	81105048	99490816	55950088	109945622	60197547	101472701	72263273	123700030	101385273	84097365	65803496	34486421	9503872	27782836	8352079	148309155	9708631	1271943	20729476	41355575	15637306	33677666	4150599	23954470	36129997	8381940
L-Pyroglutamic acid	209333413	151936560	204962576	170097071	178519156	207826024	92606511	202694408	160355975	186576127	167443218	225716124	195836384	92033131	209625105	196389341	140583097	120934731	202926573	112899989	132494024	117627806	237508444	183486857	188456566	148106711	230586407	181056000	299231895	170764316	244340018	234299789	6650145	1346962	11530239	2717037	120984694	1353682	8604868	3484093	48028185	9808257	27258746	12878953	5949825	7527105	11077348
Inosine	6766517	10162274	5506046	10544168	11499836	12540674	12812596	8607276	11594675	7594515	11058282	9710081	10636622	17350949	8878211	16330936	11095749	11294734	8996047	12696919	16647731	23674297	16118211	10743435	13606993	14429190	15969369	15186594	30448299	15081730	14448485	13234825	2513483	2477183	3069386	1579045	12308783	2160515	3521701	3460886	5574790	1246777	4592233	1309842	2767470	3022139	1783477
L-Alanyl-L-proline	2897348	4328171	2136003	4402242	5274081	2881759	2706677	1383152	412940	2238078	3303178	1410173	1963409	3683304	1735657	1687116	20255363	22054616	4935927	23301829	23702796	20977958	5951705	4235224	7173137	22186369	6976243	14272972	8153102	20642963	6050893	3803039	1301288	343330	1358806	500205	7447539	337247	2391025	383451	1952996	2341409	1518028	304743	2293977	779541	1300140
Propionylcarnitine	1262363	3048859	3630070	3439011	1056571	1783962	1103440	4084054	10369873	1037248	5017276	2196341	4394258	2890739	1279091	6893137	11531059	5478137	9165262	2815882	5492539	7874069	8508729	11961491	1974372	3555926	23063920	1046344	4331886	1414230	2723257	6228747	360882	1379333	212027	2610764	294532	1802464	760147	3312618	1527306	256912	702090	327951	321371	479763	403138
N-Acetyl-D-galactosamine	372861	3319116	452657	4553593	1750773	519497	13311155	723377	14758150	374692	2647245	424535	4069174	5385066	1080028	2820867	2629972	4781014	1656575	5252569	5348602	8316823	3275152	3977102	1936548	6596447	4689493	5596030	2550542	7581569	3129514	3468538	5677243	1345957	6592451	2561155	8418933	4524299	6730556	3350903	10203263	479263	14082013	250820	2143599	5349580	325167
L-Glutamic acid	530847833	365998392	512086035	429829678	456036566	537098013	366785218	501393522	398150208	453523789	407833215	553405059	483900541	43109722	537976809	481905077	348222602	311378873	265199302	289579275	339255921	299578263	588498573	448286881	490673076	377965484	578729822	467072269	758309117	351786045	228680089	574948101	15802210	2746739	28847375	4305091	310401331	2399369	21603843	7873098	121877809	18868874	62278459	25145715	12425029	18507599	26260809
DL-Glutamine 1	889803	677576	58698	660703	645513	493993	952665	328293	509379	737683	590022	258381	673968	635071	280320	154393	2892153	3149042	1074106	3065798	3978096	3554985	1058515	1287425	3025166	1161345	1797407	3674575	3099254	3473874	177775	732165	671002	30049	465293	409517	2866593	20139	926508	245477	578675	2783747	745578	2345432	1836052	553347	1588469
D-(+)-Tryptophan	16400569	7208482	20264935	13514196	31455042	42982905	3936666	19467975	14847430	14371864	7388694	23617284	16942785	22507641	16155756	20677132	15099478	10306212	22473319	19590827	29020573	24328290	25950284	14628613	30277697	14521971	25592688	35158289	76608820	28050129	23612135	18822987	5217830	1179363	7326966	1418763	46016755	1617501	6738309	3063970	9174919	1101669	6416723	915884	2480727	7121537	479089
