#METABOLOMICS WORKBENCH jroth002_20190717_214516 DATATRACK_ID:1774 STUDY_ID:ST001222 ANALYSIS_ID:AN002035
VERSION                          	1
CREATED_ON                       	03-03-2022
#PROJECT
PR:PROJECT_TITLE                 	Untargeted metabolomics of honey bees exposed to selenate or cadmium
PR:PROJECT_SUMMARY               	Effects of selenate and cadmium exposure on honey bees.
PR:INSTITUTE                     	University of California, Riverside
PR:LAST_NAME                     	Rothman
PR:FIRST_NAME                    	Jason
PR:ADDRESS                       	900 University Ave., Riverside, CA, 91766, USA
PR:EMAIL                         	jroth002@ucr.edu
PR:PHONE                         	9518275817
PR:DOI                           	http://dx.doi.org/10.21228/M8D98R
#STUDY
ST:STUDY_TITLE                   	Effects of selenate and cadmium exposure on the honey bee metabolome (part-II)
ST:STUDY_SUMMARY                 	We moved one frame of brood each from five healthy honey bee colonies with
ST:STUDY_SUMMARY                 	marked Italian queens and housed them in a hive body at 35°C and 50% humidity
ST:STUDY_SUMMARY                 	under constant darkness. We then allowed the bees to emerge, mixed the newly
ST:STUDY_SUMMARY                 	emerged workers (NEWs) to randomize their colony of origin and placed NEWs into
ST:STUDY_SUMMARY                 	13 cm x 10.5 cm x 6.5 cm wire cages equipped with feeders containing 35mL of
ST:STUDY_SUMMARY                 	deionized water and 35mL 50% sucrose. We also provided a pollen patty to each
ST:STUDY_SUMMARY                 	cage of bees consisting of 269g corn syrup, 113g sucrose and 113g of Bee Pro
ST:STUDY_SUMMARY                 	(Mann Lake, Hackensack, MN). To inoculate the newly emerged workers with their
ST:STUDY_SUMMARY                 	“core” microbiome, we collected 50 mL of foragers from the source hives of
ST:STUDY_SUMMARY                 	the NEWs, immobilized the bees at 4°C, aseptically dissected out the abdomens
ST:STUDY_SUMMARY                 	and macerated the whole abdomens in 50% sucrose. We added 1 mL of the resulting
ST:STUDY_SUMMARY                 	slurry to 34 mL of 50% sucrose solution and fed it to the NEWs. We allowed the
ST:STUDY_SUMMARY                 	bees to feed ad libitium on the mixture for two days before replacing the
ST:STUDY_SUMMARY                 	feeders with 50% sucrose. We allowed the bees to feed for three more days to
ST:STUDY_SUMMARY                 	fully establish a microbiome. 	Once the bees had an established microbiome, we
ST:STUDY_SUMMARY                 	prepared treatment feeding solutions of 50% sucrose (as a no metal/metalloid
ST:STUDY_SUMMARY                 	control), 50% sucrose spiked with 0.6 mg/L sodium selenate or 50% sucrose with
ST:STUDY_SUMMARY                 	0.24 mg/L cadmium chloride (Alfa Aesar, Ward Hill, MA) and pollen patties spiked
ST:STUDY_SUMMARY                 	with either 6.0 mg/L selenium or 0.46 mg/L cadmium as in Hladun et al 2015. We
ST:STUDY_SUMMARY                 	again allowed the bees to feed ad libitium. We sampled three bees from 13 cages
ST:STUDY_SUMMARY                 	after four days of continuous exposure to the above-mentioned treatments and
ST:STUDY_SUMMARY                 	immediately placed the samples on dry ice, followed by long-term storage at -80
ST:STUDY_SUMMARY                 	°C.
ST:INSTITUTE                     	University of California, Riverside
ST:LAST_NAME                     	Rothman
ST:FIRST_NAME                    	Jason
ST:ADDRESS                       	900 University Ave.
