#METABOLOMICS WORKBENCH ssah9_20220411_071736 DATATRACK_ID:3192 STUDY_ID:ST002137 ANALYSIS_ID:AN003498 PROJECT_ID:PR001353
VERSION             	1
CREATED_ON             	April 15, 2022, 8:15 am
#PROJECT
PR:PROJECT_TITLE                 	Targeted Microchip Capillary Electrophoresis-Orbitrap Mass Spectrometry
PR:PROJECT_TITLE                 	Metabolomics to Monitor Ovarian Cancer Progression.
PR:PROJECT_SUMMARY               	The lack of effective screening strategies for high-grade serous carcinoma
PR:PROJECT_SUMMARY               	(HGSC), a subtype of ovarian cancer (OC) responsible for 80% of OC related
PR:PROJECT_SUMMARY               	deaths, emphasizes the need for new diagnostic markers and a better
PR:PROJECT_SUMMARY               	understanding of disease pathogenesis. Capillary electrophoresis (CE) coupled
PR:PROJECT_SUMMARY               	with high-resolution mass spectrometry (HRMS) offers high selectivity and
PR:PROJECT_SUMMARY               	sensitivity, thereby increasing metabolite coverage and consequently enhancing
PR:PROJECT_SUMMARY               	biomarker discovery. Recent advances in CE-MS include small, chip-based CE
PR:PROJECT_SUMMARY               	systems coupled with nanoelectrospray ionization (nanoESI) to provide rapid,
PR:PROJECT_SUMMARY               	high-resolution analysis of biological specimens. Here, we describe the
PR:PROJECT_SUMMARY               	development of a targeted microchip (µ) CE-HRMS method to analyze 40 target
PR:PROJECT_SUMMARY               	metabolites in serum samples from a triple-mutant (TKO) mouse model of HGSC,
PR:PROJECT_SUMMARY               	with an acquisition time of only 3 min. Extracted ion electropherograms showed
PR:PROJECT_SUMMARY               	sharp, highly resolved peak shapes, even for structural isomers such as leucine
PR:PROJECT_SUMMARY               	and isoleucine. All analytes maintained good linearity with an average R2 of
PR:PROJECT_SUMMARY               	0.994, while detection limits were in the nM range. Thirty metabolites were
PR:PROJECT_SUMMARY               	detected in mice serum, with recoveries ranging from 78 to 120 %, indicating
PR:PROJECT_SUMMARY               	minimal ionization suppression and good accuracy. We applied the µCE-HRMS
PR:PROJECT_SUMMARY               	method to sequentially-collected serum samples from TKO and TKO-control mice.
PR:PROJECT_SUMMARY               	Time-resolved analysis revealed characteristic temporal trends for amino acids,
PR:PROJECT_SUMMARY               	nucleosides, and amino acids derivatives associated with HGSC progression.
PR:PROJECT_SUMMARY               	Comparison of the µCE-HRMS dataset with non-targeted ultra-high performance
PR:PROJECT_SUMMARY               	liquid chromatography (UHPLC) – MS results revealed identical temporal trends
PR:PROJECT_SUMMARY               	for the 5 metabolites detected on both platforms, indicating the µCE-HRMS
PR:PROJECT_SUMMARY               	method performed satisfactorily in terms of capturing metabolic reprogramming
PR:PROJECT_SUMMARY               	due to HGSC progression, while reducing the total analysis time 3-fold.
PR:INSTITUTE                     	Georgia Institute of Technology
PR:LAST_NAME                     	Sah
PR:FIRST_NAME                    	Samyukta
PR:ADDRESS                       	901 Atlantic Dr NW, Atlanta, GA, 30332, USA
PR:EMAIL                         	ssah9@gatech.edu
PR:PHONE                         	5746780124
#STUDY
ST:STUDY_TITLE                   	Targeted Microchip Capillary Electrophoresis-Orbitrap Mass Spectrometry
ST:STUDY_TITLE                   	Metabolomics to Monitor Ovarian Cancer Progression. (Ovarian cancer mouse data)
ST:STUDY_SUMMARY                 	The lack of effective screening strategies for high-grade serous carcinoma
ST:STUDY_SUMMARY                 	(HGSC), a subtype of ovarian cancer (OC) responsible for 80% of OC related
ST:STUDY_SUMMARY                 	deaths, emphasizes the need for new diagnostic markers and a better
ST:STUDY_SUMMARY                 	understanding of disease pathogenesis. Capillary electrophoresis (CE) coupled
ST:STUDY_SUMMARY                 	with high-resolution mass spectrometry (HRMS) offers high selectivity and
ST:STUDY_SUMMARY                 	sensitivity, thereby increasing metabolite coverage and consequently enhancing
ST:STUDY_SUMMARY                 	biomarker discovery. Recent advances in CE-MS include small, chip-based CE
ST:STUDY_SUMMARY                 	systems coupled with nanoelectrospray ionization (nanoESI) to provide rapid,
ST:STUDY_SUMMARY                 	high-resolution analysis of biological specimens. Here, we describe the
ST:STUDY_SUMMARY                 	development of a targeted microchip (µ) CE-HRMS method to analyze 40 target
ST:STUDY_SUMMARY                 	metabolites in serum samples from a triple-mutant (TKO) mouse model of HGSC,
ST:STUDY_SUMMARY                 	with an acquisition time of only 3 min. Extracted ion electropherograms showed
ST:STUDY_SUMMARY                 	sharp, highly resolved peak shapes, even for structural isomers such as leucine
ST:STUDY_SUMMARY                 	and isoleucine. All analytes maintained good linearity with an average R2 of
ST:STUDY_SUMMARY                 	0.994, while detection limits were in the nM range. Thirty metabolites were
ST:STUDY_SUMMARY                 	detected in mice serum, with recoveries ranging from 78 to 120 %, indicating
ST:STUDY_SUMMARY                 	minimal ionization suppression and good accuracy. We applied the µCE-HRMS
ST:STUDY_SUMMARY                 	method to sequentially-collected serum samples from TKO and TKO-control mice.
ST:STUDY_SUMMARY                 	Time-resolved analysis revealed characteristic temporal trends for amino acids,
ST:STUDY_SUMMARY                 	nucleosides, and amino acids derivatives associated with HGSC progression.
ST:STUDY_SUMMARY                 	Comparison of the µCE-HRMS dataset with non-targeted ultra-high performance
ST:STUDY_SUMMARY                 	liquid chromatography (UHPLC) – MS results revealed identical temporal trends
ST:STUDY_SUMMARY                 	for the 5 metabolites detected on both platforms, indicating the µCE-HRMS
ST:STUDY_SUMMARY                 	method performed satisfactorily in terms of capturing metabolic reprogramming
ST:STUDY_SUMMARY                 	due to HGSC progression, while reducing the total analysis time 3-fold.