Allopurinol	208379859	323495294	168268546	326662484	335361323	381498795	388201744	268273147	350196502	233828188	343335782	302021220	336568429	500043505	266783490	492381449	331783485	340754656	274950808	374186662	470413688	757201610	489953711	328287224	385773219	445709669	487357584	437601655	924773214	442129479	442251566	398819443	66562875	69544911	92852104	47401539	363417416	62544097	103849069	100512674	173023278	32523534	143686485	34233649	78013151	88352221	45597125
D-Carnitine	1019680622	530161064	1154637511	623619804	1316222992	1222110319	305459657	913406436	522132564	945451250	527176178	1103936675	732657738	937307942	788727231	833799688	620403880	201166795	714928634	458740255	924155434	443684438	732201888	339172987	1171265152	338844373	879810666	574597013	1636058156	192751643	964472475	541959013	27144921	20206611	23514484	16715576	255678020	17143293	36907311	25866859	67481057	8632084	34400103	6116189	12159948	32622450	13509008
Υ-Aminobutyric acid (GABA)	93404573	38909443	84034091	45771744	78199489	88772469	38950549	61737136	17289935	79468187	32254683	92309898	77134576	52432880	69560342	32634343	5871513	20012825	12479490	24308261	2145681	16771731	24117419	13144083	67929875	28135392	19157624	43222948	47242314	36704247	46901940	14588373	3723654	3417331	5125735	3180090	4787189	2759956	8917551	2653878	10269196	2750411	6350270	922609	2804129	5525785	6876689
DL-Glutamine 2	254065729	126099988	289057142	252035626	235003757	332283702	114227418	296821914	7147288	204591391	165393259	299045837	314755944	337993733	361403577	312872522	46784894	40908376	152423354	70504029	93108264	44379739	161153422	181255119	179845310	70861502	9006147	116042420	150347243	107306907	144799425	176972883	13203105	1170730	18768656	1738644	227534080	1169199	26730027	4057561	34560805	20615473	22877018	50756089	11782763	25249468	35099768
L-Threonine	36145091	41253751	37796925	45206366	52977792	61676282	54393982	35721809	39382811	4639271	25091783	30913872	41987833	56507483	36290267	37750402	33332018	47230008	44136238	67138453	73557919	69257200	56847581	37539259	51424615	54156350	46976746	61641036	86236955	70064062	45393679	44787779	9998293	4280417	18038496	9340734	74903191	6857904	16877590	7563967	42989693	6944668	27049190	9627292	7736437	13902156	11206682
Leucylproline 2	1961046	4161118	2565890	6323809	4178740	2140029	5374155	2844013	4283816	1916708	3699079	2412284	3426759	5315080	4234267	5273964	6068048	8100734	4878095	7545175	5754023	5605491	7154084	7973023	4764321	7218663	3307174	8468988	4231247	8874196	5542084	6001064	6768753	776670	7988734	1731125	10459976	1943723	18847267	2074023	5230944	2897189	4251556	813002	5164291	12643223	1824721
Leucylproline 3	1961046	4161118	2565890	6323809	4178740	2140029	5374155	2844013	4283816	1916708	3699079	2412284	3426759	5315080	4234267	5273964	6068048	8100734	4878095	7545175	5754023	5605491	7154084	7973023	4764321	7218663	3307174	8468988	4231247	8874196	5542084	6001064	6768753	776670	7988734	1731125	10459976	1943723	18847267	2074023	5230944	2897189	4251556	813002	5164291	12643223	1824721
Cytidine	153212	5651141	196885	6475549	1695786	305109	11766186	1661754	8145778	717509	5903721	1158952	4284021	11653402	3564332	9143512	4901753	9462756	2891518	7759185	3506071	8413816	6867795	8318672	142310	11788617	7572950	5442112	3249275	5653880	2188695	10096859	602719	150143	1178613	179279	684996	371893	1214005	355361	831802	99848	778947	570617	453912	1482168	308511
L-Norleucine	358598122	216759257	420991502	347006165	590631680	762123587	370061533	435842187	384863208	302707561	254614713	184997145	353563379	478838770	363727320	473898123	211447568	403743223	197179574	568219849	706492735	603780026	581358575	357631097	203391185	348992144	585793319	810666605	1199272950	248443403	198298603	410836565	186154407	94554861	239795501	85327093	910900166	106522847	242548604	171178369	205650251	110156047	143787130	55541737	181866124	267621064	120526737
