#METABOLOMICS WORKBENCH liu788_20221228_174024 DATATRACK_ID:3676 STUDY_ID:ST002420 ANALYSIS_ID:AN003941 PROJECT_ID:PR001558
VERSION             	1
CREATED_ON             	January 2, 2023, 8:55 pm
#PROJECT
PR:PROJECT_TITLE                 	Isobaric labeling metabolites CRC studies
PR:PROJECT_TYPE                  	MS relative quantitative analysis
PR:PROJECT_SUMMARY               	Isobaric labeling-based relative quantification for the discovery of serum
PR:PROJECT_SUMMARY               	metabolite biomarkers associated with growing early colorectal adenomas
PR:INSTITUTE                     	University of Wisconsin - Madison
PR:LAST_NAME                     	Liu
PR:FIRST_NAME                    	Yuan
PR:ADDRESS                       	777 Highland ave, Madison, Wisconsin, 53705, USA
PR:EMAIL                         	liu788@wisc.edu
PR:PHONE                         	6089606939
#STUDY
ST:STUDY_TITLE                   	Colorectal cancer isobaric labeling for metabolite quantification
ST:STUDY_SUMMARY                 	A major challenge in reducing the death rate of colorectal cancer is to screen
ST:STUDY_SUMMARY                 	patients using low-invasive testing. Blood test shows a high compliance rate
ST:STUDY_SUMMARY                 	with reduced invasiveness. In this work, a multiplex isobaric tag labeling
ST:STUDY_SUMMARY                 	strategy coupled with mass spectrometry is adopted to relatively quantify
ST:STUDY_SUMMARY                 	primary and secondary amine-containing metabolites in serum for the discovery of
ST:STUDY_SUMMARY                 	metabolite biomarkers of colorectal cancer. Serum samples from patients at
ST:STUDY_SUMMARY                 	different risk statuses and colorectal cancer growth statuses are studied.
ST:STUDY_SUMMARY                 	Metabolite identification is based on accurate mass matching and/or retention
ST:STUDY_SUMMARY                 	time of labeled metabolite standards. We quantify 40 metabolites across all the
ST:STUDY_SUMMARY                 	serum samples, including 18 metabolites validated with standards. We find
ST:STUDY_SUMMARY                 	significantly decreased levels of threonine and asparagine in the patients with
ST:STUDY_SUMMARY                 	growing adenomas or high-risk adenomas (p < 0.05). Glutamine levels decrease in
ST:STUDY_SUMMARY                 	patients with adenomas of unknown growth status or high-risk adenomas. In
ST:STUDY_SUMMARY                 	contrast, arginine levels are elevated in patients with low-risk adenoma.
ST:STUDY_SUMMARY                 	Receiver operating characteristic analysis shows high sensitivity and
ST:STUDY_SUMMARY                 	specificity of these metabolites for detecting growing adenomas. Based on these
ST:STUDY_SUMMARY                 	results, we conclude that potential metabolite biomarkers identified here
ST:STUDY_SUMMARY                 	contribute to distinguishing colorectal patients with growing adenomas from
ST:STUDY_SUMMARY                 	normal individuals and patients with unknown growth status of adenomas.
ST:INSTITUTE                     	University of Wisconsin - Madison
ST:LAST_NAME                     	Liu
ST:FIRST_NAME                    	Yuan
ST:ADDRESS                       	777 Highland ave, Madison, Wisconsin, 53705, USA
ST:EMAIL                         	liu788@wisc.edu
ST:PHONE                         	6089606939
#SUBJECT
SU:SUBJECT_TYPE                  	Human
SU:SUBJECT_SPECIES               	Homo sapiens
SU:TAXONOMY_ID                   	9606
SU:GENDER                        	Male and female
#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           	-	1_pre	classification_based on growth rate:Static | classification_based on risks:Low‐Risk	m/z of reporter ion=115; RAW_FILE_NAME=G1_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	1_post	classification_based on growth rate:Static | classification_based on risks:Low‐Risk	m/z of reporter ion=116; RAW_FILE_NAME=G1_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	2_pre	classification_based on growth rate:growing | classification_based on risks:Low‐Risk	m/z of reporter ion=117; RAW_FILE_NAME=G1_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	2_post	classification_based on growth rate:growing | classification_based on risks:Low‐Risk	m/z of reporter ion=118; RAW_FILE_NAME=G1_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	3_pre	classification_based on growth rate:Static | classification_based on risks:Low‐Risk	m/z of reporter ion=115; RAW_FILE_NAME=G2_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	3_post	classification_based on growth rate:Static | classification_based on risks:Low‐Risk	m/z of reporter ion=116; RAW_FILE_NAME=G2_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	5_pre	classification_based on growth rate:Growing | classification_based on risks:High‐risk	m/z of reporter ion=117; RAW_FILE_NAME=G2_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	5_post	classification_based on growth rate:Growing | classification_based on risks:High‐risk	m/z of reporter ion=118; RAW_FILE_NAME=G2_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	7_post	classification_based on growth rate:Growing | classification_based on risks:High‐risk	m/z of reporter ion=115; RAW_FILE_NAME=G3_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	7_pre	classification_based on growth rate:Growing | classification_based on risks:High‐risk	m/z of reporter ion=116; RAW_FILE_NAME=G3_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	8_pre	classification_based on growth rate:Growing | classification_based on risks:High‐risk	m/z of reporter ion=117; RAW_FILE_NAME=G3_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	8_post	classification_based on growth rate:Growing | classification_based on risks:High‐risk	m/z of reporter ion=118; RAW_FILE_NAME=G3_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	9_pre	classification_based on growth rate:Growing | classification_based on risks:Low‐Risk	m/z of reporter ion=115; RAW_FILE_NAME=G4_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	9_post	classification_based on growth rate:Growing | classification_based on risks:Low‐Risk	m/z of reporter ion=116; RAW_FILE_NAME=G4_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	10_pre	classification_based on growth rate:Unknown | classification_based on risks:High‐risk	m/z of reporter ion=117; RAW_FILE_NAME=G4_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	10_post	classification_based on growth rate:Unknown | classification_based on risks:High‐risk	m/z of reporter ion=118; RAW_FILE_NAME=G4_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	11_pre	classification_based on growth rate:Unknown | classification_based on risks:Low‐Risk	m/z of reporter ion=115; RAW_FILE_NAME=G5_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	11_post	classification_based on growth rate:Unknown | classification_based on risks:Low‐Risk	m/z of reporter ion=116; RAW_FILE_NAME=G5_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	12_pre	classification_based on growth rate:Unknown | classification_based on risks:High‐risk	m/z of reporter ion=117; RAW_FILE_NAME=G5_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	12_post	classification_based on growth rate:Unknown | classification_based on risks:High‐risk	m/z of reporter ion=118; RAW_FILE_NAME=G5_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	15_pre	classification_based on growth rate:Growing | classification_based on risks:High‐risk	m/z of reporter ion=115; RAW_FILE_NAME=G6_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	15_post	classification_based on growth rate:Growing | classification_based on risks:High‐risk	m/z of reporter ion=116; RAW_FILE_NAME=G6_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	18_pre	classification_based on growth rate:Growing | classification_based on risks:Low‐Risk	m/z of reporter ion=117; RAW_FILE_NAME=G6_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	18_post	classification_based on growth rate:Growing | classification_based on risks:Low‐Risk	m/z of reporter ion=118; RAW_FILE_NAME=G6_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	19_pre	classification_based on growth rate:Growing | classification_based on risks:High‐risk	m/z of reporter ion=115; RAW_FILE_NAME=G7_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	19_post	classification_based on growth rate:Growing | classification_based on risks:High‐risk	m/z of reporter ion=116; RAW_FILE_NAME=G7_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	20_pre	classification_based on growth rate:static | classification_based on risks:High‐risk	m/z of reporter ion=117; RAW_FILE_NAME=G7_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	20_post	classification_based on growth rate:static | classification_based on risks:High‐risk	m/z of reporter ion=118; RAW_FILE_NAME=G7_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	21_pre	classification_based on growth rate:Unknown | classification_based on risks:High‐risk	m/z of