#METABOLOMICS WORKBENCH SIMR_Core_Facility_20230926_225739 DATATRACK_ID:4337 STUDY_ID:ST002910 ANALYSIS_ID:AN004779 PROJECT_ID:PR001810
VERSION             	1
CREATED_ON             	October 3, 2023, 1:12 pm
#PROJECT
PR:PROJECT_TITLE                 	Identifying the impact of RelA overexpression in triple negative breast cancer
PR:PROJECT_TITLE                 	cells using mass spectroscopy-based metabolomics analysis
PR:PROJECT_TYPE                  	LC-MS/MS
PR:PROJECT_SUMMARY               	In this research study, metabolomics alterations associated with the
PR:PROJECT_SUMMARY               	overexpression of RelA in the TNBC cell line, MDA-MB-231, were investigated
PR:PROJECT_SUMMARY               	using liquid chromatography-quadrupole time-of-flight mass spectrometry
PR:PROJECT_SUMMARY               	(UHPLC-QTOF-MS) technique. UHPLC-QTOF-MS is a rapid, sensitive, high-resolution
PR:PROJECT_SUMMARY               	mass spectrometry technology that detects and identifies fragment patterns of
PR:PROJECT_SUMMARY               	compounds in complex samples. This analysis would unravel the key metabolites
PR:PROJECT_SUMMARY               	modified due to RelA overexpression, thus highlighting the altered signaling
PR:PROJECT_SUMMARY               	pathways behind the aggressiveness and chemoresistance of TNBC through NF-κB
PR:PROJECT_SUMMARY               	alteration.
PR:INSTITUTE                     	Sharjah Institute for Medical Research
PR:LAST_NAME                     	Facility
PR:FIRST_NAME                    	Core
PR:ADDRESS                       	M32, SIMR, College of Pharmacy, Health Sciences, University of Sharjah
PR:EMAIL                         	tims-tof@sharjah.ac.ae
PR:PHONE                         	065057656
#STUDY
ST:STUDY_TITLE                   	Identifying the impact of RelA overexpression in triple negative breast cancer
ST:STUDY_TITLE                   	cells using mass spectroscopy-based proteomics and metabolomics analysis
ST:STUDY_SUMMARY                 	Introduction: The incidence of chemotherapeutic resistance among breast cancer
ST:STUDY_SUMMARY                 	patients continues to rise steadily. The expression of RelA, a prominent member
ST:STUDY_SUMMARY                 	of the Rel family, is closely associated with the aggressiveness of
ST:STUDY_SUMMARY                 	triple-negative breast cancer (TNBC) and a poor prognosis for patients.
ST:STUDY_SUMMARY                 	Consequently, it is crucial to deeply investigate the molecular changes that
ST:STUDY_SUMMARY                 	underlie breast cancer progression and chemotherapy resistance associated with
ST:STUDY_SUMMARY                 	RelA overexpression. Materials and Methods: In this study, we performed a
ST:STUDY_SUMMARY                 	comprehensive quantitative analysis of proteomics and metabolomics in triple
ST:STUDY_SUMMARY                 	negative breast cancer (TNBC) cells overexpressing RelA, compared to TNBC cells
ST:STUDY_SUMMARY                 	with basal levels. State-of-the-art Trapped Ion Mobility Spectroscopy,
ST:STUDY_SUMMARY                 	Time-of-Flight Mass Spectrometry (TIMS-TOF-MS) was employed to achieve
ST:STUDY_SUMMARY                 	high-resolution analysis. Results: The results unveiled 27 significantly
ST:STUDY_SUMMARY                 	dysregulated proteins and 21 altered metabolites in MDA-MB-231 RelA cells
ST:STUDY_SUMMARY                 	relative to MDA-MB-231 cells. The upregulated proteins in MDA-MB-231 RelA cells
ST:STUDY_SUMMARY                 	include interferon-induced protein with tetratricopeptide repeats 3 (IFIT3) and
ST:STUDY_SUMMARY                 	cytochrome b5, which are involved in tumor migration and progression and
ST:STUDY_SUMMARY                 	regulation of the cellular redox system, respectively. Conversely, the adhesion
ST:STUDY_SUMMARY                 	molecule, integrin alpha-2, was downregulated in MDA-MB-231 RelA cells. In
ST:STUDY_SUMMARY                 	addition, metabolomics analysis revealed significant upregulation in the signal
ST:STUDY_SUMMARY                 	transducer, cyclic AMP, and downregulation in multiple nucleotides such as
ST:STUDY_SUMMARY                 	pyridine, adenine, and thymidine. The integrated analysis of multi-omics data
ST:STUDY_SUMMARY                 	highlighted the highest impacted pathways, including ABC transporters, arginine
ST:STUDY_SUMMARY                 	biosynthesis, purine, pyruvate, pyrimidine, glutathione, and phenylalanine
ST:STUDY_SUMMARY                 	metabolism. Conclusion: This study effectively recognized significantly
ST:STUDY_SUMMARY                 	dysregulated proteins and metabolites in MDA-MB-231 RelA cells, providing
ST:STUDY_SUMMARY                 	valuable insights into potential proteins, metabolites, and signaling pathways
ST:STUDY_SUMMARY                 	that mediate the aggressiveness of TNBC through RelA. Moreover, the multi-omics
ST:STUDY_SUMMARY                 	integrated analysis elucidated RelA role in chemotherapy resistance, tumor
ST:STUDY_SUMMARY                 	progression, migration, and invasion, which would propose potential biomarkers
ST:STUDY_SUMMARY                 	and novel therapeutic targets.