L-Aspartic acid 2	18182039	19472579	51541191	18472315	53509158	95975797	23215745	51675141	20835193	38267877	23590315	68753144	30485154	41441210	29161141	45848959	19365394	12449413	26322158	19971861	30599305	22304932	25850958	18041425	47022689	23507568	36754140	35074786	87450259	31633990	45606346	27638201	245212	220128	471508	175048	17010560	189839	342789	182668	4052746	406099	2754228	1000507	232977	412943	637529
1,5-Naphthalenediamine	5292243	3447309	5380853	4694784	6871672	9454522	6269053	5958253	5337328	4568489	2041712	5508846	4495727	7947827	6551534	5867876	4259252	4191016	5579304	5782412	7056676	5912159	7013400	4746220	7831534	4992984	6092949	8332203	10798426	8197509	6447928	5121296	1657419	314343	2132241	423607	2144439	582674	1904181	1040277	3273827	389193	2177004	394742	918564	1986609	543617
Pyridoxal	463348	744444	652846	1720246	639136	764635	1658130	396561	4939928	355328	632466	675741	1691898	1083293	403180	704256	497093	1348929	329015	863297	807331	1250541	557049	894730	612113	2336826	1063059	952923	654281	1622606	535143	922621	1791030	1421805	3160393	2545550	2228031	2957871	1914787	2248041	4490761	830971	3153288	880685	892887	2545296	823507
(S)-(+)-2-Amino-1-propanol 1	9079306	4075962	14941949	5686266	12805283	34802667	3460489	365223	195457	7058906	4668936	11792866	2870203	10112618	220285	179821	527845	476314	884277	2671020	3679027	2230000	199259	212574	2565163	717787	1093269	4660922	11282994	3740299	442475	280572	471239	1012412	982614	1694936	9574178	2703813	231101	359732	705256	316144	1610223	366242	508764	234905	332750
Lactamide	120661337	177409569	272418423	200415702	284605445	359596008	144531777	258396832	160517110	241899321	131431280	294796712	222920993	283583717	179808695	230696846	217074725	108047240	239358002	214978084	283269091	206135539	268107536	82086012	148147983	201509719	254002364	263515946	409544281	232886720	261773399	145018625	58555130	21810333	51913699	24523860	265832457	24421495	91394074	42390963	122010214	40312930	79710323	20860988	35449578	76714947	44466516
Acetohydroxamic acid	16184011	11861232	21521954	12252967	18530715	31896293	590845	16658034	1676334	17515409	12357910	23098851	15544334	21639921	10043016	17542443	11624684	12926141	11721302	8164876	14157086	9231973	15882963	7605611	13310450	13812629	14961462	16520956	28112751	13658330	15013359	10265423	2904195	572357	4822951	2267514	549745	1953992	4206338	2388102	13353655	1649028	5479235	2120300	1604315	3812161	2196513
Dihydrothymine	93743697	43689537	97826432	45549719	79620938	161349082	20572369	110454601	27574864	83058644	55936307	122501160	68758253	90570521	128758958	107876227	78970641	30060812	64997540	77074589	98809290	60337198	98847850	44976134	190211162	72999319	89102365	89650768	190891528	80796294	117906322	60453137	25399537	4164258	28665035	4309154	27012041	4683564	34415889	15560662	47003103	1407429	27953352	1960713	1974656	43389387	1774405
Acetyl-β-methylcholine	13759935	7731313	14308164	7365178	1168450	22866214	5893320	1369038	5297159	10860826	8479444	13075804	9609229	13104630	7141483	8620624	11569711	5862273	11429493	10351141	17296072	10489075	7850837	1297487	22964279	7364840	14712794	16024520	36424392	12269144	15526140	9068169	1309508	973569	1666259	915463	16467290	1095260	1869139	1449238	4400703	86104	1452398	82250	147148	1206486	133301
Diethanolamine	20581244	19535544	21051019	18595392	39213959	63865837	30888009	28900392	22731719	18781093	21227074	34908602	19991517	26663190	16457287	26308476	13315504	17265472	16566409	18809663	16943869	25154806	20750619	19032554	24444329	16192594	21907863	20577258	25786292	28131904	20445642	15081397	12825065	34884634	19544999	30010080	22286703	24730947	16134285	29921661	21132678	43908091	13748995	33725616	14887375	17789798	42400607
