#METABOLOMICS WORKBENCH KDSpencer77_20230821_115049 DATATRACK_ID:4237 STUDY_ID:ST002837 ANALYSIS_ID:AN004640 PROJECT_ID:PR001776
VERSION             	1
CREATED_ON             	August 22, 2023, 12:07 pm
#PROJECT
PR:PROJECT_TITLE                 	Bacterial tryptophan metabolites increased by prebiotic galactooligosaccharide
PR:PROJECT_TITLE                 	reduce microglial reactivity and are associated with lower anxiety-like behavior
PR:PROJECT_SUMMARY               	Prebiotic galactooligosaccharides (GOS) reduce anxiety-like behaviors in mice
PR:PROJECT_SUMMARY               	and humans. However, the biological pathways behind these behavioral changes are
PR:PROJECT_SUMMARY               	not well understood. To begin to study these pathways, C57BL/6 mice were fed a
PR:PROJECT_SUMMARY               	standard diet with or without GOS supplementation for 3 weeks prior to testing
PR:PROJECT_SUMMARY               	on the open field. After behavioral testing, colonic contents and serum were
PR:PROJECT_SUMMARY               	collected for bacteriome (16S rRNA gene sequencing, colonic contents only) and
PR:PROJECT_SUMMARY               	metabolome (UPLC-MS, colonic contents and serum data) analyses. As expected, GOS
PR:PROJECT_SUMMARY               	significantly reduced anxiety-like behavior (i.e., increased time in the center
PR:PROJECT_SUMMARY               	(p < 0.05)). Time in the center was significantly correlated with serum
PR:PROJECT_SUMMARY               	methyl-indole-3-acetate (mI3A). This metabolite, which is a methylated form of
PR:PROJECT_SUMMARY               	indole-3-acetate, is derived from bacterial metabolism of tryptophan. Sequencing
PR:PROJECT_SUMMARY               	analyses showed that GOS significantly increased Akkermansia, which is known to
PR:PROJECT_SUMMARY               	metabolize both GOS and tryptophan. To test the hypothesis that mI3A can reduce
PR:PROJECT_SUMMARY               	anxiety-like behavior and affect microglial activity, we first tested mI3A
PR:PROJECT_SUMMARY               	effects on LPS-stimulated BV2 microglia. Cells treated with mI3A produced
PR:PROJECT_SUMMARY               	significantly less CCL2 and TNF-α than vehicle-treated cells (p<.05). We then
PR:PROJECT_SUMMARY               	treated mice with an intraperitoneal injection of mI3A or vehicle control, and
PR:PROJECT_SUMMARY               	found that mice given mI3A had lower CCL2 in the prefrontal cortex and
PR:PROJECT_SUMMARY               	hippocampus, as well as a reduction in a composite behavioral score in the open
PR:PROJECT_SUMMARY               	field. Together, these data support a novel pathway through which GOS reduces
PR:PROJECT_SUMMARY               	anxiety-like behaviors in mice, and suggests that the bacterial metabolite mI3A,
PR:PROJECT_SUMMARY               	which is elevated by GOS, reduces microglial CCL2, which in turn reduces
PR:PROJECT_SUMMARY               	anxiety-like behavior.
PR:INSTITUTE                     	National Center for Advancing Translational Sciences
PR:LAST_NAME                     	Spencer
PR:FIRST_NAME                    	Kyle
PR:ADDRESS                       	9800 Medical Center Dr, Rockville, MD 20850
PR:EMAIL                         	kyle.spencer@nih.gov
PR:PHONE                         	9897084858
#STUDY
ST:STUDY_TITLE                   	Bacterial tryptophan metabolites increased by prebiotic galactooligosaccharide
ST:STUDY_TITLE                   	reduce microglial reactivity and are associated with lower anxiety-like behavior
ST:STUDY_TITLE                   	(Blood)
ST:STUDY_SUMMARY                 	Prebiotic galactooligosaccharides (GOS) reduce anxiety-like behaviors in mice
ST:STUDY_SUMMARY                 	and humans. However, the biological pathways behind these behavioral changes are
ST:STUDY_SUMMARY                 	not well understood. To begin to study these pathways, C57BL/6 mice were fed a
ST:STUDY_SUMMARY                 	standard diet with or without GOS supplementation for 3 weeks prior to testing
ST:STUDY_SUMMARY                 	on the open field. After behavioral testing, colonic contents and serum were
ST:STUDY_SUMMARY                 	collected for bacteriome (16S rRNA gene sequencing, colonic contents only) and
ST:STUDY_SUMMARY                 	metabolome (UPLC-MS, colonic contents and serum data) analyses. As expected, GOS
ST:STUDY_SUMMARY                 	significantly reduced anxiety-like behavior (i.e., increased time in the center
ST:STUDY_SUMMARY                 	(p < 0.05)). Time in the center was significantly correlated with serum
ST:STUDY_SUMMARY                 	methyl-indole-3-acetate (mI3A). This metabolite, which is a methylated form of
ST:STUDY_SUMMARY                 	indole-3-acetate, is derived from bacterial metabolism of tryptophan. Sequencing
ST:STUDY_SUMMARY                 	analyses showed that GOS significantly increased Akkermansia, which is known to
ST:STUDY_SUMMARY                 	metabolize both GOS and tryptophan. To test the hypothesis that mI3A can reduce
ST:STUDY_SUMMARY                 	anxiety-like behavior and affect microglial activity, we first tested mI3A
ST:STUDY_SUMMARY                 	effects on LPS-stimulated BV2 microglia. Cells treated with mI3A produced
ST:STUDY_SUMMARY                 	significantly less CCL2 and TNF-α than vehicle-treated cells (p<.05). We then
ST:STUDY_SUMMARY                 	treated mice with an intraperitoneal injection of mI3A or vehicle control, and
ST:STUDY_SUMMARY                 	found that mice given mI3A had lower CCL2 in the prefrontal cortex and
ST:STUDY_SUMMARY                 	hippocampus, as well as a reduction in a composite behavioral score in the open
ST:STUDY_SUMMARY                 	field. Together, these data support a novel pathway through which GOS reduces
ST:STUDY_SUMMARY                 	anxiety-like behaviors in mice, and suggests that the bacterial metabolite mI3A,
ST:STUDY_SUMMARY                 	which is elevated by GOS, reduces microglial CCL2, which in turn reduces
ST:STUDY_SUMMARY                 	anxiety-like behavior.