ST:INSTITUTE                     	Georgia Institute of Technology
ST:LAST_NAME                     	Sah
ST:FIRST_NAME                    	Samyukta
ST:ADDRESS                       	901 Atlantic Dr NW, Atlanta, GA, 30332, USA
ST:EMAIL                         	ssah9@gatech.edu
ST:PHONE                         	5746780124
#SUBJECT
SU:SUBJECT_TYPE                  	Mammal
SU:SUBJECT_SPECIES               	Mus musculus
SU:TAXONOMY_ID                   	10090
#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           	-	T13_1	pheotype:TKO	RAW_FILE_NAME=8465_1 5110; Age (weeks)=8
SUBJECT_SAMPLE_FACTORS           	-	T13_2	pheotype:TKO	RAW_FILE_NAME=8465 _2 5110; Age (weeks)=10
SUBJECT_SAMPLE_FACTORS           	-	T13_3	pheotype:TKO	RAW_FILE_NAME=8465_3 5110; Age (weeks)=12
SUBJECT_SAMPLE_FACTORS           	-	T13_4	pheotype:TKO	RAW_FILE_NAME=8465_4 5110; Age (weeks)=14
SUBJECT_SAMPLE_FACTORS           	-	T13_5	pheotype:TKO	RAW_FILE_NAME=8465_5 5110; Age (weeks)=16
SUBJECT_SAMPLE_FACTORS           	-	T13_6	pheotype:TKO	RAW_FILE_NAME=8465_6 5110; Age (weeks)=18
SUBJECT_SAMPLE_FACTORS           	-	T13_7	pheotype:TKO	RAW_FILE_NAME=8465_7 5110; Age (weeks)=20
SUBJECT_SAMPLE_FACTORS           	-	T13_8	pheotype:TKO	RAW_FILE_NAME=8465_8 5110; Age (weeks)=22
SUBJECT_SAMPLE_FACTORS           	-	T13_9	pheotype:TKO	RAW_FILE_NAME=8465_9 5110; Age (weeks)=24
SUBJECT_SAMPLE_FACTORS           	-	T11_1	pheotype:TKO	RAW_FILE_NAME=8256_1_chip5109; Age (weeks)=8
SUBJECT_SAMPLE_FACTORS           	-	T11_2	pheotype:TKO	RAW_FILE_NAME=8256_2repeatwith5110; Age (weeks)=10
SUBJECT_SAMPLE_FACTORS           	-	T11_3	pheotype:TKO	RAW_FILE_NAME=8256_3_chip5109; Age (weeks)=12
SUBJECT_SAMPLE_FACTORS           	-	T11_4	pheotype:TKO	RAW_FILE_NAME=8256_4_run2_5110; Age (weeks)=14
SUBJECT_SAMPLE_FACTORS           	-	T11_5	pheotype:TKO	RAW_FILE_NAME=8256_5_5510; Age (weeks)=16
SUBJECT_SAMPLE_FACTORS           	-	T11_6	pheotype:TKO	RAW_FILE_NAME=8256_6_chip5109; Age (weeks)=18
SUBJECT_SAMPLE_FACTORS           	-	T11_7	pheotype:TKO	RAW_FILE_NAME=8256_7_chip5109; Age (weeks)=20
SUBJECT_SAMPLE_FACTORS           	-	T11_8	pheotype:TKO	RAW_FILE_NAME=8256_8_chip5109; Age (weeks)=22
SUBJECT_SAMPLE_FACTORS           	-	T11_10	pheotype:TKO	RAW_FILE_NAME=8256_10_5110; Age (weeks)=26
SUBJECT_SAMPLE_FACTORS           	-	T11_11	pheotype:TKO	RAW_FILE_NAME=8256_11_5110; Age (weeks)=28
SUBJECT_SAMPLE_FACTORS           	-	T11_12	pheotype:TKO	RAW_FILE_NAME=8256_12_chip5109; Age (weeks)=30
SUBJECT_SAMPLE_FACTORS           	-	T11_13	pheotype:TKO	RAW_FILE_NAME=8256_13_chip5109; Age (weeks)=32
SUBJECT_SAMPLE_FACTORS           	-	T11_14	pheotype:TKO	RAW_FILE_NAME=8256_14_5110; Age (weeks)=34
SUBJECT_SAMPLE_FACTORS           	-	T11_15	pheotype:TKO	RAW_FILE_NAME=8256_15_5110; Age (weeks)=36
SUBJECT_SAMPLE_FACTORS           	-	T15_1	pheotype:TKO	RAW_FILE_NAME=TKO8472_1_run_4574; Age (weeks)=8
SUBJECT_SAMPLE_FACTORS           	-	T15_3	pheotype:TKO	RAW_FILE_NAME=TKO8472_3_run2_4574; Age (weeks)=12
SUBJECT_SAMPLE_FACTORS           	-	T15_5	pheotype:TKO	RAW_FILE_NAME=TKO8472_5_run_4574; Age (weeks)=16
SUBJECT_SAMPLE_FACTORS           	-	T15_6	pheotype:TKO	RAW_FILE_NAME=TKO8472_6_run2_4574; Age (weeks)=18
SUBJECT_SAMPLE_FACTORS           	-	T15_7	pheotype:TKO	RAW_FILE_NAME=TKO8472_7_run2_4574; Age (weeks)=20
SUBJECT_SAMPLE_FACTORS           	-	T15_8	pheotype:TKO	RAW_FILE_NAME=TKO8472_8_run_4574; Age (weeks)=22
SUBJECT_SAMPLE_FACTORS           	-	T15_9	pheotype:TKO	RAW_FILE_NAME=TKO8472_9_run_4574; Age (weeks)=24
SUBJECT_SAMPLE_FACTORS           	-	T15_10	pheotype:TKO	RAW_FILE_NAME=TKO8472_10_run_4574; Age (weeks)=26
SUBJECT_SAMPLE_FACTORS           	-	T15_11	pheotype:TKO	RAW_FILE_NAME=TKO8472_11_run_4574; Age (weeks)=28
SUBJECT_SAMPLE_FACTORS           	-	C8_1	pheotype:Control	RAW_FILE_NAME=8103_1_5110; Age (weeks)=8
SUBJECT_SAMPLE_FACTORS           	-	C8_2	pheotype:Control	RAW_FILE_NAME=8103_2 5110; Age (weeks)=10
SUBJECT_SAMPLE_FACTORS           	-	C8_3	pheotype:Control	RAW_FILE_NAME=8103_3_repeat_5110; Age (weeks)=12
SUBJECT_SAMPLE_FACTORS           	-	C8_4	pheotype:Control	RAW_FILE_NAME=8103_4_5110; Age (weeks)=14
SUBJECT_SAMPLE_FACTORS           	-	C8_5	pheotype:Control	RAW_FILE_NAME=8103_5 5110; Age (weeks)=16
SUBJECT_SAMPLE_FACTORS           	-	C8_6	pheotype:Control	RAW_FILE_NAME=8103_6_5110; Age (weeks)=18
SUBJECT_SAMPLE_FACTORS           	-	C8_7	pheotype:Control	RAW_FILE_NAME=8103_7_5110; Age (weeks)=20
SUBJECT_SAMPLE_FACTORS           	-	C8_8	pheotype:Control	RAW_FILE_NAME=8103_8 5110; Age (weeks)=22
SUBJECT_SAMPLE_FACTORS           	-	C8_9	pheotype:Control	RAW_FILE_NAME=8103_9_5110; Age (weeks)=24
SUBJECT_SAMPLE_FACTORS           	-	C8_10	pheotype:Control	RAW_FILE_NAME=8103_10_5110; Age (weeks)=26
SUBJECT_SAMPLE_FACTORS           	-	C8_11	pheotype:Control	RAW_FILE_NAME=8103_11 5110; Age (weeks)=28
SUBJECT_SAMPLE_FACTORS           	-	C8_12	pheotype:Control	RAW_FILE_NAME=8103_12_5ul samplevolume_5110; Age (weeks)=30
SUBJECT_SAMPLE_FACTORS           	-	C8_13	pheotype:Control	RAW_FILE_NAME=8103_13_5110; Age (weeks)=32
SUBJECT_SAMPLE_FACTORS           	-	C8_14	pheotype:Control	RAW_FILE_NAME=8103_14_5110; Age (weeks)=34
SUBJECT_SAMPLE_FACTORS           	-	C10_1	pheotype:Control	RAW_FILE_NAME=ctrl_8284_1_chip5110; Age (weeks)=8
SUBJECT_SAMPLE_FACTORS           	-	C10_2	pheotype:Control	RAW_FILE_NAME=ctrl_8284_2_chip5110; Age (weeks)=10
SUBJECT_SAMPLE_FACTORS           	-	C10_3	pheotype:Control	RAW_FILE_NAME=ctrl_8284_3_run2chip5110; Age (weeks)=12
SUBJECT_SAMPLE_FACTORS           	-	C10_4	pheotype:Control	RAW_FILE_NAME=ctrl_8284_4_chip5110; Age (weeks)=14
SUBJECT_SAMPLE_FACTORS           	-	C10_5	pheotype:Control	RAW_FILE_NAME=ctrl_8284_5_chip5110; Age (weeks)=16
SUBJECT_SAMPLE_FACTORS           	-	C10_6	pheotype:Control	RAW_FILE_NAME=ctrl_8284_6_chip5110; Age (weeks)=18
