#METABOLOMICS WORKBENCH SIMR_Core_Facility_20221018_005146 DATATRACK_ID:3516 STUDY_ID:ST002317 ANALYSIS_ID:AN003784 PROJECT_ID:PR001485
VERSION             	1
CREATED_ON             	October 19, 2022, 8:57 am
#PROJECT
PR:PROJECT_TITLE                 	Mass spectroscopy‑based proteomics and metabolomics analysis of
PR:PROJECT_TITLE                 	triple‑positive breast cancer cells treated with tamoxifen and/ or trastuzumab
PR:PROJECT_TYPE                  	LC-MS/MS
PR:PROJECT_SUMMARY               	HER2-enriched breast cancer with high levels of hormone receptor expression,
PR:PROJECT_SUMMARY               	known as "triple positive" breast cancer, may represent a new entity with a
PR:PROJECT_SUMMARY               	relatively favourable prognosis against which the combination of chemotherapy,
PR:PROJECT_SUMMARY               	HER-2 inhibition, and endocrine treatment may be considered overtreatment. We
PR:PROJECT_SUMMARY               	explored the effect of the anticancer drugs tamoxifen and trastuzumab, both
PR:PROJECT_SUMMARY               	separately and in combination, on the integrated proteomic and metabolic profile
PR:PROJECT_SUMMARY               	of "triple positive" breast cancer cells (BT-474). Method We employed
PR:PROJECT_SUMMARY               	ultra-high-performance liquid chromatography-quadrupole time of flight mass
PR:PROJECT_SUMMARY               	spectrometry using a Bruker timsTOF to investigate changes in BT-474 cell line
PR:PROJECT_SUMMARY               	treated with either tamoxifen, trastuzumab or a combination. Differentially
PR:PROJECT_SUMMARY               	abundant metabolites were identified using the Bruker Human Metabolome Database
PR:PROJECT_SUMMARY               	metabolite library and proteins using the Uniprot proteome for Homo sapiens
PR:PROJECT_SUMMARY               	using MetaboScape and MaxQuant, respectively, for identification and
PR:PROJECT_SUMMARY               	quantitation. Results A total of 77 proteins and 85 metabolites were found to
PR:PROJECT_SUMMARY               	significantly differ in abundance in BT-474 treated cells with tamoxifen 5
PR:PROJECT_SUMMARY               	μM/and or trastuzumab 2.5 μM. Findings suggest that by targeting important
PR:PROJECT_SUMMARY               	cellular signalling pathways which regulate cell growth, apoptosis,
PR:PROJECT_SUMMARY               	proliferation, and chemoresistance, these medicines have a considerable
PR:PROJECT_SUMMARY               	anti-growth effect in BT-474 cells. Pathways enriched for dysregulation include
PR:PROJECT_SUMMARY               	RNA splicing, neutrophil degranulation and activation, cellular redox
PR:PROJECT_SUMMARY               	homeostasis, mitochondrial transmembrane transport, ferroptosis and necroptosis,
PR:PROJECT_SUMMARY               	ABC transporters and central carbon metabolism. Conclusion Our findings in
PR:PROJECT_SUMMARY               	protein and metabolite level research revealed that anti-cancer drug therapy had
PR:PROJECT_SUMMARY               	a significant impact on the key signalling pathways and molecular processes in
PR:PROJECT_SUMMARY               	triple positive BT-474 cell lines.