ST:INSTITUTE                     	National Center for Advancing Translational Sciences
ST:LAST_NAME                     	Spencer
ST:FIRST_NAME                    	Kyle
ST:ADDRESS                       	9800 Medical Center Dr, Rockville, MD 20850
ST:EMAIL                         	kyle.spencer@nih.gov
ST:PHONE                         	9897084858
#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           	-	NACH-00271	GROUP_ID:GOS	
SUBJECT_SAMPLE_FACTORS           	-	NACH-00272	GROUP_ID:GOS	
SUBJECT_SAMPLE_FACTORS           	-	NACH-00273	GROUP_ID:GOS	
SUBJECT_SAMPLE_FACTORS           	-	NACH-00274	GROUP_ID:GOS	
SUBJECT_SAMPLE_FACTORS           	-	NACH-00275	GROUP_ID:GOS	
SUBJECT_SAMPLE_FACTORS           	-	NACH-00276	GROUP_ID:CON	
SUBJECT_SAMPLE_FACTORS           	-	NACH-00277	GROUP_ID:CON	
SUBJECT_SAMPLE_FACTORS           	-	NACH-00278	GROUP_ID:CON	
SUBJECT_SAMPLE_FACTORS           	-	NACH-00279	GROUP_ID:CON	
SUBJECT_SAMPLE_FACTORS           	-	NACH-00280	GROUP_ID:CON	
SUBJECT_SAMPLE_FACTORS           	-	NACH-00281	GROUP_ID:CON	
#COLLECTION
CO:COLLECTION_SUMMARY            	After euthanasia the serum of the mice were collected and sent to Metabolon Inc
CO:COLLECTION_SUMMARY            	for UPLC-MS Analysis.
CO:SAMPLE_TYPE                   	Blood (serum)
#TREATMENT
TR:TREATMENT_SUMMARY             	The mice were fed either a control diet or a diet supplemented with 10%
TR:TREATMENT_SUMMARY             	galactooligosaccharide for 3 weeks. After the 3 weeks had elapsed the mice were
TR:TREATMENT_SUMMARY             	euthanized and the colonic contents were collected.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	The colonic contents were sent to Metabolon who then performed the sample
SP:SAMPLEPREP_SUMMARY            	preparation prior to UPLC-MS analysis.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	HILIC (LC/MS Polar Neg)
CH:CHROMATOGRAPHY_TYPE           	HILIC
CH:INSTRUMENT_NAME               	Waters Acquity
CH:COLUMN_NAME                   	Waters Acquity BEH Amide (150 x 2.1mm, 1.7um)
CH:SOLVENT_A                     	-
CH:SOLVENT_B                     	-
CH:FLOW_GRADIENT                 	-
CH:FLOW_RATE                     	-
CH:COLUMN_TEMPERATURE            	-
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Thermo Q Exactive Orbitrap
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	Other
MS:ION_MODE                      	NEGATIVE
MS:MS_COMMENTS                   	Metabolon (LC/MS Polar)
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	Relative Abundance
MS_METABOLITE_DATA_START
Samples	NACH-00271	NACH-00272	NACH-00273	NACH-00274	NACH-00275	NACH-00276	NACH-00277	NACH-00278	NACH-00279	NACH-00280	NACH-00281
Factors	GROUP_ID:GOS	GROUP_ID:GOS	GROUP_ID:GOS	GROUP_ID:GOS	GROUP_ID:GOS	GROUP_ID:CON	GROUP_ID:CON	GROUP_ID:CON	GROUP_ID:CON	GROUP_ID:CON	GROUP_ID:CON
2,3-dihydroxyisovalerate	0.3524	1	0.8854	0.9658	1.2558	0.9297	1.064	0.9403	1.0347	1.1605	1.3819
2,6-dihydroxybenzoic acid	0.578	0.8196	0.8195	0.8451	1.009	1.137	0.9763	1	1.0077	1.2007	1.0398
2-aminoadipate	1	1.1816	1.3073	0.9325	0.6694	1.2555	0.9889	0.7893	1.0177	0.9755	1.0258
2-hydroxy-4-(methylthio)butanoic acid	1.2152	0.959	0.7173	0.8123	1.8163	1.189	1.1444	1.5166	0.9706	1	0.9619
2-hydroxyadipate	1.9462	1.2701	0.8417	0.7715	1.1181	1	1.0602	1.0157	0.6656	0.8495	0.9195
2-hydroxybutyrate/2-hydroxyisobutyrate	0.8221	1.1547	1	0.9365	1.0983	1.3549	1.1869	0.8181	1.0022	0.8744	0.8004
