#METABOLOMICS WORKBENCH KDSpencer77_20230821_110949 DATATRACK_ID:4236 STUDY_ID:ST002836 ANALYSIS_ID:AN004636 PROJECT_ID:PR001776
VERSION             	1
CREATED_ON             	August 22, 2023, 12:00 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                         	989-708-4858
#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                   	(Intestine)
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                         	989-708-4858
#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-00282	GROUP_ID:GOS	
SUBJECT_SAMPLE_FACTORS           	-	NACH-00283	GROUP_ID:GOS	
SUBJECT_SAMPLE_FACTORS           	-	NACH-00284	GROUP_ID:GOS	
SUBJECT_SAMPLE_FACTORS           	-	NACH-00285	GROUP_ID:GOS	
SUBJECT_SAMPLE_FACTORS           	-	NACH-00286	GROUP_ID:GOS	
SUBJECT_SAMPLE_FACTORS           	-	NACH-00287	GROUP_ID:GOS	
SUBJECT_SAMPLE_FACTORS           	-	NACH-00288	GROUP_ID:CON	
SUBJECT_SAMPLE_FACTORS           	-	NACH-00289	GROUP_ID:CON	
SUBJECT_SAMPLE_FACTORS           	-	NACH-00290	GROUP_ID:CON	
SUBJECT_SAMPLE_FACTORS           	-	NACH-00291	GROUP_ID:CON	
SUBJECT_SAMPLE_FACTORS           	-	NACH-00292	GROUP_ID:CON	
SUBJECT_SAMPLE_FACTORS           	-	NACH-00293	GROUP_ID:CON	
#COLLECTION
CO:COLLECTION_SUMMARY            	After euthanasia the colonic contents of the mice were collected and sent to
CO:COLLECTION_SUMMARY            	Metabolon Inc for UPLC-MS Analysis.
CO:SAMPLE_TYPE                   	Intestine
#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-00282	NACH-00283	NACH-00284	NACH-00285	NACH-00286	NACH-00287	NACH-00288	NACH-00289	NACH-00290	NACH-00291	NACH-00292	NACH-00293
Factors	GROUP_ID:GOS	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	1.0135	1.7091	0.752	0.6987	1.3988	2.8288	0.3756	0.9865	1.143	0.6164	1.3356	0.7331
2-hydroxy-4-(methylthio)butanoic acid	0.752	0.9066	2.4049	1	0.9732	2.2794	3.2909	8.0785	1.1837	0.5105	0.5105	0.8405
2-hydroxybutyrate/2-hydroxyisobutyrate	0.5742	2.6007	1.2385	0.8926	4.6774	1.0947	0.772	1.3488	0.876	0.5818	1.7659	0.9053
2-hydroxyheptanoate*	0.6229	0.6229	1.7416	0.8903	1.5609	1	0.9243	2.5918	0.6328	1.3119	0.7971	1.1481
2-hydroxylignocerate*	1.3669	0.5955	0.7005	2.1765	0.4367	2.6105	0.7942	0.8515	1.1485	1.889	0.3708	1.2574
2-isopropylmalate	0.5073	1.2822	0.5073	0.5073	1.3934	3.924	0.5073	0.7178	0.5073	0.6763	1.9832	0.6226
2R,3R-dihydroxybutyrate	0.44	2.571	0.44	0.7768	4.5544	1	0.44	0.8778	0.44	0.44	2.7856	0.44
2S,3R-dihydroxybutyrate	0.8774	1.3465	1	1.5468	3.5878	1.2719	0.5251	0.7622	1.3371	0.6655	0.5251	0.5327
3,4-dihydroxybutyrate	1.5652	0.9724	0.9103	1	1.7438	1.7831	0.5568	0.9111	1.2413	0.5568	0.5568	0.5568
3-hydroxy-3-methylglutarate	0.6795	1.6665	0.5793	1.4568	3.23	2.7987	0.3234	0.6563	0.3234	0.3234	1.3205	0.5479
3-hydroxybutyrate (BHBA)	0.8974	3.8078	0.4013	1.1844	9.2352	0.3172	1	1.129	0.7348	0.3172	4.8249	0.4225
3-hydroxyisobutyrate	0.0954	1.1091	0.2854	0.0954	2.1056	0.0954	0.0954	1.4713	0.0954	0.0954	0.8909	0.0954