ST:INSTITUTE                     	Sharjah Institute for Medical Research
ST:LAST_NAME                     	Facility
ST:FIRST_NAME                    	Core
ST:ADDRESS                       	M32, SIMR, College of Pharmacy, Health Sciences, University of Sharjah, Sharjah,
ST:ADDRESS                       	UAE, Sharjah, 000, United Arab Emirates
ST:EMAIL                         	tims-tof@sharjah.ac.ae
ST:STUDY_TYPE                    	LC/MS/MS
ST:PHONE                         	+971 6 5057656
#SUBJECT
SU:SUBJECT_TYPE                  	Cultured cells
SU:SUBJECT_SPECIES               	Homo sapiens
SU:TAXONOMY_ID                   	9606
#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           	-	MDA01-01-4738	Treatment:MDA	RAW_FILE_NAME=MDA01-01-4738.d
SUBJECT_SAMPLE_FACTORS           	-	MDA01-02-4739	Treatment:MDA	RAW_FILE_NAME=MDA01-02-4739.d
SUBJECT_SAMPLE_FACTORS           	-	MDA02-01-4740	Treatment:MDA	RAW_FILE_NAME=MDA02-01-4740.d
SUBJECT_SAMPLE_FACTORS           	-	MDA02-02-4741	Treatment:MDA	RAW_FILE_NAME=MDA02-02-4741.d
SUBJECT_SAMPLE_FACTORS           	-	MDA03-01-4742	Treatment:MDA	RAW_FILE_NAME=MDA03-01-4742.d
SUBJECT_SAMPLE_FACTORS           	-	MDA03-02-4743	Treatment:MDA	RAW_FILE_NAME=MDA03-02-4743.d
SUBJECT_SAMPLE_FACTORS           	-	REL01-01-4732	Treatment:REL	RAW_FILE_NAME=REL01-01-4732.d
SUBJECT_SAMPLE_FACTORS           	-	REL01-02-4733	Treatment:REL	RAW_FILE_NAME=REL01-02-4733.d
SUBJECT_SAMPLE_FACTORS           	-	REL02-01-4734	Treatment:REL	RAW_FILE_NAME=REL02-01-4734.d
SUBJECT_SAMPLE_FACTORS           	-	REL02-02-4735	Treatment:REL	RAW_FILE_NAME=REL02-02-4735.d
SUBJECT_SAMPLE_FACTORS           	-	REL03-01-4736	Treatment:REL	RAW_FILE_NAME=REL03-01-4736.d
SUBJECT_SAMPLE_FACTORS           	-	REL03-02-4737	Treatment:REL	RAW_FILE_NAME=REL03-02-4737.d
#COLLECTION
CO:COLLECTION_SUMMARY            	Triple negative breast cancer cell line (MDA-MB-231) was purchased from the
CO:COLLECTION_SUMMARY            	European Collection of Authenticated Cell Cultures (ECACC). MDA-MB-231 cells
CO:COLLECTION_SUMMARY            	were maintained in Dulbecco’s Modified Eagle Medium (DMEM, Sigma-Aldrich)
CO:COLLECTION_SUMMARY            	supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin.
CO:COLLECTION_SUMMARY            	All incubations were done at 37˚C in a 5% CO2 air atmosphere.