reporter ion=115; RAW_FILE_NAME=G8_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	21_post	classification_based on growth rate:Unknown | classification_based on risks:High‐risk	m/z of reporter ion=116; RAW_FILE_NAME=G8_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	23_pre	classification_based on growth rate:Growing | classification_based on risks:High‐risk	m/z of reporter ion=117; RAW_FILE_NAME=G8_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	23_post	classification_based on growth rate:Growing | classification_based on risks:High‐risk	m/z of reporter ion=118; RAW_FILE_NAME=G8_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	24_pre	classification_based on growth rate:Growing | classification_based on risks:High‐risk	m/z of reporter ion=115; RAW_FILE_NAME=G9_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	24_post	classification_based on growth rate:Growing | classification_based on risks:High‐risk	m/z of reporter ion=116; RAW_FILE_NAME=G9_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	25_pre	classification_based on growth rate:Growing | classification_based on risks:High‐risk	m/z of reporter ion=117; RAW_FILE_NAME=G9_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	25_post	classification_based on growth rate:Growing | classification_based on risks:High‐risk	m/z of reporter ion=118; RAW_FILE_NAME=G9_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	26_pre	classification_based on growth rate:Unknown | classification_based on risks:Low‐Risk	m/z of reporter ion=115; RAW_FILE_NAME=G10_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	26_post	classification_based on growth rate:Unknown | classification_based on risks:Low‐Risk	m/z of reporter ion=116; RAW_FILE_NAME=G10_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	28_pre	classification_based on growth rate:Growing | classification_based on risks:High‐risk	m/z of reporter ion=117; RAW_FILE_NAME=G10_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	28_post	classification_based on growth rate:Growing | classification_based on risks:High‐risk	m/z of reporter ion=118; RAW_FILE_NAME=G10_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	29_pre	classification_based on growth rate:Unknown | classification_based on risks:High‐risk	m/z of reporter ion=115; RAW_FILE_NAME=G11_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	29_post	classification_based on growth rate:Unknown | classification_based on risks:High‐risk	m/z of reporter ion=116; RAW_FILE_NAME=G11_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	32_pre	classification_based on growth rate:Growing | classification_based on risks:Low‐Risk	m/z of reporter ion=117; RAW_FILE_NAME=G11_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	32_post	classification_based on growth rate:Growing | classification_based on risks:Low‐Risk	m/z of reporter ion=118; RAW_FILE_NAME=G11_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	33_pre	classification_based on growth rate:Growing | classification_based on risks:High‐risk	m/z of reporter ion=115; RAW_FILE_NAME=G12_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	33_post	classification_based on growth rate:Growing | classification_based on risks:High‐risk	m/z of reporter ion=116; RAW_FILE_NAME=G12_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	N1	classification_based on growth rate:control | classification_based on risks:control	m/z of reporter ion=117; RAW_FILE_NAME=G12_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	N2	classification_based on growth rate:control | classification_based on risks:control	m/z of reporter ion=118; RAW_FILE_NAME=G12_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	N3	classification_based on growth rate:control | classification_based on risks:control	m/z of reporter ion=115; RAW_FILE_NAME=G13_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	N4	classification_based on growth rate:control | classification_based on risks:control	m/z of reporter ion=116; RAW_FILE_NAME=G13_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	N5	classification_based on growth rate:control | classification_based on risks:control	m/z of reporter ion=117; RAW_FILE_NAME=G13_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	N6	classification_based on growth rate:control | classification_based on risks:control	m/z of reporter ion=118; RAW_FILE_NAME=G13_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	N7	classification_based on growth rate:control | classification_based on risks:control	m/z of reporter ion=115; RAW_FILE_NAME=G14_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	N8	classification_based on growth rate:control | classification_based on risks:control	m/z of reporter ion=116; RAW_FILE_NAME=G14_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	N9	classification_based on growth rate:control | classification_based on risks:control	m/z of reporter ion=117; RAW_FILE_NAME=G14_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	N10	classification_based on growth rate:control | classification_based on risks:control	m/z of reporter ion=118; RAW_FILE_NAME=G14_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	N11	classification_based on growth rate:control | classification_based on risks:control	m/z of reporter ion=115; RAW_FILE_NAME=G15_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	N12	classification_based on growth rate:control | classification_based on risks:control	m/z of reporter ion=116; RAW_FILE_NAME=G15_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	N13	classification_based on growth rate:control | classification_based on risks:control	m/z of reporter ion=117; RAW_FILE_NAME=G15_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	N14	classification_based on growth rate:control | classification_based on risks:control	m/z of reporter ion=118; RAW_FILE_NAME=G15_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	N15	classification_based on growth rate:control | classification_based on risks:control	m/z of reporter ion=115; RAW_FILE_NAME=G16_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	N16	classification_based on growth rate:control | classification_based on risks:control	m/z of reporter ion=116; RAW_FILE_NAME=G16_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	N17	classification_based on growth rate:control | classification_based on risks:control	m/z of reporter ion=117; RAW_FILE_NAME=G16_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	N18	classification_based on growth rate:control | classification_based on risks:control	m/z of reporter ion=118; RAW_FILE_NAME=G16_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	N19	classification_based on growth rate:control | classification_based on risks:control	m/z of reporter ion=115; RAW_FILE_NAME=G17_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	N20	classification_based on growth rate:control | classification_based on risks:control	m/z of reporter ion=116; RAW_FILE_NAME=G17_1.mzML
#COLLECTION
CO:COLLECTION_SUMMARY            	Serum samples were thawed on ice and centrifuged at 2000 g for 10 min to remove
CO:COLLECTION_SUMMARY            	particulates and debris. Molecular weight cut-off filters (MWCO, 3 kDa,
CO:COLLECTION_SUMMARY            	Millipore Amicon Ultra, Burlington, MA) were prerinsed 3 times with optima water
CO:COLLECTION_SUMMARY            	at 14000 g for 20 min. Twenty microliter serum supernatant from each sample was
CO:COLLECTION_SUMMARY            	diluted in 380 µl water and added to the filter then centrifuged for 20 min at
CO:COLLECTION_SUMMARY            	14000 g. Flowthrough was collected. Then 400 µL water was added to the filter
CO:COLLECTION_SUMMARY            	to rinse the sample. An additional rinse step was applied. All the flowthrough
CO:COLLECTION_SUMMARY            	(about 1.2 mL) was combined and dried down in Speedvac and stored at −20 °C
CO:COLLECTION_SUMMARY            	until labeling. For normalization, a pooled serum sample from screening normal
CO:COLLECTION_SUMMARY            	individuals was also prepared. For labeling of serum samples, 40 µL activated
CO:COLLECTION_SUMMARY            	DiLeu reagents were mixed with serum flowthrough dissolved in 10 µL 0.5 M TEAB
CO:COLLECTION_SUMMARY            	and shaken for 2 h. After the reaction was quenched, 2.5 µL of each reaction
CO:COLLECTION_SUMMARY            	mixture with different DiLeu tags were combined at 1: 1: 1: 1 ratio and mixed
CO:COLLECTION_SUMMARY            	well. To compare the relative abundance of each set of 4-plex DiLeu labeled
CO:COLLECTION_SUMMARY            	metabolites, one channel from each 4-plex combination was selected and combined
CO:COLLECTION_SUMMARY            	with 118-DiLeu tag labeled pooled serum samples. Ten µL of the mixture was
CO:COLLECTION_SUMMARY            	taken for drying down and desalted using SCX Ziptips. Samples were dissolved in
CO:COLLECTION_SUMMARY            	15 µL 0.1% FA and 3 µL was injected into LC-MS for data acquisition with 3
CO:COLLECTION_SUMMARY            	technical replicates.