PR:INSTITUTE                     	Sharjah Institute for Medical Research
PR:DEPARTMENT                    	Sharjah Institute for Medical Research
PR:LABORATORY                    	Biomarker Discovery Group
PR:LAST_NAME                     	Facility
PR:FIRST_NAME                    	Core
PR:ADDRESS                       	M32, SIMR, College of Pharmacy, Health Sciences, University of Sharjah, Sharjah,
PR:ADDRESS                       	UAE, Sharjah, 000, United Arab Emirates
PR:EMAIL                         	tims-tof@sharjah.ac.ae
PR:PHONE                         	+971 6 5057656
#STUDY
ST:STUDY_TITLE                   	Mass spectroscopy‑based proteomics and metabolomics analysis of
ST:STUDY_TITLE                   	triple‑positive breast cancer cells treated with tamoxifen
ST:STUDY_SUMMARY                 	HER2-enriched breast cancer with high levels of hormone receptor expression,
ST:STUDY_SUMMARY                 	known as "triple positive" breast cancer, may represent a new entity with a
ST:STUDY_SUMMARY                 	relatively favourable prognosis against which the combination of chemotherapy,
ST:STUDY_SUMMARY                 	HER-2 inhibition, and endocrine treatment may be considered overtreatment. We
ST:STUDY_SUMMARY                 	explored the effect of the anticancer drugs tamoxifen and trastuzumab, both
ST:STUDY_SUMMARY                 	separately and in combination, on the integrated proteomic and metabolic profile
ST:STUDY_SUMMARY                 	of "triple positive" breast cancer cells (BT-474). Method We employed
ST:STUDY_SUMMARY                 	ultra-high-performance liquid chromatography-quadrupole time of flight mass
ST:STUDY_SUMMARY                 	spectrometry using a Bruker timsTOF to investigate changes in BT-474 cell line
ST:STUDY_SUMMARY                 	treated with either tamoxifen, trastuzumab or a combination. Differentially
ST:STUDY_SUMMARY                 	abundant metabolites were identified using the Bruker Human Metabolome Database
ST:STUDY_SUMMARY                 	metabolite library and proteins using the Uniprot proteome for Homo sapiens
ST:STUDY_SUMMARY                 	using MetaboScape and MaxQuant, respectively, for identification and
ST:STUDY_SUMMARY                 	quantitation. Results A total of 77 proteins and 85 metabolites were found to
ST:STUDY_SUMMARY                 	significantly differ in abundance in BT-474 treated cells with tamoxifen 5
ST:STUDY_SUMMARY                 	μM/and or trastuzumab 2.5 μM. Findings suggest that by targeting important
ST:STUDY_SUMMARY                 	cellular signalling pathways which regulate cell growth, apoptosis,
ST:STUDY_SUMMARY                 	proliferation, and chemoresistance, these medicines have a considerable
ST:STUDY_SUMMARY                 	anti-growth effect in BT-474 cells. Pathways enriched for dysregulation include
ST:STUDY_SUMMARY                 	RNA splicing, neutrophil degranulation and activation, cellular redox
ST:STUDY_SUMMARY                 	homeostasis, mitochondrial transmembrane transport, ferroptosis and necroptosis,
ST:STUDY_SUMMARY                 	ABC transporters and central carbon metabolism. Conclusion Our findings in
ST:STUDY_SUMMARY                 	protein and metabolite level research revealed that anti-cancer drug therapy had
ST:STUDY_SUMMARY                 	a significant impact on the key signalling pathways and molecular processes in
ST:STUDY_SUMMARY                 	triple positive BT-474 cell lines.
