#METABOLOMICS WORKBENCH csun0221_20230322_041603 DATATRACK_ID:3805 STUDY_ID:ST002524 ANALYSIS_ID:AN004158 PROJECT_ID:PR001626
VERSION             	1
CREATED_ON             	March 27, 2023, 8:15 am
#PROJECT
PR:PROJECT_TITLE                 	In situ microwave fixation provides an instantaneous snapshot of the brain
PR:PROJECT_TITLE                 	metabolome
PR:PROJECT_SUMMARY               	The brain metabolome directly connects to brain physiology and neuronal
PR:PROJECT_SUMMARY               	function. Brain glucose metabolism is highly heterogeneous among brain regions
PR:PROJECT_SUMMARY               	and continues postmortem. Therefore, challenges remain to capture an accurate
PR:PROJECT_SUMMARY               	snapshot of the physiological brain metabolome in healthy and diseased rodent
PR:PROJECT_SUMMARY               	models. To overcome this barrier, we employed high-power focused microwave for
PR:PROJECT_SUMMARY               	the simultaneous euthanasia and fixation of mouse brain tissue to preserve
PR:PROJECT_SUMMARY               	metabolite pools prior to surgical removal and dissection of brain regions. We
PR:PROJECT_SUMMARY               	demonstrate exhaustion of glycogen and glucose and an increase in lactate
PR:PROJECT_SUMMARY               	production during conventional rapid brain resection prior to preservation by
PR:PROJECT_SUMMARY               	liquid nitrogen that is not observed with microwave fixation. Next, microwave
PR:PROJECT_SUMMARY               	fixation was employed to define the impact of brain glucose metabolism in the
PR:PROJECT_SUMMARY               	mouse model of streptozotocin-induced type 1 diabetes. Using both total pool and
PR:PROJECT_SUMMARY               	isotope tracing analyses, we identified global glucose hypometabolism in
PR:PROJECT_SUMMARY               	multiple regions of the mouse brain, evidenced by reduced 13C enrichment into
PR:PROJECT_SUMMARY               	glycogen, glycolysis, and the TCA cycle. Reduced glucose metabolism correlated
PR:PROJECT_SUMMARY               	with a marked decrease in GLUT2 expression and several metabolic enzymes in
PR:PROJECT_SUMMARY               	unique brain regions. In conclusion, our study supports the incorporation of
PR:PROJECT_SUMMARY               	microwave fixation to study terminal brain metabolism in rodent models.
PR:INSTITUTE                     	University of Florida
PR:DEPARTMENT                    	Biochemistry and Molecular Biology
PR:LABORATORY                    	Sun lab
PR:LAST_NAME                     	Sun
PR:FIRST_NAME                    	Ramon
PR:ADDRESS                       	1200 Newell Drive, ARB
PR:EMAIL                         	ramonsun@ufl.edu
PR:PHONE                         	3522948407
#STUDY
ST:STUDY_TITLE                   	In situ microwave fixation provides an instantaneous snapshot of the brain
ST:STUDY_TITLE                   	metabolome - Part 1
ST:STUDY_SUMMARY                 	We demonstrate exhaustion of glycogen and glucose and an increase in lactate
ST:STUDY_SUMMARY                 	production during conventional rapid brain resection prior to preservation by
ST:STUDY_SUMMARY                 	liquid nitrogen that is not observed with microwave fixation. Next, microwave
ST:STUDY_SUMMARY                 	fixation was employed to define the impact of brain glucose metabolism in the
ST:STUDY_SUMMARY                 	mouse model of streptozotocin-induced type 1 diabetes. Using both total pool and
ST:STUDY_SUMMARY                 	isotope tracing analyses, we identified global glucose hypometabolism in
ST:STUDY_SUMMARY                 	multiple regions of the mouse brain, evidenced by reduced 13C enrichment into
ST:STUDY_SUMMARY                 	glycogen, glycolysis, and the TCA cycle. Reduced glucose metabolism correlated
ST:STUDY_SUMMARY                 	with a marked decrease in GLUT2 expression and several metabolic enzymes in
ST:STUDY_SUMMARY                 	unique brain regions. In conclusion, our study supports the incorporation of
ST:STUDY_SUMMARY                 	microwave fixation to study terminal brain metabolism in rodent models.