2-hydroxyphenylacetate	0.7211	0.7922	1.4082	1.1942	1.0168	1.0804	0.953	0.8084	0.9035	1	1.5297
2-isopropylmalate	0.6264	1.4726	1.3279	0.4417	1	1.1752	2.1994	0.9138	0.9041	1.1716	0.654
2-keto-3-deoxy-gluconate	0.9997	1.5409	1.444	1.1354	0.8705	0.9661	1.0003	0.7888	0.7888	0.9441	1.0146
2-oxoadipate	1.511	1.4418	1.3026	0.8688	1.4581	1.2763	0.882	1	0.9351	0.8547	0.9951
2R,3R-dihydroxybutyrate	0.7218	0.876	0.9595	0.9765	1.021	1.1437	1.0758	1	0.9699	1.0303	1.0156
2S,3R-dihydroxybutyrate	0.7503	0.9594	0.9136	1.094	1.528	1.1513	1.2804	0.9089	0.9714	1	1.0564
3,4-dihydroxybutyrate	0.7414	1.1689	0.914	1.0039	0.8666	1.0916	1.2266	1.3706	0.9035	1	0.9654
3-hydroxy-3-methylglutarate	1	1.5294	0.9526	0.9812	1.4517	1.3329	1.2964	0.8076	1.0233	0.9682	0.8504
3-hydroxyadipate*	0.1945	1.0034	0.4601	0.7576	1.0809	1.0293	1.8278	1	0.8611	1.0974	0.4374
3-hydroxybutyrate (BHBA)	1.4852	0.9433	1.1661	0.8514	1.06	1.3643	1.0195	0.9406	1	0.9648	0.6396
3-hydroxyisobutyrate	1.3798	1.4095	1.0314	0.6037	1.6522	1.3826	0.644	1	0.6125	0.7041	0.9607
3-methylglutaconate	0.9168	1.2282	1.0261	1	1.2116	1.0201	0.8732	0.8322	0.7646	0.7302	1.2419
acetylphosphate	0.5711	0.7521	1.5194	1.4486	0.6085	0.8669	0.5634	1	1.8588	1.3075	4.7926
adipate (C6-DC)	1.01	1.1546	0.5231	0.9643	1.6038	1.2206	1.4937	1	0.8818	0.9224	0.8119
allantoin	0.8788	0.9335	0.9544	1	0.9806	1.1241	1.2443	1.0161	0.9462	1.0451	1.3187
alpha-ketoglutaramate*	1.4855	1.0383	0.9929	0.9549	1.2106	0.989	1.0429	0.9135	0.974	1	1.4097
alpha-ketoglutarate	1.4652	1.4573	0.8308	1.2387	1.6083	1.3517	0.9934	0.6618	1	0.9455	0.2252
arabinose	0.1317	1.1231	1.084	0.1317	0.916	0.8981	1.2577	1.1292	0.1317	0.1317	0.883
arabitol/xylitol	0.9455	1.2658	0.9015	0.8179	1.174	1	1.5304	0.9519	0.8621	1.0593	1.0744
arabonate/xylonate	0.6573	1.4379	1	0.9666	0.9755	1.1719	1.4114	0.7319	0.9779	1.1156	1.0374
ascorbic acid 2-sulfate	0.4635	1.0625	1	0.9239	0.8721	1.1532	1.4478	0.9998	0.9497	1.0733	1.1711
ascorbic acid 3-sulfate*	0.5139	1.4068	1	0.9483	0.8932	1.5549	1.8823	0.9861	1.2124	1.3315	0.9005
caffeic acid sulfate	1.0729	3.0253	14.3167	1	1.915	0.6861	1.1429	0.8618	0.3703	0.9716	0.2321
creatine phosphate	1.0359	0.803	0.4444	1.0799	0.625	1.0773	1.0165	0.5137	1.071	0.9835	0.4444
cysteine s-sulfate	7.1239	1.0583	0.7807	1.2348	1.1608	1.0101	0.9568	0.9884	0.8713	0.8589	1
dihydrocaffeate sulfate (2)	1.2811	1.2306	0.3148	1.179	1.7307	0.3799	0.6574	0.4463	1	1.7406	0.612
dihydroorotate	2.3704	1.5889	0.4288	1.0894	1.6666	1.1384	0.9698	0.5168	1	0.9629	0.1965
erythritol	0.7142	0.726	1	0.7942	0.5654	1.0271	1.0065	1.0939	0.935	1.2148	1.6459
erythronate*	0.6131	1.0239	0.9441	0.9449	1.022	1.0201	1.0749	0.7984	0.8188	1.0188	1
ethylmalonate	0.5148	1.7837	1.0118	1.0239	1	1.202	1.0229	0.6231	0.8852	0.8858	0.7912
fructose	1.6023	1.4383	1	0.7541	1.9562	1.0551	1.0071	0.6765	0.7602	0.7578	0.972
fumarate	1.556	1.716	1.3837	0.7134	1.26	0.9987	1	0.8568	0.8877	0.893	1.237
galactitol (dulcitol)	3.1912	1	1.6817	1.277	0.7923	0.6822	0.9734	1.0772	0.3004	0.5433	2.2888
galactonate	1.2313	1.1532	1.4405	0.8308	0.9484	0.8879	1.2636	1	0.572	0.8884	1.3118
gluconate	2.1287	1.2965	1.0613	0.7339	0.9863	0.9228	1	0.8797	0.7562	1.0032	1.2403
glucose	0.4963	1.2129	0.7931	1.0547	0.9231	1.2181	1.4113	0.8903	1	1.0955	0.6246
glucuronate	2.0401	0.8493	0.9162	1	1.0118	0.8725	1.0325	1.0272	0.9324	0.9633	1.2201
glutarate (C5-DC)	1.2597	1.6802	0.8452	0.7746	1.4234	1.7839	1	0.595	1.5097	0.8744	0.6256