ST:INSTITUTE                     	University of Sharjah
ST:DEPARTMENT                    	Sharjah Institute for Medical Research
ST:LABORATORY                    	Biomarker Discovery Group
ST:LAST_NAME                     	Soares
ST:FIRST_NAME                    	Nelson
ST:ADDRESS                       	M32, SIMR, College of Pharmacy, Health Sciences, University of Sharjah
ST:EMAIL                         	nsoares@sharjah.ac.ae
ST:PHONE                         	065057656
#SUBJECT
SU:SUBJECT_TYPE                  	Human
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           	-	1A_93_1_328	Treatment:Control	RAW_FILE_NAME=1A''_93_1_328.d
SUBJECT_SAMPLE_FACTORS           	-	1A_93_1_331	Treatment:Control	RAW_FILE_NAME=1A''_93_1_331.d
SUBJECT_SAMPLE_FACTORS           	-	1A_92_1_327	Treatment:Control	RAW_FILE_NAME=1A'_92_1_327.d
SUBJECT_SAMPLE_FACTORS           	-	1A_92_1_330	Treatment:Control	RAW_FILE_NAME=1A'_92_1_330.d
SUBJECT_SAMPLE_FACTORS           	-	1A_91_1_326	Treatment:Control	RAW_FILE_NAME=1A_91_1_326.d
SUBJECT_SAMPLE_FACTORS           	-	1A_91_1_329	Treatment:Control	RAW_FILE_NAME=1A_91_1_329.d
SUBJECT_SAMPLE_FACTORS           	-	2a_-1_59_1_440	Treatment:TAMOXIFEN	RAW_FILE_NAME=2a''_-1_59_1_440.d
SUBJECT_SAMPLE_FACTORS           	-	2a_-2_59_1_441	Treatment:TAMOXIFEN	RAW_FILE_NAME=2a''_-2_59_1_441.d
SUBJECT_SAMPLE_FACTORS           	-	2a_-1_58_1_438	Treatment:TAMOXIFEN	RAW_FILE_NAME=2a'_-1_58_1_438.d
SUBJECT_SAMPLE_FACTORS           	-	2a_-2_58_1_439	Treatment:TAMOXIFEN	RAW_FILE_NAME=2a'_-2_58_1_439.d
SUBJECT_SAMPLE_FACTORS           	-	2a-1_57_1_436	Treatment:TAMOXIFEN	RAW_FILE_NAME=2a-1_57_1_436.d
SUBJECT_SAMPLE_FACTORS           	-	2a-2_57_1_437	Treatment:TAMOXIFEN	RAW_FILE_NAME=2a-2_57_1_437.d
#COLLECTION
CO:COLLECTION_SUMMARY            	The BT-474 BC cell line utilized in this study was cultured as monolayers in
CO:COLLECTION_SUMMARY            	DMEM medium supplemented with 10% fetal bovine serum and 1%
CO:COLLECTION_SUMMARY            	penicillin/streptomycin (Sigma Aldrich, St. Louis, MO, USA). All cultures were
CO:COLLECTION_SUMMARY            	incubated at 37 °C in a humidified atmosphere of 5% CO2.
CO:SAMPLE_TYPE                   	Breast cancer cells
CO:STORAGE_CONDITIONS            	Described in summary
#TREATMENT
TR:TREATMENT_SUMMARY             	Triplicate flasks were prepared for each treatment condition for each analysis
TR:TREATMENT_SUMMARY             	(metabolomic and proteomic) for a total of 24 flasks. Two million cells were
TR:TREATMENT_SUMMARY             	seeded in each 75 cm2 tissue culture flask and incubated for 24 h. The cells
TR:TREATMENT_SUMMARY             	were then treated with Tamoxifen (5 μM) and/or Trastuzumab (2.5 μM) for 24 h.
TR:TREATMENT_SUMMARY             	These concentrations correspond to the IC50 of these compounds with BT-474
TR:TREATMENT_SUMMARY             	cells, as determined by cytotoxicity assays (data not shown). Control cells were
TR:TREATMENT_SUMMARY             	treated with vehicle (dimethyl sulfoxide (DMSO) at 0.5% for 24 h. Following the
TR:TREATMENT_SUMMARY             	incubation period, cells were collected by trypsinization and washed twice with
TR:TREATMENT_SUMMARY             	phosphate-buffered saline solution (PBS) before re-suspending in 1 mL 1 × PBS
TR:TREATMENT_SUMMARY             	for further analysis. Finally, cells were collected as pellets by centrifugation
TR:TREATMENT_SUMMARY             	at 1200 rounds per minute (rpm) for 10 min at room temperature. To negate the
TR:TREATMENT_SUMMARY             	effect of Circadian rhythms on the response of cells to treatment, cells were
TR:TREATMENT_SUMMARY             	kept under the same conditions during the entire incubation period and the cell
TR:TREATMENT_SUMMARY             	collection was done concurrently for all samples. In addition, the same number
TR:TREATMENT_SUMMARY             	of cells were used for each sample to avoid the effect of variation in cell
TR:TREATMENT_SUMMARY             	numbers.