CO:SAMPLE_TYPE                   	Breast cancer cells
#TREATMENT
TR:TREATMENT_SUMMARY             	P65 plasmid tagged with GFP (Addgene plasmid #23255) was used for transfecting
TR:TREATMENT_SUMMARY             	MDA-MB-231 cells. Transfection was performed as described previously [8]. In
TR:TREATMENT_SUMMARY             	brief, following the seeding of MDA-MB-231 cells in 6 well plates, cells were
TR:TREATMENT_SUMMARY             	transfected with 2.5 μg of RelA/P65 plasmid using lipofectamine 3000 prepared
TR:TREATMENT_SUMMARY             	in Opti-MEM I media (Invitrogen Carlsbad, CA, USA). After cells incubated for 6
TR:TREATMENT_SUMMARY             	h, RPMI-1640 culture media was added to replace Opti-MEM I media. Cells were
TR:TREATMENT_SUMMARY             	selected using Genticine antibiotic (G418) (InvivoGen, USA, #ant-gn-1).
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	MDA-MB-231 and MDA-MB-231 RelA cells were cultured in T75 flasks until reaching
SP:SAMPLEPREP_SUMMARY            	70% confluency. Cells were detached using trypsin, and centrifugation was
SP:SAMPLEPREP_SUMMARY            	performed to collect the cell pellets. A total of 3×106 cells were collected
SP:SAMPLEPREP_SUMMARY            	and stored at -80 ℃ in PBS buffer for further analysis. Cell pellets were
SP:SAMPLEPREP_SUMMARY            	prepared in triplicates for metabolomics and proteomics analyses.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	For metabolomics analysis, a Hamilton Intensity Solo 2 C18 column (100 mm × 2.1
CH:CHROMATOGRAPHY_SUMMARY        	mm, 1.8 μm beads) was kept at 35 °C. For metabolomics, 10 μL of each sample
CH:CHROMATOGRAPHY_SUMMARY        	was injected twice and eluted using a 30-min gradient
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Bruker Elute
CH:COLUMN_NAME                   	Hamilton Intensity Solo 2 C18 (100 mm x 2.1 mm, 1.8um)
CH:SOLVENT_A                     	100% water; 0.1% formic acid
CH:SOLVENT_B                     	100% acetonitrile; 0.1% formic acid
CH:FLOW_GRADIENT                 	a 30-min gradient as follows: 1% ACN was kept for 2 minutes before ramping to
CH:FLOW_GRADIENT                 	99% ACN for 15 minutes, then holding at 99% ACN for 3 minutes before
CH:FLOW_GRADIENT                 	re-equilibrating to 1% ACN for 10 minutes
CH:FLOW_RATE                     	For elution 250 μL/min and 350 μL/min for re-equilibration
CH:COLUMN_TEMPERATURE            	35
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Bruker timsTOF
MS:INSTRUMENT_TYPE               	QTOF
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	POSITIVE
MS:MS_COMMENTS                   	Using an Apollo II electrospray ionization (ESI) source or an CaptiveSpray ion
MS:MS_COMMENTS                   	source, and a timsTOF (Bruker, Bremen, Germany), the MS analysis was carried
MS:MS_COMMENTS                   	out. For metabolomics, the nebulizer pressure was adjusted to 2.2 bar, the
MS:MS_COMMENTS                   	drying temperature to 220°C, and the gas flow rate to 10 L/min. 4500 V was the
MS:MS_COMMENTS                   	capillary voltage, while 500 V was the end plate offset and the scan range for
MS:MS_COMMENTS                   	metabolomics was 20-1300 m/z. The instrument was operated in auto-MS/MS mode and
MS:MS_COMMENTS                   	the acquisition was performed using the positive mode at 12 Hz. The width of the
MS:MS_COMMENTS                   	precursor ion was ±0.5, the cycle time was 0.5 sec., and the threshold was 400
MS:MS_COMMENTS                   	counts per thousand. Active exclusion was excluded after 3 spectra and released