glycerate	0.7954	0.9515	0.9474	1	1.0626	1.0847	1.0721	0.9276	1.0271	0.7942	1.2286
glycerophosphoethanolamine	0.7401	1.0065	1.105	0.8857	0.9755	1	1.0186	0.8088	1.0984	0.9925	2.025
glycerophosphoglycerol	1.2305	1.0395	1.8444	0.5754	0.9465	0.9296	1	0.7973	1.0239	0.8412	1.3181
glycerophosphoinositol*	0.9144	1.0873	0.8366	1.3638	0.9824	0.8417	1.1137	0.8293	1.1876	1	1.7667
glycolate (hydroxyacetate)	2.0611	1.2041	1.5324	0.3771	1.0617	1.0376	0.9103	0.8276	0.3959	0.9624	0.3771
gulonate*	0.6986	1.4936	1.0727	0.8043	1.8243	1	1.378	0.9043	0.8429	1.2411	0.5976
hydantoin-5-propionate	0.4859	1.9597	0.5834	0.9072	2.4082	1	0.8166	1.001	1.2335	0.8306	2.1538
hypotaurine	2.5542	1.0168	1.0181	0.9199	0.8052	0.6914	1.0845	0.9721	0.7309	1	1.3283
isocitric lactone	1.0205	1.5672	1.1328	0.9535	1	1.0623	0.9946	0.8376	0.7459	0.8527	1.5146
lactate	0.8514	1.2576	1	1.0441	1.0328	0.9789	1.0069	0.7955	1.0261	0.9823	0.8984
malate	1.3517	1.5953	1.4631	0.7944	0.9463	1.1005	0.8914	0.7502	1	0.9058	1.4472
maleate	1.4897	1.9382	1.0045	0.9689	1.225	1.1788	0.8546	0.6459	0.8497	1	0.9567
malonate	1	1.0658	0.8955	1.2293	1.018	0.9477	0.8821	0.8306	1.0681	0.7839	1.0449
mannitol/sorbitol	0.8706	1.2859	0.9852	1.1348	1	1.0642	1.3409	0.8731	0.921	1.2153	0.8422
mannonate*	0.7852	1.1244	1.0457	0.8941	0.9367	0.9318	1.4023	1	0.8458	1.2075	1.1434
mannose	0.6586	1	0.6402	1.1919	0.7917	1.323	1.7268	1.348	0.9617	1.1871	0.5685
methylmalonate (MMA)	0.8892	1.4549	0.9913	0.9173	1.139	1.0357	1.2646	0.8347	1	0.9786	1.0069
methylsuccinate	0.94	1.2648	0.9259	0.7999	1	1.1399	1.4543	0.8386	1.0652	1.1174	0.6948
mevalonate	0.9253	0.827	0.8573	0.8813	1.2093	1.1698	1.0163	1.0668	1	0.849	1.2481
myo-inositol	0.7705	1.0278	1.0064	1.2287	0.9791	0.8931	0.9822	0.8099	1.0659	1	1.8224
N-acetylalanine	1.0372	0.9326	0.9614	1	0.9518	1.1374	1.1654	1.0551	0.9265	0.9257	1.0676
N-acetylaspartate (NAA)	0.8453	0.9977	1.5422	1.227	0.7333	1.1977	0.9601	1	1.3008	0.9548	2.5979
N-acetylglucosamine 6-phosphate	2.1018	1	2.3027	0.8573	0.8537	0.7222	0.8978	1.0137	1.2786	0.973	2.6943
N-acetylglycine	0.5333	1.0818	0.9441	0.9927	1.1042	1.0202	1.045	0.9381	0.8473	1	1.1794
N-acetylneuraminate	0.8721	0.9522	1.133	0.9642	0.9016	1.1102	1	0.9864	1.1206	1.1869	1.8118
N-acetylserine	0.8514	1	0.8966	1.0099	1.1184	0.9517	1.099	1.0251	0.982	0.975	1.2404
N-formylanthranilic acid	2.0542	0.8359	1.5464	1.1509	1.1864	0.8896	0.5404	0.8028	0.7974	1	2.7305
N2,N5-diacetylornithine	1.0136	1.0247	0.8684	1.1829	1.1445	0.9006	0.9864	0.7902	0.5466	0.5466	1.1168
N2-acetyllysine	0.499	1.4246	1.2056	0.875	0.9435	1.5033	1.3141	0.9271	1	1.392	0.839
N6-acetyllysine	0.7603	0.9763	1.0394	0.7839	1.0113	1.2079	0.9974	1.0794	0.9141	1.0352	1
orotate	1.6725	0.9977	1.3682	1.0994	0.7404	1.1103	0.7895	0.8122	1	0.85	1.5385
orotidine	0.9007	1.0826	1	1.0654	0.726	1.057	0.9877	0.7126	1.0111	0.9855	1.4486
pentose acid*	0.6848	1.2211	1.0086	0.6848	0.9914	0.6848	1.13	0.6848	0.6848	0.6848	0.7009
perfluorooctanesulfonate (PFOS)	1.4219	1.1989	1.0154	0.834	0.8664	1.1853	1	0.7663	1.0352	0.9649	0.774
pimelate (C7-DC)	0.3783	0.3761	0.5395	0.3761	1.2026	1.1445	1.5082	0.912	1.088	1.1266	0.4106
pyruvate	1.3509	1.306	0.7708	1	0.9822	1.072	1.0538	0.7881	0.9734	1.0117	0.595
ribitol	1.0257	1.4196	0.917	1	1.1079	0.974	1.2754	0.8954	0.895	0.9592	1.093
ribonate	0.6724	1.279	0.9821	0.7795	1	1.5607	1.684	1.0459	0.9937	1.1375	0.8842
