#METABOLOMICS WORKBENCH juliehaines_20230821_092741 DATATRACK_ID:4234 STUDY_ID:ST002828 ANALYSIS_ID:AN004618 PROJECT_ID:PR001770
VERSION             	1
CREATED_ON             	August 22, 2023, 7:06 am
#PROJECT
PR:PROJECT_TITLE                 	Role of the Afferent Lymph as an Immunological Conduit to Analyze Tissue
PR:PROJECT_TITLE                 	Antigenic and Inflammatory Load - metabolomics
PR:PROJECT_SUMMARY               	The intestinal barrier, and the gut-associated lymphoid tissue, are the
PR:PROJECT_SUMMARY               	interface of the host-microbiome/pathogens interactions and, its disruption, has
PR:PROJECT_SUMMARY               	been associated with a series of inflammatory, autoimmune, and degenerative
PR:PROJECT_SUMMARY               	diseases. Herein we performed the analysis of the pre-nodal mesenteric lymph at
PR:PROJECT_SUMMARY               	steady state and following disruption of the intestinal barrier to map the
PR:PROJECT_SUMMARY               	antigenic and pro-inflammatory load in both mice and human subjects with
PR:PROJECT_SUMMARY               	inflammatory bowel disease. The antigenic signature of gastrointestinal tissue
PR:PROJECT_SUMMARY               	inflammation was reflected in the mesenteric nodal dendritic-cells
PR:PROJECT_SUMMARY               	(DC)-MHCII-eluted immunopeptidome, in a tissue specific manner, when compared to
PR:PROJECT_SUMMARY               	the DC-eluted-MHCII-peptidome from cervical lymph nodes. Pro-inflammatory and
PR:PROJECT_SUMMARY               	microbiome-derived by-products, such as amino acids, deoxy sugars, component of
PR:PROJECT_SUMMARY               	the bacterial wall and other immunomodulators of the gut-brain axis, were found
PR:PROJECT_SUMMARY               	in the afferent lymph following damage to the gut epithelium. Our data points to
PR:PROJECT_SUMMARY               	the relevance of the lymphatic fluid to probe the tissue-specific antigenic and
PR:PROJECT_SUMMARY               	inflammatory load transported to the draining lymph node.
PR:INSTITUTE                     	University of Colorado Denver
PR:LABORATORY                    	Lab of Angelo D'Alessandro in collaboration with lab of Laura Santambrogio
PR:LABORATORY                    	(Weill Cornell)
PR:LAST_NAME                     	Haines
PR:FIRST_NAME                    	Julie
PR:ADDRESS                       	12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
PR:EMAIL                         	julie.haines@cuanschutz.edu
PR:PHONE                         	3037243339
#STUDY
ST:STUDY_TITLE                   	Role of the Afferent Lymph as an Immunological Conduit to Analyze Tissue
ST:STUDY_TITLE                   	Antigenic and Inflammatory Load - metabolomics
ST:STUDY_SUMMARY                 	The intestinal barrier, and the gut-associated lymphoid tissue, are the
ST:STUDY_SUMMARY                 	interface of the host-microbiome/pathogens interactions and, its disruption, has
ST:STUDY_SUMMARY                 	been associated with a series of inflammatory, autoimmune, and degenerative
ST:STUDY_SUMMARY                 	diseases. Herein we performed the analysis of the pre-nodal mesenteric lymph at
ST:STUDY_SUMMARY                 	steady state and following disruption of the intestinal barrier to map the
ST:STUDY_SUMMARY                 	antigenic and pro-inflammatory load in both mice and human subjects with
ST:STUDY_SUMMARY                 	inflammatory bowel disease. The antigenic signature of gastrointestinal tissue
ST:STUDY_SUMMARY                 	inflammation was reflected in the mesenteric nodal dendritic-cells
ST:STUDY_SUMMARY                 	(DC)-MHCII-eluted immunopeptidome, in a tissue specific manner, when compared to
ST:STUDY_SUMMARY                 	the DC-eluted-MHCII-peptidome from cervical lymph nodes. Pro-inflammatory and
ST:STUDY_SUMMARY                 	microbiome-derived by-products, such as amino acids, deoxy sugars, component of
ST:STUDY_SUMMARY                 	the bacterial wall and other immunomodulators of the gut-brain axis, were found
ST:STUDY_SUMMARY                 	in the afferent lymph following damage to the gut epithelium. Our data points to
ST:STUDY_SUMMARY                 	the relevance of the lymphatic fluid to probe the tissue-specific antigenic and
ST:STUDY_SUMMARY                 	inflammatory load transported to the draining lymph node.