MS:MS_COMMENTS                   	after 0.2 min. For MS2 acquisition the data dependent acquisition (DDA) was
MS:MS_COMMENTS                   	used, and the collision energy stepping fluctuated between 100 and 250% set at
MS:MS_COMMENTS                   	20 eV. In the first 0.3 minutes of each LC-MS/MS run, sodium formate was
MS:MS_COMMENTS                   	injected as an external calibrant during data processing
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	AU
MS_METABOLITE_DATA_START
Samples	MDA01-01-4738	MDA01-02-4739	MDA02-01-4740	MDA02-02-4741	MDA03-01-4742	MDA03-02-4743	REL01-01-4732	REL01-02-4733	REL02-01-4734	REL02-02-4735	REL03-01-4736	REL03-02-4737
Factors	Treatment:MDA	Treatment:MDA	Treatment:MDA	Treatment:MDA	Treatment:MDA	Treatment:MDA	Treatment:REL	Treatment:REL	Treatment:REL	Treatment:REL	Treatment:REL	Treatment:REL
1,3,5-Trimethoxybenzene	19536	21258	20526	19084	23666	18764	16618	20160	17824	18416	21184	18170
2,3-Diaminopropionic acid	34758	32284	22936	22482	28102	28216	28052	27444	20436	17370	20054	19246
2,4-Diaminobutyric acid	36100	34090	36330	35046	30444	29070	37706	39052	35928	34518	32812	34114
2,5-Furandicarboxylic acid	3774	3360	3950	4624	0	5050	1412	1950	1492	1404	1412	1350
2-Furoylglycine	1824	1768	1780	1780	2468	2722	712	548	626	546	704	630
2-Hydroxycaproic acid	71132	70546	66648	68914	71500	72502	74658	74114	73914	74628	71936	73044
2-Phenylbutyric acid	32156	30014	30064	29996	31132	28844	29650	33866	24742	30364	30726	32040
2-Pyrrolidinone	1330	1690	1722	1356	1958	1422	1832	1722	1704	1318	2112	2942
3,4,5-Trimethoxycinnamic acid	1870	1498	1544	2094	1934	1706	1822	1464	1888	1880	1666	1474
3,4-Dihydroxyphenylglycol	2730	2256	2618	2110	2326	1938	2904	2658	2944	2512	2356	2306
3-Aminoisobutanoic acid	544304	561050	537876	555952	548846	538140	549716	562972	566700	550350	546710	544724
3-Hexenedioic acid	8588	2778	12054	6720	10206	9502	5926	6554	8176	6490	5880	4784
3-Hydroxymandelic acid	2982	2718	2912	2578	3102	2678	2802	2964	3030	2484	2986	3454
3-Methyl-2-oxovaleric acid	1934	1866	2154	1726	1846	1656	1792	2112	1718	2238	2216	1994
3-Methyladipic acid	19358	17120	15556	16614	20674	21306	19440	19648	18770	18846	21402	24684
3-Methylindole	3630	3438	3748	4252	3516	3892	3888	3862	3964	3586	3738	3452
4-Aminophenol	242	384	370	110	286	0	596	346	640	1098	24954	25620
4-Hydroxyproline	1948	1834	2228	2048	2348	1900	1898	2144	2102	2278	2190	1898
5-Aminolevulinic acid	2256	2400	1652	2084	2166	1774	2538	2154	2346	1912	1730	1872
5-Hydroxy-L-tryptophan	1618	1940	1858	1650	1928	1856	794	620	862	932	416	584
5-Hydroxytryptophol	2722	3062	3164	2706	3072	3114	1226	1800	1800	1458	2136	1888
7,8-Dihydrobiopterin	1048	1082	970	1018	1228	1306	766	764	810	990	1032	936
Acetaminophen	1322	1800	730	674	1208	1144	2206	2092	3356	4850	159748	165178
Adenine	20398	21152	24962	21376	23870	22074	12392	0	13144	13082	11696	11738
Adenosine monophosphate	13750	11864	12902	13622	14818	13342	8340	8640	10580	10998	7598	5196
ADP	58620	30396	60048	61118	73950	73780	46944	47752	39448	47548	47808	46750
Alpha-hydroxyhippuric acid	1496	1124	1754	1588	1606	1682	1176	1366	1304	1462	1602	1184
Alpha-N-phenylacetyl-L-glutamine	2998	2964	3176	3090	2922	3826	1880	1920	1304	1768	1504	1246
Aminoadipic acid	6336	5832	6632	4594	8638	8184	2462	2022	1720	2178	2486	1746
Androstenedione	14560	12202	12816	14380	12952	12538	9670	9428	11474	8630	11264	10722
Androsterone	25068	24796	25198	25672	23970	24960	27158	25952	25732	25664	24662	25340
Azythromycin	181520	169600	173172	182806	164936	162222	165618	166792	168738	165624	177490	174624
Benzaldehyde	10360	10832	11078	9844	10776	10456	11370	12270	13080	10816	11230	13484