CO:COLLECTION_PROTOCOL_FILENAME  	methods.txt
CO:SAMPLE_TYPE                   	Blood (serum)
#TREATMENT
TR:TREATMENT_SUMMARY             	Among the 23 patients carrying adenomas, 15 patients were classified
TR:TREATMENT_SUMMARY             	histologically as high-risk and the remaining 8 patients were classified as
TR:TREATMENT_SUMMARY             	low-risk cases. In the high-risk cases, 10 were classified as growing by
TR:TREATMENT_SUMMARY             	longitudinal CTC analysis, one as static, and 4 as unknown growth status.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Serum samples were thawed on ice and centrifuged at 2000 g for 10 min to remove
SP:SAMPLEPREP_SUMMARY            	particulates and debris. Molecular weight cut-off filters (MWCO, 3 kDa,
SP:SAMPLEPREP_SUMMARY            	Millipore Amicon Ultra, Burlington, MA) were prerinsed 3 times with optima water
SP:SAMPLEPREP_SUMMARY            	at 14000 g for 20 min. Twenty microliter serum supernatant from each sample was
SP:SAMPLEPREP_SUMMARY            	diluted in 380 µl water and added to the filter then centrifuged for 20 min at
SP:SAMPLEPREP_SUMMARY            	14000 g. Flowthrough was collected. Then 400 µL water was added to the filter
SP:SAMPLEPREP_SUMMARY            	to rinse the sample. An additional rinse step was applied. All the flowthrough
SP:SAMPLEPREP_SUMMARY            	(about 1.2 mL) was combined and dried down in Speedvac and stored at −20 °C
SP:SAMPLEPREP_SUMMARY            	until labeling. For normalization, a pooled serum sample from screening normal
SP:SAMPLEPREP_SUMMARY            	individuals was also prepared. For labeling of serum samples, 40 µL activated
SP:SAMPLEPREP_SUMMARY            	DiLeu reagents were mixed with serum flowthrough dissolved in 10 µL 0.5 M TEAB
SP:SAMPLEPREP_SUMMARY            	and shaken for 2 h. After the reaction was quenched, 2.5 µL of each reaction
SP:SAMPLEPREP_SUMMARY            	mixture with different DiLeu tags were combined at 1: 1: 1: 1 ratio and mixed
SP:SAMPLEPREP_SUMMARY            	well. To compare the relative abundance of each set of 4-plex DiLeu labeled
SP:SAMPLEPREP_SUMMARY            	metabolites, one channel from each 4-plex combination was selected and combined
SP:SAMPLEPREP_SUMMARY            	with 118-DiLeu tag labeled pooled serum samples. Ten µL of the mixture was
SP:SAMPLEPREP_SUMMARY            	taken for drying down and desalted using SCX Ziptips. Samples were dissolved in
SP:SAMPLEPREP_SUMMARY            	15 µL 0.1% FA and 3 µL was injected into LC-MS for data acquisition with 3
SP:SAMPLEPREP_SUMMARY            	technical replicates.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	Four-plex pooled samples were reconstituted in 0.1% formic acid before
CH:CHROMATOGRAPHY_SUMMARY        	injection. The HPLC-MS/MS analysis was conducted using a Waters nanoACQUITY UPLC
CH:CHROMATOGRAPHY_SUMMARY        	coupled with Thermo Q Exactive Orbitrap MS. The separation column was in-house
CH:CHROMATOGRAPHY_SUMMARY        	made with an emitter tip and dimensions of 75 μm inner diameter × 15 cm
CH:CHROMATOGRAPHY_SUMMARY        	length. The column was packed with 1.7 μm, 150 Å, ethylene-bridged-hybrid
CH:CHROMATOGRAPHY_SUMMARY        	(BEH) C18 material (Waters, Milford, MA). Mobile phase A was water containing
CH:CHROMATOGRAPHY_SUMMARY        	0.1% formic acid, and mobile phase B was acetonitrile containing 0.1% formic
CH:CHROMATOGRAPHY_SUMMARY        	acid. The flow rate was set as 0.3 μL/min, and the LC gradient was 55 min and
CH:CHROMATOGRAPHY_SUMMARY        	set as follows: 0−10 min, 3%-30% solvent B; 10−30 min, 30−80% B; 30-30.5
CH:CHROMATOGRAPHY_SUMMARY        	min, 80%-95% B; 30.5− 40.5 min, 95% B; 40.5−41 min, 95%-3% B; 41-55 min, 3%
CH:CHROMATOGRAPHY_SUMMARY        	B. Positive ionization mode was used and full MS scans were acquired from m/z
CH:CHROMATOGRAPHY_SUMMARY        	180 to 800 at a resolution of 60 k, automatic gain control (AGC) was set as 5x
CH:CHROMATOGRAPHY_SUMMARY        	10^5, and a maximum injection time as set as 30 ms. The top 20 precursors were
CH:CHROMATOGRAPHY_SUMMARY        	selected for normalized collision energy (NCE) dissociation (NCE = 30) with an
CH:CHROMATOGRAPHY_SUMMARY        	isolation window of m/z 1, fixed first m/z 110, dynamic exclusion of 5 seconds,
CH:CHROMATOGRAPHY_SUMMARY        	charge exclusion of >2, and a resolution of 35 k.
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Waters NanoAcquity
CH:COLUMN_NAME                   	Waters ACQUITY UPLC BEH C18 (150 mm x 75 μm, 1.7 μm, 150 Å)
CH:SOLVENT_A                     	100% water; 0.01% formic acid
CH:SOLVENT_B                     	100% acetonitrile; 0.01% formic acid
CH:FLOW_GRADIENT                 	LC gradient was 55 min and set as follows: 0−10 min, 3%-30% solvent B; 10−30
CH:FLOW_GRADIENT                 	min, 30−80% B; 30-30.5 min, 80%-95% B; 30.5− 40.5 min, 95% B; 40.5−41 min,
CH:FLOW_GRADIENT                 	95%-3% B; 41-55 min, 3% B.
CH:FLOW_RATE                     	0.3 μL/min
CH:COLUMN_TEMPERATURE            	Room temperature
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Thermo Q Exactive Orbitrap
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	POSITIVE
MS:MS_COMMENTS                   	Positive ionization mode was used and full MS scans were acquired from m/z 180
MS:MS_COMMENTS                   	to 800 at a resolution of 60 k, automatic gain control (AGC) was set as 5x 10^5,
MS:MS_COMMENTS                   	and a maximum injection time as set as 30 ms. The top 20 precursors were
MS:MS_COMMENTS                   	selected for normalized collision energy (NCE) dissociation (NCE = 30) with an
MS:MS_COMMENTS                   	isolation window of m/z 1, fixed first m/z 110, dynamic exclusion of 5 seconds,
MS:MS_COMMENTS                   	charge exclusion of >2, and a resolution of 35 k.
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	1
MS_METABOLITE_DATA_START
Samples	1_pre	1_post	2_pre	2_post	3_pre	3_post	5_pre	5_post	7_post	7_pre	8_pre	8_post	9_pre	9_post	10_pre	10_post	11_pre	11_post	12_pre	12_post	15_pre	15_post	18_pre	18_post	19_pre	19_post	20_pre	20_post	21_pre	21_post	23_pre	23_post	24_pre	24_post	25_pre	25_post	26_pre	26_post	28_pre	28_post	29_pre	29_post	32_pre	32_post	33_pre	33_post	N1	N2	N3	N4	N5	N6	N7	N8	N9	N10	N11	N12	N13	N14	N15	N16	N17	N18	N19	N20
Factors	classification_based on growth rate:Static | classification_based on risks:Low‐Risk	classification_based on growth rate:Static | classification_based on risks:Low‐Risk	classification_based on growth rate:growing | classification_based on risks:Low‐Risk	classification_based on growth rate:growing | classification_based on risks:Low‐Risk	classification_based on growth rate:Static | classification_based on risks:Low‐Risk	classification_based on growth rate:Static | classification_based on risks:Low‐Risk	classification_based on growth rate:Growing | classification_based on risks:High‐risk	classification_based on growth rate:Growing | classification_based on risks:High‐risk	classification_based on growth rate:Growing | classification_based on risks:High‐risk	classification_based on growth rate:Growing | classification_based on risks:High‐risk	classification_based on growth rate:Growing | classification_based on risks:High‐risk	classification_based on growth rate:Growing | classification_based on risks:High‐risk	classification_based on growth rate:Growing | classification_based on risks:Low‐Risk	classification_based on growth rate:Growing | classification_based on risks:Low‐Risk	classification_based on growth rate:Unknown | classification_based on risks:High‐risk	classification_based on growth rate:Unknown | classification_based on risks:High‐risk	classification_based on growth rate:Unknown | classification_based on risks:Low‐Risk	classification_based on growth rate:Unknown | classification_based on risks:Low‐Risk	classification_based on growth rate:Unknown | classification_based on risks:High‐risk	classification_based on growth rate:Unknown | classification_based on risks:High‐risk	classification_based on growth rate:Growing | classification_based on risks:High‐risk	classification_based on growth rate:Growing | classification_based on risks:High‐risk	classification_based on growth rate:Growing | classification_based on risks:Low‐Risk	classification_based on growth rate:Growing | classification_based on risks:Low‐Risk	classification_based on growth rate:Growing | classification_based on risks:High‐risk	classification_based on growth rate:Growing | classification_based on risks:High‐risk	classification_based on growth rate:static | classification_based on risks:High‐risk	classification_based on growth rate:static | classification_based on risks:High‐risk	classification_based on growth rate:Unknown | classification_based on risks:High‐risk	classification_based on growth rate:Unknown | classification_based on risks:High‐risk	classification_based on growth rate:Growing | classification_based on risks:High‐risk	classification_based on growth rate:Growing | classification_based on risks:High‐risk	classification_based on growth rate:Growing | classification_based on risks:High‐risk	classification_based on growth rate:Growing | classification_based on risks:High‐risk	classification_based on growth rate:Growing | classification_based on risks:High‐risk	classification_based on growth rate:Growing | classification_based on risks:High‐risk	classification_based on growth rate:Unknown | classification_based on risks:Low‐Risk	classification_based on growth rate:Unknown | classification_based on risks:Low‐Risk	classification_based on growth rate:Growing | classification_based on risks:High‐risk	classification_based on growth rate:Growing | classification_based on risks:High‐risk	classification_based on growth rate:Unknown | classification_based on risks:High‐risk	classification_based on growth rate:Unknown | classification_based on risks:High‐risk	classification_based on growth rate:Growing | classification_based on risks:Low‐Risk	classification_based on growth rate:Growing | classification_based on risks:Low‐Risk	classification_based on growth rate:Growing | classification_based on risks:High‐risk	classification_based on growth rate:Growing | classification_based on risks:High‐risk	classification_based on growth rate:control | classification_based on risks:control	classification_based on growth rate:control | classification_based on risks:control	classification_based on growth rate:control | classification_based on risks:control	classification_based on growth rate:control | classification_based on risks:control	classification_based on growth rate:control | classification_based on risks:control	classification_based on growth rate:control | classification_based on risks:control	classification_based on growth rate:control | classification_based on risks:control	classification_based on growth rate:control | classification_based on risks:control	classification_based on growth rate:control | classification_based on risks:control	classification_based on growth rate:control | classification_based on risks:control	classification_based on growth rate:control | classification_based on risks:control	classification_based on growth rate:control | classification_based on risks:control	classification_based on growth rate:control | classification_based on risks:control	classification_based on growth rate:control | classification_based on risks:control	classification_based on growth rate:control | classification_based on risks:control	classification_based on growth rate:control | classification_based on risks:control	classification_based on growth rate:control | classification_based on risks:control	classification_based on growth rate:control | classification_based on risks:control	classification_based on growth rate:control | classification_based on risks:control	classification_based on growth rate:control | classification_based on risks:control
Alanine	1.16120383	1.541312393	0.960753195	1.011355783	0.923854206	1.616502585	0.781319761	0.878116919	1.344688884	1.534836034	1.069712892	1.415490176	0.848232395	1.404198124	1.057847399	1.256550407	1.131388256	1.463615276	0.74450952	1.21270013	0.994244935	1.012599259	1.025757503	1.352270557	1.307522895	1.477792794	1.192831143	1.931217308	0.929065045	1.367765931	1.153576987	1.694841133	0.695645166	1.302282223	1.299572981	1.219913019	0.820863909	1.01871839	0.944736447	1.848377654	0.949450267	1.826836791	1.008737938	1.298883205	0.944928893	0.985666911	1.244819042	0.995648811	1.029865407	0.879527761	1.016855416	1.106554201	0.846076928	1.001341058	0.867095297	0.976371607	1.022614812	1.065325242	0.963514477	1.16391038	1.147828038	0.942473131	1.138554917	0.861599602	0.703659568	0.943104623
Proline	1.336526389	2.640566926	1.037674779	2.256864035	0.925197407	1.739768983	0.700186278	0.922404091	1.368174483	1.191896254	1.133431888	2.087105592	1.141230697	2.216940259	0.820429456	1.288424081	1.195531037	1.719521256	0.676888228	1.428227635	0.948305896	1.234132055	0.967056196	2.00381555	1.100988266	1.639148424	1.003878888	2.368224123	1.105198363	2.237481632	1.144742932	1.971642093	0.867110816	2.408114627	1.167955975	1.990696981	1.339787824	1.374587201	0.596287592	1.174570457	0.751289681	1.577793044	0.542751307	0.797504506	0.866590797	0.937861576	0.981964523	0.793557307	0.885408274	1.237257065	1.695645392	0.787111033	1.01243777	1.000029488	1.179838126	0.854294826	1.13478122	1.013085593	1.171189472	0.968313755	0.866544569	1.121408052	0.951906076	0.842419862	0.705906551	1.118313691
Valine	1.158540365	1.50971809	0.788299979	1.278440746	0.809408065	0.968954663	0.881869247	0.91463845	1.20045018	1.04909552	0.972029349	1.085020418	0.767157625	1.027616989	0.851758402	0.879210366	1.055993779	1.293399281	0.846197538	0.912543779	0.647241587	0.958336524	0.772710096	1.125266613	1.10689331	1.651206658	1.182379248	1.389277999	0.832243111	1.137687637	0.731966604	0.947583575	0.773218864	1.244840162	1.020317216	1.082473707	1.097760569	0.899796026	1.009059741	1.105077087	0.683020015	1.104477958	0.660552077	0.694585178	0.687503283	0.849544116	1.104488788	1.092756383	0.830738805	0.910215451	1.506220749	0.929905307	0.843483143	0.829347153	1.503096733	1.010726077	0.950280736	0.903015002	0.858051123	1.019184519	0.797859322	1.074722034	1.450802044	0.889321932	0.658924412	0.980828909
Threonine	0.889863734	1.016477624	0.821118293	0.898210213	0.950447321	1.011349867	0.680173492	0.881890069	1.130641657	0.948736408	0.749529692	1.011206336	0.614531493	1.029218826	0.745901382	0.931598148	0.695955643	0.893492698	0.840481345	1.345015535	0.691448544	0.770163501	0.691819335	0.982310822	0.749457875	0.814429871	1.232840203	1.692529547	0.861435502	1.007537778	0.864598625	1.10753809	0.619542005	0.94491576	0.810027114	0.734914362	0.787977298	0.766888622	0.713504515	1.029843359	0.676770509	1.118438922	0.765816357	1.028203763	0.696407228	0.949058271	0.899731144	0.920692026	0.953902515	0.777717814	0.9805961	1.076084798	0.739114976	0.942294563	0.830541385	1.047490671	1.269737322	0.917940086	1.05357226	0.815766054	0.749557393	0.855166703	0.879243379	0.879448012	0.462266277	0.774135346