SUBJECT_SAMPLE_FACTORS           	-	C10_7	pheotype:Control	RAW_FILE_NAME=ctrl_8284_7_chip5110; Age (weeks)=20
SUBJECT_SAMPLE_FACTORS           	-	C10_8	pheotype:Control	RAW_FILE_NAME=ctrl_8284_8_chip5110; Age (weeks)=22
SUBJECT_SAMPLE_FACTORS           	-	C10_9	pheotype:Control	RAW_FILE_NAME=ctrl_8284_9_chip5110; Age (weeks)=24
SUBJECT_SAMPLE_FACTORS           	-	C10_10	pheotype:Control	RAW_FILE_NAME=ctrl_8284_10_chip5110; Age (weeks)=26
SUBJECT_SAMPLE_FACTORS           	-	C10_11	pheotype:Control	RAW_FILE_NAME=ctrl_8284_11_chip5110; Age (weeks)=28
SUBJECT_SAMPLE_FACTORS           	-	C10_12	pheotype:Control	RAW_FILE_NAME=ctrl_8284_12_chip5110; Age (weeks)=30
SUBJECT_SAMPLE_FACTORS           	-	C13_1	pheotype:Control	RAW_FILE_NAME=ctrl8512_1_4574; Age (weeks)=8
SUBJECT_SAMPLE_FACTORS           	-	C13_2	pheotype:Control	RAW_FILE_NAME=ctrl8512_2_4574; Age (weeks)=10
SUBJECT_SAMPLE_FACTORS           	-	C13_3	pheotype:Control	RAW_FILE_NAME=ctrl8512_3_4574; Age (weeks)=12
SUBJECT_SAMPLE_FACTORS           	-	C13_4	pheotype:Control	RAW_FILE_NAME=ctrl8512_4_4574; Age (weeks)=14
SUBJECT_SAMPLE_FACTORS           	-	C13_5	pheotype:Control	RAW_FILE_NAME=ctrl8512_5_4574; Age (weeks)=16
SUBJECT_SAMPLE_FACTORS           	-	C13_6	pheotype:Control	RAW_FILE_NAME=ctrl8512_6_4574; Age (weeks)=18
SUBJECT_SAMPLE_FACTORS           	-	C13_7	pheotype:Control	RAW_FILE_NAME=ctrl8512_7_4574; Age (weeks)=20
SUBJECT_SAMPLE_FACTORS           	-	C13_8	pheotype:Control	RAW_FILE_NAME=ctrl8512_8_4574; Age (weeks)=22
SUBJECT_SAMPLE_FACTORS           	-	C13_9	pheotype:Control	RAW_FILE_NAME=ctrl8512_9_4574; Age (weeks)=24
SUBJECT_SAMPLE_FACTORS           	-	C13_10	pheotype:Control	RAW_FILE_NAME=ctrl8512_10_4574; Age (weeks)=26
SUBJECT_SAMPLE_FACTORS           	-	C13_11	pheotype:Control	RAW_FILE_NAME=ctrl8512_11_4574; Age (weeks)=28
SUBJECT_SAMPLE_FACTORS           	-	C13_12	pheotype:Control	RAW_FILE_NAME=ctrl8512_12_4574; Age (weeks)=30
SUBJECT_SAMPLE_FACTORS           	-	C13_13	pheotype:Control	RAW_FILE_NAME=ctrl8512_13_4574; Age (weeks)=32
#COLLECTION
CO:COLLECTION_SUMMARY            	TKO p53LSL R172H/+ Dicer1flox/flox Ptenflox/flox Amhr2cre/+ mice were generated
CO:COLLECTION_SUMMARY            	by mating p53LSL-R172H/+Dicer1flox/floxPtenflox/flox female mice with
CO:COLLECTION_SUMMARY            	Dicer1flox/floxPtenflox/floxAmhr2cre/+ male mice. For TKO controls,
CO:COLLECTION_SUMMARY            	p53LSL-R172H/+Dicer1flox/floxPtenflox/flox were generated. TKO control mice
CO:COLLECTION_SUMMARY            	carry the same genetic makeup as TKO mice but do not develop HGSC. A sequential
CO:COLLECTION_SUMMARY            	serum collection protocol was conducted to collect samples from TKO and TKO
CO:COLLECTION_SUMMARY            	controls every two weeks starting at 8 weeks of age until a humane end point for
CO:COLLECTION_SUMMARY            	sacrifice or development of ascites. TKO mice were sacrificed at Indiana
CO:COLLECTION_SUMMARY            	University School of Medicine in accordance with animal protocol (21124)
CO:COLLECTION_SUMMARY            	approved by the IACUC.
CO:SAMPLE_TYPE                   	Blood (serum)
#TREATMENT
TR:TREATMENT_SUMMARY             	TKO p53LSL R172H/+ Dicer1flox/flox Ptenflox/flox Amhr2cre/+ mice were generated
TR:TREATMENT_SUMMARY             	by mating p53LSL-R172H/+Dicer1flox/floxPtenflox/flox female mice with
TR:TREATMENT_SUMMARY             	Dicer1flox/floxPtenflox/floxAmhr2cre/+ male mice. For TKO controls,
TR:TREATMENT_SUMMARY             	p53LSL-R172H/+Dicer1flox/floxPtenflox/flox were generated. TKO control mice
TR:TREATMENT_SUMMARY             	carry the same genetic makeup as TKO mice but do not develop HGSC. A sequential
TR:TREATMENT_SUMMARY             	serum collection protocol was conducted to collect samples from TKO and TKO
TR:TREATMENT_SUMMARY             	controls every two weeks starting at 8 weeks of age until a humane end point for
TR:TREATMENT_SUMMARY             	sacrifice or development of ascites. TKO mice were sacrificed at Indiana
TR:TREATMENT_SUMMARY             	University School of Medicine in accordance with animal protocol (21124)
TR:TREATMENT_SUMMARY             	approved by the IACUC.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	An extraction solvent consisting of a mixture of isotopically labeled internal
SP:SAMPLEPREP_SUMMARY            	standards including 808 µM 13C6 arginine and 212 µM 13C methionine D3 was
SP:SAMPLEPREP_SUMMARY            	added to methanol in a 1:60 ratio and stored at 4 °C until further use. Serum
SP:SAMPLEPREP_SUMMARY            	samples were thawed on ice, followed by protein precipitation in a 10 µl serum
SP:SAMPLEPREP_SUMMARY            	aliquot with extraction solvent in a 1:3 ratio. Samples were vortexed for 30 s
SP:SAMPLEPREP_SUMMARY            	and centrifuged at 13,000 rpm for 7 min. To enhance CE peak shape, the resulting
SP:SAMPLEPREP_SUMMARY            	supernatant was diluted in a 1:4 ratio with the sample diluent containing 133 mM
SP:SAMPLEPREP_SUMMARY            	ammonium acetate, 0.1% formic acid and 1 µM 13C phenylalanine.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_TYPE           	CE
CH:INSTRUMENT_NAME               	ZipChip (908 Devices)
CH:COLUMN_NAME                   	HS chip
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Thermo Exactive Plus Orbitrap
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	POSITIVE
MS:MS_COMMENTS                   	µCE-HRMS analyses were performed using the microchip ZipChip® capillary
MS:MS_COMMENTS                   	electrophoresis system (classic interface, 908 Devices, Boston MA), coupled to a
MS:MS_COMMENTS                   	high-resolution accurate mass Q Exactive plus mass spectrometer (Thermo Fisher
MS:MS_COMMENTS                   	Scientific, MA). µCE-HRMS separations used ZipChip® HS chips (10 cm channel).