Benzene	10832	10686	9412	12088	8538	8788	10336	10380	13074	10912	10248	10858
Butanone	1016	1248	1260	874	1268	1262	1018	1350	900	1050	1262	1496
Cinnamic acid	17432	17952	18972	17884	20132	20118	17160	17298	17910	16630	16244	15762
Cortisol	53266	48544	47262	48938	46176	39258	56254	55190	48582	54112	53048	54076
Creatine	20948	17656	20750	18038	22408	20982	7374	0	7882	0	0	6658
Cyclic AMP	8042	7452	198	6366	336	5996	43714	42586	39910	35344	188	39506
Cytosine	148	1732	524	912	186	3174	2010	444	1472	1396	174	914
D-Alanine	5546	5144	6276	6650	5928	6176	2444	2998	2582	2346	3108	2444
D-Glucurono-6,3-lactone	3528	4636	4530	4650	2928	4132	5074	4946	5138	5260	4502	4540
D-Serine	7152	5000	6314	3144	3932	2500	5916	6644	5430	5082	5232	4682
Deoxyinosine	15562	16242	18138	17424	15060	15196	12414	11274	12916	11520	10256	11182
DGTP	0	15800	12678	13116	10686	9776	22744	19926	16422	21212	19658	21126
Diacetyl	4804	5210	4252	3658	3694	3682	2576	3588	2922	2624	3270	4102
Digoxin	5474	3284	5970	4116	4430	4986	6850	6080	6672	5524	6282	5730
DL-2-aminooctanoic acid	13166	13920	15836	14346	17006	16738	9780	10658	10730	9386	8478	7864
DUMP	29566	20552	35176	22602	31724	26570	16456	20838	18484	16818	30820	20502
Gamma-Glutamylcysteine	4854	3452	5074	5102	3766	4112	8560	8314	6748	6282	6496	5054
Gluconolactone	88636	87540	56688	76444	72342	96772	131046	115068	123072	117120	81892	93370
Glucosamine	2736	156	3582	3564	3850	3390	1700	1060	1526	1864	1582	2308
Glutathione	396620	387090	271042	357106	340944	453000	571980	523020	564466	542078	393610	489346
Glycerol	15126	17680	16104	11798	15358	17200	18148	17452	17916	18212	21116	18142
Glycerophosphocholine	11020	8356	12916	10532	14160	11740	15972	21524	17212	18878	18208	16004
Guaifenesin	1354	1196	1256	1232	1448	892	1144	1328	1104	1720	982	1078
Guanine	52112	0	65706	46786	91904	90604	17428	18140	21372	16964	17872	16158
Guanosine 5'-diphosphate	57416	47070	53282	49312	46826	50408	30962	34442	28398	28962	27796	27398
Guanosine 5'-triphosphate	13640	10392	14958	14122	13718	12464	12322	13238	12098	12378	13048	11050
Hypoxanthine	276328	280002	245760	273128	322816	405854	76786	68474	76972	71676	60012	74056
Indole	13162	11756	12198	13032	12268	12546	13140	12758	11810	11358	11044	11286
Indoleacetic acid	23844	23158	23444	20542	21334	24162	14634	13180	10478	12134	9906	10420
Indoleacrylic acid	209306	201470	210452	207792	216238	214392	217132	218516	202754	193928	177954	189026
Isobutyric acid	92648	76012	92184	89454	101814	97830	106830	98314	105614	108296	102222	94658
Isovalerylcarnitine	6818	6362	7548	6666	7184	6698	10790	11462	10818	12556	10388	10630
L-Acetylcarnitine	154006	15278	195518	121484	164174	177800	89302	98300	152104	99736	82170	77378
L-Arginine	1636	1268	1766	2004	1914	1498	1310	1318	1460	1426	1250	1090
L-Carnitine	16456	13764	19310	17464	18276	17940	12484	14386	11638	12952	12872	11150
L-Glutamic acid	10908	10450	11888	12030	12970	11774	13168	13360	11538	11522	12648	11540
L-Glutamine	1436	1290	1280	1214	1254	1174	1166	1458	1250	1618	1538	1120
L-Histidine	1498	1386	1818	1904	1568	1238	1426	1680	1812	1496	1222	1112
L-Malic acid	6798	7560	0	7106	262	11022	1670	928	1114	768	0	1010
L-Methionine	19186	5398	19838	14002	12516	12920	15466	14076	5934	10316	16834	19962
L-Norleucine	13814	14442	14736	16482	15170	13478	13318	12748	11780	11614	12678	13190
L-Phenylalanine	207054	195482	211418	208102	211160	217940	193648	193570	174392	174274	185172	166680
L-Threonine	15788	16038	15248	15534	15512	15064	17044	15474	14682	16030	14818	0
L-Tryptophan	32508	34754	39536	38334	39124	38010	38598	40102	36778	34272	35028	34990
L-Valine	10972	10238	12184	9846	12782	13330	10326	9900	10206	10464	9614	8448