ribose	2.8238	0.2468	0.4636	2.5476	0.6316	0.3368	0.2468	1.4899	1	1.1057	0.2468
ribulonate/xylulonate/lyxonate*	0.565	1.1807	0.8087	1	0.8978	1.1581	1.294	0.8673	1.0353	1.0847	0.7148
ribulose/xylulose	0.708	0.7998	1	1.1558	0.708	0.708	0.708	1.1127	0.991	1.1492	0.708
S-methylcysteine	1.1221	0.6976	0.686	0.7748	1.1553	1.1669	0.836	1.2822	0.686	1.1153	0.8847
salicylate	1.5522	0.934	1.1883	1.1174	1.0643	0.9357	0.7439	0.6683	0.411	0.411	1.7571
sedoheptulose	0.9952	1.0773	0.8526	0.5925	1.8136	0.6096	1.0048	0.735	0.5925	1.0167	1.1822
sphingosine 1-phosphate	1.4439	0.8902	0.8282	1.0862	1	0.9438	1.0512	0.9337	1.118	1.0868	0.7913
succinate	0.8431	1.1858	0.8619	0.8425	1.0161	1.1377	1.1361	0.7878	1.077	1	0.8001
sucrose	0.8221	2.4538	1.62	0.9201	3.7637	0.9832	1.5164	0.9398	0.8933	1	2.5381
taurine	0.9042	1	1.0503	1.0044	0.8177	0.9444	1.0232	0.8529	1.0007	0.9068	1.4653
threonate	0.5704	1.1033	1.0811	0.7084	1	1.114	1.1924	0.769	0.8307	1.0143	0.7907
tricarballylate	0.0733	3.0085	2.6125	1.1375	3.5927	0.6248	1	0.4856	0.733	1.0063	0.5161
xylose	0.6976	0.9141	0.9142	0.6976	1.0113	1.2889	0.9887	1.064	0.6976	0.6976	1.1408
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	SUPER_PATHWAY	SUB_PATHWAY	COMP_ID	PLATFORM	CHEMICAL_ID	RI	MASS	CAS	PUBCHEM	CHEMSPIDER	KEGG	HMDB_ID
2,3-dihydroxyisovalerate	Xenobiotics	Food Component/Plant	38276	LC/MS Polar	100002417	1050	133.05063	1756-18-9	677	657	C04039	HMDB12141
2,6-dihydroxybenzoic acid	Xenobiotics	Drug - Topical Agents	63042	LC/MS Polar	100020837	729	153.01933	303-07-1	9338	8974		HMDB13676
2-aminoadipate	Amino Acid	Lysine Metabolism	6146	LC/MS Polar	381	3500	160.06153	542-32-5;1118-90-7	469	456	C00956	HMDB00510
2-hydroxy-4-(methylthio)butanoic acid	Amino Acid	Methionine, Cysteine, SAM and Taurine Metabolism	63739	LC/MS Polar	100021372	1065	149.02778	583-91-5
2-hydroxyadipate	Lipid	Fatty Acid, Dicarboxylate	31934	LC/MS Polar	100001153	3000	161.04554	18294-85-4	193530	167943	C02360	HMDB00321
2-hydroxybutyrate/2-hydroxyisobutyrate	Amino Acid	Glutathione Metabolism	52281	LC/MS Polar	100008928	1258	103.04006
2-hydroxyphenylacetate	Amino Acid	Phenylalanine Metabolism	1432	LC/MS Polar	235	858.6	151.04006	614-75-5	11970	11476	C05852	HMDB00669
2-isopropylmalate	Xenobiotics	Food Component/Plant	15667	LC/MS Polar	100000409	2605	175.06119	3237-44-3	77	76	C02504	HMDB00402
2-keto-3-deoxy-gluconate	Xenobiotics	Food Component/Plant	48141	LC/MS Polar	100005946	1900	177.04046	17510-99-5	161227	141630	C00204	HMDB01353
2-oxoadipate	Amino Acid	Lysine Metabolism	38347	LC/MS Polar	92	2635	159.0299	3184-35-8	71	70	C00322	HMDB00225
2R,3R-dihydroxybutyrate	Lipid	Fatty Acid, Dihydroxy	63109	LC/MS Polar	100002367	1543	119.03498	759-06-8	13120901	17215960		HMDB00498
2S,3R-dihydroxybutyrate	Lipid	Fatty Acid, Dihydroxy	62950	LC/MS Polar	100002284	1660	119.03498	5057-93-2	10964471	9139682		HMDB02453
3,4-dihydroxybutyrate	Lipid	Fatty Acid, Dihydroxy	62952	LC/MS Polar	100001730	2204	119.03498	51267-44-8
3-hydroxy-3-methylglutarate	Lipid	Mevalonate Metabolism	531	LC/MS Polar	112	2850	161.04555	503-49-1	1662	4573695	C03761	HMDB00355
3-hydroxyadipate*	Lipid	Fatty Acid, Dicarboxylate	62069	LC/MS Polar	100004396	3131	161.04554		151913	133894		HMDB00345
3-hydroxybutyrate (BHBA)	Lipid	Ketone Bodies	542	LC/MS Polar	254	1480	103.04007	625-72-9	441	428	C01089	HMDB00357
3-hydroxyisobutyrate	Amino Acid	Leucine, Isoleucine and Valine Metabolism	1549	LC/MS Polar	111	1619	103.04007	2068-83-9	87	10140307	C06001	HMDB00336