3-hydroxysebacate	0.2996	0.4538	1.0432	1.7432	0.811	4.597	0.9541	0.8354	1.5673	1.1738	0.9568	1.3728
3-hydroxysuberate	0.5453	0.6974	0.8694	0.8298	0.2269	2.5593	1.2734	1.11	1.531	1.2377	0.89	1.8607
3-hydroxyvalerate	0.5097	2.391	0.5097	0.5097	3.8952	0.5097	0.6316	1.5431	0.7393	0.6584	1.7896	1
3-methyl-2-oxobutyrate	1.3907	0.6937	0.8356	1.2184	0.6156	2.3118	0.7728	1.5136	1.9612	1.001	0.7151	0.999
6-oxopiperidine-2-carboxylate	0.4946	1.0523	0.366	0.366	1.3355	1.5318	0.366	0.4349	0.9477	1.4302	1.2875	0.8336
allantoin	0.2129	3.6148	0.4853	18.6256	6.7469	0.3088	0.3411	1.5147	0.1985	2.2336	2.4319	0.1961
allo-threonine	2.2458	0.2881	1.173	1	0.2881	2.6492	0.2881	0.2881	0.2881	0.9056	0.2881	0.9288
alpha-hydroxyisovalerate	0.5515	2.0107	0.9011	1.0097	4.0832	0.9903	1.1469	1.8037	0.8678	0.8131	2.2399	0.9307
alpha-ketoglutaramate*	0.9331	0.5434	0.8261	1.047	1.1252	1.4586	0.6649	1.8934	1.1193	0.9837	0.8048	1.0163
alpha-ketoglutarate	1.4511	0.508	0.5878	1.188	0.7094	3.1462	0.915	0.6267	2.3349	1.2779	0.5	1.085
arabinose	0.5085	0.5085	0.5085	4.7183	0.5085	14.3838	0.7828	0.5085	0.5085	1.2172	0.5702	0.5085
arabonate/xylonate	0.5679	2.5626	0.6123	1.9403	7.644	1.1714	0.4598	1.1673	0.3887	0.8327	2.105	0.4452
ascorbic acid 3-sulfate*	0.1333	1	0.1333	10.3107	3.1906	0.2026	0.1333	0.1333	0.1333	1.3112	0.5534	0.1333
beta-hydroxyisovalerate	0.9104	1.3766	1.5978	0.7708	2.1193	1.0866	0.671	2.8701	0.8153	0.5317	1.1757	0.9134
dimethylmalonic acid	1.4187	1.2243	1.0842	0.9158	0.6216	1.6094	0.6231	0.4714	0.3693	1.5305	0.7984	1.2702
erythronate*	0.7058	2.215	0.6341	1.9476	6.0356	1.505	0.3947	0.7027	0.3598	1.2942	1.6439	0.586
ethylmalonate	1.1858	1.9682	0.6732	4.7713	4.4255	5.1485	0.5025	0.3496	0.5085	0.4057	1.6308	0.8142
fructose	1.3102	3.1282	0.5628	1.1066	3.6847	1.8446	0.5304	0.6698	0.9911	1.0089	0.8957	0.5813
fucose	0.6382	4.2551	0.9479	2.2451	12.2988	0.651	0.7824	1.2256	0.6262	1.0521	6.3151	0.5724
fumarate	0.7448	2.9546	1.2552	3.4251	8.9181	2.3228	0.5725	0.479	0.3284	0.4607	1.8215	0.3981
galactitol (dulcitol)	1.1258	2.4907	0.3059	6.2612	4.8613	2.092	0.3831	0.5584	0.8708	0.8742	1.3209	0.4757
galactonate	0.089	2.4694	0.089	1.4278	6.5931	0.8433	0.089	0.1224	0.089	0.2911	1.1567	0.089
gluconate	0.5757	5.9376	0.1632	0.7528	30.652	2.5345	0.1632	0.1632	0.1632	0.3579	1.2472	0.1632
glucose	2.1993	0.7002	1.0156	1.2303	0.5407	1.7994	0.977	0.6565	1.3943	1.2646	0.654	0.9844
glucose 6-phosphate	0.0425	0.0425	0.0425	0.0425	1.9575	0.0425	0.0425	0.0425	0.0425	0.0425	0.0425	0.0425
glucuronate	0.8138	2.0929	0.715	1.9313	3.2336	1.3823	0.6763	1.2066	0.8128	0.7198	1.1862	0.6685
glutarate (C5-DC)	1.5409	1.2545	0.8367	5.6123	3.3564	8.9382	0.7658	0.6367	0.9926	0.8408	1.0074	0.5503
glycerate	1.1456	2.8604	0.6968	1.521	4.0046	3.0453	0.3484	0.7001	0.6923	0.8544	1.8103	0.7418
glycerophosphoglycerol	0.6534	3.9729	0.9606	1.0394	3.9679	4.0978	0.432	2.0604	0.628	0.6913	10.2247	0.5341