ST:INSTITUTE                     	University of Florida
ST:LAST_NAME                     	Sun
ST:FIRST_NAME                    	Ramon
ST:ADDRESS                       	1200 Newell Drive, ARB
ST:EMAIL                         	ramonsun@ufl.edu
ST:PHONE                         	3522948407
#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           	-	Hipp CP 1	Tissue Region:Hippocampus | preservation method:cryopreservation	Treatment=none; RAW_FILE_NAME=steady state CP Hipp_1_polar.CDF
SUBJECT_SAMPLE_FACTORS           	-	Hipp CP 2	Tissue Region:Hippocampus | preservation method:cryopreservation	Treatment=none; RAW_FILE_NAME=steady state CP Hipp_2_polar.CDF
SUBJECT_SAMPLE_FACTORS           	-	Hipp CP 3	Tissue Region:Hippocampus | preservation method:cryopreservation	Treatment=none; RAW_FILE_NAME=steady state CP Hipp_3_polar.CDF
SUBJECT_SAMPLE_FACTORS           	-	DVC CP 1	Tissue Region:dorsal vagal complex | preservation method:cryopreservation	Treatment=none; RAW_FILE_NAME=steady state CP DVC_1_polar.CDF
SUBJECT_SAMPLE_FACTORS           	-	DVC CP 2	Tissue Region:dorsal vagal complex | preservation method:cryopreservation	Treatment=none; RAW_FILE_NAME=steady state CP DVC_2_polar.CDF
SUBJECT_SAMPLE_FACTORS           	-	DVC CP 3	Tissue Region:dorsal vagal complex | preservation method:cryopreservation	Treatment=none; RAW_FILE_NAME=steady state CP DVC_3_polar.CDF
SUBJECT_SAMPLE_FACTORS           	-	CTX CP 1	Tissue Region:cortex | preservation method:cryopreservation	Treatment=none; RAW_FILE_NAME=steady state CP CTX_1_polar.CDF
SUBJECT_SAMPLE_FACTORS           	-	CTX CP 2	Tissue Region:cortex | preservation method:cryopreservation	Treatment=none; RAW_FILE_NAME=steady state CP CTX_2_polar.CDF
SUBJECT_SAMPLE_FACTORS           	-	CTX CP 3	Tissue Region:cortex | preservation method:cryopreservation	Treatment=none; RAW_FILE_NAME=steady state CP CTX_3_polar.CDF
SUBJECT_SAMPLE_FACTORS           	-	Hipp HPFM 1	Tissue Region:Hippocampus | preservation method:focused microwave	Treatment=none; RAW_FILE_NAME=steady state FM Hipp_1_polar.CDF
SUBJECT_SAMPLE_FACTORS           	-	Hipp HPFM 2	Tissue Region:Hippocampus | preservation method:focused microwave	Treatment=none; RAW_FILE_NAME=steady state FM Hipp_2_polar.CDF
SUBJECT_SAMPLE_FACTORS           	-	Hipp HPFM 3	Tissue Region:Hippocampus | preservation method:focused microwave	Treatment=none; RAW_FILE_NAME=steady state FM Hipp_3_polar.CDF
SUBJECT_SAMPLE_FACTORS           	-	DVC HPFM 1	Tissue Region:dorsal vagal complex | preservation method:focused microwave	Treatment=none; RAW_FILE_NAME=steady state FM DVC_1_polar.CDF
SUBJECT_SAMPLE_FACTORS           	-	DVC HPFM 2	Tissue Region:dorsal vagal complex | preservation method:focused microwave	Treatment=none; RAW_FILE_NAME=steady state FM DVC_2_polar.CDF
SUBJECT_SAMPLE_FACTORS           	-	DVC HPFM 3	Tissue Region:dorsal vagal complex | preservation method:focused microwave	Treatment=none; RAW_FILE_NAME=steady state FM DVC_3_polar.CDF
SUBJECT_SAMPLE_FACTORS           	-	CTX HPFM 1	Tissue Region:cortex | preservation method:focused microwave	Treatment=none; RAW_FILE_NAME=steady state FM CTX_1_polar.CDF
SUBJECT_SAMPLE_FACTORS           	-	CTX HPFM 2	Tissue Region:cortex | preservation method:focused microwave	Treatment=none; RAW_FILE_NAME=steady state FM CTX_2_polar.CDF
SUBJECT_SAMPLE_FACTORS           	-	CTX HPFM 3	Tissue Region:cortex | preservation method:focused microwave	Treatment=none; RAW_FILE_NAME=steady state FM CTX_3_polar.CDF
#COLLECTION
CO:COLLECTION_SUMMARY            	Mice were euthanized by microwave fixation system at 5kW for 0.6 seconds
CO:COLLECTION_SUMMARY            	(MMW-05, Muromachi Kikai Company, Japan). Brain regions (HIPP, CTX, DVC), as
CO:COLLECTION_SUMMARY            	well as muscle and liver, were dissected postmortem.