TR:TREATMENT                     	Drugs
TR:TREATMENT_COMPOUND            	Tamoxifen and/or Trastuzumab
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Sample metabolite extraction A volume of 1 mL of the extraction solvent
SP:SAMPLEPREP_SUMMARY            	(methanol+0.1% formic acid) was added to the cell pellets to quench cells. The
SP:SAMPLEPREP_SUMMARY            	cells were then vortexed for 2 min to ensure the quantitative extraction of the
SP:SAMPLEPREP_SUMMARY            	metabolites and stored on ice for 1 h, during which the samples were vortexed
SP:SAMPLEPREP_SUMMARY            	every 15 min. After this, the insoluble cell matrices were collected and
SP:SAMPLEPREP_SUMMARY            	transferred to centrifuge tubes, intermittent ultrasonication using the COPLEY
SP:SAMPLEPREP_SUMMARY            	sonicator (QSONICA SONICATOR, USA) under 30% amplifier and for 30 s with an ice
SP:SAMPLEPREP_SUMMARY            	bath employed throughout the process. Following that, cells debris were then
SP:SAMPLEPREP_SUMMARY            	centrifuged (15,000 rpm, 10 min, − 4 °C) and the sample supernatants were
SP:SAMPLEPREP_SUMMARY            	collected and transferred to LC vials for drying in the EZ-2 Plus
SP:SAMPLEPREP_SUMMARY            	(GeneVac-Ipswich, UK) at 37±1 °C. Dried samples were resuspended with 200 µL
SP:SAMPLEPREP_SUMMARY            	(water+0.1% formic acid), and vortexed for 2 min. Finally, the samples were
SP:SAMPLEPREP_SUMMARY            	filtered using a hydrophilic Nylon Syringe Filter of 0.45 µm pore size and
SP:SAMPLEPREP_SUMMARY            	analyzed by Q-TOF MS
SP:PROCESSING_STORAGE_CONDITIONS 	Described in summary
SP:EXTRACT_STORAGE               	Described in summary
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	Samples were chromatographically separated by inline reversed-phase
CH:CHROMATOGRAPHY_SUMMARY        	chromatography using the Elute HPG 1300 pumps and Elute Autosampler (Bruker,
CH:CHROMATOGRAPHY_SUMMARY        	Darmstadt, Germany) with solvent A 0.1% FA in HPLC grade water and solvent B
CH:CHROMATOGRAPHY_SUMMARY        	0.1% FA in ACN. A Hamilton Intensity Solo 2 C18 column (100 mm × 2.1 mm, 1.8
CH:CHROMATOGRAPHY_SUMMARY        	μm beads) was maintained at 35 ℃ (metabolomics analyses).
CH:CHROMATOGRAPHY_TYPE           	Reversed phase LC
CH:INSTRUMENT_NAME               	Bruker Elute HPG 1300
CH:COLUMN_NAME                   	Hamilton Intensity Solo 2 C18
CH:FLOW_GRADIENT                 	1%B to 99%B in 15 min
CH:FLOW_RATE                     	250 uL/min
CH:COLUMN_TEMPERATURE            	35
CH:METHODS_FILENAME              	.
CH:SOLVENT_A                     	Water (0.1% Formic Acid)
CH:SOLVENT_B                     	ACN (0.1% Formic Acid)
CH:METHODS_ID                    	.
CH:COLUMN_PRESSURE               	.
CH:INJECTION_TEMPERATURE         	.
CH:INTERNAL_STANDARD             	.
CH:INTERNAL_STANDARD_MT          	.
CH:RETENTION_INDEX               	.
CH:RETENTION_TIME                	.
CH:SAMPLE_INJECTION              	.
CH:SAMPLING_CONE                 	.
CH:ANALYTICAL_TIME               	.
CH:CAPILLARY_VOLTAGE             	.
CH:MIGRATION_TIME                	.
CH:OVEN_TEMPERATURE              	35C
CH:PRECONDITIONING               	.
CH:RUNNING_BUFFER                	.
