#METABOLOMICS WORKBENCH rifathamoudi_20190708_054947 DATATRACK_ID:1759 STUDY_ID:ST001265 ANALYSIS_ID:AN002102 PROJECT_ID:PR000850
VERSION             	1
CREATED_ON             	October 9, 2019, 11:43 am
#PROJECT
PR:PROJECT_TITLE                 	Comparative metabolomics of MCF-7 breast cancer cells using different extraction
PR:PROJECT_TITLE                 	solvents assessed by mass spectroscopy
PR:PROJECT_TYPE                  	Metabolomics using Mass Spectrometry
PR:PROJECT_SUMMARY               	Metabolic profiling of cancer cells can play an important role in revealing the
PR:PROJECT_SUMMARY               	molecular bases of cancer development and progression. In this work, gas
PR:PROJECT_SUMMARY               	chromatography coupled with mass spectrometry (GC-MS) was employed for the
PR:PROJECT_SUMMARY               	determination of signatures found in ER+/PR+ breast cancer cells derived from
PR:PROJECT_SUMMARY               	MCF-7 using different extraction solvents including: A, formic acid in water; B,
PR:PROJECT_SUMMARY               	ammonium hydroxide in water; C, ethyl acetate; D, methanol: water (1:1, v/v);
PR:PROJECT_SUMMARY               	and E, acetonitrile: water (1:1, v/v). The greatest extraction rate and
PR:PROJECT_SUMMARY               	diversity of metabolites occurs with extraction solvents A and E. Extraction
PR:PROJECT_SUMMARY               	solvent D showed moderate extraction efficiency, whereas extraction solvent B
PR:PROJECT_SUMMARY               	and C showed inferior metabolite diversity. Metabolite set enrichment analysis
PR:PROJECT_SUMMARY               	results showed energy production pathways to be key in MCF-7 cell lines. This
PR:PROJECT_SUMMARY               	study showed that mass spectrometry could identify key metabolites associated
PR:PROJECT_SUMMARY               	with cancers. The highest enriched pathways were related to energy production as
PR:PROJECT_SUMMARY               	well as Warburg effect pathways, which may shed light on how energy metabolism
PR:PROJECT_SUMMARY               	has been hijacked to encourage tumour progression and eventually metastasis in
PR:PROJECT_SUMMARY               	breast cancer.
PR:INSTITUTE                     	University of Sharjah
PR:DEPARTMENT                    	Sharjah Institute for Medical Research
PR:LABORATORY                    	Mohammad Semreen
PR:LAST_NAME                     	Hamoudi
PR:FIRST_NAME                    	Rifat
PR:ADDRESS                       	College of Medicine, University of Sharjah
PR:EMAIL                         	rhamoudi@sharjah.ac.ae
PR:PHONE                         	567154756
PR:FUNDING_SOURCE                	Al-Jalila Foundation (Grant No: AJF201741), Breast Cancer Now (Grant No:
PR:FUNDING_SOURCE                	2014MaySP323) and University of Sharjah
PR:PROJECT_COMMENTS              	Multidisciplinary based on mass spectrometry and bioinformatics
#STUDY
ST:STUDY_TITLE                   	Comparative metabolomics of MCF-7 breast cancer cells using different extraction
ST:STUDY_TITLE                   	solvents assessed by mass spectroscopy
ST:STUDY_TYPE                    	Analysing metabolomics using GC Mass Spectroscopy
ST:STUDY_SUMMARY                 	Metabolic profiling of cancer cells can play a vital role in revealing the
ST:STUDY_SUMMARY                 	molecular bases of cancer development and progression. In this study, gas
ST:STUDY_SUMMARY                 	chromatography coupled with mass spectrometry (GC-MS) was employed for the
ST:STUDY_SUMMARY                 	determination of signatures found in ER+/ PR+ breast cancer cells derived from
ST:STUDY_SUMMARY                 	MCF-7 using different extraction solvents including: A, formic acid in water; B,
ST:STUDY_SUMMARY                 	ammonium hydroxide in water; C, ethyl acetate; D, methanol: water (1:1, v/v);
ST:STUDY_SUMMARY                 	and E, acetonitrile: water (1:1, v/v). The greatest extraction rate and
ST:STUDY_SUMMARY                 	diversity of metabolites occurs with extraction solvents A and E. Extraction
ST:STUDY_SUMMARY                 	solvent D showed moderate extraction efficiency, whereas extraction solvent B
ST:STUDY_SUMMARY                 	and C showed inferior metabolite diversity. Metabolite set enrichment analysis
ST:STUDY_SUMMARY                 	results showed energy production pathways to be key in MCF-7 cell lines. This
ST:STUDY_SUMMARY                 	study showed that mass spectrometry could identify key metabolites associated
ST:STUDY_SUMMARY                 	with cancers. The highest enriched pathways were related to energy production as
ST:STUDY_SUMMARY                 	well as Warburg effect pathways, which may shed light on how energy metabolism
ST:STUDY_SUMMARY                 	has been hijacked to encourage tumour progression and eventually metastasis in
ST:STUDY_SUMMARY                 	breast cancer.