homoserine	5.0184	0.5454	0.3826	0.9454	1.8309	3.2948	1.0546	0.3744	0.5989	0.3744	0.3744	1.1781
inositol 1-phosphate (I1P)	0.0868	0.0868	0.0868	0.0868	1.9132	0.0868	0.0868	0.0868	0.0868	0.0868	0.0868	0.0868
lactate	0.6604	8.4979	1.2019	1.3658	15.9483	0.7714	0.512	2.8747	0.6178	0.4943	5.7396	0.7981
maltose	1.3748	0.2755	1.5432	0.9988	0.2755	1.5847	1.329	0.7255	0.9673	0.7093	0.2755	1.0012
mannitol/sorbitol	2.8528	2.129	0.6767	1.2986	3.7155	2.2402	0.552	0.4951	1.0675	0.9325	0.6673	0.5509
mannose	1.3064	1.8268	0.9512	1.1002	2.732	1.7971	0.348	0.6908	0.6459	0.9511	1.0488	0.6183
methylmalonate (MMA)	1.663	3.3306	0.8555	8.4437	6.8266	5.8193	0.9953	0.5165	0.7264	1.0047	0.935	0.9189
methylsuccinate	0.8874	1.5797	0.7069	3.9373	2.8081	4.6206	0.7711	0.767	0.7594	1.443	1.1126	0.7115
mevalonate	0.5809	1.2527	0.9359	1	2.2611	1.0189	0.7193	0.5809	0.5809	0.5809	1.4389	0.8258
myo-inositol	0.7106	10.9189	1.0088	1.6569	7.9812	0.8722	0.7706	6.477	0.7565	0.9912	5.435	0.9471
N-acetylalanine	1.2536	5.0868	0.8347	0.8776	7.5523	1.2014	0.5902	1.1224	0.6853	0.8368	3.4826	0.6033
N-acetylaspartate (NAA)	0.4815	1.0039	0.2422	0.9961	2.0137	2.1334	0.3992	0.2632	0.5516	5.3774	2.9918	2.561
N-carbamoylaspartate	1.5059	2.7519	0.6275	1.7199	11.588	6.2427	0.2145	0.0888	0.393	0.8377	1.1623	0.4138
N-formylanthranilic acid	1.3841	0.9947	0.851	2.1937	1.2689	1.0123	0.9547	1.3595	0.9666	1.0053	0.9411	0.8893
orotate	0.9472	5.4108	0.6764	3.3426	8.9319	2.4726	0.3772	0.537	0.4687	1.0528	1.5288	0.3674
orotidine	0.146	5.3582	0.146	1.3076	6.5653	0.196	0.1527	0.6924	0.146	0.146	9.0091	0.146
pantoate	6.3489	0.4635	1.0598	1.2913	0.4146	0.948	1.0248	0.9752	0.676	1.6026	0.8246	1.1094
pimelate (C7-DC)	0.7625	0.7625	0.7625	0.7625	0.7625	0.8664	1.0066	0.8117	1.3211	1.2989	1.5149	0.9934
pyruvate	0.9292	0.6458	1.0708	0.4908	0.3556	1.1543	1.2572	1.1749	1.4095	0.6164	0.7959	1.2114
quinate	0.933	0.8667	1	0.8461	0.3643	3.108	0.4559	0.3643	1.032	1.5818	1.773	1.5555
ribonate	1.3591	1.0503	0.4599	1.8854	5.0351	0.8371	0.3579	0.3579	0.3579	0.9497	0.3579	0.3579
ribose	1.8274	0.9908	0.7689	1.6574	0.6898	3.0821	1.0173	0.5311	1.0092	1.3554	0.5582	0.7816
ribulonate/xylulonate/lyxonate*	0.4113	7.2892	0.312	1.8237	13.22	0.7815	0.2809	1.2185	0.2809	0.5999	4.8951	0.2809
ribulose/xylulose	1.4899	1.5549	0.5756	1.0124	0.7955	2.0878	0.6567	2.8017	0.8643	0.5876	0.9973	1.0027
salicylate	0.6728	0.6728	0.852	0.6728	1.148	0.6728	1.2135	1.2551	0.6728	0.7873	0.6728	0.6728
sedoheptulose	1.1353	1.8667	0.5287	0.8865	3.0028	1.7371	0.4642	1.1135	0.5666	0.6893	1.4054	0.5175
suberate (C8-DC)	0.1882	0.2561	0.4475	0.4299	0.3364	0.6013	1.3987	1.6288	1.7735	2.1375	1.7485	1.5239
succinate	2.1406	0.2673	2.0375	14.4029	0.5459	24.3643	1.2633	0.3141	1.3159	0.7367	0.3325	0.7334
succinimide	1.0447	0.4915	1.2397	1.0286	0.6697	1.5732	0.93	1.0099	0.9901	0.9319	0.5249	1.39
succinoyltaurine	1.2542	0.6977	0.66	1.6223	0.7164	3.0546	1.0654	0.5186	1.5422	0.8106	0.9346	1.4666
sucrose	0.9595	0.4474	1.3088	0.5954	0.441	6.9654	0.8413	1.0405	2.5615	0.441	0.441	1.6215