CH:RUNNING_VOLTAGE               	.
CH:SHEATH_LIQUID                 	.
CH:TIME_PROGRAM                  	.
CH:TRANSFERLINE_TEMPERATURE      	.
CH:WASHING_BUFFER                	.
CH:WEAK_WASH_SOLVENT_NAME        	.
CH:WEAK_WASH_VOLUME              	.
CH:STRONG_WASH_SOLVENT_NAME      	.
CH:STRONG_WASH_VOLUME            	.
CH:TARGET_SAMPLE_TEMPERATURE     	.
CH:SAMPLE_LOOP_SIZE              	.
CH:SAMPLE_SYRINGE_SIZE           	.
CH:RANDOMIZATION_ORDER           	.
CH:CHROMATOGRAPHY_COMMENTS       	.
#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                   	The MS analysis was performed using a timsTOF (Bruker, Darmstadt, Germany) with
MS:MS_COMMENTS                   	Apollo II electrosprayionization (ESI) source. The drying gas was set to flow at
MS:MS_COMMENTS                   	10 L/min and the drying temperature to 220℃ and the nebulizer pressure to 2.2
MS:MS_COMMENTS                   	bar. The capillary voltage was 4500 V and the end plate offset 500 V. For
MS:MS_COMMENTS                   	metabolomics 20–1300 m/z. The instrument was operated in auto-MS/MS mode. For
MS:MS_COMMENTS                   	metabolomics the collision energy was set to 20 eV, the cycle time to 0.5 s with
MS:MS_COMMENTS                   	a relative minimum intensity threshold of 400 counts per thousand and a target
MS:MS_COMMENTS                   	intensity of 20,000. Sodium formate was injected as an external calibrant in the
MS:MS_COMMENTS                   	first 0.3 min of each LC–MS/MS run.
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	AU
MS_METABOLITE_DATA_START
Samples	1A_93_1_328	1A_93_1_331	1A_92_1_327	1A_92_1_330	1A_91_1_326	1A_91_1_329	2a_-1_59_1_440	2a_-2_59_1_441	2a_-1_58_1_438	2a_-2_58_1_439	2a-1_57_1_436	2a-2_57_1_437
Factors	Treatment:Control	Treatment:Control	Treatment:Control	Treatment:Control	Treatment:Control	Treatment:Control	Treatment:TAMOXIFEN	Treatment:TAMOXIFEN	Treatment:TAMOXIFEN	Treatment:TAMOXIFEN	Treatment:TAMOXIFEN	Treatment:TAMOXIFEN
L-Arginine	1678	2032	1866	1960	2126	2186	2364	2094	2440	1524	1810	2042
L-Proline	254	464	480	314	254	96	1072	1566	700	694	1108	798
L-Acetylcarnitine	0	0	0	0	0	0	16922	15334	14726	15112	21838	21154
Adenosine monophosphate	96	112	90	0	540	108	37020	34826	33290	39654	45808	44182
Cytidine	0	0	0	0	0	0	4216	2906	2462	3958	4378	4090
Pyridine	0	0	0	0	0	0	1066	1130	736	868	1324	590
L-Valine	880	1616	1600	1414	1554	1900	0	0	0	0	216	0
Adenosine 3_,5_-diphosphate	17774	39590	31048	42806	8724	46536	210	318	166	374	386	262
Guanosine monophosphate	4552	7114	7072	7174	20838	6828	120	138	112	188	222	100
Deoxyadenosine monophosphate	1242	1606	1114	1896	0	1944	0	0	0	0	0	0