ST:INSTITUTE                     	University of Colorado Denver
ST:LAST_NAME                     	Haines
ST:FIRST_NAME                    	Julie
ST:ADDRESS                       	12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
ST:EMAIL                         	julie.haines@cuanschutz.edu
ST:PHONE                         	3037243339
#SUBJECT
SU:SUBJECT_TYPE                  	Mammal
SU:SUBJECT_SPECIES               	Mus musculus
#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	1A	Factor:control afferent	RAW_FILE_NAME=F2-48-01
SUBJECT_SAMPLE_FACTORS           	2	2A	Factor:control afferent	RAW_FILE_NAME=F2-48-02
SUBJECT_SAMPLE_FACTORS           	9	9A	Factor:control afferent	RAW_FILE_NAME=F2-48-03
SUBJECT_SAMPLE_FACTORS           	13	13A	Factor:control afferent	RAW_FILE_NAME=F2-48-04
SUBJECT_SAMPLE_FACTORS           	14	14A	Factor:control afferent	RAW_FILE_NAME=F2-48-05
SUBJECT_SAMPLE_FACTORS           	1	1E	Factor:control efferent	RAW_FILE_NAME=F2-48-06
SUBJECT_SAMPLE_FACTORS           	2	2E	Factor:control efferent	RAW_FILE_NAME=F2-48-07
SUBJECT_SAMPLE_FACTORS           	9	9E	Factor:control efferent	RAW_FILE_NAME=F2-48-08
SUBJECT_SAMPLE_FACTORS           	13	13E	Factor:control efferent	RAW_FILE_NAME=F2-48-09
SUBJECT_SAMPLE_FACTORS           	14	14E	Factor:control efferent	RAW_FILE_NAME=F2-48-10
SUBJECT_SAMPLE_FACTORS           	3	3A	Factor:DSS afferent	RAW_FILE_NAME=F2-48-11
SUBJECT_SAMPLE_FACTORS           	4	4A	Factor:DSS afferent	RAW_FILE_NAME=F2-48-12
SUBJECT_SAMPLE_FACTORS           	5	5A	Factor:DSS afferent	RAW_FILE_NAME=F2-48-13
SUBJECT_SAMPLE_FACTORS           	6	6A	Factor:DSS afferent	RAW_FILE_NAME=F2-48-14
SUBJECT_SAMPLE_FACTORS           	7	7A	Factor:DSS afferent	RAW_FILE_NAME=F2-48-15
SUBJECT_SAMPLE_FACTORS           	16	16A	Factor:DSS afferent	RAW_FILE_NAME=F2-48-16
SUBJECT_SAMPLE_FACTORS           	3	3E	Factor:DSS efferent	RAW_FILE_NAME=F2-48-17
SUBJECT_SAMPLE_FACTORS           	4	4E	Factor:DSS efferent	RAW_FILE_NAME=F2-48-18
SUBJECT_SAMPLE_FACTORS           	5	5E	Factor:DSS efferent	RAW_FILE_NAME=F2-48-19
SUBJECT_SAMPLE_FACTORS           	6	6E	Factor:DSS efferent	RAW_FILE_NAME=F2-48-20
SUBJECT_SAMPLE_FACTORS           	7	7E	Factor:DSS efferent	RAW_FILE_NAME=F2-48-21
SUBJECT_SAMPLE_FACTORS           	16	16E	Factor:DSS efferent	RAW_FILE_NAME=F2-48-22
#COLLECTION
CO:COLLECTION_SUMMARY            	Mice (C57BL/6J) mice were obtained from Jackson Laboratories and housed at Weill
CO:COLLECTION_SUMMARY            	Cornell Medicine animal facilities. The mice were anesthetized using ketamine
CO:COLLECTION_SUMMARY            	(80 mg/kg) and Xylazine (10 mg/kg), injected intraperitonially. Cannulation is
CO:COLLECTION_SUMMARY            	performed 15-20 minutes post anesthetizing the mice, when the mice is no longer
CO:COLLECTION_SUMMARY            	able to feel toe pinch. Mice are placed face up with paws stretched and pinned.