ST:INSTITUTE                     	Sharjah Institute for Medical Research
ST:DEPARTMENT                    	Clinical Science
ST:LAST_NAME                     	Hamoudi
ST:FIRST_NAME                    	Rifat
ST:ADDRESS                       	College of Medicine, University of Sharjah
ST:EMAIL                         	rhamoudi@sharjah.ac.ae
ST:PHONE                         	567154756
ST:TOTAL_SUBJECTS                	Five different extractions
ST:STUDY_COMMENTS                	MCF-7 cell line
#SUBJECT
SU:SUBJECT_TYPE                  	Cultured cells
SU:SUBJECT_SPECIES               	Homo sapiens
SU:TAXONOMY_ID                   	9606
SU:GENDER                        	Female
SU:CELL_BIOSOURCE_OR_SUPPLIER    	ATCC
SU:CELL_STRAIN_DETAILS           	MCF-7
SU:CELL_PASSAGE_NUMBER           	30
SU:SPECIES_GROUP                 	Homo Sapien
#FACTORS
#SUBJECT_SAMPLE_FACTORS:         	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data
SUBJECT_SAMPLE_FACTORS           	Breast cancer	FA	Exctraction:0.2% Formic acid in water	
SUBJECT_SAMPLE_FACTORS           	Breast cancer	NH4OH	Exctraction:0.2% Ammonium hydroxide in water	
SUBJECT_SAMPLE_FACTORS           	Breast cancer	MeOH-H2O	Exctraction:Methanol/Water	
SUBJECT_SAMPLE_FACTORS           	Breast cancer	ACN-H2O	Exctraction:Acetonitrile/Water	
SUBJECT_SAMPLE_FACTORS           	Breast cancer	EthAcet	Exctraction:Ethyl acetate	
#COLLECTION
CO:COLLECTION_SUMMARY            	The breast adenocarcinoma cells (MCF-7) were cultured in DMEM medium
CO:COLLECTION_SUMMARY            	supplemented with 10% fetal calf serum and 1% penicillin/streptomycin solution
CO:COLLECTION_SUMMARY            	and incubated at 37oC in an atmosphere of 5% CO2. In preparation for an
CO:COLLECTION_SUMMARY            	experiment, 3 × 106 cells were cultured in each of 15 tissue culture flasks
CO:COLLECTION_SUMMARY            	(175 cm2) and cells were incubated for three days. When the cells reach 80%
CO:COLLECTION_SUMMARY            	confluency, they were collected by trypsinization, counted in a cell counter and
CO:COLLECTION_SUMMARY            	each 3 ×106 cells were suspended in 1 ml phosphate-buffered saline in an
CO:COLLECTION_SUMMARY            	Eppendorf tube.
CO:SAMPLE_TYPE                   	Breast cancer cells
CO:COLLECTION_METHOD             	trypsinization protocol
CO:STORAGE_CONDITIONS            	-80℃
#TREATMENT
TR:TREATMENT_SUMMARY             	The aim of this study was to investigate the solvent extraction efficiency, so
TR:TREATMENT_SUMMARY             	there was no cell treatment
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	A total of 5 triplicated MCF-7 cell culture samples were provided in Eppendorf
SP:SAMPLEPREP_SUMMARY            	vials dissolved in PBS, and stored at 4 °C for preservation purposes. Samples
SP:SAMPLEPREP_SUMMARY            	were centrifuged at 13000 rpm for 10 minutes at -4 °C. Supernatant was
SP:SAMPLEPREP_SUMMARY            	discarded, and cell pellets, each containing 3 million cells, were subjected to
SP:SAMPLEPREP_SUMMARY            	metabolomics analysis. In order to evaluate the influence of the solvents on the
SP:SAMPLEPREP_SUMMARY            	extraction rate, samples were divided in five different extraction groups: A,
SP:SAMPLEPREP_SUMMARY            	0.2 % formic acid in water; B, 0.2 % ammonium hydroxide in water; C, ethyl
SP:SAMPLEPREP_SUMMARY            	acetate; D, methanol/water (1:1, v/v); and E, acetonitrile/water (1:1, v/v).