CO:SAMPLE_TYPE                   	Brain
#TREATMENT
TR:TREATMENT_SUMMARY             	We designed a two-arm study, where euthanasia occurs through either 1) direct
TR:TREATMENT_SUMMARY             	decapitation or 2) focused microwave (Fig. 1). In both arms, we performed rapid
TR:TREATMENT_SUMMARY             	dissection of the brain followed by cryopreservation in liquid nitrogen. The
TR:TREATMENT_SUMMARY             	major difference between the two arms is the tissue fixation or enzyme
TR:TREATMENT_SUMMARY             	inactivation step occurs either during cryopreservation for decapitation (CP,
TR:TREATMENT_SUMMARY             	~90 seconds) or focused microwave (FM, ~0.6 second).
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Brains were removed immediately post-mortem, and washed once with PBS, twice
SP:SAMPLEPREP_SUMMARY            	with diH2O, blotted dry, and snap frozen in liquid nitrogen. Other set of brains
SP:SAMPLEPREP_SUMMARY            	were snap frozen after microwave fixation as described above. The frozen tissues
SP:SAMPLEPREP_SUMMARY            	were pulverized to 10 μm particles in liquid N2 using a Freezer/Mill
SP:SAMPLEPREP_SUMMARY            	Cryogenic Grinder (SPEX SamplePrep). Brain regions were extracted with 1ml of
SP:SAMPLEPREP_SUMMARY            	50% methanol in the grinder, while for muscle and liver twenty milligrams of
SP:SAMPLEPREP_SUMMARY            	each pulverized tissue were extracted in 1ml of 50% methanol and separated into
SP:SAMPLEPREP_SUMMARY            	polar (aqueous layer), and protein/DNA/RNA/glycogen pellet. The polar fraction
SP:SAMPLEPREP_SUMMARY            	was dried at 10-3 mBar using a SpeedVac (Thermo) followed by derivatization.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_TYPE           	GC
CH:INSTRUMENT_NAME               	Agilent 7890A
CH:COLUMN_NAME                   	Agilent HP5-MS (30m x 0.25mm, 0.25 um)
CH:SOLVENT_A                     	na
CH:SOLVENT_B                     	na
CH:FLOW_GRADIENT                 	na
CH:FLOW_RATE                     	0.69 ml/min
CH:COLUMN_TEMPERATURE            	60-325
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Agilent 5975C
MS:INSTRUMENT_TYPE               	Single quadrupole
MS:MS_TYPE                       	EI
MS:ION_MODE                      	POSITIVE
MS:MS_COMMENTS                   	GCMS protocols were similar to those described previously with a modified
MS:MS_COMMENTS                   	temperature gradient was used for GC: Initial temperature was 130degC, held for
MS:MS_COMMENTS                   	4 minutes, rising at 6degC/minutes to 243degC, rising at 60degC/minutes to
MS:MS_COMMENTS                   	280degC, held for 2 minutes. The electron ionization (EI) energy was set to 70
MS:MS_COMMENTS                   	eV. Scan (m/z:50-800) and full scan mode were used for metabolomics analysis.