Isoleucine	1.247357619	1.498185547	0.855850237	1.771697753	0.829673432	0.924718768	0.835798656	0.704642633	1.79611668	1.071934847	1.046527424	1.054442217	0.806373837	1.044515532	0.768012452	0.686476804	1.294284428	1.242640855	1.095672404	0.986491194	0.774003754	1.18677858	0.92874512	1.124539315	1.094823593	2.311489417	1.45246086	1.440667526	0.841364137	1.204898383	0.735246561	0.916974798	0.854241865	1.398591124	1.262618243	1.042164517	0.940710806	0.926400472	0.965133036	1.057569092	0.391267426	0.913184805	0.555209421	0.504492426	0.698836467	0.718492759	1.097855326	1.225805552	0.779524865	0.843418707	1.854665924	0.802620636	1.141071587	0.796909163	1.54405519	0.903059918	0.911645836	0.966767252	0.710468572	0.921249341	0.796036986	1.029213536	1.609917273	0.81580401	0.618928319	1.124872245
Leucine	1.080148273	1.104693052	0.794830277	1.412267232	0.79959544	0.853673141	0.767355194	0.669663226	1.251153474	1.147370441	0.890560259	0.843081796	0.675142536	0.733007111	0.693475557	0.576891312	0.891915677	0.956532786	0.685266343	0.581228511	0.669955644	0.778817502	0.749020032	0.888241118	0.974105516	1.622116392	1.236070874	1.027322365	0.833894988	1.138865587	0.634340183	0.729857806	0.705968733	0.968766139	0.947454947	0.826393196	1.050595388	0.926846005	1.013588571	0.89138846	0.621096073	1.027471311	0.661852211	0.65385658	0.756547222	0.708855768	1.077060658	1.131719274	0.757701117	0.766363638	1.427941472	0.810761272	0.809661658	0.792032487	1.474718318	0.964892357	1.000994274	0.917808637	0.84027278	1.065634337	0.762784518	1.021415899	1.619057229	0.843246686	0.605200404	0.971623168
Asparagine	0.704489008	0.94122583	0.778178987	1.361654087	0.472972495	0.888534646	0.62037891	0.90623364	1.047090239	0.931711479	0.671637057	0.775798936	0.599301587	0.913123295	0.729180055	0.732006393	0.704803156	0.732938922	0.461169162	0.659694619	0.656662173	0.615934364	0.715027191	1.218287728	0.513354812	0.789065927	0.692969971	0.781426724	0.497870326	0.522722937	0.545570498	0.644001855	0.478079618	0.690836676	0.559283154	0.432394208	0.687040726	0.48071495	0.581462079	0.698252434	0.405305913	0.664164548	0.608903947	0.628389081	0.788986105	0.593129002	0.734282733	0.904417014	0.654763724	0.74782771	1.059023277	1.012273051	0.687021554	0.806779672	1.119772423	1.001037599	0.908622237	0.589591642	0.75255757	0.878071106	0.639133041	0.737540686	0.894871932	0.866464451	0.512083264	0.85585779
Glutamine	0.833718722	0.792128742	1.040874697	1.194218454	0.688203311	0.792710949	0.768262835	0.825792326	0.807764316	1.004535971	0.825138642	0.85313372	0.780701067	1.014521608	0.811430836	0.78142007	0.773539624	0.771070616	0.601882859	0.729831365	0.76602158	0.720672892	0.801485042	0.908022543	0.768693186	1.000967876	0.948827907	0.898590743	0.78576636	0.842814884	0.974002407	0.987454403	0.80018714	0.956616342	0.857241246	0.70583399	0.842939278	0.805284586	0.876850604	0.912087419	0.717360671	0.954602955	1.086759395	1.21026381	0.831675891	1.033362529	0.84565554	1.174244744	0.820942726	1.081469024	1.090678615	1.012522438	0.8045053	0.947143078	0.978568947	1.102809554	0.971115583	0.929613278	0.938163585	0.944351463	0.901519291	0.820901483	1.092214379	0.898746699	0.606147157	0.919509096
Methionine	1.004958986	0.981274702	1.133318001	1.221209145	0.959375171	0.996734505	0.823691191	0.972105317	1.326361817	1.093614279	0.965555385	1.038213691	0.820583508	1.070869046	0.919978188	0.799836147	1.081709231	1.068518202	0.855857087	1.190658413	1.015046822	0.9485401	0.899484269	1.227071689	1.14161554	1.589038504	1.447320499	1.349532595	1.056783989	1.387355404	0.913707137	1.437006125	0.87294919	1.564390416	1.237583299	0.655644402	0.873165372	0.906675792	0.908785446	1.251810079	0.777902511	1.035217887	0.844393955	0.935800632	1.190495502	1.120619788	1.185483071	1.002486317	0.874342044	0.885155401	1.356472392	1.161447849	0.687177478	0.881940797	1.220133659	1.123537151	0.957741422	0.988928675	0.80534276	1.138844805	0.950867349	0.963753683	1.113449577	0.984095473	0.556804005	0.839485877
Lysine	0.977127366	0.887156361	1.205468147	1.082957382	1.200051029	1.253596996	0.768631708	0.892219711	1.322145341	1.138283781	0.867988872	0.943497339	0.878039865	1.294770828	1.006396291	1.192645009	0.947694696	0.913115122	0.85283416	0.971298411	0.730683614	0.907049226	0.779063877	0.959998332	1.234974385	1.424871037	1.644499445	1.454617292	1.007357017	1.280694934	1.105445043	1.283038221	0.7483129	1.136149212	1.03160325	0.902899603	1.013783519	0.937367533	1.082731458	1.064034826	0.909972155	1.102823571	1.390383223	1.269404466	0.648562284	1.019055804	1.194153418	0.997109543	1.014353329	0.859228519	0.977268184	0.982761391	0.827388372	0.933099451	1.004775928	1.192261651	0.834362795	0.783875164	0.954184751	1.284173913	0.860064348	1.036431867	1.215032017	0.831915692	0.556948932	0.838575095
Glutamic acid	1.005384947	1.437671727	1.170309092	1.78884251	0.776053269	1.367553371	0.940061257	0.96830186	1.038319104	1.212544778	0.820853543	0.775325575	1.183045659	1.157708585	0.987503404	1.267923432	1.761570966	2.105389042	0.730772522	0.92717845	0.595637076	0.882283665	0.856311634	1.191901903	0.760533544	1.599800187	0.922184373	0.988341469	0.894783609	0.945575873	0.912846788	1.020348838	1.035372786	1.651884493	1.036566227	1.124772131	1.389117938	1.387252688	1.579259377	1.446265902	0.686417768	0.937287338	0.871752634	0.894582562	0.83746754	0.549131492	1.002679092	1.146349055	0.416125478	0.434122796	1.023143497	0.66042482	0.630337092	0.697534013	1.985547341	1.030706031	0.82769092	0.674260126	0.682998757	0.86433036	0.324864991	1.466677903	0.902830994	0.593845615	0.48311252	1.121580992
Histidine	0.969581033	0.991751156	0.933360343	1.145434067	0.832355233	0.986743673	0.732028974	0.802997427	0.988123279	1.073662714	0.831294016	0.80913376	0.808905245	1.018393453	0.936848842	0.890983921	0.850696785	0.935502008	0.667830576	0.755890251	0.804614529	0.900696261	0.798301589	0.894455384	1.03001054	1.149993017	1.104755242	1.1989466	0.965187864	1.01904223	0.868878385	0.973038012	0.801871106	1.129567173	1.024112058	0.930926366	0.937820262	1.033110127	1.017520658	1.019616065	0.798402089	1.125678227	0.75370208	0.870923856	0.895112502	1.170058301	0.934731498	0.996911201	1.051008943	0.870239303	0.957785337	1.080978564	0.759424038	1.082390795	0.92397807	1.135756777	1.066628519	0.834108774	1.037422227	1.045484345	1.003111164	0.990424484	1.032446516	0.910882705	0.652541308	0.854130588
Phenylalanine	1.056759651	1.27791581	1.049947862	1.422976331	0.785178154	0.827667861	0.76524886	0.785569693	0.998220357	0.891906724	0.972032274	0.925467142	0.851110934	0.907396095	0.814065361	0.76321364	0.97693477	0.913798822	0.835637488	0.841762537	0.755127708	0.843004625	0.93888494	1.156577686	1.04909054	1.223109504	1.127015045	1.059400328	1.059721548	1.250856801	1.062358859	1.322868877	0.651122884	1.021122708	1.11909508	0.98555025	0.947813752	1.154835296	1.21122647	1.534966071	0.733789857	0.927455455	0.835709827	0.838874098	0.968688733	0.96363933	1.031123591	1.152489483	0.865404881	0.930199715	1.052554749	1.09434161	0.833018442	0.910789361	1.193439925	0.834079216	1.01575356	0.891895168	0.986476171	1.203343742	0.915826707	0.920583525	1.136593057	0.893727614	0.696356194	0.923639523
Arginine	1.514047159	1.900178724	1.40581971	1.842961988	1.379789724	1.724855249	0.906976059	1.023468892	1.406398307	1.162495389	1.091578819	1.2695913	1.183784017	1.545296669	0.951843256	1.200883534	1.222588962	1.544812206	1.094376916	1.302375372	1.050027567	1.517239425	0.957454885	1.368704789	1.304796639	2.180146454	1.572265251	1.830472491	1.307095368	1.587915981	1.100641359	1.552140871	1.065078502	1.560753221	1.454357831	1.261183875	1.312529255	1.738425864	1.263563099	1.43162118	1.099608321	1.69197834	1.515734588	1.618290071	0.758915202	1.430663551	0.60586777	1.243175866	1.407735139	0.972277185	1.115533538	1.102986129	0.943925429	0.684949676	0.852484593	1.125868625	1.090645515	1.048975672	1.053896395	1.301368518	1.286304132	1.158313455	0.868938474	1.113137955	0.828011102	0.797908108