SP:SAMPLEPREP_SUMMARY            	Briefly, 300 µL of the extraction solvent was added to 3 million cell pellets,
SP:SAMPLEPREP_SUMMARY            	then vortexed for 2 minutes. All samples have been stored in ice for 1 hour,
SP:SAMPLEPREP_SUMMARY            	during which samples have been vortexed every 15 minutes. After this, cell
SP:SAMPLEPREP_SUMMARY            	insoluble matrix was centrifuged (13000 rpm, 10 minutes, -4 °C). Supernatant
SP:SAMPLEPREP_SUMMARY            	was collected then dried using EZ-2 Plus (GeneVac, Ipswich, UK) at 37 ± 1 °C.
SP:SAMPLEPREP_SUMMARY            	To detect all amino acids and metabolites, all samples were derivatized with
SP:SAMPLEPREP_SUMMARY            	methoxyamine hydrochloride and MSTFA + 1% TMCS prior to injection to GC-MS.
SP:PROCESSING_STORAGE_CONDITIONS 	Described in summary
SP:EXTRACTION_METHOD             	Direct extraction
SP:EXTRACT_ENRICHMENT            	described in Summary
SP:EXTRACT_STORAGE               	Described in summary
SP:SAMPLE_DERIVATIZATION         	methoxyamine hydrochloride and MSTFA + 1% TMCS
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	A GC/MS-QP 2010 Ultra System (Shimadzu, Kyoto, Japan) was employed for the
CH:CHROMATOGRAPHY_SUMMARY        	metabolomic analysis, along with LabSolutions GC-MS software (ver). A Restek
CH:CHROMATOGRAPHY_SUMMARY        	Rtx®-5ms column (30.0 m × 0.25 mm, 0.25 µm) was utilized for separation of
CH:CHROMATOGRAPHY_SUMMARY        	the metabolites. Helium (99.9%) was used as the carrier gas at the flow rate of
CH:CHROMATOGRAPHY_SUMMARY        	1.0 mL/min. The initial oven temperature was set at 60 °C and was held at this
CH:CHROMATOGRAPHY_SUMMARY        	temperature for 2 minutes, then raised to 310 °C by 50 °C/min and held at this
CH:CHROMATOGRAPHY_SUMMARY        	temperature during the analysis. Both the auxiliary temperature at the interface
CH:CHROMATOGRAPHY_SUMMARY        	and the ionization temperature were 250 °C. Metabolites were analysed in full
CH:CHROMATOGRAPHY_SUMMARY        	scan mode within the range of 50 – 650 amu. Total volume of 10 µL was
CH:CHROMATOGRAPHY_SUMMARY        	injected in splitless mode, by employing AOC-20i Auto Injector (Shimadzu, Kyoto,
CH:CHROMATOGRAPHY_SUMMARY        	Japan). GC total ion chromatograms (TIC) and fragmentation patterns of the
CH:CHROMATOGRAPHY_SUMMARY        	metabolites identified using NIST/EPA/NIH Mass Spectral Library (NIST 14). Run
CH:CHROMATOGRAPHY_SUMMARY        	time for each sample was 43.67 min.