Maltitol	34496	29434	32372	31066	32692	32704	31092	32498	32462	34064	33390	28174
Methylhippuric acid	2656	3166	2844	3178	3456	2846	1204	1304	1572	798	1234	1222
Mevalonic acid	7440	8970	6970	8524	5052	6572	9396	7142	9242	9786	9438	8158
N,N-Dimethylformamide	1270	1040	1108	1294	1074	1030	924	1020	864	1152	336	1024
N-Acetyl-D-glucosamine	1446	1568	1786	1426	1646	1474	708	638	538	740	750	516
N-Acetyl-L-alanine	10966	11110	10622	11300	9430	10348	11830	12232	11072	10646	12494	12236
N-Acetylserotonin	3080	2232	1756	1966	1994	2198	2814	3154	3144	2664	2890	2750
N-Methylnicotinamide	11850	10530	13908	11654	14314	13006	12852	13444	12092	12604	11780	12440
Niacinamide	141922	166158	115244	151756	121116	161340	83516	65322	81284	72014	50162	66664
Normetanephrine	16322	20140	20832	19184	21422	21856	7632	7590	6348	6046	6310	7046
Oxypurinol	11848	10026	11472	12450	9504	9290	658	842	1050	622	776	724
Pantothenic acid	240858	249574	257608	261970	274960	268096	91038	92914	80794	82472	84408	80188
PC(16:0/16:0)	5462	5004	4120	3732	2704	2648	1530	1328	738	3378	2332	2042
Phenylacetaldehyde	14214	14954	15600	15296	14200	15646	14670	15204	15354	14906	14328	16072
Phenylacetic acid	2864	3606	3222	3384	3678	3090	4008	4064	3062	3294	3014	3500
Phenylacetylglycine	4708	5026	3908	4550	4798	5532	6120	4868	5712	5988	5598	5554
Phosphocreatine	67410	63114	67454	71570	69072	66018	70094	68704	76810	71394	74846	82378
Phosphoric acid	2620	2460	2288	2334	2598	2328	2766	2758	2382	2628	2528	2360
Propionic acid	45482	66912	53944	62290	54914	59156	52424	57296	57324	55588	59868	50714
Pyridine	70416	77144	51044	70524	49930	72214	37134	31876	35664	33710	22532	27344
Pyroglutamic acid	78150	117144	89948	94408	80336	85012	98866	97854	105284	106418	95104	110784
Pyruvaldehyde	858	972	920	854	900	912	720	768	1086	744	974	1054
Quinic acid	6324	7784	9412	8228	12746	8054	6454	5898	5220	4866	5936	4776
Rhamnose	4106	3780	3956	3838	3426	3978	3924	3780	3992	3710	3832	3540
Saccharopine	3058	0	3162	2636	2980	3248	2824	0	3750	2826	3140	2744
Spermine	83862	61560	74026	58594	78820	77602	57160	72498	60504	64816	67106	60774
Sphingosine	2762	2422	2672	2756	4836	7620	4708	5210	3500	3438	2664	2744
Stearic acid	2176	1818	1516	1810	2054	1568	1580	1930	1760	1536	1562	1994
Succinic acid	10644	8122	11134	10958	16000	16806	192	394	438	564	362	368
Succinylacetone	36864	32008	34498	39136	33256	34124	35328	41750	39540	42622	33600	35406
Syringic acid	2826	2588	1930	1718	2108	1710	2296	2658	2510	3058	2052	2550
Tartaric acid	16426	19520	14486	18180	13292	15136	18384	16170	20340	17408	14324	15754
Taurocholic acid	1308	1088	1330	936	1618	1036	1322	1576	1090	1972	1556	0
Thymidine	4092	4836	4872	4802	6188	6150	2928	2906	1772	1858	2190	2442
Tiglylglycine	1242	1704	1328	1338	1504	1536	1708	1272	1484	1356	1284	1316
Tolnaftate	1688	2014	1956	1854	2334	2480	2576	2498	2010	1744	2416	2594
Trehalose	1188	1022	1470	1254	1134	1038	1298	1274	972	794	952	1078
Uracil	7048	0	6704	4686	7378	7860	2244	1702	1196	1806	1688	2422
Urea	37452	38834	41110	36772	35682	37372	47238	47148	43182	48526	35622	45614
Ureidosuccinic acid	3842	4020	4254	4846	3794	4682	4612	5412	4730	5344	4782	4260
Uric acid	191140	226102	172846	219808	160432	174752	208734	184784	221764	215668	163210	200624
Uridine	15914	15188	16934	16996	15018	15930	20234	20340	16712	17036	16500	19160
Urocanic acid	832	194	836	1104	1128	966	940	1126	1090	986	762	1134
Verapamil	1650	1472	1448	1444	0	1610	1618	1784	1750	1590	1468	1598
Xanthosine	3934	3394	4166	3510	3686	2880	0	390	448	332	214	234
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	Molecular Formula	RT [min]	m/z meas.