MS:MS_COMMENTS                   	Mass spectra were translated to relative metabolite abundance using the
MS:MS_COMMENTS                   	Automated Mass Spectral Deconvolution and Identification System (AMDIS) software
MS:MS_COMMENTS                   	matched to the FiehnLib metabolomics library (available through Agilent) for
MS:MS_COMMENTS                   	retention time and fragmentation pattern matching with a confidence score of >
MS:MS_COMMENTS                   	80 (Fiehn, 2016; Fiehn et al., 2000; Kind et al., 2009). Data was further
MS:MS_COMMENTS                   	analyzed using the Data Extraction for Stable Isotope-labelled Metabolites
MS:MS_COMMENTS                   	(DEXSI) software package. Untargeted metabolomics data was normalized to total
MS:MS_COMMENTS                   	ion chromatogram. For glucose tracer raw data was exported and correction for
MS:MS_COMMENTS                   	natural abundance was done by IsoCorrectoR. Fractional enrichment of each
MS:MS_COMMENTS                   	metabolite was calculated as the relative abundance of each isotopologue
MS:MS_COMMENTS                   	relative to the sum of all other isotopologues. Mean enrichment was calculated
MS:MS_COMMENTS                   	as sum of fractional enrichment of labeled isotopologues (M1, M2, M3…). For
MS:MS_COMMENTS                   	principal component analysis, pathway impact analysis the online tool
MS:MS_COMMENTS                   	Metaboanlyst was used (https://www.metaboanalyst.ca/). Data was auto scaled and
MS:MS_COMMENTS                   	log transformed.
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	Relative abundance
MS_METABOLITE_DATA_START
Samples	DVC CP 1	DVC CP 2	DVC CP 3	DVC HPFM 1	DVC HPFM 2	DVC HPFM 3	CTX CP 1	CTX CP 2	CTX CP 3	CTX HPFM 1	CTX HPFM 2	CTX HPFM 3	Hipp CP 1	Hipp CP 2	Hipp CP 3	Hipp HPFM 1	Hipp HPFM 2	Hipp HPFM 3
Factors	Tissue Region:dorsal vagal complex | preservation method:cryopreservation	Tissue Region:dorsal vagal complex | preservation method:cryopreservation	Tissue Region:dorsal vagal complex | preservation method:cryopreservation	Tissue Region:dorsal vagal complex | preservation method:focused microwave	Tissue Region:dorsal vagal complex | preservation method:focused microwave	Tissue Region:dorsal vagal complex | preservation method:focused microwave	Tissue Region:cortex | preservation method:cryopreservation	Tissue Region:cortex | preservation method:cryopreservation	Tissue Region:cortex | preservation method:cryopreservation	Tissue Region:cortex | preservation method:focused microwave	Tissue Region:cortex | preservation method:focused microwave	Tissue Region:cortex | preservation method:focused microwave	Tissue Region:Hippocampus | preservation method:cryopreservation	Tissue Region:Hippocampus | preservation method:cryopreservation	Tissue Region:Hippocampus | preservation method:cryopreservation	Tissue Region:Hippocampus | preservation method:focused microwave	Tissue Region:Hippocampus | preservation method:focused microwave	Tissue Region:Hippocampus | preservation method:focused microwave
Manose	0.010435942	0.010472475	0.010692125	0.015203427	0.011911179	0.011574778	0.00217767	0.014516691	0.007964214	0.008359658	0.008087995	0.010816991	0.010548992	0.007923781	0.008277298	0.009500033	0.009183908	0.011259771
Galactose	0.025275727	0.021288516	0.019613125	0.045739246	0.023302418	0.027842158	0.004473681	0.023992967	0.020207022	0.019754996	0.019285243	0.022285376	0.011686057	0.010962945	0.010970679	0.00925647	0.015610512	0.012377846