CH:CHROMATOGRAPHY_TYPE           	GC
CH:INSTRUMENT_NAME               	Shimadzu GCMS-QP2010 Ultra
CH:COLUMN_NAME                   	Restek Rtx-5Sil MS (30 x 0.25mm, 0.25um)
CH:FLOW_RATE                     	1
CH:COLUMN_TEMPERATURE            	60
CH:METHODS_FILENAME              	PAH scan
CH:SOLVENT_A                     	Pyridine
CH:COLUMN_PRESSURE               	57.4
CH:INJECTION_TEMPERATURE         	250
CH:RETENTION_TIME                	many retention time
CH:SAMPLE_INJECTION              	1
CH:ANALYTICAL_TIME               	43.67
CH:OVEN_TEMPERATURE              	60
CH:TIME_PROGRAM                  	43.67
CH:TRANSFERLINE_TEMPERATURE      	250
CH:STRONG_WASH_SOLVENT_NAME      	Acetone
CH:STRONG_WASH_VOLUME            	10 uL
CH:SAMPLE_SYRINGE_SIZE           	10 uL
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
AN:LABORATORY_NAME               	Integrated Analytical lab
AN:OPERATOR_NAME                 	Muath
AN:DETECTOR_TYPE                 	MS
AN:SOFTWARE_VERSION              	GCMS solution
AN:DATA_FORMAT                   	qgd
#MS
MS:INSTRUMENT_NAME               	Shimadzu QP2010 Ultra
MS:INSTRUMENT_TYPE               	Single quadrupole
MS:MS_TYPE                       	EI
MS:ION_MODE                      	POSITIVE
MS:MS_COMMENTS                   	A GC/MS-QP 2010 Ultra System (Shimadzu, Kyoto, Japan) was employed for the
MS:MS_COMMENTS                   	metabolomic analysis, along with LabSolutions GC-MS software (ver). A Restek
MS:MS_COMMENTS                   	Rtx®-5ms column (30.0 m × 0.25 mm, 0.25 µm) was utilized for separation of
MS:MS_COMMENTS                   	the metabolites. Helium (99.9%) was used as the carrier gas at the flow rate of
MS:MS_COMMENTS                   	1.0 mL/min. The initial oven temperature was set at 60 °C and was held at this
MS:MS_COMMENTS                   	temperature for 2 minutes, then raised to 310 °C by 50 °C/min and held at this
MS:MS_COMMENTS                   	temperature during the analysis. Both the auxiliary temperature at the interface
MS:MS_COMMENTS                   	and the ionization temperature were 250 °C. Metabolites were analysed in full
MS:MS_COMMENTS                   	scan mode within the range of 50 – 650 amu. Total volume of 10 µL was
MS:MS_COMMENTS                   	injected in splitless mode, by employing AOC-20i Auto Injector (Shimadzu, Kyoto,
MS:MS_COMMENTS                   	Japan). GC total ion chromatograms (TIC) and fragmentation patterns of the
MS:MS_COMMENTS                   	metabolites identified using NIST/EPA/NIH Mass Spectral Library (NIST 14). Run
MS:MS_COMMENTS                   	time for each sample was 43.67 min.
MS:GAS_PRESSURE                  	57.4 kPa
MS:HELIUM_FLOW                   	1 ml/min
MS:ION_SOURCE_TEMPERATURE        	250
MS:IONIZATION_ENERGY             	70
MS:SOURCE_TEMPERATURE            	250
MS:SCANNING_RANGE                	50-650
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	Peak Area
MS_METABOLITE_DATA_START
Samples	NH4OH	EthAcet	FA	MeOH-H2O	ACN-H2O
Factors	Exctraction:0.2% Ammonium hydroxide in water	Exctraction:Ethyl acetate	Exctraction:0.2% Formic acid in water	Exctraction:Methanol/Water	Exctraction:Acetonitrile/Water
Boric acid, 3TMS derivative	0	384617821	23321619	0	23321619