threonate	0.6337	12.792	0.6621	2.4043	25.0327	0.9932	0.7459	5.2472	0.5534	1.0068	10.5204	0.6922
xylose	1.7042	0.834	1.0937	7.8987	0.2332	12.1085	0.4935	0.4333	1.2328	3.284	0.2077	0.9063
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	SUPER_PATHWAY	SUB_PATHWAY	CHEMICAL_ID	RI	MASS	CAS	PUBCHEM	CHEMSPIDER	KEGG	HMDB_ID
2,3-dihydroxyisovalerate	Xenobiotics	Food Component/Plant	100002417	1050	133.05063	1756-18-9	677	657	C04039	HMDB12141
2-hydroxy-4-(methylthio)butanoic acid	Amino Acid	Methionine, Cysteine, SAM and Taurine Metabolism	100021372	1065	149.02778	583-91-5
2-hydroxybutyrate/2-hydroxyisobutyrate	Amino Acid	Glutathione Metabolism	100008928	1258	103.04006
2-hydroxyheptanoate*	Lipid	Fatty Acid, Monohydroxy	100019960	925	145.08701		2750949	2032206
2-hydroxylignocerate*	Lipid	Fatty Acid, Monohydroxy	100019894	710	383.35307		102430	92507	C17873	HMDB39540
2-isopropylmalate	Xenobiotics	Food Component/Plant	100000409	2605	175.06119	3237-44-3	77	76	C02504	HMDB00402
2R,3R-dihydroxybutyrate	Lipid	Fatty Acid, Dihydroxy	100002367	1543	119.03498	759-06-8		17215960		HMDB00498
2S,3R-dihydroxybutyrate	Lipid	Fatty Acid, Dihydroxy	100002284	1660	119.03498	5057-93-2	10964471	9139682		HMDB02453
3,4-dihydroxybutyrate	Lipid	Fatty Acid, Dihydroxy	100001730	2204	119.03498	51267-44-8
3-hydroxy-3-methylglutarate	Lipid	Mevalonate Metabolism	112	2850	161.04555	503-49-1	1662	4573695	C03761	HMDB00355
3-hydroxybutyrate (BHBA)	Lipid	Ketone Bodies	254	1480	103.04007	625-72-9	441	428	C01089	HMDB00357
3-hydroxyisobutyrate	Amino Acid	Leucine, Isoleucine and Valine Metabolism	111	1619	103.04007	2068-83-9	87	10140307	C06001	HMDB00336
3-hydroxysebacate	Lipid	Fatty Acid, Monohydroxy	100001145	2411	217.10814	73141-46-5	3017884	2285525		HMDB00350
3-hydroxysuberate	Lipid	Fatty Acid, Monohydroxy	100001143	3000	189.07684	73141-47-6	22328017	13498677		HMDB00325
3-hydroxyvalerate	Lipid	Fatty Acid, Monohydroxy	100002459	1241	117.05571	10237-77-1	107802	96952		HMDB00531
3-methyl-2-oxobutyrate	Amino Acid	Leucine, Isoleucine and Valine Metabolism	100000936	911	115.04006	3715-29-5	49	48	C00141	HMDB00019
6-oxopiperidine-2-carboxylate	Amino Acid	Lysine Metabolism	100004499	1692	142.05096	34622-39-4	3014237	2282737		HMDB61705
allantoin	Nucleotide	Purine Metabolism, (Hypo)Xanthine/Inosine containing	1002	1672	157.03671	97-59-6	204	199	C02350	HMDB00462
allo-threonine	Amino Acid	Glycine, Serine and Threonine Metabolism	100000272	2511.1	118.05096	28954-12-3	99289	89699	C05519	HMDB04041
alpha-hydroxyisovalerate	Amino Acid	Leucine, Isoleucine and Valine Metabolism	100001300	1052	117.05571	600-37-3	99823	90190		HMDB00407
alpha-ketoglutaramate*	Amino Acid	Glutamate Metabolism	100002397	1916	144.03023	18465-19-5
alpha-ketoglutarate	Energy	TCA Cycle	93	2700	145.01425	305-72-6;328-50-7;22202-68-2	51	50	C00026	HMDB00208
arabinose	Carbohydrate	Pentose Metabolism	828	1804.5	195.05103	28697-53-2	66308	59687	C00216	HMDB00646