ST:EMAIL                         	jroth002@ucr.edu
ST:PHONE                         	9518275817
ST:SUBMIT_DATE                   	2019-07-17
#SUBJECT
SU:SUBJECT_TYPE                  	Invertebrate
SU:SUBJECT_SPECIES               	Apis mellifera
SU:TAXONOMY_ID                   	7460
#SUBJECT_SAMPLE_FACTORS:         	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data
SUBJECT_SAMPLE_FACTORS           	-	cadmium-1	Factor:Cadmium	
SUBJECT_SAMPLE_FACTORS           	-	cadmium-2	Factor:Cadmium	
SUBJECT_SAMPLE_FACTORS           	-	cadmium-3	Factor:Cadmium	
SUBJECT_SAMPLE_FACTORS           	-	cadmium-4	Factor:Cadmium	
SUBJECT_SAMPLE_FACTORS           	-	control-1	Factor:Control	
SUBJECT_SAMPLE_FACTORS           	-	control-2	Factor:Control	
SUBJECT_SAMPLE_FACTORS           	-	control-3	Factor:Control	
SUBJECT_SAMPLE_FACTORS           	-	control-4	Factor:Control	
SUBJECT_SAMPLE_FACTORS           	-	QC-1	Factor:QCPool	
SUBJECT_SAMPLE_FACTORS           	-	QC-2	Factor:QCPool	
SUBJECT_SAMPLE_FACTORS           	-	QC-3	Factor:QCPool	
SUBJECT_SAMPLE_FACTORS           	-	selenium-1	Factor:Selenate	
SUBJECT_SAMPLE_FACTORS           	-	selenium-2	Factor:Selenate	
SUBJECT_SAMPLE_FACTORS           	-	selenium-3	Factor:Selenate	
SUBJECT_SAMPLE_FACTORS           	-	selenium-4	Factor:Selenate	
SUBJECT_SAMPLE_FACTORS           	-	selenium-5	Factor:Selenate	
#COLLECTION
CO:COLLECTION_SUMMARY            	We sampled three bees from 13 cages after four days of continuous exposure to
CO:COLLECTION_SUMMARY            	the treatments and immediately placed the samples on dry ice, followed by
CO:COLLECTION_SUMMARY            	long-term storage at -80 °C.
CO:SAMPLE_TYPE                   	Insect tissue
CO:STORAGE_CONDITIONS            	Described in summary
#TREATMENT
TR:TREATMENT_SUMMARY             	Once the bees had an established microbiome, we prepared treatment feeding
TR:TREATMENT_SUMMARY             	solutions of 50% sucrose (as a no metal/metalloid control), 50% sucrose spiked
TR:TREATMENT_SUMMARY             	with 0.6 mg/L sodium selenate or 50% sucrose with 0.24 mg/L cadmium chloride
TR:TREATMENT_SUMMARY             	(Alfa Aesar, Ward Hill, MA) and pollen patties spiked with either 6.0 mg/L
TR:TREATMENT_SUMMARY             	selenium or 0.46 mg/L cadmium and allowed the bees to feed ad libitium.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Samples were freeze-dried, homogenized, and extracted with 30:30:20:20
SP:SAMPLEPREP_SUMMARY            	acetonitrile:methanol:water:isopropanol. Samples were then sonicated, vortexed,
SP:SAMPLEPREP_SUMMARY            	and centrifuged before analysis.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	We analyzed targeted, polar primary metabolites on a Xevo TQ-XS triple
CH:CHROMATOGRAPHY_SUMMARY        	quadrupole mass spectrometer (Waters) coupled to an I-class UPLC system (Waters)
CH:CHROMATOGRAPHY_SUMMARY        	in the UC Riverside Metabolomics Core Facility. We used a ZIC-pHILIC column (2.1
CH:CHROMATOGRAPHY_SUMMARY        	x 150 mm, 5 µM) (EMD Millipore) for separations with the following mobile
CH:CHROMATOGRAPHY_SUMMARY        	phases:(A) water with 15 mM ammonium bicarbonate adjusted to pH 9.6 with
CH:CHROMATOGRAPHY_SUMMARY        	ammonium hydroxide and (B) acetonitrile. We set the flow rate to 200 µL/min and
CH:CHROMATOGRAPHY_SUMMARY        	held the column 50° C with an injection volume of 1 µL. We used the following
CH:CHROMATOGRAPHY_SUMMARY        	gradient: 0 min, 90% B; 1.5 min, 90% B; 16 min, 20% B; 18 min, 20% B; 20 min,
CH:CHROMATOGRAPHY_SUMMARY        	90% B; 28 min, 90% B.