Lactic Acid, 2TMS derivative	21342253	0	135115766	228585341	155779670
Glycolic acid, 2TMS derivative	0	0	225444	266851	278303
1H-Azonine, octahydro-1-nitroso-	0	0	0	710926	998300
L-Valine, TMS derivative	0	0	2448318	1184688	1473123
Alanine, 2TMS derivative	0	0	4899056	1696802	2167166
Ethylene glycol, 2TMS derivative	0	0	0	0	3805783
Glycine, 2TMS derivative	0	0	2082755	1032492	1615541
3-Pyridinol, TMS derivative	0	0	730440	0	970561
Silane, [(1-methoxy-1,3-propanediyl)bis(oxy)]bis[trimethyl-	0	7024276	0	0	14886287
L-Leucine, TMS derivative	0	400788	11211540	5302656	4401764
4-Hydroxybutanoic acid, 2TMS derivative	0	0	0	0	596333
L-Proline, TMS derivative	0	0	4646866	0	2484408
L-Isoleucine, TMS derivative	0	664503	4412840	5206709	5495186
Trisiloxane, 1,1,1,5,5,5-hexamethyl-3,3-bis[(trimethylsilyl)oxy]-	0	0	3735642	4673349	2831229
N-Acetylaminoethanol, TMS derivative	0	0	0	0	1169597
L-Valine, 2TMS derivative	0	0	0	0	833561
Benzoic Acid, TMS derivative	0	0	0	0	357662
3-Ethyl-3-methylheptadecane	0	0	0	0	216819
Diethylene glycol, 2TMS derivative	0	0	879981	0	508831
4-Aminobutanoic acid, 3TMS derivative	0	0	0	0	15822091
Glycerol, 3TMS derivative	9011155	4358175	0	97130865	37655907
L-Threonine, 2TMS derivative	0	0	2599095	2599095	2587250
3-Aminoisobutyric acid, 3TMS derivative	0	992691	2686471	0	3447730
Butanedioic acid, 2TMS derivative	0	845520	4746815	4151836	4654901
Cyclohexasiloxane, dodecamethyl-	8928220	2261580	2425750	0	973227
2-Butenedioic acid, (E)-, 2TMS derivative	0	582703	1528801	1871099	2498902
Glyceric acid, 3TMS derivative	0	677598	1205776	862986	1261658
Myristic acid, TMS derivative	0	0	0	0	967775
Ethylphosphonic acid, 2TMS derivative	0	0	2199159	2564961	2825696
6-Dimethyl(trimethylsilyl)silyloxytetradecane	0	0	157392	0	578909
Thiodiglycol, 2TMS derivative	1784213	0	10610738	10115886	8177157
.beta.-Alanine, 3TMS derivative	0	0	0	0	1011004
L-Aspartic acid, 2TMS derivative	0	0	13323823	0	11095137
2,4'-Bipyridine	0	0	0	0	1093634
L-Hydroxyproline, (E)-, 2TMS derivative	0	0	8449003	0	5809140
Succinic acid, 3,5-difluorophenyl 2-(dimethylamino)ethyl ester	0	0	8494069	0	12240640
1-Hexanol, 5-methyl-2-(1-methylethyl)-	0	0	0	0	170485
Malic acid, 3TMS derivative	0	0	32721975	17389740	30825026
Pyroglutamic acid, TMS derivative	0	0	5508543	9582395	8720110
(2-Ethoxyethoxy)acetic acid, TMS derivative	0	0	3588884	0	3591291
L-5-Oxoproline, , 2TMS derivative	0	20064518	21049984	2630123	18654026
Acetoin, TMS derivative	0	0	0	0	2021585
DL-Phenylalanine, TMS derivative	0	0	7999166	0	2458644
2,3,4-Trihydroxybutyric acid tetrakis	0	0	4707336	5996779	6242033
Pentanedioic acid, 2-[(trimethylsilyl)oxy]-, bis(trimethylsilyl) ester	0	0	734124	0	1354400
L-Proline, 2TMS derivative	0	0	1226575	0	1990273
Amphetamine, 2TMS derivative	0	0	0	0	1098751
Pimelic acid, 2TMS derivative	0	348700	0	0	546659
Triethanolamine, 3TMS derivative	0	0	13106735	12976871	10835385
Dodecanoic acid, TMS derivative	0	821367	0	0	2405771
2,2'-Sulfinyldiethanol, 2TMS derivative	0	0	0	985497	952592
Propanoic acid, 3-[methyl[2-oxo-2-[(trimethylsilyl)oxy]ethyl]amino]-3-oxo-, trimethylsilyl ester	0	0	0	985497	9441883
Levoglucosan, 3TMS derivative	0	0	2171042	0	2158786
Erythro-Pentonic acid, 2-deoxy-3,4,5-tris-O-(trimethylsilyl)-, trimethylsilyl ester	0	0	2934564	2455636	3302572