CO:COLLECTION_SUMMARY            	Incision is made at the abdominal area, exposing the underlying gut and
CO:COLLECTION_SUMMARY            	intestinal tissues. Pre-nodal / post-nodal lymphatic vessels are visualized and
CO:COLLECTION_SUMMARY            	identified. The pre-nodal lymphatic vessels run from intestines to mesenteric
CO:COLLECTION_SUMMARY            	lymph nodes, while post-nodal lymphatic runs behind the mesenteric lymph nodes
CO:COLLECTION_SUMMARY            	and can be reached by clearing tissues. Fat tissues is carefully removed or move
CO:COLLECTION_SUMMARY            	aside using forceps and Q-tips. Care is taken to keep the tissues hydrated using
CO:COLLECTION_SUMMARY            	1X PBS. Upon creating a small tear/cut on the lymphatic vessel, using a glass
CO:COLLECTION_SUMMARY            	pipette (tip diameter of 40-60 uM) lymph is collected. This pipette is then
CO:COLLECTION_SUMMARY            	flushed into a microcentrifuge containing 1X PBS and 2X protease inhibitor
CO:COLLECTION_SUMMARY            	(Thermofisher Scientific (A32963).
CO:SAMPLE_TYPE                   	Lymph fluid
#TREATMENT
TR:TREATMENT_SUMMARY             	C57Bl6 mice are given access to either sterile water or water supplemented with
TR:TREATMENT_SUMMARY             	40-50 kDa dextran sulfate sodium (DSS) for 7 days.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Frozen mouse lymph samples were provided as 3 uL lymph fluid in PBS to a final
SP:SAMPLEPREP_SUMMARY            	volume of 300 uL. Specimens were thawed on ice then 22.2 uL was transferred to a
SP:SAMPLEPREP_SUMMARY            	tube containing 200 uL of ice-cold 5:3:2 methanol:acetonitrile:water. Samples
SP:SAMPLEPREP_SUMMARY            	were agitated 30 min in a cold room then spun at 12,000 g, 10 min, 4 degrees C.
SP:SAMPLEPREP_SUMMARY            	Two 100 uL aliquots of supernatant from each tube were transferred into new
SP:SAMPLEPREP_SUMMARY            	(separate) tubes and dried under vacuum at room temperature. One set was
SP:SAMPLEPREP_SUMMARY            	resuspended in 100 uL of 0.1% formic acid for positive mode acquisition. The
SP:SAMPLEPREP_SUMMARY            	other set was resuspended in 100 uL of 1 mM ammonium acetate for negative mode
SP:SAMPLEPREP_SUMMARY            	acquisition.
SP:PROCESSING_STORAGE_CONDITIONS 	4℃
SP:EXTRACT_STORAGE               	-80℃
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	Positive C18
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Thermo Vanquish
CH:COLUMN_NAME                   	Phenomenex Kinetex C18 (150 x 2.1mm,1.7um)
CH:SOLVENT_A                     	100% water; 0.1% formic acid
CH:SOLVENT_B                     	100% acetonitrile; 0.1% formic acid
CH:FLOW_GRADIENT                 	0-0.5 min 5% B, 0.5-1.1 min 5-95% B, 1.1-2.75 min hold at 95% B, 2.75-3 min
CH:FLOW_GRADIENT                 	95-5% B, 3-5 min hold at 5% B
CH:FLOW_RATE                     	450 uL/min
CH:COLUMN_TEMPERATURE            	45
CH:SAMPLE_INJECTION              	10 uL
#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                   	Resolution 70,000, scan range 65-900 m/z, maximum injection time 200 ms,
MS:MS_COMMENTS                   	microscans 2, automatic gain control (AGC) 3 x 10^6 ions, source voltage 4.0 kV,
MS:MS_COMMENTS                   	capillary temperature 320 C, and sheath gas 45, auxiliary gas 15, and sweep gas
MS:MS_COMMENTS                   	0 (all nitrogen). Data converted to mzXML using RawConverter. Metabolites were
MS:MS_COMMENTS                   	annotated and integrated using Maven in conjunction with the KEGG database.