Glycogen	0.005872717	0.00899458	0.006747361	0.024176504	0.021635213	0.018455493	0.003750258	0.016366225	0.019172433	0.018339658	0.025495709	0.046635998	0.013463711	0.009758752	0.010684171	0.018932934	0.016675306	0.030014445
pyruvic acid	0.000308258	0.000607398	0.000442347	0.00093522	0.000464663	0.000708118	0.002486254	0.001690865	0.001350697	0.001342771	0.000590428	0.000809441	0.000880383	0.000920602	0.0007226	0.000847491	0.000671858	0.000423446
lactic acid	0.022077891	0.027168278	0.024653787	0.017397187	0.006788244	0.017693073	0.028081826	0.029219541	0.025144677	0.013350146	0.01149181	0.007198851	0.024139211	0.025750243	0.025760009	0.010827892	0.008199008	0.010578496
glycolic acid	0.000152877	8.39E-05	7.61E-05	0.000148336	7.69E-05	0.000213219	9.56E-05	0.000135828	0.000103478	0.000111064	9.11E-05	7.41E-05	7.91E-05	0.000104505	8.86E-05	0.000121095	0.00010359	0.000110409
alanine	0.001484444	0.00151673	0.001631587	0.001814108	0.001907489	0.003409782	0.004562244	0.004331798	0.003886858	0.005597943	0.005573035	0.004562322	0.004407218	0.005397563	0.005177448	0.003938087	0.004289393	0.004577717
2-ketoisocaproic acid	0.003491027	0.001306106	0.001657751	0.002439814	0.001524389	0.001614422	0.000644378	0.000865595	0.000865372	0.00078724	0.001048271	0.000917403	0.000928479	0.000908326	0.000816143	0.001055787	0.000857503	0.001331886
valine	0.006809821	0.039491321	0.061263383	0.018869485	0.024800322	0.013514087	0.002225503	0.003613154	0.004016378	0.007873344	0.004140893	0.002703473	0.014269092	0.008305884	0.012521382	0.009065874	0.012810116	0.009430212
leucine	0.000229875	0.001335842	0.001066593	0.00133036	0.001016172	0.001800998	0.002316093	0.002672437	0.002493891	0.004161839	0.002385037	0.001949574	0.002699865	0.002288731	0.002484867	0.00349489	0.003040849	0.00210966
phosphoric acid	0.358868294	0.221480856	0.2567181	0.224347895	0.185683832	0.239350666	0.07979435	0.083047451	0.111014015	0.059838983	0.11935189	0.083440136	0.093084112	0.10559553	0.09319361	0.087857037	0.093374496	0.256972896
isoleucine	0.000229875	0.001335842	0.001066593	0.00133036	0.001016172	0.001800998	0.002316093	0.002672437	0.002493891	0.004161839	0.002385037	0.001949574	0.002699865	0.002288731	0.002484867	0.00349489	0.003040849	0.00210966
proline	4.76E-05	0.000525105	0.000444849	0.000423775	0.000416495	0.000384516	0.001040526	0.00104517	0.000862352	0.001045706	0.000845017	0.000802419	0.000953146	0.000930764	0.001048588	0.000997675	0.001019692	0.000563504
glycine	0.107160092	0.117876568	0.1185953	0.123117263	0.117617267	0.130005042	0.05172909	0.050762558	0.046065075	0.051369454	0.050625726	0.048411121	0.04981273	0.049543478	0.053289941	0.048935777	0.046904392	0.051561721
succinate	0.001897332	0.002043981	0.002076195	0.000946353	0.000622535	0.000864299	0.002090297	0.002682915	0.002103964	0.001590906	0.001162742	0.001102639	0.002854033	0.003217217	0.002846607	0.000936486	0.000961916	0.001317345
fumaric acid	0.001650437	0.002915264	0.002285516	0.002856088	0.003125398	0.004780582	0.021903574	0.021453037	0.016550065	0.010009545	0.008535168	0.006265214	0.007650201	0.007357184	0.010084828	0.00467717	0.004658621	0.004260745
serine	0.000174688	0.00303756	0.00201733	0.002372529	0.002340752	0.003019655	0.012064588	0.013574663	0.00999622	0.019241582	0.012600198	0.014917368	0.013290776	0.01696602	0.01629018	0.017554615	0.015482797	0.008958417
threonine	0.000376794	0.001780864	0.001401288	0.001531319	0.001784459	0.001825309	0.004634082	0.004729485	0.003490789	0.006941363	0.003192974	0.00383141	0.00511591	0.004757465	0.004415774	0.005403777	0.005272326	0.002821714