1,3,5-Trimethoxybenzene	C9H12O3	26.59	57.03381
2,3-Diaminopropionic acid	C3H8N2O2	13.9	105.07
2,4-Diaminobutyric acid	C4H10N2O2	14.94	119.08554
2,5-Furandicarboxylic acid	C6H4O5	2.59	157.011
2-Furoylglycine	C7H7NO4	2.75	170.04262
2-Hydroxycaproic acid	C6H12O3	16.35	133.08602
2-Phenylbutyric acid	C10H12O2	11.75	165.09124
2-Pyrrolidinone	C4H7NO	3.44	86.06032
3,4,5-Trimethoxycinnamic acid	C12H14O5	10.25	239.08943
3,4-Dihydroxyphenylglycol	C8H10O4	13.29	171.06313
3-Aminoisobutanoic acid	C4H9NO2	8.01	104.06248
3-Hexenedioic acid	C6H8O4	3.1	145.04944
3-Hydroxymandelic acid	C8H8O4	6.19	169.05584
3-Methyl-2-oxovaleric acid	C6H10O3	2.99	131.07029
3-Methyladipic acid	C7H12O4	7.36	199.0427
3-Methylindole	C9H9N	7.36	132.08083
4-Aminophenol	C6H7NO	7.44	110.06017
4-Hydroxyproline	C5H9NO3	0.77	132.06566
5-Aminolevulinic acid	C5H9NO3	1.04	132.06568
5-Hydroxy-L-tryptophan	C11H12N2O3	6.01	221.09262
5-Hydroxytryptophol	C10H11NO2	3	178.08637
7,8-Dihydrobiopterin	C9H13N5O3	6.67	240.10227
Acetaminophen	C8H9NO2	7.44	152.07077
Adenine	C5H5N5	7.1	136.06187
Adenosine monophosphate	C10H14N5O7P	3.04	348.07061
ADP	C10H15N5O10P2	2.83	428.03664
Alpha-hydroxyhippuric acid	C9H9NO4	2.04	196.06419
Alpha-N-phenylacetyl-L-glutamine	C13H16N2O4	1.99	265.11215
Aminoadipic acid	C6H11NO4	2.86	162.07585
Androstenedione	C19H26O2	10.77	287.19692
Androsterone	C19H30O2	17.59	291.23245
Azythromycin	C38H72N2O12	16.67	771.4878
Benzaldehyde	C7H6O	13.85	107.04923
Benzene	C6H6	3.37	79.05457
Butanone	C4H8O	7.51	73.06527
Cinnamic acid	C9H8O2	6.58	149.05992
Cortisol	C21H30O5	15.31	363.21478
Creatine	C4H9N3O2	2.03	132.07647
Cyclic AMP	C10H12N5O6P	6.01	330.05997
Cytosine	C4H5N3O	5.93	112.05049
D-Alanine	C3H7NO2	2.05	90.05517
D-Glucurono-6,3-lactone	C6H8O6	3.35	177.03314
D-Serine	C3H7NO3	7.17	106.05012
Deoxyinosine	C10H12N4O4	7	253.1009
DGTP	C10H16N5O13P3	2.98	508.00313
Diacetyl	C4H6O2	6.88	87.04435
Digoxin	C41H64O14	14.82	391.22019
DL-2-aminooctanoic acid	C8H17NO2	2.72	160.13345
DUMP	C9H13N2O8P	6.23	309.05765
Gamma-Glutamylcysteine	C8H14N2O5S	7.47	251.07013
Gluconolactone	C6H10O6	3.37	179.04855
Glucosamine	C6H13NO5	2.82	180.08797
Glutathione	C10H17N3O6S	3.37	308.09041
Glycerol	C3H8O3	2.87	93.05468
Glycerophosphocholine	C8H20NO6P	1.71	258.11042
Guaifenesin	C10H14O4	10.81	199.09475
Guanine	C5H5N5O	3.09	152.05668
Guanosine 5'-diphosphate	C10H15N5O11P2	3.06	444.03164
Guanosine 5'-triphosphate	C10H16N5O14P3	3.06	523.99806