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	peak area
MS_METABOLITE_DATA_START
Samples	1A	2A	9A	13A	14A	1E	2E	9E	13E	14E	3A	4A	5A	6A	7A	16A	3E	4E	5E	6E	7E	16E
Factors	Factor:control afferent	Factor:control afferent	Factor:control afferent	Factor:control afferent	Factor:control afferent	Factor:control efferent	Factor:control efferent	Factor:control efferent	Factor:control efferent	Factor:control efferent	Factor:DSS afferent	Factor:DSS afferent	Factor:DSS afferent	Factor:DSS afferent	Factor:DSS afferent	Factor:DSS afferent	Factor:DSS efferent	Factor:DSS efferent	Factor:DSS efferent	Factor:DSS efferent	Factor:DSS efferent	Factor:DSS efferent
L-alanine	336911	959552	1240419	352099	324197	394554	484684	635010	714468	481985	448135	1436968	853079	3345703	1289646	1143992	405443	346124	348294	1773950	420335	1698662
L-arginine	89686	119153	90670	168884	449544	552136	226096	415958	453407	616174	270833	173560	45701	444629	155727	189248	510481	505344	267470	315074	402808	546503
L-glutamate	6744	9387	24374	34911	24498	23128	8715	33691	37161	62214	41982	31766	5287	32890	28968	17484	19469	20953	17074	22360	27242	20517
L-glutamine	31748	17644	105504	97882	278253	400118	211443	464878	663465	651133	120004	47089	18880	192842	15414	141210	382847	344599	229589	215008	252268	280814
L-histidine	46514	67231	43821	61418	68977	113079	66745	120641	110553	108009	122540	107140	30100	102015	121255	35771	103185	83039	70458	89162	105308	90713
L-leucine	499319	609361	547714	1284777	2069418	2292583	1213640	2892696	2572324	3118463	1165541	806131	229448	2824941	474897	545641	1830898	1723862	881180	1692669	1295037	2184291
L-lysine	49009	45359	37974	79148	311097	441122	158604	244113	409500	405031	169910	66773	27161	405026	51853	100206	298039	305223	174595	286392	259047	348907
L-methionine	54053	79996	74132	124770	736704	655159	263570	744732	590539	702898	169694	54658	18217	696820	41396	117484	322570	301161	221963	337057	253562	537290
L-phenylalanine	181116	389440	320271	634351	951578	1537759	860499	1803314	1153894	1461097	762497	343686	99477	1960539	260960	371748	1072635	846692	591840	1198733	994825	1275824
L-proline	522990	515576	383948	2493225	557234	825796	514948	833909	1307273	864679	640422	509837	300990	811291	504606	575289	971680	971355	699980	734787	875847	1106531
L-serine	64384	70406	47235	22921	27007	70559	53982	59476	61374	98696	101743	140164	40628	78816	175765	44253	89146	58804	88702	104171	93066	77156
L-threonine	30922	37369	13151	27307	51076	127847	43456	91979	120337	212483	82864	67517	35874	88886	96105	73299	111678	79466	79891	110519	114724	137438
L-tyrosine	223675	404934	281232	319745	1335603	1729768	776564	1496394	697450	1334456	795796	496736	114999	2032332	321559	369323	872103	722195	679033	1088138	728257	1222889
L-valine	708582	850838	769415	1783934	2681832	2361832	1547617	2641116	2226384	2523690	971650	767367	659297	2720196	698114	848737	2438654	2774266	1484615	1409668	2316892	2355411
AMP	11343	12676	21683	27557	115566	51334	25172	77827	29097	65088	35697	8296	2684	32005	3600	18129	10960	18927	21746	16077	29186	20907
Adenine	108804	117999	120768	27009	27539	114766	110620	165825	40412	30938	101881	126142	111620	132895	34332	117194	119853	114167	113678	126994	38052	112216
dCMP	14303	19464	12921	4833	4577	9769	15180	17544	5950	6900		30409	27166	19950	15577	24593	10482	15501	19074	22269	9055	19903