methionine	4.73E-05	4.32E-05	2.43E-05	7.00E-05	6.11E-05	0.000115252	6.47E-05	9.55E-05	6.27E-05	0.000108208	9.36E-05	7.43E-05	7.41E-05	6.52E-05	7.17E-05	6.72E-05	6.78E-05	9.94E-05
beta-alanine	0.000461144	0.00064587	0.00062013	0.000580354	0.000701082	0.00078863	0.001704804	0.001432775	0.001069694	0.001628508	0.000969792	0.001059489	0.001409574	0.001128577	0.001214419	0.001232866	0.001475738	0.000882295
nicotinamide	0.00143656	0.001258506	0.000807252	0.000742062	0.000675505	0.002092767	0.002571255	0.002601429	0.00178854	0.001774457	0.003879005	0.002063544	0.002385256	0.001947009	0.001996921	0.000951547	0.001644142	0.003446575
malic acid	0.001600414	0.002335147	0.002028495	0.001841032	0.001975458	0.003056205	0.004782028	0.005247951	0.004793344	0.004914409	0.005199622	0.003846175	0.003527419	0.003690636	0.004105393	0.002587534	0.002735559	0.003540883
aspartic acid	0.041215766	0.088147094	0.074962558	0.058685679	0.088866723	0.085826334	0.089374138	0.087204397	0.097482944	0.083752639	0.095242603	0.089147458	0.075010143	0.074818769	0.074956229	0.075477762	0.079866553	0.060459562
GABA	0.067615191	0.078475639	0.073300585	0.072403628	0.089867455	0.084178165	0.084821742	0.092289202	0.091034315	0.071059164	0.123159044	0.091491224	0.095416039	0.09965066	0.096152455	0.077379531	0.082141965	0.120425378
Phenylalanine	0.000266547	0.000206226	0.000207399	0.000374883	0.000254757	0.000558104	2.98E-05	6.29E-05	0.000172984	3.71E-05	0.000326672	0.000132353	8.40E-05	5.70E-05	0.000208067	9.57E-05	5.33E-05	0.000556481
threonic acid	0.000134945	0.00018188	0.000137279	0.000199109	0.000269361	0.000322355	0.00103596	0.000896078	0.000718293	0.0012117	0.000626505	0.000792733	0.000700033	0.000654204	0.0006593	0.000679984	0.000810132	0.000462495
creatinine	0.018248226	0.018129364	0.014273196	0.008189567	0.008133553	0.004960322	0.012916282	0.014019508	0.01477695	0.010588018	0.016190803	0.010897221	0.016808445	0.01658047	0.015856911	0.008827153	0.009781549	0.012704375
2-oxoglutaric acid	4.09E-05	4.81E-05	6.10E-05	0.000847344	0.001063343	0.000830294	6.44E-05	6.49E-05	3.92E-05	0.002998672	0.000362533	0.001626527	9.62E-05	8.50E-05	9.63E-05	0.001958168	0.001660764	0.00038108
hypotaurine	0.000115733	0.000201107	0.000164957	0.000228255	0.000257359	0.000178707	0.001014962	0.000864445	0.000684511	0.00098557	0.000505613	0.00055218	0.001510341	0.001045015	0.001046898	0.000850213	0.000497307	0.00059368
phosphoenolpyruvate	4.05E-05	6.61E-05	5.05E-05	0.000120579	0.000120589	0.000318812	5.28E-05	8.06E-05	5.63E-05	0.000463538	6.76E-05	0.000178301	1.37E-05	1.56E-05	2.43E-05	0.000261294	0.000484418	0.00012974
glutamic acid	0.08072289	0.104875886	0.089961343	0.07257764	0.079991225	0.088887734	0.131335637	0.125858309	0.164081764	0.105515645	0.152400557	0.154613589	0.165190848	0.164798688	0.16302313	0.144749001	0.148310412	0.108922271
N-acetyl-L-aspartic acid	0.046660137	0.057663377	0.053648731	0.066935387	0.062951908	0.060568782	0.086616557	0.083723784	0.078878654	0.090694595	0.07403521	0.078875732	0.068482762	0.065898433	0.067101346	0.066419481	0.067565047	0.059461138
pyrophosphate	0.001132961	0.006997577	0.006000914	0.023856372	0.021775116	0.005267231	0.008520584	0.009394065	0.009103856	0.036878348	0.008571751	0.037260361	0.010366502	0.009604512	0.010474696	0.042548175	0.040833281	0.006193723
taurine	0.000101931	0.007070934	0.000828055	0.004312244	0.017135356	0.000413469	0.077485714	0.062538221	0.041009432	0.050715682	0.038988688	0.059310323	0.076951117	0.073034359	0.064179716	0.060635796	0.076024468	0.017775151