CH:INSTRUMENT_NAME               	Waters Acquity I-Class
CH:COLUMN_NAME                   	SeQuant ZIC- pHILIC (150 x 2.1mm, 5um)
CH:COLUMN_TEMPERATURE            	50
CH:FLOW_GRADIENT                 	0 min, 90% B; 1.5 min, 90% B; 16 min, 20% B; 18 min, 20% B; 20 min, 90% B; 28
CH:FLOW_GRADIENT                 	min, 90% B
CH:FLOW_RATE                     	200 ul/min
CH:SOLVENT_A                     	water with 15 mM ammonium bicarbonate adjusted to pH 9.6 with ammonium hydroxide
CH:SOLVENT_B                     	acetonitrile
CH:CHROMATOGRAPHY_TYPE           	HILIC
#ANALYSIS
AN:LABORATORY_NAME               	UC Riverside Metabolomics Core Facility
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Waters Xevo-TQ-S
MS:INSTRUMENT_TYPE               	Triple quadrupole
MS:MS_TYPE                       	ESI
MS:MS_COMMENTS                   	We operated the MS in selected reaction monitoring mode with source and
MS:MS_COMMENTS                   	desolvation temperatures at 150° C and 500° C, respectively, with the
MS:MS_COMMENTS                   	desolvation gas set to 1000 L/hr, cone gas set to 150 L/hr and collision gas set
MS:MS_COMMENTS                   	to 0.15 mL/min. We used nitrogen gas for each step, except for the collision
MS:MS_COMMENTS                   	gas, which was argon, and we set capillary voltage to 1 kV in positive ion mode
MS:MS_COMMENTS                   	and 2 kV in negative ion mode. Similar to the untargeted methods, we generated a
MS:MS_COMMENTS                   	quality control sample by pooling equal aliquots of each sample and analyzed the
MS:MS_COMMENTS                   	QC sample every 3-4 injections to monitor system stability and performance. We
MS:MS_COMMENTS                   	processed the metabolite data (peak picking, alignment, deconvolution,
MS:MS_COMMENTS                   	integration, normalization, and spectral matching) with Progenesis Qi software
MS:MS_COMMENTS                   	(Nonlinear Dynamics, Durham, NC). We used Skyline software (MacLean et al. 2010)
MS:MS_COMMENTS                   	for data processing our targeted metabolites.
MS:ION_MODE                      	POSITIVE
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS         	Peak area
MS_METABOLITE_DATA_START
Samples	cadmium-1	cadmium-2	cadmium-3	cadmium-4	control-1	control-2	control-3	control-4	QC-1	QC-2	QC-3	selenium-1	selenium-2	selenium-3	selenium-4	selenium-5
Factors	Factor:Cadmium	Factor:Cadmium	Factor:Cadmium	Factor:Cadmium	Factor:Control	Factor:Control	Factor:Control	Factor:Control	Factor:QCPool	Factor:QCPool	Factor:QCPool	Factor:Selenate	Factor:Selenate	Factor:Selenate	Factor:Selenate	Factor:Selenate	
1-Methylhistidine	263252944.0000	316945544.0000	178695024.0000	280649008.0000	5559376.0000	202516548.0000	102806498.0000	103508032.0000	215361956.0000	254135304.0000	250746564.0000	321845008.0000	351780632.0000	235634576.0000	316772792.0000	317686504.0000
3-Methylhistidine	43549618.0000	61807760.0000	34539358.0000	67041872.0000	915674.0000	36489133.0000	21184244.0000	35561236.0000	45301300.0000	47202308.0000	45711564.0000	61310482.0000	52781978.0000	41609810.0000	66107848.0000	58441392.0000
3-Ureidopropionic acid	84507712.0000	87167192.0000	38448040.0000	88899940.0000	1435374.0000	67095184.0000	17540143.0000	67136486.0000	72080200.0000	76030548.0000	76731146.0000	76265296.0000	105446932.0000	69044280.0000	108267284.0000	107073232.0000
4-Guanidinobutyric acid	279639752.0000	206428688.0000	218052952.0000	295723032.0000	4997766.0000	364411360.0000	46615754.0000	225042584.0000	219767272.0000	236946856.0000	240983864.0000	179161568.0000	223942744.0000	202931128.0000	358175792.0000	243550216.0000