Thymidine	28223	41064	69933	15473	31094	20507	29964	19608	3964	16926	5109	8318	44571	20896	13154	13863	54019	10456	8397	14662	118782	5555
Uracil	63359	86072	51047	238827	82117	40127	72629	53563	54896	45675	47220	80158	71242	87249	72075	71915	46504	28482	54087	84395	128312	182891
Hypoxanthine	46478	193483	46900	78041	267763	50634	107620	68828	28149	549722	717350	40286	26014	532565	62275	216285	32015	31729	50644	40824	48662	238025
Xanthine	15788	41038	21030	33673	77588	10454	42709	18274	5091	116014	122314	13474	2851	91542	14219	51861	10162	7030	22034	11405	14847	109376
Allantoate	19013	31914	8275	8441	36684	26298	22406	26442	31212	28270	76211	42367	12648	58410	24187	9102	25589	30546	39430	46678	34436	31121
(S)(+)-Allantoin	3905			253157	21307	37422	6361	16176	41821	21759	1922	1563		24416			34447	40924	11600	9621	17315	17842
Urate	5836	19312	1177	24301	156411	193199	98883	37065	107071	227950	65665	10617	2448	94011	14120	42981	48602	41001	95078	58407	47099	298797
Pyridoxal	8435	6568	7139	837905	13219	10595	7952	5545	7460	10093	5005	5578	2551	11498	4631	17825	13938	9880	7283	2534	9322	7799
Nicotinamide	380267	307073	312071	436927	809757	627888	404427	632902	616772	647464	532188	274880	252415	670455	243305	417301	626323	595143	421792	320709	372139	678401
5-Phosphoribosylamine	194853	241590	92684	79590	113672	344840	317638	409343	92526	111527	266706	334443	373722	300292	37501	248265	267011	341719	348231	256816	150393	217797
UDP-glucose	267511	298342	234470	77777	76305	365064	392393	236505	164712	151147	299192	276875	256782	260068	227952	237428	346544	346852	303581	235450	245579	346293
NAD+	22919	7831	20168	3377	4468	15858	7374	9231	4723	3729	35655	20733	7918	10116	4801	16736	14607	6441	18894	14194	11977	16568
5-Oxoproline	768120	972251	633441	1215732	450952	522976	406627	542833	393310	624968	1498670	1765487	370247	2048148	1211160	393080	829185	382396	419799	819855	644447	956602
gamma-L-Glutamylputrescine	7655	15942	11132	4737		1367	9391	6785	4939	3640	9988	9154	10855	8950	12798	9401	5227	6475	5137	11451	4533	15289
S-Adenosyl-L-homocysteine	45494	68582	50537	42588	123768	96503	70487	61152	208164	162462	91733	80741	67173	65136	29827	62543	146007	138255	59390	67779	55722	73805
Ornithine	54688	88420	35892	40315	67777	68867	61053	66978	64409	72114	127436	89483	46278	153978	53380	37623	77228	75021	77188	125585	82174	77026
L-Citrulline	18158	3638	23938	21380	41381	61753	25364	27947	89241	49547	37032	79132	1590	52980	151571	19706	105631	122445	40280	43185	103939	82090
Putrescine	11523	22940	4776	21342	3930	5679	4215	4842	8818	7930	9466	75132	3177	13719	4632	4478	8310	4176	7692	7540	5962	5979
Spermidine	10084	34725	8601	56647	22116	23664	20133	25462	34798	44114	34341	6399	6323	52264	1819	27794	9664	20333	22401	21288	16483	20099
Creatine	988375	2249219	1374351	4035261	9885598	6198658	3504468	5048454	6638278	5474952	10637930	1213355	595109	9998647	1833451	4555434	5060868	5781714	5308535	3557033	6573500	7673320
Creatinine	49996	90370	73324	75108	125091	206601	100714	111985	158992	151205	100601	45403	29022	106816	41309	51807	164084	173045	142593	65401	111921	147637
N-Acetylornithine	13480	10650	9815	44989	33562	83846	20206	47156	80595	37040	18744	13546	7683	35133	6400	11221	52775	47420	34301	17162	29896	33585