ribose	4.03E-05	4.44E-05	5.30E-05	2.60E-05	3.97E-05	0.000346238	0.000228545	0.000347477	0.000220165	0.000174197	0.000468556	0.000236827	0.000207735	0.000168587	0.000146872	9.51E-05	0.00029584	0.000373584
3-phosphoglyceric acid	0.000125694	0.00017423	0.000111703	5.72E-05	5.00E-05	0.000140703	0.001445507	0.001648296	0.001336943	0.000398606	0.000780097	0.00048774	0.0013965	0.001116171	0.001427994	0.000116776	0.000340216	0.000586639
ribitol	4.42E-05	4.73E-05	3.05E-05	4.07E-05	4.62E-05	7.32E-05	0.000421773	0.000325875	0.000294471	0.000417423	0.00017667	0.0002626	0.000270463	0.000202017	0.000251068	0.000222886	0.000251415	0.000128887
dihydroxyacetone phosphate	0.000188585	0.000341187	0.000166364	8.82E-05	4.00E-05	0.000770398	0.002884513	0.003298958	0.002711345	0.000611603	0.002014705	0.001020648	0.002533134	0.002302323	0.002489419	0.000204112	0.000527356	0.001172982
glycerol 1-phosphate	0.01175456	0.024210043	0.018437986	0.014245217	0.017414669	0.010517925	0.030424848	0.031874756	0.02946488	0.029044032	0.018387458	0.023883357	0.028250117	0.027888934	0.028974244	0.025761215	0.023591618	0.012565553
glutamine	0.000375139	0.00144996	0.000994328	0.000666808	0.000607843	0.000204956	0.002703508	0.002802865	0.001806793	0.003612464	0.00173602	0.003368746	0.004244762	0.003149323	0.003343192	0.003035943	0.003120868	0.00065587
citric acid	0.001919528	0.002917815	0.002188564	0.005894473	0.006692353	0.007825804	0.011279118	0.012213001	0.009546636	0.020178892	0.011973123	0.013741596	0.00879747	0.007726215	0.008117215	0.012834663	0.013704944	0.008827034
dehydroascorbic acid	0.003157776	0.00464745	0.003733967	0.004354619	0.005580314	0.003958757	0.016911297	0.016120997	0.015595975	0.017261237	0.012712523	0.014881454	0.017455758	0.016346099	0.017484837	0.014383654	0.01239882	0.010916876
adenine	0.000444061	0.000694515	0.000601992	0.000522958	0.000594676	0.000683061	0.001682624	0.001499714	0.001073304	0.002681201	0.00173518	0.001326367	0.001227927	0.001097336	0.001196574	0.00117539	0.001493456	0.001345484
tyrosine	0.000299273	0.000774872	0.000533118	0.0006678	0.000628045	0.001548437	0.001135149	0.001832283	0.001432783	0.00174748	0.001552961	0.001360356	0.000995018	0.001426496	0.001301344	0.001471658	0.001552729	0.001499145
fructose	0.000224952	0.000529738	0.000294828	0.001406742	0.000854645	0.000821023	0.000360417	0.000454726	0.000373561	0.002133118	0.000737137	0.001131045	0.000501802	0.00059894	0.000484944	0.002005884	0.001889034	0.001125739
glucose	0.000500208	0.001595295	0.000796674	0.030420254	0.036359354	0.025249066	0.00122551	0.000668295	0.000406797	0.049540369	0.015329849	0.042233898	0.001118919	0.000538505	0.003271773	0.04861942	0.045009989	0.020224492
lysine	0.000314806	0.000302904	0.000246265	0.000387634	0.000274782	0.000204359	0.000751951	0.0009052	0.000658115	0.001398619	0.000569846	0.00058635	0.001104091	0.000998339	0.001022551	0.000790953	0.000697994	0.00034681
sorbitol	0.000245013	0.00046811	0.00043416	0.0004058	0.000426756	0.000616911	0.001966513	0.002099537	0.001853666	0.002799061	0.00145714	0.00179466	0.002121599	0.002462045	0.002535349	0.001698224	0.001406133	0.001167087
pantothenic acid	0.000138739	0.00022386	0.000179532	0.000230527	0.00029092	0.00027172	0.000782845	0.000711803	0.000589636	0.000987148	0.00038354	0.000545518	0.00083214	0.000642961	0.000751698	0.000729405	0.000742465	0.000345866