arabonate/xylonate	Carbohydrate	Pentose Metabolism	100006115	2700	165.04046
ascorbic acid 3-sulfate*	Cofactors and Vitamins	Ascorbate and Aldarate Metabolism	100020409	2025	254.98146		11425365	9600241
beta-hydroxyisovalerate	Amino Acid	Leucine, Isoleucine and Valine Metabolism	1442	1149.8	117.05572	625-08-1	69362	62571		HMDB00754
dimethylmalonic acid	Lipid	Fatty Acid, Dicarboxylate	100004251	2702	131.03498	595-46-0	11686	11195		HMDB02001
erythronate*	Carbohydrate	Aminosugar Metabolism	100001320	2186	135.02989	88759-55-1	2781043	2061231		HMDB00613
ethylmalonate	Amino Acid	Leucine, Isoleucine and Valine Metabolism	2054	2785	131.03498	601-75-2	11756	11263		HMDB00622
fructose	Carbohydrate	Fructose, Mannose and Galactose Metabolism	878	2065	225.06159	57-48-7	5984	5764	C00095	HMDB00660
fucose	Carbohydrate	Aminosugar Metabolism	918	1626.4	209.0667	2438-80-4	17106	2299091	C01018	HMDB00174
fumarate	Energy	TCA Cycle	330	3084	115.00368	100-17-8	444972	10197150	C00122	HMDB00134
galactitol (dulcitol)	Carbohydrate	Fructose, Mannose and Galactose Metabolism	1003	2360	181.07176	608-66-2	11850	11357	C01697	HMDB00107
galactonate	Carbohydrate	Fructose, Mannose and Galactose Metabolism	100001026	3085	195.05102	299-28-5	128869	114198	C00880	HMDB00565
gluconate	Xenobiotics	Food Component/Plant	338	2922	195.05102	527-07-1	10690	10240	C00257	HMDB00625
glucose	Carbohydrate	Glycolysis, Gluconeogenesis, and Pyruvate Metabolism	572	2342	225.06159	50-99-7	5793	71358	C00031	HMDB00122
glucose 6-phosphate	Carbohydrate	Glycolysis, Gluconeogenesis, and Pyruvate Metabolism	291	4537	259.02244	103192-55-8	5958	19952394	C00668	HMDB01401
glucuronate	Carbohydrate	Aminosugar Metabolism	100000257	3233.3	193.03537	207300-70-7	444791	58552	C00191	HMDB00127
glutarate (C5-DC)	Lipid	Fatty Acid, Dicarboxylate	339	2900	131.03498	110-94-1	743	3618533	C00489	HMDB00661
glycerate	Carbohydrate	Glycolysis, Gluconeogenesis, and Pyruvate Metabolism	1052	2070.4	105.01933	600-19-1	752	732	C00258	HMDB00139
glycerophosphoglycerol	Lipid	Glycerolipid Metabolism	100001619	2430	245.04317		439964	388990	C03274
homoserine	Amino Acid	Glycine, Serine and Threonine Metabolism	100000458	2875.8	118.05096	672-15-1	12647	12126	C00263	HMDB00719
inositol 1-phosphate (I1P)	Lipid	Inositol Metabolism	370	4785	259.02244	573-35-3;106032-59-1	440194	17215924	C01177	HMDB00213
lactate	Carbohydrate	Glycolysis, Gluconeogenesis, and Pyruvate Metabolism	482	1468.8	89.02442	79-33-4	612	592	C00186	HMDB00190
maltose	Carbohydrate	Glycogen Metabolism	913	3329.4	387.11442	6363-53-7	10991489	388469	C00208	HMDB00163
mannitol/sorbitol	Carbohydrate	Fructose, Mannose and Galactose Metabolism	100001740	2320	181.07176		5780	440	C00794	HMDB00247
mannose	Carbohydrate	Fructose, Mannose and Galactose Metabolism	803	2200	225.06159	3458-28-4	18950	141983	C00159	HMDB00169
methylmalonate (MMA)	Lipid	Fatty Acid Metabolism (also BCAA Metabolism)	418	3078.4	117.01933	516-05-2	487	473	C02170	HMDB00202