Pyrrolidonecarboxylic acid	18106	18506	17446	19446	8362	20722	0	0	0	84	0	0
L-Tyrosine	12872	11892	12656	13046	0	13076	236	88	0	226	230	736
Doxylamine	0	0	144	0	254	0	1358	696	1144	0	666	1056
Metoprolol	308	220	272	0	724	756	5626	6786	6228	5900	5314	6724
Phenylacetylglycine	0	0	0	0	0	0	1088	1216	614	1074	882	972
Phosphoric acid	1586	1102	1888	1440	3812	1768	198	96	190	84	0	148
Phenylacetaldehyde	2254	2582	2434	2752	1974	2418	166	0	0	0	0	0
Aspartyl-lysine	3888	5036	4410	4516	3372	4272	0	0	0	118	102	0
3-Methylindole	574	618	1984	1656	1912	1620	0	126	0	0	388	226
Spermine	43390	42606	37734	42484	47050	36178	5648	7588	1676	5248	11416	12500
Glycerophosphocholine	5096	5574	6720	8274	4964	7782	3660	3172	2910	2964	3806	3290
L-Glutamic acid	556	1196	1142	1388	574	1368	400	296	370	338	448	144
Creatine	2148	2514	3084	2904	2892	3080	1980	2500	2046	0	2382	2062
Cytidine monophosphate	2464	2586	2556	2718	2094	3350	908	1332	1646	1820	1162	1688
Uridine diphosphate-N-acetylglucosamine	106	0	0	0	0	0	1354	1550	5148	2210	2822	2948
1-Methyladenosine	114	92	0	166	118	0	2710	2160	1302	2988	3434	2574
Trimethylamine	334	130	0	0	0	118	1552	1028	1074	1648	1960	1420
Omeprazole	0	0	0	0	0	0	1516	998	946	1292	1472	1758
Pyroglutamic acid	204	98	346	330	0	246	15044	10904	7672	10296	11064	11146
Uridine 5_-monophosphate	3466	4380	4728	4634	4004	5194	0	96	0	88	0	92
Glutathione	472754	387742	463994	435460	469518	487416	650	1944	248	426	2716	2766
L-Leucine	2060	1970	1452	1704	2652	1440	360	188	0	200	98	700
Hypoxanthine	0	0	0	0	120	0	46316	37342	206	42056	57578	55478
Inosine	0	0	0	0	0	0	10214	10754	0	12528	19082	18274
Guanosine	0	0	0	0	0	0	19106	14832	0	14496	22470	21688
Guanine	64	140	536	152	648	238	33852	27898	184	30206	36924	29376
Adenosine	67130	60510	70696	59562	252	111320	478	444	190	492	0	0
L-Phenylalanine	0	0	0	0	0	0	27126	24854	15106	19084	28346	27108
Beta-Alanine	0	0	0	0	0	0	4032	2184	1018	1802	2204	2512
5_-Methylthioadenosine	0	0	0	0	0	114	3976	4758	2364	3622	4302	4136
L-Tryptophan	98	172	160	0	174	106	31150	30366	15438	22716	29358	32010
2-Ethyl-2-hydroxybutyric acid	1274	1288	1106	1086	1482	908	188	290	480	206	558	292
Isovalerylcarnitine	0	0	112	0	0	0	7120	6496	3630	5714	4116	4598
Sphingosine	17858	14684	16256	13172	11402	15478	100	0	150	242	0	98
PC_16:0_16:0_	0	0	0	0	0	0	1962	2382	2890	3370	2526	1610
PC_18:1_9Z_18:1_9Z_	0	0	0	0	0	0	9510	8784	12142	5972	28374	10916
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	Bucket label	RT	m/z	Formula
L-Arginine	174.11170 Da 68.25 s	1.14	175.11898	C6H14N4O2
L-Proline	115.06312 Da 94.80 s	1.58	116.07039	C5H9NO2
L-Acetylcarnitine	203.11597 Da 96.03 s	1.6	204.12325	C9H17NO4
Adenosine monophosphate	347.06408 Da 96.87 s	1.61	348.07136	C10H14N5O7P