palmitic acid	0.001788976	0.001493236	0.001906926	0.002136232	0.002472451	0.005355757	0.006472065	0.00814053	0.005290083	0.009017182	0.006013899	0.005698859	0.003142145	0.004571388	0.004512478	0.005778552	0.00468959	0.005126984
myo-inositol	0.208190362	0.15762105	0.169610168	0.209444553	0.166269469	0.167867064	0.108252964	0.110872363	0.117030079	0.103402756	0.119920924	0.102960898	0.128824782	0.125600008	0.127397274	0.115710234	0.109099354	0.138591396
ribose-5-phosphate	0.000187344	0.00031743	0.000238029	0.000180421	0.000186323	0.000930825	0.002302003	0.002475986	0.002106668	0.000810323	0.002282521	0.001108662	0.001445324	0.001365133	0.001426879	0.000582601	0.000919872	0.002543217
tryptophan	7.81E-05	0.000185713	0.000116126	0.000153469	0.0001516	0.000185044	0.000555751	0.000671393	0.000561624	0.000821405	0.000467167	0.000540864	0.000470978	0.000393827	0.000461692	0.000590799	0.000582343	0.000253458
stearic acid	0.00047225	0.000397904	0.000415537	0.000564173	0.000929752	0.001220872	0.003224309	0.003838309	0.002667202	0.003845605	0.002603956	0.002456124	0.001535805	0.001871715	0.001984667	0.002435345	0.001966875	0.001754733
fructose 6-phosphate	5.26E-06	2.89E-05	1.23E-05	4.91E-06	1.33E-05	1.68E-05	0.000187931	0.000302223	0.000270298	4.35E-05	4.03E-05	3.63E-05	0.000176261	0.000208838	0.000186624	3.80E-05	3.60E-05	3.56E-05
glucose-6-phosphate	4.22E-05	0.000455202	0.000263359	0.000677518	0.000916545	0.001407602	0.000678117	0.001456481	0.001351614	0.006752271	0.001777454	0.005178077	0.003564154	0.003438169	0.004975824	0.005957201	0.004766772	0.002490345
adenosine monophosphate	6.38E-05	0.001669744	0.000793398	0.000837363	0.002128484	0.000670839	0.036580477	0.034175645	0.025756307	0.035293467	0.010358641	0.016365634	0.023471769	0.020369344	0.02409436	0.019996607	0.014225658	0.003790814
cholesterol	0.00435888	0.010404873	0.005190379	0.012907236	0.032927537	0.008270439	0.048027881	0.051265601	0.040571533	0.050093476	0.04062056	0.044291278	0.02510794	0.027843714	0.029354092	0.048934196	0.038982365	0.029848144
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	PubChem ID	KEGG
Manose	18950	C00936
Galactose	439357	C00984
Glycogen	439177	C00182
pyruvic acid	1060	C00022
lactic acid	61503	C00186
glycolic acid	757	C03547
alanine	5950	C00041
2-ketoisocaproic acid	70	C00233
valine	6287	C00183
leucine	6106	C00123
phosphoric acid	57424078	C00009
isoleucine	6306	C00407
proline	145742	C00148
glycine	750	C00037
succinate	1110	C00042
fumaric acid	444972	C00122
serine	5951	C00716
threonine	6288	C00188
methionine	6137	C00073
beta-alanine	239	C00099
nicotinamide	936	C00153
malic acid	525	C03668
aspartic acid	5960	C00049
GABA	223130	C00334
Phenylalanine	6140	C00079
threonic acid	151152	C01620
creatinine	588	C00791
2-oxoglutaric acid	51	C00026
hypotaurine	107812	C00519
phosphoenolpyruvate	1005	C00074
glutamic acid	33032	C00025
N-acetyl-L-aspartic acid	65065	C01042
pyrophosphate	644102	C00013
taurine	1123	C00245
ribose	5779	C00121
3-phosphoglyceric acid	724	C00597
ribitol	NA	C00474
dihydroxyacetone phosphate	668	C00111
glycerol 1-phosphate	439162	C00093
glutamine	5961	C00064
citric acid	311	C00158
dehydroascorbic acid	7786	C05422
adenine	190	C00147
tyrosine	6057	C00082
fructose	439709	C02336
glucose	5793	C00221
lysine	5962	C00047
sorbitol	5780	C00794
pantothenic acid	6613	C00864
palmitic acid	985	C00249
myo-inositol	NA	C00137
ribose-5-phosphate	439167	C00117
tryptophan	6305	C00078
stearic acid	5281	C01530
fructose 6-phosphate	69507	C00085
glucose-6-phosphate	5958	C00092
adenosine monophosphate	6083	C00020
cholesterol	5997	C00187
METABOLITES_END
#END