Tyrosine	0.949323972	1.790457173	0.97200512	1.132622394	0.876780665	1.292330291	1.071231309	1.048849903	1.554003622	1.702858216	1.134714887	1.242011787	0.866530618	1.509130499	1.126115336	0.997073892	0.849974578	1.152682629	1.10429463	1.286517749	0.915406016	1.337303882	1.077007308	1.497404468	0.951266762	1.748082303	1.430733832	1.123406951	1.097123414	2.311582695	1.35607785	2.046057078	0.69816899	1.801598016	1.302432827	1.081757333	0.982868867	1.368873802	1.19191681	1.736952714	0.732612404	1.392078746	0.926302208	1.065435123	0.898116938	1.206300787	1.278222372	1.098640834	0.811784827	0.884219529	1.224073235	1.044315492	0.798361407	0.928911365	1.596371235	1.318916236	0.801014004	0.960519548	1.387503023	1.547677981	1.093743105	1.438945781	1.460140376	1.305593166	0.616102753	0.946195305
Tryptophan	1.124203794	0.906299559	1.669516232	1.457236892	1.827130625	1.260058992	2.167956103	1.43396881	1.186456567	1.352653251	2.543879582	0.867464003	1.035953409	0.715831177	2.061422033	0.592017385	2.1219143	0.63068094	3.795408529	1.167960656	0.859958775	0.742464331	1.291710943	0.77586731	0.756135634	0.610112536	1.253411523	0.498038035	2.513043688	0.553741297	0.734668307	0.726741782	0.711440287	0.372176312	0.753310687	0.397496558	0.624169582	0.661155955	2.110402246	0.864498682	0.827700119	0.585243271	0.878806469	0.514612878	1.360984831	0.640806245	1.082132913	2.223505127	1.161476361	1.131081616	0.88092622	0.863947397	0.530413559	0.871944359	1.27796844	0.715349873	0.991092133	1.44402244	1.108732936	1.018501046	0.651902731	1.03656861	0.971225416	1.140306567	0.677072983	0.629763486
Kynurenine	1.02515871	1.027981042	1.82768788	1.883262133	1.340763829	1.123920596	1.213562428	0.990332552	0.846126336	1.233825057	2.423633453	0.879637522	1.630377014	0.975877442	2.356685899	1.024548031	1.408684048	0.679142834	1.379305241	0.652417467	0.739474376	0.718649141	1.227614964	0.990513566	0.662052664	0.500004099	1.157258775	0.60847255	2.479748166	0.641989814	1.178686677	1.074984165	0.682052541	0.421523	0.936878603	0.579149544	0.646781437	0.643714133	1.916280569	0.982020834	0.842561733	0.492101698	0.99659473	0.887045716	0.983990447	0.587513838	1.028664046	1.758551298	0.914865638	0.788624555	0.665752312	0.695116879	0.660615735	0.962733223	1.703192377	0.555330117	1.021085539	1.069365148	0.821743437	0.873980344	0.703087845	0.938894733	0.806722013	0.872293693	0.782714939	0.719347354
Cystine	0.018588033	0.017864314	0.032403313	0.075012478	0.026494545	0.023922466	0.009526075	0.009156319	0.039435528	0.039533061	0.015598834	0.018097716	0.014555404	0.021852345	0.01325859	0.006911315	0.067874752	0.026592607	0.015332129	0.011648062	0.009676458	0.012839971	0.012629662	0.019348719	0.022305642	0.035080046	0.024929391	0.021842814	0.03483502	0.035494099	0.074853534	0.04135223	0.02840594	0.034907185	0.082074836	0.055011429	0.062456484	0.023425266	0.023351727	0.026248876	0.022055068	0.012896469	1.510451372	1.394235261	1.211240843	1.161699193	1.248995649	1.351201662	0.751318368	0.75027945	0.865011219	0.953911637	0.963831855	0.815089357	1.325816202	1.129819335	0.951168867	0.574598489	1.063928461	1.161050509	0.668715221	0.974060384	0.960742027	1.033868425	0.844504373	1.196140988
Creatinine	0.940916019	0.929545201	1.220732897	1.731832834	0.952499885	0.888881055	0.697539651	1.144556629	1.49217934	1.33257506	0.702361862	0.811838069	0.900963423	1.149468484	0.875189431	1.052847157	1.011394463	1.105759251	0.607811048	0.947410739	0.989223057	0.975022535	1.021343116	1.165921014	1.005964347	0.89356217	0.806834685	1.002534472	1.542613825	1.155347876	0.880918424	1.087984605	0.950466737	0.960932775	1.30200807	1.204680376	1.004857964	0.931517873	0.620887662	0.735171428	0.763327047	0.711175564	0.742256109	0.913690962	0.941896953	0.770864475	0.909854164	1.140690569	1.181611985	1.020400173	0.957070513	1.282144649	1.210753123	1.630752685	1.150321351	1.140787444	1.10080699	1.255200794	0.701695256	1.120497684	1.188985609	1.226490916	1.044260754	0.808094144	0.938365682	1.21676501
Creatine; Beta-Guanidinopropionic acid	0.712515482	0.769846399	1.210021761	1.548657784	0.60242028	0.67074953	0.550685162	0.633109978	1.283564179	1.265185004	0.70697493	0.793071789	0.686484114	0.882006102	0.741733293	0.790326811	0.806282926	0.708296967	0.674578109	0.789743531	0.728210381	0.664668988	0.988954439	0.976909357	0.762352882	0.717197417	0.895183162	0.91196732	1.122445666	0.975853418	0.717618346	0.772566491	0.608340654	0.669435738	1.08175843	0.975641067	0.792835294	0.76086081	0.684009395	0.768051367	0.564905594	0.543081938	0.659390639	0.788069929	0.600866944	0.637447264	0.907708409	1.026735612	0.904625455	0.77021997	0.775733576	1.053604064	0.987870387	1.385898579	0.973519299	0.972068828	0.802377077	1.034699346	0.698585402	1.046344199	1.014815327	1.061121313	1.167343821	0.84221396	0.819088845	1.066476986
2-Hydroxy-L-methionine	1.071189814	0.963831627	1.169420226	1.452585544	1.050379887	1.026556807	0.852996647	1.006692402	1.5610365	1.179770839	0.939657421	1.024530249	0.799762836	0.990040223	0.834560369	0.733809343	1.141507361	0.967204863	0.797043078	1.126237349	1.025800311	0.954948771	0.855692933	1.227168574	1.162704396	1.370655331	1.481888665	1.441901485	1.082251189	1.377881815	0.887337088	1.554077239	0.82660864	1.421292882	1.169577698	0.624459841	0.949562374	0.961129912	0.935783095	1.331693607	0.883155901	1.145362802	0.88659668	0.992928109	0.900727245	0.90243697	1.088437241	0.990144397	1.022135595	0.899869625	1.312709353	1.182989587	0.964806085	1.012993731	1.281575066	1.279514663	1.119507438	1.056715961	0.929007996	1.259757393	0.951005732	0.908240972	1.08948846	0.908761108	0.574584507	0.84505558
Citrulline	1.016536885	0.996293952	1.714654253	2.374679673	0.937502475	0.900139518	0.852558427	0.94920863	1.747598145	0.98643909	0.923144973	1.344775892	0.870883786	1.14078576	0.931639485	1.04493485	0.919703702	1.407080401	0.701557532	1.2206049	1.194507757	1.853166255	1.000918815	1.12917019	0.824186264	1.103666053	1.181911552	1.562373854	0.999909454	0.905247696	1.146156244	1.178671165	0.686746999	1.037899777	0.943784808	1.240060446	1.310416602	1.076626473	1.350848033	1.404709425	0.929259636	1.124528393	1.37204008	1.411795605	0.86308815	1.220759976	0.968566019	1.364992192	0.696473595	0.909303566	0.983600224	1.046980074	1.124124974	0.684886175	1.201403642	1.076184648	0.974721273	0.728987763	1.36857588	0.888530315	0.994491926	1.123591527	0.878441604	1.112752527	0.761217862	0.970093455
5-Hydroxy-L-tryptophan	1.094157977	0.678732636	1.273874219	1.101060505	1.885579697	1.161530616	2.010498356	1.369459799	1.748383305	1.761809413	2.418361668	1.007109341	1.081505285	0.89338846	1.575005592	0.480711703	2.318737418	0.683950965	3.969682743	1.2215659	0.96191113	0.738323987	1.335804485	0.809264723	0.835511876	0.819849662	1.230497946	0.498850176	2.434614778	0.591824188	0.714944318	0.669307544	0.892483789	0.494411445	0.76847243	0.464440127	0.602839922	0.624275782	2.034368473	0.854681129	0.692548733	0.574013212	0.668065854	0.379827425	1.479414393	0.652058559	1.038477233	1.844543056	1.113326538	1.022225722	0.789296845	0.774659417	0.592013997	0.892085958	1.254741122	0.718784698	1.227881738	1.590970326	1.15516405	1.064760858	0.998077942	1.16744208	0.926923723	1.159726166	0.715887028	0.649781954