MS:MS_COMMENTS                   	All experiments were performed in positive ionization mode in the 50-500 m/z
MS:MS_COMMENTS                   	range at a mass resolution setting of 17,500. The capillary temperature was set
MS:MS_COMMENTS                   	to 200 °C and the sheath gas flow rate was 2 psi. The automatic gain control
MS:MS_COMMENTS                   	(AGC) target value was set to 3E6 and the maximum injection time was 20 ms. Data
MS:MS_COMMENTS                   	were acquired using Xcalibur 3.0 (Thermo Scientific) and were imported to
MS:MS_COMMENTS                   	Skyline software14 for peak picking and integration. The peak picking procedure
MS:MS_COMMENTS                   	used the analyte accurate m/z and migration time. Peak areas obtained from
MS:MS_COMMENTS                   	Skyline were exported as spreadsheets for further analysis. Quantitation was
MS:MS_COMMENTS                   	performed with the analyte peak areas relative to the peak area of one of the
MS:MS_COMMENTS                   	three isotopically labeled internal standards (13C6 arginine, 13C methionine D3
MS:MS_COMMENTS                   	and 13C phenylalanine) chosen based on migration time similarities
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	Micromolars 
MS_METABOLITE_DATA_START
Samples	T11_1	T11_2	T11_3	T11_4	T11_5	T11_6	T11_7	T11_8	T11_10	T11_11	T11_12	T11_13	T11_14	T11_15	T13_1	T13_2	T13_3	T13_4	T13_5	T13_6	T13_7	T13_8	T13_9	C8_1	C8_2	C8_3	C8_4	C8_5	C8_6	C8_7	C8_8	C8_9	C8_10	C8_11	C8_12	C8_13	C8_14	C10_1	C10_2	C10_3	C10_4	C10_5	C10_6	C10_7	C10_8	C10_9	C10_10	C10_11	C10_12	T15_1	T15_3	T15_5	T15_6	T15_7	T15_8	T15_9	T15_10	T15_11	C13_1	C13_2	C13_3	C13_4	C13_5	C13_6	C13_7	C13_8	C13_9	C13_10	C13_11	C13_12	C13_13
Factors	pheotype:TKO	pheotype:TKO	pheotype:TKO	pheotype:TKO	pheotype:TKO	pheotype:TKO	pheotype:TKO	pheotype:TKO	pheotype:TKO	pheotype:TKO	pheotype:TKO	pheotype:TKO	pheotype:TKO	pheotype:TKO	pheotype:TKO	pheotype:TKO	pheotype:TKO	pheotype:TKO	pheotype:TKO	pheotype:TKO	pheotype:TKO	pheotype:TKO	pheotype:TKO	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:TKO	pheotype:TKO	pheotype:TKO	pheotype:TKO	pheotype:TKO	pheotype:TKO	pheotype:TKO	pheotype:TKO	pheotype:TKO	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control	pheotype:Control
Alanine	16.08042421	18.76082853	13.71557187	11.5784383	28.32931746	14.71174298	14.49108647	15.12982504	12.10368714	15.30137234	10.25044023	15.04765625	13.43412898	19.5920378	10.93671638	11.19196448	10.88506657	10.7436622	13.40006414	10.36323088	9.308366261	13.63693376	22.85199986	15.08073538	14.87746856	10.36747466	12.64029737	10.70072793	9.639706751	10.52168504	10.93175619	12.62501865	12.68743139	17.08340705	15.17888922	13.39720936	19.51723984	14.10045433	11.99987873	9.799119407	14.03079165	9.549657528	10.58376168	10.87373964	15.22542536	10.75757334	14.02098222	17.93033825	10.92361523	15.34764517	24.20531869	16.75501336	14.94750589	20.80568285	23.17698772	18.66900087	23.93771645	13.99607233	13.26865578	18.41759798	13.75967642	13.63779972	17.28198686	19.15920933	18.01859561	11.85396476	15.07152708	16.59816792	16.50931667	24.57785623	30.74785469
aminoisobutyric acid	0.575606135	0.573637403	0.318756985	0.402923825	0.846681767	0.632751219	0.320131723	0.647948924	0.474880651	0.446280898	0.447988805	0.419684441	0.64573538	0.595767784	0.2352547	0.306049452	0.437215502	0.33473729	0.441397717	0.287759981	0.433195309	0.570995033	0.42377746	0.598609878	0.579068746	0.345514344	0.614163646	0.347237411	0.52786038	0.450512529	0.515319983	0.453362884	0.43467079	0.719651197	0.569606317	0.383664793	0.314477736	0.480859557	0.584074977	0.392349731	1.036698395	0.34083762	0.443684989	0.43239809	0.400814709	0.678344374	0.492636228	0.461227052	0.397234695	0.594966761	0.948462961	0.664897724	0.346066655	0.680678666	0.945583314	0.373498069	0.626156148	0.50985908	0.399775605	0.54823296	0.592308261	0.381199894	0.491151114	0.563779151	0.496881103	0.514857946	0.366431987	0.347090794	0.371818506	1.185408171	0.528298891
arginine	19.24043364	16.61033908	14.87189025	10.87665055	14.28891981	14.10771033	12.80617077	17.40507766	16.35001654	11.85322257	8.513420078	12.34306932	11.485376	8.41131829	12.80308911	14.4922945	14.3816955	14.91784699	15.30342589	15.64190188	15.99773282	5.166498685	2.830547778	19.29280255	14.12216677	13.24707668	14.33410332	13.72847572	14.28909321	14.07560623	11.8394456	11.54686916	15.10186381	18.73261129	17.69309246	11.05379777	8.22200764	14.99713148	11.00429623	13.45626094	13.17380137	10.86054115	12.93390943	13.23437257	16.56930343	12.95471892	13.38618841	12.5276797	5.559270043	13.70717918	13.20707482	11.79528925	15.42261491	12.70839371	12.93401965	9.316087749	10.87706179	5.500801956	14.04981548	18.91053821	15.36409639	18.95159227	15.03975789	20.31972111	16.04111446	13.86940164	12.34353213	13.92212432	16.46433074	18.53198451	15.8235983
asparagine	3.688708142	2.828896398	2.767664121	2.668590342	2.137309274	3.589798322	4.141794337	2.543847367	2.34098397	1.292422131	2.726910159	1.798488423	1.97683492	1.04065591	2.750262098	2.715549451	2.775957545	2.643238046	3.596989825	2.638407042	2.55383956	2.338937216	3.20576386	3.528896148	3.267475037	3.207322085	2.516042353	2.641910381	2.287840996	3.455724383	2.899982591	3.00313235	2.926976026	3.730094635	3.912423013	2.415139343	3.644584009	2.492763131	2.569627223	2.883205365	2.803972181	2.803547492	1.694353922	2.553788155	3.758593347	3.52369394	2.759610841	2.388314719	3.629223928	1.304574416	1.343917477	1.129598197	1.808738088	1.501537521	0.990558865	1.415173665	1.576079999	0.82942891	3.162945909	2.001661459	3.411676809	1.622957109	2.957825003	3.730527404	2.950138366	2.231517122	3.377098815	2.594875561	2.995986966	2.350889341	4.261020355