Cytidine	243.08577 Da 97.25 s	1.62	244.09304	C9H13N3O5
Pyridine	79.04255 Da 107.09 s	1.78	80.04983	C5H5N
L-Valine	117.07895 Da 118.12 s	1.97	118.08623	C5H11NO2
Adenosine 3_,5_-diphosphate	427.03111 Da 145.94 s	2.43	428.03838	C10H15N5O10P2
Guanosine monophosphate	363.05925 Da 149.86 s	2.5	364.06653	C10H14N5O8P
Deoxyadenosine monophosphate	331.06886 Da 152.48 s	2.54	332.07614	C10H14N5O6P
Pyrrolidonecarboxylic acid	129.04249 Da 163.68 s	2.73	130.04976	C5H7NO3
L-Tyrosine	181.07404 Da 233.80 s	3.9	182.08132	C9H11NO3
Doxylamine	270.17124 Da 372.87 s	6.21	271.17852	C17H22N2O
Metoprolol	267.18348 Da 450.22 s	7.5	268.19076	C15H25NO3
Phenylacetylglycine	193.07427 Da 455.64 s	7.59	194.08155	C10H11NO3
Phosphoric acid	97.97680 Da 583.94 s	9.73	98.98407	H3O4P
Phenylacetaldehyde	120.05739 Da 776.22 s	12.94	121.06466	C8H8O
Aspartyl-lysine	261.13684 Da 789.33 s	13.16	262.14411	C10H19N3O5
3-Methylindole	131.07341 Da 889.82 s	14.83	132.08069	C9H9N
Spermine	202.21584 Da 57.30 s	0.95	203.22312	C10H26N4
Glycerophosphocholine	257.10296 Da 74.01 s	1.23	258.11023	C8H20NO6P
L-Glutamic acid	147.05315 Da 78.68 s	1.31	148.06043	C5H9NO4
Creatine	131.06934 Da 81.54 s	1.36	132.07661	C4H9N3O2
Cytidine monophosphate	323.05212 Da 85.38 s	1.42	324.0594	C9H14N3O8P
Uridine diphosphate-N-acetylglucosamine	607.08422 Da 95.47 s	1.59	608.0915	C17H27N3O17P2
1-Methyladenosine	281.11293 Da 96.96 s	1.62	282.12021	C11H15N5O4
Trimethylamine	59.07376 Da 96.06 s	1.6	60.08103	C3H9N
Omeprazole	345.11832 Da 98.27 s	1.64	346.12559	C17H19N3O3S
Pyroglutamic acid	129.04263 Da 103.43 s	1.72	130.04985	C5H7NO3
Uridine 5_-monophosphate	324.03634 Da 134.02 s	2.23	325.04361	C9H13N2O9P
Glutathione	307.08436 Da 148.37 s	2.47	308.09163	C10H17N3O6S
L-Leucine	131.09482 Da 213.47 s	3.56	132.1021	C6H13NO2
Hypoxanthine	136.03838 Da 285.18 s	4.75	137.04565	C5H4N4O
Inosine	268.08034 Da 285.40 s	4.76	269.08761	C10H12N4O5
Guanosine	283.09106 Da 285.52 s	4.76	284.09834	C10H13N5O5
Guanine	151.04925 Da 285.51 s	4.76	152.05652	C5H5N5O
Adenosine	267.09682 Da 304.38 s	5.07	268.1041	C10H13N5O4
L-Phenylalanine	165.07899 Da 313.32 s	5.22	166.08626	C9H11NO2
Beta-Alanine	89.04761 Da 335.29 s	5.59	90.05488	C3H7NO2
5_-Methylthioadenosine	297.09013 Da 356.85 s	5.95	298.09741	C11H15N5O3S
L-Tryptophan	204.09035 Da 366.10 s	6.1	205.09763	C11H12N2O2
2-Ethyl-2-hydroxybutyric acid	132.07829 Da 379.55 s	6.33	133.08557	C6H12O3
Isovalerylcarnitine	245.16271 Da 389.28 s	6.49	246.16999	C12H23NO4
Sphingosine	299.28329 Da 752.77 s	12.55	282.27997	C18H37NO2
PC_16:0_16:0_	733.56294 Da 871.19 s	14.52	734.57022	C40H80NO8P
PC_18:1_9Z_18:1_9Z_	785.59742 Da 1155.44 s	19.26	786.6047	C44H84NO8P
METABOLITES_END
#END