N-Succinyl-L-glutamate	478308	419072	107539	158420	162140	399716	421276	570987	167986	153016	472135	498473	659591	568260	81348	518555	459360	576606	419003	439589	158828	328962
N-Succinyl-L-glutamate 5-semialdehyde	564858	552260	707423	468577	81782	716794	540823	778546	461132	152916	608471	537657	264506	570773	277972	372439	584895	640003	344994	649665	1980391	697149
5-Aminopentanoate	708582	850838	769415	1783934	2681832	2361832	1547617	2641116	2226384	2523690	971650	767367	659297	2720196	698114	848737	2438654	2774266	1484615	1409668	2316892	2355411
Pantothenol	5933	6069	11297	86856	37948	24758	11582	22130	13890	39796	17227	6762	1817	22228	5154	13364	38515	31948	14475	12438	10893	17708
3-Methyldioxyindole	29278	19761	20100	269299	44136	48979	24865	26312	33347	49844	14498	15801	11834	25850	18130	12420	31053	23425	19294	19307	36531	22352
Indole	1204274	1127445	891799	378569	511943	1187016	1039503	1884754	422892	515130	1502183	1534194	1459784	1583649	459257	1357699	1015360	1633908	1255514	1235944	631719	856543
6-Hydroxykynurenic acid	37698840	35494980	10646400	11681790	15904470	38843510	38945120	52951410	13533040	14971380	42680370	40264550	47525340	47166050	6241190	42569410	33669820	48352320	36634380	36407480	17707100	22701660
L-Tryptophanamide	95551		10865	17864		11603	50124	37997		24051	4457	18348				62640	9405	80388	89788	8051		36341
kynurenine	3962		1808	6263	19065	18204	2130	6701	5846	12505	1857	2418	1301	8713	1936	1472	7732	12223	5682	3966	3412	6364
N-formyl kynurenine	17333	9734	6897	5506	7472	16732	2270	7623	5907	3590	12231	6814	3190	14995			5673	7451	9987	4692	6714	10466
Anthranilate	154606	124990	114574	7403172	183991	235763	114263	187511	176871	127939	80632	85713	87900	119885	78603	128551	176148	179080	147508	76874	128906	114533
Picolinic acid	53593	66299	74106	1082494	59837	67813	75559	65799	220787	69191	82078	143427	55095	218488	72914	78020	63855	85952	72947	58392	107401	75561
g-Oxalo-crotonate	30518	29090	44281	8792	22271	44734	40744	36220	12543	56009	60952	9942	5656	49991	7259	31483	54880	42454	30080	37607	25649	48685
Formyl-5-hydroxy-kynurenamine	3962		1808	6263	19065	18204	2130	6701	5846	12505	1857	2418	1301	8713	1936	1472	7732	12223	5682	3966	3412	6364
8-Methoxykynurenate	53420	60500	75401	555763	117021	1301447	67150	42166	732107	283882	188353	94255	53381	54461	97768	57797	1270759	725721	66203	83345	1566689	56454
Serotonin	16596	17766	16724	59513	7766	10779	10486	12095	14432	7387	24779	11989	7373	10910	5484	7456	13126	2837	7530	10722	10805	13168
Acetylcholine	400797	327960	455177	314197	415203	399188	406066	367397	398080	383659	358406	299234	216837	452063	222010	356859	389752	360533	360982	337510	378964	426931
acetyl-carnitine (acyl-C2)	185966	206300	374901	694223	2271266	1436034	808058	2187794	1835257	1685338	780442	259795	54129	2446666	113618	548924	2176304	2151293	631783	1118352	1013759	1357207
propionyl-carnitine (acyl-C3)	10116	12569	19394	25996	43974	20759	26885	54678	45081	41629	16063	2895	2944	61257	2802	16177	33559	27776	14121	33229	26892	37705
acyl-C5-OH (hydroxyisovalerylcarnitine)	63009	44206	48848	81258	30938	29950	47971	56824	53345	41311	50671	42417	48709	206258	40437	84497	46040	59941	59578	24564	53779	94940
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	KEGG ID	m/z	r.t.