betaine	1.769384597	2.910908991	1.762129525	6.496322971	4.799293106	1.919907498	1.91684566	1.540513586	9.180400835	5.480705114	1.391396499	2.945347106	3.860296506	3.498995134	1.415951708	1.202560523	0.950416617	1.655081602	1.846056737	1.596626204	1.55935038	1.24383835	1.74470881	1.676048134	1.454717353	1.369614039	1.201177285	1.012494297	0.994293488	0.983733482	0.843660769	1.340566471	1.401357051	2.285542191	2.642499787	2.332707448	2.095616161	3.436719149	1.219584199	1.051887186	2.916629596	0.980129506	1.779789118	2.268514154	2.633794818	1.617797073	2.623022042	4.885046261	2.080374909	3.966679418	4.621150873	5.543386395	4.465331568	6.915296215	8.049037851	3.491152647	7.253912475	9.087434296	1.338096894	9.503401728	2.066067188	9.563319448	3.904798367	2.767641314	5.9908234	5.555557242	3.493244146	3.287507056	4.052941796	11.84947178	6.242409511
carnitine	0.863073578	1.11478802	0.840745024	3.262712299	2.098179779	1.345111786	0.54887101	1.053142336	2.505360161	3.258532004	0.476578126	1.488881692	2.336169268	1.446560798	1.108625777	0.943152841	0.577286031	0.840112394	1.020428154	0.832237438	0.818025534	0.855308198	1.341353328	1.088484595	1.220276128	1.19809699	0.73607917	1.031067589	1.039806521	0.813232613	0.856098682	1.278733118	1.127823833	1.184211916	1.580129897	1.284454693	1.450469364	1.753285288	0.709418709	0.869947785	1.248547122	0.862411148	0.737685233	1.23595939	1.136006968	0.795925976	1.721053721	2.185188185	1.084689301	1.487740107	1.773388031	2.154320448	1.724096811	2.368757993	3.093080489	1.89245736	2.564525593	3.181795358	1.051671216	2.44654383	0.649904945	1.921462416	1.556432442	0.929453691	1.450359482	1.823063538	1.644777335	1.345642725	1.320035302	1.773045133	1.782166685
choline	2.448178509	2.083823523	4.626035763	4.989261269	2.979539781	3.833252507	1.601656843	2.610517325	4.700765347	2.578112956	1.5685056	2.99439603	2.724646082	1.38390739	4.387249146	4.292131101	2.092218514	2.145569909	3.470962744	4.724924028	2.669731903	2.258026408	2.777675957	4.034588166	4.313050469	7.669750956	2.713258	4.915460898	4.332521477	3.573480048	2.245835194	3.130840477	5.12447913	3.187820506	3.858073522	2.742398859	2.525454535	7.133389348	2.557741471	4.825587725	4.280419269	4.03763082	1.610439355	4.29522665	4.376029061	3.622362616	3.785793599	3.72922623	1.411545216	2.540843664	2.588587123	3.175823782	2.561918043	3.217746769	3.013592909	2.109188688	3.694278526	3.208208846	4.681615247	3.428013444	1.649105215	5.677086576	2.396992119	2.960974152	3.37224506	3.092859924	5.2947465	2.378665933	2.236438706	2.195189598	2.590756246
dimethylglycine	0.398925217	0.330036138	0.42478352	0.59682379	0.638164726	0.332192658	0.38068652	0.259916108	0.52990033	0.467078377	0.238977488	0.461700446	0.435808665	0.597847606	0.250816792	0.21844387	0.255544434	0.280354512	0.388004889	0.292635467	0.410727382	0.732024426	0.428594596	0.347099868	0.343056183	0.278655348	0.25724187	0.220324401	0.187398579	0.201704268	0.209761585	0.2516359	0.252214345	0.309178552	0.416606709	0.249399984	0.318532717	0.320204885	0.29227449	0.18912849	0.289218086	0.206623989	0.313728744	0.334173819	0.286142242	0.22592525	0.305526044	0.274359186	0.30882704	0.378491915	0.458904826	0.574543809	0.439206592	0.581091314	0.7890682	0.426515866	0.5350901	0.569899554	0.195701019	0.497545794	0.298779324	0.658974251	0.270058146	0.375116597	0.22318418	0.280260645	0.249561534	0.24492683	0.254328151	0.500854273	0.36683583
glutamine	18.55464106	27.14706159	17.24863571	27.14156329	37.81659899	25.83154874	28.12831108	18.57260693	23.63771911	30.27768793	32.17207247	21.80293432	26.50137845	27.7790462	21.01189931	13.91969696	11.9826899	18.20545333	17.58311094	15.35900614	17.93197696	11.00605373	22.31367361	24.76792079	26.06505978	18.94134233	20.12096475	22.52854859	21.97495598	16.81367422	20.05751197	20.81148941	21.46827977	27.75994634	22.64357192	20.44578531	24.94298666	22.03782921	16.71442139	17.57190597	23.09397354	15.97633592	14.06942025	20.38353813	18.60739537	29.45908692	18.23693528	21.54853568	17.28855637	19.97572487	16.68976282	16.26778384	17.37859535	20.48027076	21.73222907	17.98249576	27.81534566	19.20012303	21.89009647	18.53746797	15.07899799	22.90561643	19.30897936	20.76242605	25.39159347	21.53866475	21.60152298	15.55651392	20.50068781	28.00202071	29.24954668
glycine	15.73508264	24.20034715	15.01904231	23.49023862	44.35571798	16.14658767	21.50816279	11.56413988	16.2805663	19.61072139	21.33250448	18.52127577	21.79430022	21.37929352	14.06023233	13.46444944	11.22035294	11.86340832	19.17273096	12.64827287	13.17298699	10.23839675	23.2601085	18.03900458	20.15626302	17.72108051	11.36133626	13.64037585	12.13431726	14.00743365	15.59239421	14.59228374	17.35389491	15.26675719	19.81912427	13.41193005	16.11012415	16.04894556	13.13688242	14.31655811	16.13019817	11.92866637	9.55137395	13.99981695	13.19865126	18.50284385	15.26703744	18.32572736	8.643360418	24.81490977	27.00608976	24.98412354	20.89304366	31.26310028	29.49752535	25.07999343	35.37420834	18.65323916	17.96258349	24.77635547	14.52194741	18.11755734	20.23488338	17.75109467	18.74623532	21.61381863	19.65234843	20.2049805	19.24193549	25.46529202	29.93508213
histidine	4.775858854	3.362162962	3.501452279	3.615921131	5.12953853	4.380018215	5.655939369	4.063415376	4.408196459	3.668595883	3.458024409	3.700884355	3.513819925	3.264740296	3.16116962	3.076329708	3.784779391	3.70279635	3.530808979	3.650137837	4.428308848	2.552397867	1.554394908	4.272236361	3.318208858	3.057380419	3.498890452	3.419670915	2.518545301	3.103088338	2.937924671	3.697152895	3.50093495	5.444044126	4.723144968	3.797500123	3.493649978	1.505362424	2.793339388	2.84488149	3.482798101	3.648966028	3.694856176	2.916938683	3.738549265	3.083648522	3.590773614	3.293786436	3.220839351	2.512895659	2.742288057	3.138219337	3.836183999	3.628394424	3.661556744	2.7411813	3.080668408	1.833451971	2.914325434	4.944090044	4.019865424	3.942501122	3.859858075	4.547236509	5.126994693	3.497720067	3.513127046	4.516288317	3.735581399	4.831649346	4.144812398