3-methylglutaconate	Amino Acid	Leucine, Isoleucine and Valine Metabolism	57747	LC/MS Polar	100002458	2700	99.04518	5746-90-7	1551553	1267861		HMDB00522
acetylphosphate	Energy	Oxidative Phosphorylation	15488	LC/MS Polar	1211	4013.9	138.98018	94249-01-1	186	181	C00227	HMDB01494
adipate (C6-DC)	Lipid	Fatty Acid, Dicarboxylate	21134	LC/MS Polar	100000863	3000	145.05063	124-04-9	196	191	C06104	HMDB00448
allantoin	Nucleotide	Purine Metabolism, (Hypo)Xanthine/Inosine containing	1107	LC/MS Polar	1002	1672	157.03671	97-59-6	204	199	C02350	HMDB00462
alpha-ketoglutaramate*	Amino Acid	Glutamate Metabolism	62101	LC/MS Polar	100002397	1916	144.03023	18465-19-5	48
alpha-ketoglutarate	Energy	TCA Cycle	528	LC/MS Polar	93	2700	145.01425	305-72-6;328-50-7;22202-68-2	51	50	C00026	HMDB00208
arabinose	Carbohydrate	Pentose Metabolism	575	LC/MS Polar	828	1804.5	195.05103	28697-53-2	66308	59687	C00216	HMDB00646
arabitol/xylitol	Carbohydrate	Pentose Metabolism	48885	LC/MS Polar	100006430	1932.4	151.0612		6912		C01904
arabonate/xylonate	Carbohydrate	Pentose Metabolism	48255	LC/MS Polar	100006115	2700	165.04046
ascorbic acid 2-sulfate	Cofactors and Vitamins	Ascorbate and Aldarate Metabolism	63590	LC/MS Polar	100009329	3042	254.98162		86484	77998
ascorbic acid 3-sulfate*	Cofactors and Vitamins	Ascorbate and Aldarate Metabolism	63687	LC/MS Polar	100020409	2025	254.98146		11425365	9600241
caffeic acid sulfate	Xenobiotics	Food Component/Plant	53254	LC/MS Polar	100009337	1307	258.99179			30777608		HMDB41708
creatine phosphate	Amino Acid	Creatine Metabolism	33951	LC/MS Polar	100000112	3831	210.02853	922-32-7	9548602	567	C02305	HMDB01511
cysteine s-sulfate	Amino Acid	Methionine, Cysteine, SAM and Taurine Metabolism	22176	LC/MS Polar	100000808	2273	199.96928	1637-71-4	115015	102939	C05824	HMDB00731
dihydrocaffeate sulfate (2)	Xenobiotics	Food Component/Plant	62524	LC/MS Polar	100020822	1328	261.00744		49844181
dihydroorotate	Nucleotide	Pyrimidine Metabolism, Orotate containing	601	LC/MS Polar	923	1910	157.02548	155-54-4	648	628	C00337	HMDB03349
erythritol	Xenobiotics	Food Component/Plant	20699	LC/MS Polar	100000846	1491	167.05611	149-32-6	222285	13835932	C00503	HMDB02994
erythronate*	Carbohydrate	Aminosugar Metabolism	42420	LC/MS Polar	100001320	2186	135.02989	88759-55-1	2781043	2061231		HMDB00613
ethylmalonate	Amino Acid	Leucine, Isoleucine and Valine Metabolism	15765	LC/MS Polar	2054	2785	131.03498	601-75-2	11756	11263		HMDB00622
fructose	Carbohydrate	Fructose, Mannose and Galactose Metabolism	577	LC/MS Polar	878	2065	179.05611	57-48-7	5984	5764	C00095	HMDB00660
fumarate	Energy	TCA Cycle	1643	LC/MS Polar	330	3084	115.00368	100-17-8	444972	10197150	C00122	HMDB00134
galactitol (dulcitol)	Carbohydrate	Fructose, Mannose and Galactose Metabolism	1117	LC/MS Polar	1003	2360	181.07176	608-66-2	11850	11357	C01697	HMDB00107
galactonate	Carbohydrate	Fructose, Mannose and Galactose Metabolism	27719	LC/MS Polar	100001026	3085	195.05102	299-28-5	128869	114198	C00880	HMDB00565
gluconate	Xenobiotics	Food Component/Plant	587	LC/MS Polar	338	2922	195.05102	527-07-1	10690	10240	C00257	HMDB00625
glucose	Carbohydrate	Glycolysis, Gluconeogenesis, and Pyruvate Metabolism	20488	LC/MS Polar	572	2342	179.05611	50-99-7	5793	71358	C00031	HMDB00122
glucuronate	Carbohydrate	Aminosugar Metabolism	15443	LC/MS Polar	100000257	3233.3	193.03537	207300-70-7	444791	58552	C00191	HMDB00127