L-alanine	C00041	90.0550	1.05
L-arginine	C00062	175.1189	0.63
L-glutamate	C00025	148.0603	0.67
L-glutamine	C00064	147.0764	0.68
L-histidine	C00135	156.0767	0.62
L-leucine	C00123	132.1019	1.02
L-lysine	C00047	147.1128	0.61
L-methionine	C00073	150.0573	0.90
L-phenylalanine	C00079	166.0863	1.41
L-proline	C00148	116.0705	0.70
L-serine	C00065	106.0498	0.65
L-threonine	C00188	120.0655	0.66
L-tyrosine	C00082	182.0811	0.91
L-valine	C00183	118.0861	0.70
AMP	C00020	348.0699	0.82
Adenine	C00147	136.0625	1.06
dCMP	C00239	308.0647	0.71
Thymidine	C00214	243.0990	1.74
Uracil	C00106	113.0355	0.59
Hypoxanthine	C00262	137.0459	0.90
Xanthine	C00385	153.0406	0.90
Allantoate	C00499	177.0614	0.62
(S)(+)-Allantoin	C02350	159.0513	0.68
Urate	C00366	169.0355	0.89
Pyridoxal	C00250	168.0655	0.94
Nicotinamide	C00153	123.0552	0.88
5-Phosphoribosylamine	C03090	230.0440	1.76
UDP-glucose	C00029	567.0608	0.66
NAD+	C00003	664.1158	1.83
5-Oxoproline	C01879	130.0499	0.89
gamma-L-Glutamylputrescine	C15699	218.1535	2.03
S-Adenosyl-L-homocysteine	C00021	385.1337	0.68
Ornithine	C01602	133.0972	0.61
L-Citrulline	C00327	176.1030	0.68
Putrescine	C00134	89.1072	0.57
Spermidine	C00315	146.1651	0.59
Creatine	C00300	132.0767	0.68
Creatinine	C00791	114.0662	0.68
N-Acetylornithine	C00437	175.1077	0.71
N-Succinyl-L-glutamate	C05931	248.0742	1.76
N-Succinyl-L-glutamate 5-semialdehyde	C05932	232.0799	1.76
5-Aminopentanoate	C00431	118.0861	0.71
Pantothenol	C00864	220.1177	1.67
3-Methyldioxyindole	C05834	146.0600	1.74
Indole	C00463	118.0649	1.76
6-Hydroxykynurenic acid	C08480	206.0445	1.76
L-Tryptophanamide	C00977	204.1130	1.79
kynurenine	C00328	209.0920	1.40
N-formyl kynurenine	C02700	237.0857	1.63
Anthranilate	C00108	138.0545	0.67
Picolinic acid	C10164	124.0391	0.59
g-Oxalo-crotonate	C03453	159.0276	0.79
Formyl-5-hydroxy-kynurenamine	C05647	209.0920	1.40
8-Methoxykynurenate	C05830	220.0617	1.77
Serotonin	C00780	177.1021	0.89
Acetylcholine	C01996	146.1176	0.87
acetyl-carnitine (acyl-C2)	C02571	204.1229	0.90
propionyl-carnitine (acyl-C3)	C03017	218.1385	1.08
acyl-C5-OH (hydroxyisovalerylcarnitine)	ac107	262.1644	1.88
METABOLITES_END
#END