hydroxyproline	0.421494155	0.300698502	0.354529457	0.433373001	0.443361893	0.329827069	0.642426998	0.25950243	0.282058914	0.2758623	0.281245975	0.389099629	0.31318205	0.401237815	0.529519994	0.39544219	0.235813017	0.282410737	0.314759867	0.254997909	0.260843472	0.229276217	0.505906426	0.618015725	0.579365227	1.092734955	0.358035984	0.361590584	45.45518828	0.244951663	0.257177636	0.311215026	0.281587053	0.331809427	0.350702216	0.308284316	0.350044065	0.432473136	0.333965716	0.320674617	0.261714794	0.262772548	0.269469851	0.281019084	0.287596663	0.296056477	0.238352424	0.268418951	0.282276507	0.455659068	0.281211606	0.208389058	0.235055284	0.31270095	0.249942801	0.256949267	0.402890642	0.367543907	0.435026029	0.387956628	0.243182043	0.201818996	0.246286485	0.206885526	0.228492137	0.23317374	0.237125433	0.182152494	0.159149507	0.253079841	0.262089262
methionine	10.63146155	6.865774055	6.878265351	3.486073396	5.266652291	6.239809506	6.355499326	6.838463743	8.280670334	6.002704906	3.144540556	4.029952914	5.152810095	4.213242019	4.246457581	6.541986086	6.723226755	7.801871832	7.852475459	6.707872478	8.993853586	4.97157134	6.121519236	7.195896298	4.642245982	4.077780599	4.754471932	4.090792372	4.147334605	6.031121448	4.06268173	4.42540673	5.318991121	9.672950801	7.968539352	5.458989244	7.738366991	5.358729605	3.756250192	4.238566612	5.176329881	4.733863475	5.357906934	4.589017968	7.734507354	3.781895158	5.605949791	5.515365022	4.603331126	3.51198358	6.253137288	4.102211433	6.943514371	7.05818031	4.820372881	4.683566418	3.56228176	3.451497747	4.447478818	7.339613698	9.305945608	6.572342068	8.136155902	10.07303016	9.938710292	6.452805935	6.100892881	5.18107347	9.431682095	10.84338239	9.301126683
phenylalanine	5.77094485	4.338657481	5.361295128	4.958543693	8.038935977	4.836652821	4.908334247	4.80905832	5.516443053	4.901508007	3.70141122	5.544827323	4.246951892	4.998450736	3.574696252	3.522360994	4.264042354	4.323081825	5.007691049	4.02493643	4.678834909	4.736368615	2.577093602	4.115692688	4.645549417	3.709973318	3.859285748	4.010264228	3.532210438	3.391176723	3.573112271	3.901984675	4.297954387	5.359436305	4.864105058	3.763172754	3.973641645	3.3300647	3.491189557	2.988503176	3.844137646	3.141423221	3.821219586	3.910368745	3.907601954	2.900180524	3.970329887	3.620198549	3.466998389	2.79506468	3.708328612	2.864962968	4.232008411	4.190961572	3.815852955	3.208328776	3.492256761	2.514149558	3.764557945	4.666687062	4.154028207	4.257351313	4.630451176	4.896791069	4.586407095	4.204614612	4.037257014	3.912661872	4.19234648	4.625235091	4.833796855
proline	8.424061611	6.949416132	7.381197636	6.037179288	9.791222357	6.587270954	8.274794549	5.938504691	7.959670919	5.475197542	4.004127164	6.36429342	5.256326583	6.524670739	4.271609764	5.086595442	5.663797677	5.441165058	7.093349357	5.37471101	7.124381843	6.254252507	8.477089168	6.978831994	5.707200593	4.546779547	5.293867532	4.110556035	4.278065454	3.916859508	4.27464276	5.064339774	6.535825118	8.310494593	7.172035261	5.362114478	6.908722186	6.291416935	5.111684044	3.833437246	4.747254667	3.69871575	5.062567463	4.164951969	6.359021074	3.740697484	5.542275725	4.530368293	4.328832337	3.441467599	5.993931032	3.430668352	5.84319511	5.257884175	4.692081748	3.882130485	6.381343514	4.312148186	4.358061957	7.216605503	7.620784859	6.891982756	6.283018244	10.35129168	7.060735387	5.169528635	5.046625283	4.65715982	7.27792768	8.21812742	9.197827632
serine	4.535726015	3.180379562	3.758127869	3.466844962	2.886914325	4.693204321	4.399093018	3.726630331	3.01370686	1.730719524	3.093293722	2.92364587	2.395662535	1.022484333	2.813189605	3.469454985	3.519327606	2.955094345	4.517029026	3.940335767	3.491496554	1.697739806	1.516125575	4.074140829	3.275264885	4.192168585	3.164086552	3.08175332	2.659713224	3.805666941	2.827293836	3.605896408	3.971002973	4.034119508	5.183617803	2.979899174	3.30818804	1.749573106	2.614704531	3.236152755	3.348892894	3.303225167	2.254646769	2.9978539	4.101827031	3.444069349	3.385794684	3.486772958	3.366944451	1.859211455	2.007165818	1.60376841	2.192405824	2.12222511	1.776227965	1.81839978	2.37211536	0.767336344	3.405062091	2.358043023	3.826807275	2.174585406	4.239063869	4.925140169	3.356885145	2.578342424	4.322606283	3.179390156	3.606841672	3.229704109	3.297421752
threonine	17.14252886	14.2607961	13.35688654	12.97757234	15.99713821	14.4312699	13.76985382	12.91445932	15.62499998	12.81922535	7.902164114	8.936814294	14.31927669	6.675400558	10.23084095	12.42637298	12.94063642	13.08895491	15.27813139	12.07548177	14.5850844	9.854141253	10.86139662	15.1438714	11.93822613	11.47922744	9.997791876	11.12737417	11.46482781	12.20897901	10.89190849	12.21148751	13.05655271	14.86788072	18.91510718	9.550065304	11.21631255	10.16382791	9.019942935	11.2992155	13.35747889	11.3297902	10.02879087	11.53145129	16.94446575	10.51019519	12.78812762	11.06265178	9.173053275	10.63264633	12.29263814	11.06029591	14.26455365	13.01826133	12.13013038	10.95340007	9.119068247	8.061010956	11.44097773	16.74721959	13.70332805	11.49879107	17.27714581	17.94447676	16.3946456	11.96176507	14.80744394	14.84355578	14.32145013	18.01474568	15.86679278