glutarate (C5-DC)	Lipid	Fatty Acid, Dicarboxylate	396	LC/MS Polar	339	2900	131.03498	110-94-1	743	3618533	C00489	HMDB00661
glycerate	Carbohydrate	Glycolysis, Gluconeogenesis, and Pyruvate Metabolism	1572	LC/MS Polar	1052	2070.4	105.01933	600-19-1	752	732	C00258	HMDB00139
glycerophosphoethanolamine	Lipid	Phospholipid Metabolism	37455	LC/MS Polar	100001620	3075	214.04859	33049-08-0	123874	110410	C01233	HMDB00114
glycerophosphoglycerol	Lipid	Glycerolipid Metabolism	48857	LC/MS Polar	100001619	2430	245.04317		439964	388990	C03274
glycerophosphoinositol*	Lipid	Phospholipid Metabolism	47155	LC/MS Polar	100001621	3850	333.05922	16824-65-0	167572
glycolate (hydroxyacetate)	Xenobiotics	Chemical	15737	LC/MS Polar	203	1790.4	159.01073	79-14-1	757	737	C00160	HMDB00115
gulonate*	Cofactors and Vitamins	Ascorbate and Aldarate Metabolism	46957	LC/MS Polar	100001586	2750	195.05102	20246-53-1	152304	7969943	C00257	HMDB03290
hydantoin-5-propionate	Amino Acid	Histidine Metabolism	40473	LC/MS Polar	100002514	2080	171.04113	5624-26-0	782	761	C05565	HMDB01212
hypotaurine	Amino Acid	Methionine, Cysteine, SAM and Taurine Metabolism	590	LC/MS Polar	358	2763.4	108.01247	300-84-5	107812	96959	C00519	HMDB00965
isocitric lactone	Energy	TCA Cycle	54724	LC/MS Polar	100009239	2970	173.00916	4702-32-3	98259	88729
lactate	Carbohydrate	Glycolysis, Gluconeogenesis, and Pyruvate Metabolism	527	LC/MS Polar	482	1468.8	89.02442	79-33-4	612	592	C00186	HMDB00190
malate	Energy	TCA Cycle	1303	LC/MS Polar	409	3343.1	133.01425	6915-15-7	525	510	C00149	HMDB00156
maleate	Lipid	Fatty Acid, Dicarboxylate	20676	LC/MS Polar	100000707	2640	115.00368	110-16-7	444266	392248	C01384	HMDB00176
malonate	Lipid	Fatty Acid Synthesis	15872	LC/MS Polar	818	3447	103.00368	141-82-2;26522-22-85-0	867	844	C00383	HMDB00691
mannitol/sorbitol	Carbohydrate	Fructose, Mannose and Galactose Metabolism	46142	LC/MS Polar	100001740	2320	181.07176		5780	440	C00794	HMDB00247
mannonate*	Xenobiotics	Food Component/Plant	62864	LC/MS Polar	100003695	2736	195.05102	642-99-9	3246006	2496757
mannose	Carbohydrate	Fructose, Mannose and Galactose Metabolism	584	LC/MS Polar	803	2200	179.05611	3458-28-4	18950	141983	C00159	HMDB00169
methylmalonate (MMA)	Lipid	Fatty Acid Metabolism (also BCAA Metabolism)	1496	LC/MS Polar	418	3078.4	117.01933	516-05-2	487	473	C02170	HMDB00202
methylsuccinate	Amino Acid	Leucine, Isoleucine and Valine Metabolism	15745	LC/MS Polar	2051	2800	131.03498	498-21-5	10349	9922		HMDB01844
mevalonate	Lipid	Mevalonate Metabolism	39583	LC/MS Polar	30	1462	147.06628	96949-03-0	439230	388367	C02104	HMDB00227
myo-inositol	Lipid	Inositol Metabolism	1124	LC/MS Polar	363	3506.3	225.06159	87-89-8	892	10239179	C00137	HMDB00211
N-acetylalanine	Amino Acid	Alanine and Aspartate Metabolism	1585	LC/MS Polar	1110	1564.8	130.05096	97-69-8	88064	79449	C02847	HMDB00766
N-acetylaspartate (NAA)	Amino Acid	Alanine and Aspartate Metabolism	22185	LC/MS Polar	100000787	3143	174.04079	997-55-7;997-55-7	65065	58576	C01042	HMDB00812
N-acetylglucosamine 6-phosphate	Carbohydrate	Aminosugar Metabolism	15107	LC/MS Polar	1213	4114	300.04899	102029-88-9	439219	8216065	C00357	HMDB02817
N-acetylglycine	Amino Acid	Glycine, Serine and Threonine Metabolism	27710	LC/MS Polar	100001006	1780	116.03531	543-24-8	10972	10507		HMDB00532
N-acetylneuraminate	Carbohydrate	Aminosugar Metabolism	32377	LC/MS Polar	1162	3072.6	308.0987	131-48-6	439197	10292217	C00270	HMDB00230