Nonadecane	0	0	0	727665	941706
Phosphoric acid, bis(trimethylsilyl) 2,3-bis[(trimethylsilyl)oxy]propyl ester	0	1494950	24335810	11923779	18037128
Phosphorylethanolamine, 4TMS derivative	0	0	0	0	7396066
Citric acid, 4TMS derivative	0	0	8083869	0	6263400
D-(+)-Ribono-1,4-lactone, 3TMS derivative	0	0	0	60622872	62588114
Deoxyglucose, 4TMS derivative	0	0	0	0	26071453
Sulfurous acid, decyl 2-propyl ester	0	0	0	0	503838
.beta.-D-Galactofuranoside, ethyl 2,3,5,6-tetrakis-O-(trimethylsilyl)-	0	0	0	0	469232
D-Pinitol, pentakis(trimethylsilyl) ether	14634505	0	0	45401248	38510171
D-(-)-Tagatose, pentakis(trimethylsilyl) ether, methyloxime (anti)	0	0	0	0	3568082
Glucopyranose, 5TMS derivative	0	23.356	21798540	23.386	13472478
Pentadecanoic acid, TMS derivative	0	0	0	0	1278013
d-Glucose, 2,3,4,5,6-pentakis-O-(trimethylsilyl)-, o-methyloxyme, (1E)-	0	0	22356914	0	9635809
L-Tyrosine, 3TMS derivative	0	913608	0	0	2263508
d-Mannose, 2,3,4,5,6-pentakis-O-(trimethylsilyl)-, o-methyloxyme, (1E)-	3278546	0	0	0	3010342
Diisooctyl phthalate	0	0	0	0	4019977
Palmitic Acid, TMS derivative	48574221	21900783	172160405	390605918	306689365
D-Gluconic acid, 6TMS derivative	0	0	5370617	0	4991403
2(1H)-Pyrimidinone, 1-[2,3-bis-O-(trimethylsilyl)-.beta.-D-ribofuranosyl]-4-(trimethylsiloxy)-, 5'-[bis(trimethylsilyl) phospha	0	0	8018536	0	3972490
Scyllo-Inositol, 6TMS derivative	0	0	0	616640	1783065
Octadecane, 5-methyl-	0	0	0	0	1439130
Heptadecanoic acid, TMS derivative	0	0	8018536	0	932062
Octadecane-1,2-diol, 2TMS derivative	0	5427308	9552163	8676122	7414257
Cyclopentanemethanol, .alpha.-cyclohexyl-2-nitro-	0	0	0	0	1437515
2-Methyltetracosane	0	0	0	2711590	1089644
Spermine, 6TMS derivative	0	0	27.304	0	1872030
Heptadecyl acetate	4307192	0	0	14309998	4140022
9,12-Octadecadienoic acid (Z,Z)-, TMS derivative	0	0	21675714	20762372	20037127
Oleic Acid, (Z)-, TMS derivative	0	4107097	9953575	9608737	7384438
Stearic acid, TMS derivative	36067511	16141552	130881548	263133932	222010162
N,N-Dimethyldodecanamide	7140680	0	0	8369157	4800988
18-Methyl-nonadecanol, trimethylsilyl ether	0	0	0	385256	1002285
D-Galactofuranose, 1,2,3,5-tetrakis-O-(trimethylsilyl)-, bis(trimethylsilyl) phosphate	0	0	7305680	0	4247381
d-Glucose, 2,3,4,5-tetrakis-O-(trimethylsilyl)-, o-methyloxime, 6-[bis(trimethylsilyl) phosphate]	0	0	2295652	0	2390988
1-Monomyristin, 2TMS derivative	0	0	20073580	16199139	14360122
Sebacic acid, 2TMS derivative	0	0	0	0	2221057
Octasiloxane, 1,1,3,3,5,5,7,7,9,9,11,11,13,13,15,15-hexadecamethyl-	15057648	3216188	10707270	12003544	5396325
Eicosanoic acid, TMS	0	0	0	0	6318356
Phenol, 2,2'-methylenebis[6-(1,1-dimethylethyl)-4-methyl-	53356073	0	0	11653162	1520808
Myo-Inositol, 1,3,4,5,6-pentakis-O-(trimethylsilyl)-, bis(trimethylsilyl) phosphate	0	0	11647390	6007297	7284716
2-Methyl-6-tert-butylphenol, TMS derivative	108732894	0	61793387	95799753	56096509
benzene, 1,1'-methylenebis[3-(1,1-dimethylethyl)-5-methyl-4-[(trimethylsilyl)oxy]-	0	0	28567659	20333567	42673697
1-Monopalmitin, 2TMS derivative	0	430815	748222288	655278547	725688268
5-Methyluridine, 3TMS derivative	0	0	0	0	2665893
Adenosine, 4TMS derivative	0	0	2851465	0	10346826
Heptadecanoic acid, glycerine-(1)-monoester, bis-O-trimethylsilyl-	0	0	2675410	2047385	1889228