tryptophan	7.186217972	4.658495995	7.292301698	6.77773402	7.398376837	6.950836316	5.316060271	6.236683476	7.022269396	5.942870129	4.116887055	4.858174555	7.492293294	2.163640862	5.677938999	5.35483017	6.19870067	7.613486237	7.560849316	6.763773186	8.377775679	5.178419067	3.628105196	5.42972991	5.837999361	6.162734826	4.69738714	5.949185698	6.068477116	7.753738656	6.202055648	5.927473649	5.504073824	7.785879416	8.667477634	4.190471557	3.998350253	4.079779789	4.345257859	4.714235783	6.90531512	5.884214915	5.842745205	5.786132878	7.055201116	5.818034861	6.233489038	4.000863929	5.237474429	4.134270104	4.355620095	6.85280987	6.500996676	8.328324911	6.111604775	5.842703604	3.958569727	3.347369167	3.486130282	9.660166652	5.291366848	4.995495045	5.8003228	6.132360962	7.432010258	7.517836172	6.001630447	8.647460523	5.231063734	10.46127267	7.021611062
tyrosine	11.41753962	3.863966561	8.530508147	7.676790807	7.793179005	6.56243866	8.548933618	6.563900881	10.29441917	5.977674862	4.294164164	4.363604708	6.885049276	3.285438355	5.991042588	5.81926449	6.589060021	9.552517016	14.41949727	9.3420584	9.876154444	5.050979634	5.359440155	8.269592684	6.725249886	4.649103428	3.927136821	5.438151725	5.717304972	7.108520261	5.625455445	6.052831426	6.07568262	7.073287203	8.440583702	4.496248631	4.725993067	5.609290793	4.218961627	5.126906784	4.962267994	6.971752048	6.361360951	6.936828703	9.907582045	3.80709566	6.363205508	5.578686727	5.220162564	4.276513338	5.947932693	5.698544009	12.08556547	9.562923271	7.50834411	8.036413677	4.569848684	4.823660672	5.031756292	11.83468052	11.7030398	6.54077268	9.427218912	7.573244299	8.003140088	8.166502926	9.559308992	10.8926002	6.381753347	12.96122861	11.58210053
valine	20.22867027	12.52659302	14.77916916	16.44358209	17.217587	15.20230103	9.30603411	16.27196003	18.5658482	15.45146196	6.457613694	9.822364611	15.42054444	15.47824114	11.37052811	9.468753194	13.68234619	14.87809617	21.03523226	15.50251433	19.33653881	10.78358239	10.82452202	16.44718056	14.64708842	13.93408101	12.03550488	13.71493632	13.9411414	11.76892919	12.73758217	12.71869832	15.31457764	17.17529424	17.43583505	12.27025532	11.28639268	11.68101948	12.22644878	11.94875335	16.85643489	13.27610557	14.53880824	14.18661158	16.14847345	13.55479358	15.81663909	13.34618251	12.21502954	15.35345832	21.06576744	28.54864231	20.80009478	28.69958936	32.84055388	15.07506736	23.06626116	38.60716155	13.06631056	22.77738324	19.186975	15.47780714	22.89307449	26.53127013	21.10511229	17.36272845	14.70281795	19.87855461	17.04740042	31.68258523	23.23990275
nicotinamide	0.741261021	0.712532634	0.740082737	0.251841912	0.827633313	0.661152319	0.702661034	0.692108802	1.82384348	1.648039979	0.457598984	0.547532154	0.91123309	0.473722236	0.670966146	0.425272325	0.802235987	0.715858956	0.893661297	1.000793282	0.73467588	0.414020106	0.55053396	1.102456061	0.999682531	1.469929937	0.718968664	0.765200975	0.67109942	0.65498653	0.611274625	0.801461467	0.963547184	0.90071391	0.87376492	0.867301528	0.642061799	0.753780431	0.824747047	0.646631082	0.815558212	0.820326292	0.745304635	0.85327543	0.943354494	0.696371181	0.9009421	1.0059846	0.717439418	0.465099185	0.911446601	1.153594583	1.244726393	1.101745939	1.469802273	1.302513146	1.031819279	1.543989215	0.762390382	0.251841912	0.728412213	1.206182629	0.97351929	0.875289329	0.923648588	0.781282427	0.950569228	1.160207186	0.856340187	1.187873689	0.778239079
aspartic acid	0.748969389	0.130239229	1.123884279	0.392707188	0.111073401	0.99713097	0.898380009	0.62889331	0.565565863	0.07951096	0.569346778	0.588616859	0.287936227	0.173093242	0.73445769	0.480655391	0.452871174	0.495406606	1.624069706	0.956165467	0.639505832	0.481870998	0.199474192	1.304157202	0.907933721	1.634914411	0.659335458	0.766354415	0.868213767	0.55059683	0.345600705	0.803104912	1.274806986	0.703739662	1.260040816	0.560039669	0.226274887	1.031573511	0.566645322	0.841335147	0.604466233	0.836178657	0.198881535	0.566364085	1.50153104	0.402613264	0.705323291	0.507498514	0.115324453	0.239574387	0.122994856	0.323772567	0.315319965	0.133069849	0.096019373	0.139166823	0.44216292	0.627637294	0.616882403	0.506250805	0.377705209	0.237817629	0.353743758	0.696374802	0.493844354	0.055853628	0.567609208	0.159509357	0.183146816	0.102757362	0.033289976
aminoadipic acid	0.236111331	0.289225319	0.237380704	0.328259473	0.434311749	0.308167141	0.189832208	0.234532834	0.245762944	0.345468452	0.365110137	0.193108801	0.244735398	0.235621924	0.215195965	0.224362732	0.241273228	0.243651889	0.209658236	0.195450323	0.24774427	0.145540229	0.207617655	0.327420287	0.33589667	0.279316595	0.237893108	0.219083945	0.245645959	0.203436748	0.222595166	0.207399285	0.263308195	0.271796362	0.295670975	0.159688417	0.234368634	0.287315463	0.231432344	0.252391903	0.326126937	0.179180789	0.172740864	0.250666315	0.245299951	0.260833056	0.267658065	0.180305532	0.19185744	0.198395197	0.311551154	0.229356472	0.217658744	0.255288629	0.267774746	0.208676587	0.246219997	0.206281884	0.329370119	0.338062693	0.151780389	0.381448351	0.261061201	0.329154998	0.351014296	0.263982645	0.259487704	0.212709535	0.263000768	0.401324122	0.399310534
ornithine	5.768994113	4.805229407	4.684124614	6.280338272	4.921379766	4.59199045	5.026916771	5.030792942	6.671382614	4.59798138	2.656442768	3.29369748	4.997882795	4.249609219	3.077259874	4.201809608	4.721623932	4.585158342	7.913001467	5.523222003	7.276158817	9.732669511	4.620721825	6.241350217	3.83631101	4.783707595	3.96534171	3.64593811	3.912572424	4.777479105	3.060700749	3.440637854	5.676787363	5.616215168	5.890707509	3.497820228	4.042407219	3.918118927	3.740690992	4.311114209	4.62825169	3.368434298	3.432054067	3.87620658	6.152474187	4.304751989	4.71098385	4.7451957	4.017320079	2.759846887	3.606747768	3.854450785	4.644104501	4.761325977	3.481977226	4.10333599	3.363043757	3.083396742	3.949741006	6.56727502	5.254533471	6.441125895	4.61035009	6.656397892	6.381847307	4.846768665	4.258216452	4.293228545	6.19932904	8.069281417	8.45816812