Hypoxanthine	C5H4N4O	3.37	137.04561
Indole	C8H7N	7.36	118.06516
Indoleacetic acid	C10H9NO2	11.8	176.07088
Indoleacrylic acid	C11H9NO2	7.36	188.07079
Isobutyric acid	C4H8O2	16.33	89.06004
Isovalerylcarnitine	C12H23NO4	7.6	246.17044
L-Acetylcarnitine	C9H17NO4	3.09	204.12312
L-Arginine	C6H14N4O2	1.81	175.11933
L-Carnitine	C7H15NO3	1.92	162.11265
L-Glutamic acid	C5H9NO4	1.96	148.06057
L-Glutamine	C5H10N2O3	3.02	147.08438
L-Histidine	C6H9N3O2	9.45	156.08073
L-Malic acid	C4H6O5	6	135.02992
L-Methionine	C5H11NO2S	3.35	150.05814
L-Norleucine	C6H13NO2	10.77	132.10196
L-Phenylalanine	C9H11NO2	6.58	166.08629
L-Threonine	C4H9NO3	8.02	120.0571
L-Tryptophan	C11H12N2O2	7.36	205.09755
L-Valine	C5H11NO2	2.73	118.08622
Maltitol	C12H24O11	2.02	345.13936
Methylhippuric acid	C10H11NO3	9.18	194.08154
Mevalonic acid	C6H12O4	3.37	149.08099
N,N-Dimethylformamide	C3H7NO	6.39	74.06052
N-Acetyl-D-glucosamine	C8H15NO6	2.12	222.09771
N-Acetyl-L-alanine	C5H9NO3	2.34	132.06557
N-Acetylserotonin	C12H14N2O2	7.56	219.11152
N-Methylnicotinamide	C7H8N2O	1.93	137.07087
Niacinamide	C6H6N2O	3.38	123.05518
Normetanephrine	C9H13NO3	6.9	184.09713
Oxypurinol	C5H4N4O2	6.28	153.04069
Pantothenic acid	C9H17NO5	6.9	220.11813
PC(16:0/16:0)	C40H80NO8P	21.41	734.56945
Phenylacetaldehyde	C8H8O	17.73	121.06485
Phenylacetic acid	C8H8O2	12.17	137.05988
Phenylacetylglycine	C10H11NO3	6.59	194.07912
Phosphocreatine	C4H10N3O5P	13.85	212.03786
Phosphoric acid	H3O4P	2.09	98.98431
Propionic acid	C3H6O2	2.01	75.04438
Pyridine	C5H5N	3.37	80.04983
Pyroglutamic acid	C5H7NO3	3.35	130.04979
Pyruvaldehyde	C3H4O2	3.39	73.02868
Quinic acid	C7H12O6	6.06	215.05301
Rhamnose	C6H12O5	2.02	165.07591
Saccharopine	C11H20N2O6	13.22	277.14169
Spermine	C10H26N4	1.44	203.22332
Sphingosine	C18H37NO2	14.61	300.29009
Stearic acid	C18H36O2	0.94	285.27925
Succinic acid	C4H6O4	3	119.03518
Succinylacetone	C7H10O4	10.83	159.0653
Syringic acid	C9H10O5	7.85	199.05805
Tartaric acid	C4H6O6	3.37	151.0213
Taurocholic acid	C26H45NO7S	12.85	516.29056
Thymidine	C10H14N2O5	10.13	243.0881
Tiglylglycine	C7H11NO3	21.85	158.08139
Tolnaftate	C19H17NOS	9.64	308.11172
Trehalose	C12H22O11	9.78	343.12906
Uracil	C4H4N2O2	3.27	113.03451
Urea	CH4N2O	13.85	61.04007
Ureidosuccinic acid	C5H8N2O5	14.6	177.05496
Uric acid	C5H4N4O3	3.37	169.03173
Uridine	C9H12N2O6	14.6	245.07886
Urocanic acid	C6H6N2O2	0.47	139.05025
Verapamil	C27H38N2O4	8.46	455.28677
Xanthosine	C10H12N4O6	6.28	285.08347
METABOLITES_END
#END