N-acetylserine	Amino Acid	Glycine, Serine and Threonine Metabolism	37076	LC/MS Polar	100001851	2000	146.04588	97-14-3	65249	58744		HMDB02931
N-formylanthranilic acid	Amino Acid	Tryptophan Metabolism	43549	LC/MS Polar	100004561	900	164.03531	3342-77-6	101399	91624	C05653	HMDB04089
N2,N5-diacetylornithine	Amino Acid	Urea cycle; Arginine and Proline Metabolism	43591	LC/MS Polar	100004575	1650	215.10373	39825-23-5	65977	8573834
N2-acetyllysine	Amino Acid	Lysine Metabolism	36751	LC/MS Polar	100001721	3372.6	187.10881	1946-82-3	92907	83869	C12989	HMDB00446
N6-acetyllysine	Amino Acid	Lysine Metabolism	36752	LC/MS Polar	100001734	2612.6	187.10881	692-04-6	92832	83801	C02727	HMDB00206
orotate	Nucleotide	Pyrimidine Metabolism, Orotate containing	1505	LC/MS Polar	445	1638.1	155.00983	50887-69-9	967	942	C00295	HMDB00226
orotidine	Nucleotide	Pyrimidine Metabolism, Orotate containing	35172	LC/MS Polar	100001416	2250	287.05209	314-50-1	92751	83729		HMDB00788
pentose acid*	Partially Characterized Molecules	Partially Characterized Molecules	63264	LC/MS Polar	100021467	2180	165.04046
perfluorooctanesulfonate (PFOS)	Xenobiotics	Chemical	57564	LC/MS Polar	100010955	662	498.93021	1763-23-1	74483	67068	C18142	HMDB59586
pimelate (C7-DC)	Lipid	Fatty Acid, Dicarboxylate	15704	LC/MS Polar	100000101	2745	159.06628	111-16-0	385	376	C02656	HMDB00857
pyruvate	Carbohydrate	Glycolysis, Gluconeogenesis, and Pyruvate Metabolism	48990	LC/MS Polar	823	1083	87.00877	127-17-3	1060	96901	C00022	HMDB00243
ribitol	Carbohydrate	Pentose Metabolism	15772	LC/MS Polar	100000406	1789.6	151.06119	488-81-3	6912	10254628	C00474	HMDB00508
ribonate	Carbohydrate	Pentose Metabolism	27731	LC/MS Polar	100001007	2425	165.04046	8/3/5336	5460677	4574162	C01685	HMDB00867
ribose	Carbohydrate	Pentose Metabolism	1471	LC/MS Polar	914	1508.2	195.05103	50-69-1	5779	4470639	C00121	HMDB00283
ribulonate/xylulonate/lyxonate*	Carbohydrate	Pentose Metabolism	61858	LC/MS Polar	100019968	2350	165.04046
ribulose/xylulose	Carbohydrate	Pentose Metabolism	54671	LC/MS Polar	100006122	1400	195.05102		5289590
S-methylcysteine	Amino Acid	Methionine, Cysteine, SAM and Taurine Metabolism	39592	LC/MS Polar	100002749	1942	134.02812	1187-84-4	24417	22826		HMDB02108
salicylate	Xenobiotics	Drug - Topical Agents	1515	LC/MS Polar	501	802	137.02422	69-72-7	338	331	C00805	HMDB01895
sedoheptulose	Carbohydrate	Pentose Metabolism	53237	LC/MS Polar	100002619	2255	209.06667	3019-74-7	5459879	4573620		HMDB03219
sphingosine 1-phosphate	Lipid	Sphingosines	34445	LC/MS Polar	100000626	1850	378.24148	26993-30-6	5283560	4446673	C06124	HMDB00277
succinate	Energy	TCA Cycle	1437	LC/MS Polar	252	3149	117.01933	110-15-6	1110	1078	C00042	HMDB00254
sucrose	Carbohydrate	Disaccharides and Oligosaccharides	1519	LC/MS Polar	935	3092	387.11442	57-50-1	5988	5768	C00089	HMDB00258
taurine	Amino Acid	Methionine, Cysteine, SAM and Taurine Metabolism	2125	LC/MS Polar	512	2120	124.00739	107-35-7	1123	1091	C00245	HMDB00251
threonate	Cofactors and Vitamins	Ascorbate and Aldarate Metabolism	27738	LC/MS Polar	100001022	2384	135.02989	70753-61-6	151152	133224	C01620	HMDB00943
tricarballylate	Energy	TCA Cycle	15729	LC/MS Polar	2053	4200	175.02481	99-14-9	14925	14220	C19806	HMDB31193
xylose	Carbohydrate	Pentose Metabolism	15581	LC/MS Polar	826	1727	195.05103	609-06-3	135191	85957	C00181	HMDB00098
METABOLITES_END
#END