citrulline	5.115554613	3.417125155	4.80040965	5.133253077	5.360590467	5.300798026	5.983710368	4.744940776	5.952472723	4.669210102	3.776740367	3.221832688	4.96414744	3.479625471	4.657326421	3.436989378	4.776886062	5.033719477	7.888080987	7.20618782	6.92613546	2.57449202	4.783378155	5.478900899	5.234114628	4.62576099	3.773301657	5.282920501	5.414870756	4.275811775	4.921100876	4.765574226	5.744016202	6.153125093	7.333497312	4.413138579	4.931993546	4.526416977	3.317890851	5.423517332	4.715438726	4.869334745	3.803386104	4.689619392	6.67507647	4.331508373	4.561362457	4.894908814	3.446573373	3.333326183	4.251462178	4.682151451	6.783939808	4.843870139	5.337374819	4.805979152	3.66688842	3.285293086	4.808879749	8.43188885	4.804107588	4.893000705	5.956293299	5.317597901	7.541315917	6.658343077	7.47343424	5.137755607	6.390575379	8.626580233	7.592672597
isoleucine	8.475562461	7.491824946	6.606809147	6.962279494	13.01007535	6.763219923	4.941290202	6.420895133	8.743289857	7.746495427	4.735568509	7.2343546	8.237054593	7.331817315	5.46055981	4.981654081	5.74327065	5.805836169	7.37274442	5.902236775	7.190522727	5.902929956	7.670150776	6.880033834	8.768580543	5.546376529	6.347935055	6.913597382	6.255183665	4.328231275	5.480014384	5.022477076	7.454917986	7.243734555	6.350027123	5.720536519	5.182297723	6.663024625	5.049896479	4.859256914	7.133783426	4.670102598	5.847459595	6.189062642	5.749593316	6.228288647	7.172411293	6.269105216	5.235709436	6.389563948	7.931284872	4.243170007	7.122850368	7.434315052	8.059339027	5.940831706	6.406229048	5.135386324	5.520348724	6.650816288	6.269578832	7.644537064	8.051380055	9.166326379	7.289218243	7.175051163	5.483227753	6.868042021	6.420288524	9.598702844	9.122026335
leucine	19.0627748	11.19535928	14.031338	14.2297843	13.84043102	14.50169194	8.970911007	13.739325	18.5762584	13.98066203	4.603496826	8.370897782	11.50519154	15.05623481	10.15561848	8.367687163	11.93034602	14.15731885	16.75748259	15.38265508	17.11848372	7.777725133	7.890640777	15.07175551	10.9249199	10.93813103	9.61997263	10.87369863	10.65668599	10.96904244	8.895921405	9.858159053	12.90487504	16.18444598	14.018215	10.77816048	10.74312557	8.592765651	9.020372221	8.943205647	13.49615158	11.26254172	12.72360497	10.70541802	15.52639305	12.22716838	13.53178206	13.77717875	9.107490448	10.26199123	15.97571156	11.8224516	20.29461294	17.49095506	16.54191648	11.44508019	10.1652022	8.765780336	9.491810025	21.08136965	15.92589726	14.89352647	20.16882537	20.83782047	19.03911695	13.6315787	10.00930694	15.94185611	13.77634492	24.02310032	17.18435857
lysine	16.12581978	24.75258201	13.66433259	26.01787381	41.41635442	12.46289043	13.07048832	15.34983698	26.61393555	36.44501633	12.41955307	15.25759387	24.86880015	37.95994138	13.28469979	15.1848215	10.44580233	12.58748145	12.69868657	10.35201311	15.04935748	10.35779893	17.42165694	18.05638214	17.89927827	13.5607255	12.99750507	14.21339612	13.54344511	12.36474494	11.49446374	13.17390499	15.45826503	18.66283997	19.5856239	13.01843925	14.93460079	17.93698808	10.95822928	11.99556504	17.1874942	10.10898619	13.49569716	15.31624292	13.68221992	16.41773501	17.70806001	19.14127448	9.098396134	22.16954871	27.27712983	19.79316141	21.86553314	29.19267097	30.23196031	21.83767077	23.45963858	27.47846327	14.74919361	32.36066454	10.66773349	28.16393459	13.65367143	15.00668381	17.90634609	21.31213745	15.26096468	17.98684471	15.15519923	33.1416481	26.87364804
glutamic acid	3.32011988	3.569222475	5.119017362	7.536974947	6.309619571	4.63282954	5.248077222	3.340965179	4.213293563	3.475355071	4.176134917	3.311105637	2.82518527	1.721919349	3.752654648	2.530576691	2.328438936	2.761049746	4.931420562	4.356120008	2.393552852	1.4337738	1.687781412	7.565157533	8.129088515	11.59682478	3.912534454	5.06485436	5.773145873	2.728741591	2.856607629	4.905978303	9.309183714	3.444101499	5.423662519	2.956129238	2.082612412	7.75612645	4.03332669	5.266750634	5.787082109	4.132584815	1.874079873	4.320482646	5.860901945	2.824889941	4.051174922	3.612216649	1.211497044	2.45223378	3.417897544	3.466637797	3.071658656	3.130880837	3.328083428	3.12562358	5.240647617	1.425535158	4.385977638	4.842733964	2.227612023	4.447373967	3.004912231	2.978105452	3.095424827	2.059228088	4.218549271	2.565250308	2.131307139	2.017402424	1.597295852
cytidine	0.051568217	0.1592782	0.067943656	0.138307206	0.18752704	0.080117216	0.099116365	0.064842355	0.079113699	0.086259275	0.128968727	0.128412725	0.10881286	0.111936445	0.092143191	0.089392387	0.062014364	0.037455695	0.127971579	0.072121311	0.024029208	0.119900868	0.08023904	0.12186907	0.152366622	0.169670516	0.077956964	0.121413942	0.12631864	0.07136683	0.054156305	0.07371814	0.110691765	0.044783878	0.079935161	0.05995361	0.043067112	0.173167445	0.080708908	0.076288178	0.092078742	0.064607048	0.019959015	0.116880798	0.078954687	0.052929053	0.066944344	0.084502313	0.025982328	0.101827141	0.092248189	0.085311709	0.055348967	0.075644008	0.096155025	0.105157092	0.186548324	0.076638967	0.120757826	0.08219634	0.011459261	0.06842513	0.061549308	0.028455892	0.035263159	0.064981844	0.092455583	0.071098098	0.042759255	0.03453903	0.01912587
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	m/z	Migration time (min)
Alanine	90.0553	1.2
aminoisobutyric acid	104.0709	1.3
arginine	175.119	1
asparagine	133.0608	1.6
betaine	118.0864	1.9
carnitine	162.1123	1.2
choline	104.1073	0.9
dimethylglycine	104.0709	1.8
glutamine	147.0726	1.7
glycine	76.0397	1.2
histidine	156.0767	1.1
hydroxyproline	132.0655	2
methionine	150.0582	1.5
phenylalanine	166.0862	1.6
proline	116.0707	1.6
serine	106.0501	1.5
threonine	120.0657	1.5
tryptophan	205.0971	1.6
tyrosine	182.0812	1.5
valine	118.0864	1.4
nicotinamide	123.0556	1.5
aspartic acid	134.0448	1.8
aminoadipic acid	162.0759	1.7
ornithine	133.0971	1
citrulline	176.103	1.7
isoleucine	132.1018	1.4
leucine	132.1018	1.4
lysine	147.1126	1
glutamic acid	148.0603	1.7
cytidine	244.0927	1.5
METABOLITES_END
#END