Phenylalanyl-Serine	2.646039315	1.721330629	0.4322927	0.329404796	0.679409028	1.210855598	1.204553724	1.28235275	1.418439352	1.905341594	0.34833959	0.342588188	0.537738084	1.498806048	2.055104621	2.001328151	0.456547752	0.420351722	0.934489363	1.027450914	1.428632923	1.546268797	0.470738464	1.2882296	0.958453468	0.478881424	0.910598779	1.112904979	1.488618977	0.784618641	0.389719474	0.135291753	0.531732033	0.216265335	1.145209701	1.420506232	0.946438433	1.334107675	1.189026954	1.31144559	0.759541365	0.507226771	1.154257158	2.329807261	0.133899668	0.118291322	0.524760148	0.929889557	0.877019394	1.34802236	0.64874785	1.396296161	0.987048352	1.787098034	1.173909098	0.680738871	0.681134579	1.131436458	0.600546074	1.12339092	0.895089954	1.28933916	0.954092236	0.703592321	0.904009291	1.207288676
Norophthalmic acid	1.287756718	0.966233551	1.766834055	2.100610659	1.450005119	1.217158064	1.531815044	1.596851318	1.300175585	1.224542843	1.233090399	1.435252953	1.463369231	1.57785516	1.251125041	1.243353342	1.001577239	0.939888428	0.825109265	1.13085379	1.530900034	1.307430828	1.228612231	1.127239654	1.326689724	1.5997603	1.796023905	2.165249914	0.999742584	1.20391319	1.615893078	1.870352214	2.019599036	2.108800016	1.213378668	1.236046836	1.389248704	1.171703664	1.531531082	1.665504761	1.121546915	1.753878434	0.89427964	1.152306955	0.794981741	0.852518102	1.048724056	1.52786083	0.735758859	1.040953471	0.979399345	0.895173343	0.807660861	0.979073184	1.060361485	1.525951096	0.74805074	1.042039815	0.775775838	0.716722372	0.927574609	0.853822844	1.323855556	0.774311916	0.545505787	0.711948642
Fructosyl-lysine	0.882045091	0.628823342	0.829757601	1.220237137	0.348784974	1.096917904	0.477260081	0.821423595	1.360590747	1.042614858	0.768684377	0.817950108	0.798661256	1.076618756	0.72378356	0.702472198	0.684257477	0.393652367	0.558212212	0.756332221	0.604158896	0.26301474	0.624410901	1.048924462	0.107377673	1.295786314	0.146287416	0.101909833	0.07083174	0.07029907	0.078321202	0.041259362	0.584054386	0.221447354	0.230264379	0.07045856	0.215349469	0.129570848	0.270927472	0.123332104	0.041623453	0.200335624	0.156335412	0.115159301	0.527779748	0.064794095	0.077694422	0.578083006	0.848078816	0.666319072	1.108337192	1.252755575	0.599865013	1.045274344	1.065532047	1.158568529	0.963353461	0.523101414	0.875995081	1.206155053	0.954452861	1.011693698	0.986577423	0.876833053	0.587191591	0.79377559
LysoPE(0:0/15:0)	0.699498758	1.129871249	0.565886612	1.619620014	0.204813141	1.22237379	0.508059068	0.873435286	1.434987685	1.10388161	0.744202244	0.937714462	0.61405618	0.982240094	0.712462904	0.708741708	0.776416034	0.922392516	0.604992945	1.143198952	0.768745291	0.552139666	0.607250861	1.408326391	0.182525754	1.962462159	0.278859257	0.280302941	0.283632263	0.268942652	0.332232238	0.336330905	0.468299798	0.362580809	0.251912609	0.1593478	0.331669126	0.211500104	0.40001912	0.30852642	0.06749402	0.238426773	0.23601501	0.197889885	0.746492505	0.179961948	0.246317166	1.39624102	0.412150726	0.730516085	0.740374692	0.991859647	0.53354987	0.92109984	1.200534535	1.083949765	1.191879486	0.47120909	0.888479917	0.948223251	0.560086951	1.217489294	0.929546495	0.934951208	0.45662529	1.054410035
LysoPE(0:0/16:0)	0.748791302	0.670824987	0.722446446	1.071215134	0.337157841	1.018461673	0.395195644	0.7295444	1.186522579	1.01076224	0.76105433	0.77595384	0.775383901	1.115092837	0.768518424	0.742542273	0.684997193	0.468217932	0.559764036	0.730141784	0.580413538	0.292807959	0.622169369	1.025954232	0.163974349	1.023384941	0.169042864	0.134357165	0.097859539	0.097043615	0.114953918	0.05167141	0.525812435	0.279587249	0.248709958	0.090377346	0.274910871	0.167299207	0.283640962	0.164190837	0.060282184	0.255802117	0.192624676	0.142279358	0.77404248	0.170591954	0.167997233	0.779939198	0.775233329	0.623643174	1.039258848	1.176261793	0.528046531	0.959548709	0.934114359	1.041232483	0.973311197	0.59646947	0.7904331	1.315235196	0.890382375	0.986756315	1.006479459	0.87141822	0.543790723	0.781036988
Glutamylleucine	1.724180367	1.688837652	1.789808794	2.68948037	2.12953996	1.828697579	1.742332497	1.847737148	2.741971301	1.544018261	1.726743561	1.740930593	2.259496969	2.355519235	2.188494063	2.389469303	2.105476083	2.484041454	1.814562703	1.738091978	1.997996463	2.521859942	1.580106434	1.999667567	2.855695036	6.009179353	3.77160675	4.551456301	1.74577126	3.171744688	1.996328631	2.645138462	2.54943116	3.331042491	2.173620186	2.521937663	2.529113513	2.758969422	2.915969542	3.094113635	2.543407639	3.49055411	1.172117637	1.301639342	0.958867598	1.294074715	1.426336873	1.613333744	1.163762283	1.001507713	2.034498396	1.042561875	1.104761901	1.025722217	1.489469766	1.374923537	1.561859794	1.287627357	0.814732147	1.103530214	1.180198706	1.563178357	1.406364436	0.936933593	1.020856651	1.313992082
Phenylalanylglutamic acid	1.324004943	0.95157531	1.092474208	1.32161715	1.484881769	1.200895487	1.179398547	1.335769566	2.777019994	1.052582455	1.017392123	1.089214693	1.82297383	1.602843362	1.127423163	1.257354508	1.456974072	1.258096243	1.164573242	1.339980532	1.393495703	1.358074115	1.234453536	1.711908105	1.733793101	1.156421102	1.047535301	1.046528777	1.24279408	1.297425069	1.120680406	1.631112663	1.264244956	1.370305402	1.228316396	1.351807027	1.303083475	1.619348811	1.928735891	2.498818248	1.494255558	1.360866644	1.467752576	1.48146697	0.774440079	0.64842647	1.1053021	0.969940627	1.914730775	0.791428275	1.210251316	1.053421329	1.301785519	1.090060804	1.66390825	1.03157985	3.392391162	1.337588418	0.867866857	0.958493253	1.253184332	1.065694474	1.05170401	0.772940932	0.792674666	0.822808741
Phenylalanylphenylalanine	0.959472731	0.681475469	0.234728616	0.331752378	0.497035438	1.051696698	0.42412368	0.450356373	0.328248682	0.678038782	0.305025625	0.118435428	0.535704456	0.587523314	1.405441547	0.597616528	0.489780225	0.35770028	0.604796503	0.598947344	0.747407319	0.526499441	0.14841083	0.782145187	0.347994262	0.093841213	0.503629672	0.706881603	0.957045662	0.169999455	0.556234868	0.106931283	0.279339545	0.521370617	0.459609789	0.992662523	1.834990246	0.462796103	1.302198888	1.854871516	0.534711978	0.397407352	0.622160752	0.843472307	0.474338217	0.132727081	0.755967909	1.930409975	1.164520111	1.090526906	0.442820081	0.718264492	0.926931961	1.251160135	1.706890532	0.55316278	1.353643755	0.8953424	0.660903915	1.816736988	0.2097353	1.084647398	0.886652834	0.828807253	0.70912056	1.639205633
Ornithine	0.84586356	1.528511184	0.790876414	1.364345368	0.658698451	1.306355254	0.803564918	0.968350431	1.532555317	1.180544416	0.767407605	0.988542044	0.628544619	0.9995379	0.819904969	0.910134598	0.93646003	1.284663023	0.637304784	0.888923602	0.906074586	1.531188035	0.605015857	1.17082231	0.668699952	0.930232776	0.807289398	1.088127167	0.83026206	1.079538111	0.650122647	0.985649188	0.517376289	1.370560611	0.598523289	0.933315218	1.42259051	1.205382986	0.997767	1.762207881	0.732306112	0.856938315	1.1048022	1.286715107	0.846451313	1.740730547	1.741861849	1.754141653	0.546230559	1.068309227	0.759503945	0.914678651	0.925724405	1.002160316	1.764622974	1.43937434	1.075433956	0.598176367	0.921996907	0.934406853	0.517929643	1.078352138	1.049742749	0.882180057	0.592272874	1.481129141
Aminoadipic acid	1.16269128	1.247264005	3.319058681	3.94961083	1.089267501	1.477822023	2.816491722	2.826554934	1.565461536	2.363065327	1.199874128	1.081123955	1.570126241	2.345266883	1.634673728	1.778055096	3.196736994	3.372985304	2.075960528	2.269619073	0.748908835	1.077876808	2.361644935	2.751195073	0.966404762	0.999088867	0.571156246	0.360987221	1.421288401	1.976832279	1.181813569	0.83385919	1.394114976	1.841364802	1.871824732	1.348799291	1.117750077	1.688814715	0.752477444	0.44762377	0.843688271	0.989030027	1.618966804	1.793807915	0.557172551	0.567579051	0.979646454	1.426012706	0.742389821	0.55470988	1.880974634	1.266097109	1.688618119	1.170603613	2.356093745	2.407302759	0.623711733	1.28843966	0.618120871	0.847013809	1.060460698	1.232706269	1.315322205	1.064027063	1.073552501	2.877372034