(S)-(+)-2-Amino-1-propanol 2	31409826	21091330	47225800	32139134	92649561	185576971	9950368	32758433	35096633	39226982	25877153	97280405	47473754	48366005	23220218	87008903	16165252	12629545	15417801	17255076	24199016	26482186	33688418	21934028	54337100	32289809	59934986	65326286	40875926	50912644	40088070	36220677	2405873	3732234	2556162	3200984	8099997	3230771	1900372	1753552	2435837	2359226	2077736	2639763	2053245	2520010	2706692
N-Methyl-D-aspartic acid (NMDA)	1944921	1610444	2398535	2180246	1356188	1968627	1501864	2465887	2675926	1605082	3209036	2176995	1465400	3231312	2147035	2698142	667379	465603	1246182	389520	404157	513616	1047343	553281	550313	618942	247547	450937	273521	479838	1003610	562640	176918	661411	249488	802181	662845	314931	225351	403834	679174	517119	244493	418793	1313083	346665	565310
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	Formula	Molecular Weight	RT [min]	Library Match Factor (mzCloud)	Library Match Factor (mzVault)
Cuminaldehyde	C10 H12 O	148.08866	0.805	58.8
Sphingosine (d18:1)	C18 H37 N O2	299.28234	1.118	59.9	55.2
Urocanic acid	C6 H6 N2 O2	138.04271	1.138	68.4	56.7
DL-Lysine	C6 H14 N2 O2	146.1053	5.902		64.8
N-Benzylformamide	C8 H9 N O	135.06812	2.243	65.5	66.3
Acetyl-L-carnitine	C9 H17 N O4	203.11569	2.539	76.8	67.4
Pyrogallol	C6 H6 O3	126.03151	2.141		75.3
Guanine 1	C5 H5 N5 O	151.04902	2.068	88.9	76.4
4-Hydroxybenzaldehyde	C7 H6 O2	122.03647	2.247	68.9	76.4
Pipecolic acid	C6 H11 N O2	129.07873	5.9		78.4
Valine	C5 H11 N O2	117.07857	2.301	85.7	83.6
Trigonelline	C7 H7 N O2	137.04741	2.145	89.3	84.1
Creatinine	C4 H7 N3 O	113.05861	1.579	83.9	84.5
Nicotinamide	C6 H6 N2 O	122.04766	1.197	87.3	87.5
Taurine	C2 H7 N O3 S	125.01431	2.177	86.7	88.1
Leucylproline 1	C11 H20 N2 O3	228.14735	1.534	85.9	88.3
Guanine 2	C5 H5 N5 O	151.04912	1.847	91.3	88.7
L-2-Aminoadipic acid 1	C6 H11 N O4	161.06857	2.159	89.5	88.8
D-(+)-Proline	C5 H9 N O2	115.06281	2.639	85.6	90.6
L-Aspartic acid 1	C4 H7 N O4	133.03715	1.984	92.2	90.7
L-2-Aminoadipic acid 2	C6 H11 N O4	161.0686	1.657	89.8	91
trans-3-Indoleacrylic acid	C11 H9 N O2	187.06324	1.783	86.4	91.7
Methionine	C5 H11 N O2 S	149.05073	2.123	83.1	92.5
2-Hydroxyphenylalanine	C9 H11 N O3	181.07373	2.247	89.5	92.5
L-Pyroglutamic acid	C5 H7 N O3	129.04227	1.931	88.8	93
Inosine	C10 H12 N4 O5	268.08058	1.73	91.7	94
L-Alanyl-L-proline	C8 H14 N2 O3	186.10035	2.447	92.7	94.1
Propionylcarnitine	C10 H19 N O4	217.13135	2.173	90	95.2
N-Acetyl-D-galactosamine	C8 H15 N O6	221.08988	2.139	93.9	96.2
L-Glutamic acid	C5 H9 N O4	147.05276	1.939	88.9	97.1
DL-Glutamine 1	C5 H10 N2 O3	146.06896	2.017	76	97.2
D-(+)-Tryptophan	C11 H12 N2 O2	204.08979	1.786	95.8	97.4
Allopurinol	C5 H4 N4 O	136.03807	1.731	88.1	97.5
D-Carnitine	C7 H15 N O3	161.10484	3.466	97.4	97.6
Υ-Aminobutyric acid (GABA)	C4 H9 N O2	103.06305	3.872	77.7	97.6
DL-Glutamine 2	C5 H10 N2 O3	146.06891	4.39	92	98.6
L-Threonine	C4 H9 N O3	119.05802	4.389	92.1	98.7
Leucylproline 2	C11 H20 N2 O3	228.14739	1.286	91.7	99.3
Leucylproline 3	C11 H20 N2 O3	228.14739	1.286	91.7	99.3
Cytidine	C9 H13 N3 O5	243.08538	1.948	87.7	99.7
L-Norleucine	C6 H13 N O2	131.09422	1.978	89.7	100
L-Aspartic acid 2	C4 H7 N O4	133.03714	2.056	91.6	100
1,5-Naphthalenediamine	C10 H10 N2	158.08416	1.78	81.4
Pyridoxal	C8 H9 N O3	167.05809	2.144	52.4
(S)-(+)-2-Amino-1-propanol 1	C3 H9 N O	75.06815	3.199	52.7
Lactamide	C3 H7 N O2	89.04737	3.254	84.3
Acetohydroxamic acid	C2 H5 N O2	75.03181	3.548	68.4
Dihydrothymine	C5 H8 N2 O2	128.05833	3.56	50.6
Acetyl-β-methylcholine	C8 H17 N O2	159.12572	4.356	83.8
Diethanolamine	C4 H11 N O2	105.07874	7.11	85.4
(S)-(+)-2-Amino-1-propanol 2	C3 H9 N O	75.06816	9.332	54
N-Methyl-D-aspartic acid (NMDA)	C5 H9 N O4	147.05291	1.127	75.5
METABOLITES_END
#END