1,1,1,5,7,7,7-Heptamethyl-3,3-bis(trimethylsiloxy)tetrasiloxane	0	0	8216347	7121779	6856043
2-Monostearin, 2TMS derivative	0	0	3587058	2952262	1864294
Glycerol monostearate, 2TMS derivative	71396128	0	352315760	302379487	355277040
9-Octadecenamide, (Z)-	22103423	0	0	0	53146417
2,3-Dihydroxypropyl icosanoate, 2TMS derivative	0	0	4720541	3980275	3565644
Cholesterol, TMS derivative	17827559	70835367	3066638	12102866	8293218
Stigmasterol, TMS derivative	0	0	658466	800923	674722
Stigmast-5-ene, 3.beta.-(trimethylsiloxy)-, (24S)-	0	0	2194388	2708608	2036593
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	Pubchem_Derivative	Pubchem ID
Boric acid, 3TMS derivative	78020	7628
Lactic Acid, 2TMS derivative	519450	107689
Glycolic acid, 2TMS derivative	118503	757
1H-Azonine, octahydro-1-nitroso-	30361	13287582
L-Valine, TMS derivative	22211754	6287
Alanine, 2TMS derivative	11424808	5950
Ethylene glycol, 2TMS derivative	81858	174
Glycine, 2TMS derivative	530141	750
3-Pyridinol, TMS derivative	592532	7971
Silane, [(1-methoxy-1,3-propanediyl)bis(oxy)]bis[trimethyl-	553103	23953
L-Leucine, TMS derivative	12028662	6106
4-Hydroxybutanoic acid, 2TMS derivative	589994	7302
L-Proline, TMS derivative	13514636	145742
L-Isoleucine, TMS derivative	19003238	6306
Trisiloxane, 1,1,1,5,5,5-hexamethyl-3,3-bis[(trimethylsilyl)oxy]-	19086	17860093
N-Acetylaminoethanol, TMS derivative	15644713	14960462
L-Valine, 2TMS derivative	11108121	6287
Benzoic Acid, TMS derivative	74955	243
3-Ethyl-3-methylheptadecane	11536486	11536486
Diethylene glycol, 2TMS derivative	552860	8117
4-Aminobutanoic acid, 3TMS derivative	119	119
Glycerol, 3TMS derivative	522285	753
L-Threonine, 2TMS derivative	91696554	6288
3-Aminoisobutyric acid, 3TMS derivative	553272	64956
Butanedioic acid, 2TMS derivative	520988	1110
Cyclohexasiloxane, dodecamethyl-	10911	9903713
2-Butenedioic acid, (E)-, 2TMS derivative	5353016	444266
Glyceric acid, 3TMS derivative	520886	439194
Myristic acid, TMS derivative	519592	11005
Ethylphosphonic acid, 2TMS derivative	519266	204482
6-Dimethyl(trimethylsilyl)silyloxytetradecane	590061	590061
Thiodiglycol, 2TMS derivative	519784	5447
.beta.-Alanine, 3TMS derivative	521597	239
L-Aspartic acid, 2TMS derivative	85830684	5960
2,4'-Bipyridine	68488	68488
L-Hydroxyproline, (E)-, 2TMS derivative	594032	5810
Succinic acid, 3,5-difluorophenyl 2-(dimethylamino)ethyl ester	91693828	1110
1-Hexanol, 5-methyl-2-(1-methylethyl)-	96291	8103
Malic acid, 3TMS derivative	522155	525
Pyroglutamic acid, TMS derivative	14038131	7405
(2-Ethoxyethoxy)acetic acid, TMS derivative	552152	192850
L-5-Oxoproline, , 2TMS derivative	7405	7405
Acetoin, TMS derivative	12739199	179
DL-Phenylalanine, TMS derivative	577739	994
2,3,4-Trihydroxybutyric acid tetrakis	439535	439535
Pentanedioic acid, 2-[(trimethylsilyl)oxy]-, bis(trimethylsilyl) ester	521646	743
L-Proline, 2TMS derivative	522501	145742
Amphetamine, 2TMS derivative	604670	3007
Pimelic acid, 2TMS derivative	521645	385
Triethanolamine, 3TMS derivative	123394	7618
Dodecanoic acid, TMS derivative	521640	3893
2,2'-Sulfinyldiethanol, 2TMS derivative	552844	18330
Propanoic acid, 3-[methyl[2-oxo-2-[(trimethylsilyl)oxy]ethyl]amino]-3-oxo-, trimethylsilyl ester	552467	1032
Levoglucosan, 3TMS derivative	91696777	2724705