Amino (2S)-2,5-diamino-5-oxopentanoate	0.738329499	1.075879767	1.314480283	1.612864677	0.62416811	1.237850273	1.037649089	1.120665666	1.013973354	1.40263515	0.952593579	0.669430155	0.935428868	1.493331224	1.189849074	0.636747654	1.24551775	2.422368592	1.401201724	1.103341304	0.668393093	0.799348942	1.057311012	1.239677695	0.771798663	1.256968211	1.167995945	0.972748368	0.903249371	1.105073337	0.952550174	0.818581188	0.845769871	1.305240826	1.16870311	0.831212193	0.959785273	0.880818843	0.932270182	0.751688547	0.631961304	0.97819069	1.225953004	1.012782263	0.931969089	1.148954243	1.163624284	1.258480439	0.721061762	0.93923775	1.20700822	0.852569043	0.803544051	1.022012612	1.327173051	1.088669737	0.886903513	1.073419954	0.954136502	0.879475294	1.063281181	0.987433421	1.003154796	0.736812798	0.604000952	1.180819606
1-Methylhistidine	0.696265582	0.687954025	1.702365625	3.250794192	0.746440752	1.228542841	0.778715977	0.984823924	2.486453211	1.175546342	0.898155398	0.766097266	1.157064344	1.354966546	0.943205968	0.901708307	1.187578736	0.908135871	0.715151063	0.85019251	0.724597256	1.312540452	0.804495356	1.222881487	0.85091494	1.076851493	0.932633666	1.188549998	1.23218904	0.743153775	1.786417774	1.835520816	0.687318356	0.908735686	2.261507513	1.282076151	1.302822351	1.059247812	0.933364644	0.629922889	0.659102902	0.734567484	0.700268875	0.970222713	0.590789033	0.540592441	1.162981593	0.811500605	1.14621363	0.530263679	0.949074451	0.88964313	1.371656078	0.904816541	1.624478263	0.898147357	0.868593417	0.786764141	0.855438008	1.154741787	0.91284415	1.039786577	1.325731315	0.911178658	0.931362283	1.120581528
Nitrotyrosine	0.980939605	1.742271798	0.979879121	1.116608471	0.882492748	1.173093817	1.062227086	0.997072066	1.485114104	1.612534384	1.127864387	1.034936111	1.044789069	1.943799054	1.562308971	1.166017649	0.954433306	1.29486925	1.236184375	1.486902836	0.872661833	1.226633933	1.071478238	1.505285026	0.976505598	1.527298732	1.484606787	1.124225118	1.077137098	2.093836129	1.341323497	1.893055434	0.743679689	1.619938589	1.336103801	1.019648761	1.007972501	1.275230624	1.258038386	1.826527551	0.554318808	1.46379187	0.699074655	1.107745665	0.919116429	1.125319786	1.338044725	1.096085269	0.907724362	1.018421209	1.413672046	1.212946121	0.94937371	1.002456365	1.718714083	1.453955411	0.713606073	0.772999006	1.247808455	1.320423695	1.060047043	1.348266013	1.422456376	1.290978701	0.63844503	0.882639346
Uridine	0.66545585	1.353996451	1.133583604	1.149662258	1.021709396	1.871674487	1.390189089	1.158266033	0.478697313	1.378143111	0.844042041	0.769556341	0.661911796	1.039953726	0.858180581	0.694283982	0.793221625	1.262267216	0.77146446	0.655540301	0.981125149	1.098270154	0.919006763	0.767166954	0.792565964	0.724215411	1.05520865	0.534339851	2.863022991	1.445422157	2.239421686	1.15981645	0.581819052	0.962678496	1.052966897	0.7808717	0.823209393	1.726210857	0.905370059	0.87651967	0.349431192	1.007948881	0.473153972	0.48136373	0.866551346	1.075013704	1.051700038	1.666728616	0.776986839	1.419877306	0.869884166	0.959130363	1.698403162	1.590781619	1.47227795	1.398291175	0.622414373	1.185295381	0.889412088	1.265094243	1.167971904	1.552328284	1.179710458	1.337977449	0.988432401	1.251122707
Gamma-Glutamyltyrosine	0.823808212	1.463311242	0.960436648	0.983747794	0.293803245	1.125387655	1.085304746	0.921642476	0.436920329	0.634519804	0.231444281	0.200706049	0.558641461	0.934503215	0.874059004	0.81569027	0.230358893	0.308259784	0.433605397	0.491465009	0.863290962	0.723007948	0.200503304	1.272827781	0.206465021	0.220626389	0.918611041	0.721432088	1.829134981	0.584164398	0.51612101	0.123055988	0.206184325	0.349960189	0.475243165	0.725558117	1.133554661	1.369523196	1.663095435	1.857886746	0.944776608	1.243530047	1.206636632	1.227324603	0.252839067	0.283703362	1.047268432	1.570125345	1.123506796	1.825483802	1.627204114	2.34267933	1.166580795	1.213439434	0.919214744	0.992338517	0.561743424	0.827623849	1.304729397	1.229701235	0.936612349	0.918498545	1.062633471	1.129374023	1.113762433	1.107648479
N-Fructosyl phenylalanine	0.881206596	1.376856861	0.691326188	1.840938213	0.276399857	0.850824286	0.317407947	0.571416079	1.359012875	1.474004332	0.856669107	1.012488874	0.755289986	0.872190664	0.756376132	0.662865024	0.804626011	0.757453527	0.700290869	0.830868427	0.56857864	0.347970211	0.874714871	1.483264894	0.310915475	1.153049688	0.236524932	0.211009404	0.185165261	0.182016985	0.189696157	0.134615007	0.512175472	0.283380216	0.252358229	0.142530214	0.303155769	0.310238076	0.3757694	0.347877436	0.178616411	0.330341851	0.266702379	0.219631131	0.707630567	0.174009105	0.216899388	0.749491564	0.778706656	0.666754465	1.014637021	1.129989827	0.548981	0.977670827	1.070500673	0.894137475	0.849328054	0.615864438	0.706008201	1.265689089	0.761004225	0.814731479	0.987469621	0.849712758	0.52869285	0.875754436
N-(1-Deoxy-1-fructosyl)tyrosine	0.835579214	1.803422516	0.588360677	1.331470252	0.29438744	1.176001221	0.556677153	0.923871832	1.570544073	1.645097628	1.047389692	1.209649729	0.773940131	1.37920836	1.028775551	0.833259315	0.844525008	1.069753198	1.004419158	1.204453298	0.679091341	0.475348747	0.922281346	1.769326394	0.251483283	1.342261787	0.239819791	0.184252588	0.244592944	0.352827644	0.303087187	0.202411149	0.595116468	0.583066885	0.333215264	0.179338968	0.323158754	0.360011494	0.400977423	0.409180375	0.109097938	0.456036	0.263169605	0.231419743	0.762816836	0.213682652	0.254426592	0.764601082	0.855667835	0.825823421	1.481414182	1.407066269	0.577178253	1.014482739	1.397234049	1.334592346	0.684918804	0.540782926	1.032189932	1.547871143	0.856199079	1.112457896	1.205382203	1.165294797	0.525355028	0.887023121
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	KEGG	PubChem ID	quantitated m/z
1-Methylhistidine	C01152	92105	169.0839
2-Hydroxy-L-methionine		45082566	165.0454
5-Hydroxy-L-tryptophan	C00643	439280	220.0845
Alanine	C00041	5950	89.0471
Amino (2S)-2,5-diamino-5-oxopentanoate		22417575	161.0795
Aminoadipic acid	C00956	469	161.0682
Arginine	C00062	6322	174.1107
Asparagine	C00152	6267	132.0531
Citrulline	C00327	9750	175.0953
Creatine; Beta-Guanidinopropionic acid	C00300	586	131.0689
Creatinine	C00791	588	113.0585
Cystine	C00491	67678	240.0239
Fructosyl-lysine		9839580	308.1573
Gamma-Glutamyltyrosine		94340	310.1163
Glutamic acid	C00025	33032	147.0525
Glutamine	C00064	5961	146.0687
Glutamylleucine		9856500	260.1369
Histidine	C00135	6274	155.0692
Isoleucine	C00407	6306	131.0942
Kynurenine	C00328	846	208.0854
Leucine	C00123	6106	131.0942
Lysine	C00047	5962	146.1053
LysoPE(0:0/15:0)		53480921	439.2685
LysoPE(0:0/16:0)		53480922	453.2843
Methionine	C00073	6137	149.0497
N-(1-Deoxy-1-fructosyl)tyrosine		131752248	343.1265
N-Fructosyl phenylalanine		85152457	327.1313
Nitrotyrosine		9834478	226.0588
Norophthalmic acid		5489007	275.1109
Ornithine	C00077	6262	132.0898
Phenylalanine	C00079	6140	165.0786
Phenylalanylglutamic acid		151134	294.1212
Phenylalanylphenylalanine		65104	312.1472
Phenylalanyl-Serine		193508	252.1107
Proline	C00148	145742	115.0628
Threonine	C00188	6288	119.0579
Tryptophan	C00078	6305	204.0898
Tyrosine	C00082	6057	181.0734
Uridine	C00299	6029	244.0689
Valine	C00183	6287	117.0785
METABOLITES_END
#END