methylsuccinate	Amino Acid	Leucine, Isoleucine and Valine Metabolism	2051	2800	131.03498	498-21-5	10349	9922		HMDB01844
mevalonate	Lipid	Mevalonate Metabolism	30	1462	147.06628	96949-03-0	439230	388367	C02104	HMDB00227
myo-inositol	Lipid	Inositol Metabolism	363	3506.3	225.06159	87-89-8	892	10239179	C00137	HMDB00211
N-acetylalanine	Amino Acid	Alanine and Aspartate Metabolism	1110	1564.8	130.05096	97-69-8	88064	79449	C02847	HMDB00766
N-acetylaspartate (NAA)	Amino Acid	Alanine and Aspartate Metabolism	100000787	3143	174.04079	997-55-7;997-55-7	65065	58576	C01042	HMDB00812
N-carbamoylaspartate	Nucleotide	Pyrimidine Metabolism, Orotate containing	1108	3800	175.03604	923-37-5	93072	4573859	C00438	HMDB00828
N-formylanthranilic acid	Amino Acid	Tryptophan Metabolism	100004561	900	164.03531	3342-77-6	101399	91624	C05653	HMDB04089
orotate	Nucleotide	Pyrimidine Metabolism, Orotate containing	445	1638.1	155.00983	50887-69-9	967	942	C00295	HMDB00226
orotidine	Nucleotide	Pyrimidine Metabolism, Orotate containing	100001416	2250	287.05209	314-50-1	92751	83729		HMDB00788
pantoate	Cofactors and Vitamins	Pantothenate and CoA Metabolism	100002613	1100	147.06628	470-29-1	439251	388387	C00522
pimelate (C7-DC)	Lipid	Fatty Acid, Dicarboxylate	100000101	2745	159.06628	111-16-0	385	376	C02656	HMDB00857
pyruvate	Carbohydrate	Glycolysis, Gluconeogenesis, and Pyruvate Metabolism	823	1083	87.00877	127-17-3	1060	96901	C00022	HMDB00243
quinate	Xenobiotics	Food Component/Plant	100000442	2432.9	191.05611	77-95-2	6508	10246715	C00296	HMDB03072
ribonate	Carbohydrate	Pentose Metabolism	100001007	2425	165.04046	8/3/5336	5460677	4574162	C01685	HMDB00867
ribose	Carbohydrate	Pentose Metabolism	914	1508.2	195.05103	50-69-1	5779	4470639	C00121	HMDB00283
ribulonate/xylulonate/lyxonate*	Carbohydrate	Pentose Metabolism	100019968	2350	165.04046
ribulose/xylulose	Carbohydrate	Pentose Metabolism	100006122	1400	195.05102		5289590
salicylate	Xenobiotics	Drug - Topical Agents	501	802	137.02422	69-72-7	338	331	C00805	HMDB01895
sedoheptulose	Carbohydrate	Pentose Metabolism	100002619	2255	209.06667	3019-74-7	5459879	4573620		HMDB03219
suberate (C8-DC)	Lipid	Fatty Acid, Dicarboxylate	100000016	2577.6	173.08193	505-48-6	10457	10025	C08278	HMDB00893
succinate	Energy	TCA Cycle	252	3149	117.01933	110-15-6	1110	1078	C00042	HMDB00254
succinimide	Xenobiotics	Chemical	100003696	2155	116.03532	123-56-8	11439	10955	C07273
succinoyltaurine	Amino Acid	Methionine, Cysteine, SAM and Taurine Metabolism	100021220	2572	224.02343			20509456
sucrose	Carbohydrate	Disaccharides and Oligosaccharides	935	3092	387.11442	57-50-1	5988	5768	C00089	HMDB00258
threonate	Cofactors and Vitamins	Ascorbate and Aldarate Metabolism	100001022	2384	135.02989	70753-61-6	151152	133224	C01620	HMDB00943
xylose	Carbohydrate	Pentose Metabolism	826	1727	195.05103	609-06-3	135191	85957	C00181	HMDB00098
METABOLITES_END
#END