Erythro-Pentonic acid, 2-deoxy-3,4,5-tris-O-(trimethylsilyl)-, trimethylsilyl ester	528638	90658608
Nonadecane	12401	12401
Phosphoric acid, bis(trimethylsilyl) 2,3-bis[(trimethylsilyl)oxy]propyl ester	553774	1004
Phosphorylethanolamine, 4TMS derivative	553761	1015
Citric acid, 4TMS derivative	553132	311
D-(+)-Ribono-1,4-lactone, 3TMS derivative	111064	111064
Deoxyglucose, 4TMS derivative	553409	108223
Sulfurous acid, decyl 2-propyl ester	6420364	1100
.beta.-D-Galactofuranoside, ethyl 2,3,5,6-tetrakis-O-(trimethylsilyl)-	553474	22215147
D-Pinitol, pentakis(trimethylsilyl) ether	14534973	164619
D-(-)-Tagatose, pentakis(trimethylsilyl) ether, methyloxime (anti)	519639	92092
Pentadecanoic acid, TMS derivative	523679	13849
d-Glucose, 2,3,4,5,6-pentakis-O-(trimethylsilyl)-, o-methyloxyme, (1E)-	522260	522260
L-Tyrosine, 3TMS derivative	14189425	6057
d-Mannose, 2,3,4,5,6-pentakis-O-(trimethylsilyl)-, o-methyloxyme, (1E)-	629230	5378916
Diisooctyl phthalate	33934	33934
Glucopyranose, 5TMS derivative	13587619	5793
Palmitic Acid, TMS derivative	521638	985
D-Gluconic acid, 6TMS derivative	520681	10690
2(1H)-Pyrimidinone, 1-[2,3-bis-O-(trimethylsilyl)-.beta.-D-ribofuranosyl]-4-(trimethylsiloxy)-, 5'-[bis(trimethylsilyl) phospha	91696709	68401
Scyllo-Inositol, 6TMS derivative	520232	892
Octadecane, 5-methyl-	520183	11635
Heptadecanoic acid, TMS derivative	521635	10465
Octadecane-1,2-diol, 2TMS derivative	91742674	89314
Cyclopentanemethanol, .alpha.-cyclohexyl-2-nitro-	565562	77195
2-Methyltetracosane	527459	527459
Spermine, 6TMS derivative	599851	1103
Heptadecyl acetate	69967	69967
9,12-Octadecadienoic acid (Z,Z)-, TMS derivative	5352430	5379173
Oleic Acid, (Z)-, TMS derivative	5366433	445639
Stearic acid, TMS derivative	87777	5281
N,N-Dimethyldodecanamide	18159	18159
18-Methyl-nonadecanol, trimethylsilyl ether	91743660	91743660
D-Galactofuranose, 1,2,3,5-tetrakis-O-(trimethylsilyl)-, bis(trimethylsilyl) phosphate	553776	553776
d-Glucose, 2,3,4,5-tetrakis-O-(trimethylsilyl)-, o-methyloxime, 6-[bis(trimethylsilyl) phosphate]	90473467	90473467
1-Monomyristin, 2TMS derivative	631192	79050
Sebacic acid, 2TMS derivative	519558	5192
Octasiloxane, 1,1,3,3,5,5,7,7,9,9,11,11,13,13,15,15-hexadecamethyl-	6329087	6329087
Eicosanoic acid, TMS	521648	10467
Phenol, 2,2'-methylenebis[6-(1,1-dimethylethyl)-4-methyl-	8398	8398
Myo-Inositol, 1,3,4,5,6-pentakis-O-(trimethylsilyl)-, bis(trimethylsilyl) phosphate	553143	553143
2-Methyl-6-tert-butylphenol, TMS derivative	91696816	16678
benzene, 1,1'-methylenebis[3-(1,1-dimethylethyl)-5-methyl-4-[(trimethylsilyl)oxy]-	91697106	91697106
1-Monopalmitin, 2TMS derivative	552033	3084463
5-Methyluridine, 3TMS derivative	524921	445408
Adenosine, 4TMS derivative	14216913	60961
Heptadecanoic acid, glycerine-(1)-monoester, bis-O-trimethylsilyl-	633500	633500
1,1,1,5,7,7,7-Heptamethyl-3,3-bis(trimethylsiloxy)tetrasiloxane	6329081	6329081
2-Monostearin, 2TMS derivative	582132	79075
Glycerol monostearate, 2TMS derivative	553936	79075
9-Octadecenamide, (Z)-	5283387	5283387
2,3-Dihydroxypropyl icosanoate, 2TMS derivative	537899	3246944
Cholesterol, TMS derivative	102173	11025495
Stigmasterol, TMS derivative	91698162	5280794
Stigmast-5-ene, 3.beta.-(trimethylsiloxy)-, (24S)-	91714198	21771614
METABOLITES_END
#END