4-Hydroxyproline	499968768.0000	532841488.0000	315682248.0000	591461488.0000	9859152.0000	339429352.0000	213363832.0000	287902240.0000	457008128.0000	524318784.0000	525106384.0000	681653168.0000	590824320.0000	519047920.0000	719506688.0000	975017856.0000
5-Aminovaleric acid	58846846.0000	348093536.0000	106342432.0000	34621060.0000	1103228.0000	181462640.0000	111563308.0000	15992851.0000	130400868.0000	135186928.0000	139777204.0000	289576752.0000	8939076.0000	190271080.0000	71826766.0000	154353000.0000
Acetylcarnitine	4841894.0000	3641693.0000	6791303.0000	16182105.0000	105419.0000	18819630.0000	6260578.0000	9053719.0000	16739181.0000	17765739.0000	17961577.0000	1732480.0000	9543815.0000	2111756.0000	2379071.0000	141052220.0000
Acetylcholine	30824605.0000	14187324.0000	35679290.0000	5096144.0000	361652.0000	64227605.0000	4617769.0000	15816621.0000	18736034.0000	21273413.0000	22231963.0000	22961137.0000	19625731.0000	16342146.0000	31355458.0000	11820632.0000
Adenine	448441360.0000	419827768.0000	309674032.0000	445279504.0000	14827968.0000	416841044.0000	186624678.0000	277894988.0000	433872200.0000	463918328.0000	463824400.0000	615844464.0000	456585304.0000	353418196.0000	561487192.0000	802794912.0000
Alanine	17830200.0000	13446471.0000	13717046.0000	11543198.0000	883011.0000	25363354.0000	4443757.0000	7223348.0000	13543401.0000	14424247.0000	14538851.0000	15613031.0000	24787940.0000	16147119.0000	10846997.0000	16716743.0000
AMP	129991591.0000	101290524.0000	67589778.0000	132771374.0000	7985203.0000	55795455.0000	45059902.0000	76657208.0000	85783467.0000	98783635.0000	104431280.0000	104750372.0000	110417716.0000	66832974.0000	124461580.0000	124538435.0000
Arginine	630655576.0000	431583264.0000	518200256.0000	395509912.0000	31916449.0000	244715696.0000	307145480.0000	274379124.0000	561396488.0000	567883032.0000	559843136.0000	322782544.0000	1760287392.0000	343406360.0000	642686872.0000	420758056.0000
beta-Alanine	10236848.0000	7822781.0000	6639827.0000	9036926.0000	317761.0000	6812237.0000	3685034.0000	7411909.0000	7046872.0000	7346391.0000	7834735.0000	6260699.0000	9343166.0000	5702330.0000	8566503.0000	6800910.0000
Betaine	559996048.0000	648026148.0000	446050968.0000	605839676.0000	14035395.0000	698207048.0000	229836138.0000	438855708.0000	538434152.0000	586835380.0000	585149532.0000	599185900.0000	895577776.0000	437988348.0000	774473428.0000	800850548.0000
Carnitine	38499562.0000	17309309.0000	12087404.0000	29417697.0000	230961.0000	23399652.0000	6563114.0000	31428322.0000	22256313.0000	23954092.0000	23719887.0000	11082539.0000	44977630.0000	14074445.0000	28581195.0000	38771741.0000
Carnosine	1707114.0000	1317248.0000	577352.0000	2095946.0000	76806.0000	488909.0000	538664.0000	1051785.0000	1063553.0000	1087315.0000	1144218.0000	651670.0000	1594613.0000	636040.0000	1401222.0000	1104724.0000
CDP-choline	820974.0000	673087.0000	434318.0000	895817.0000	8221.0000	2490124.0000	215047.0000	509860.0000	769964.0000	768126.0000	895782.0000	546612.0000	830038.0000	394600.0000	1010513.0000	1083304.0000
Choline	2330734784.0000	1853013152.0000	1048746624.0000	1232567896.0000	51046802.0000	2634924896.0000	960889728.0000	1520629232.0000	1789088480.0000	1880776128.0000	1887180064.0000	1893952768.0000	2162427120.0000	1605864912.0000	2864513440.0000	2274464192.0000
Citrulline	3445904.0000	9093350.0000	1944638.0000	4531690.0000	413093.0000	13047816.0000	1516878.0000	5062675.0000	5852583.0000	6394216.0000	6741532.0000	5261464.0000	4404350.0000	6419713.0000	3822850.0000	12831227.0000
Dimethylarginine	122355152.0000	126222490.0000	83977937.0000	612338816.0000	5909222.0000	108960730.0000	218334368.0000	511251520.0000	305744024.0000	327754416.0000	327880296.0000	138100766.0000	755729152.0000	100996532.0000	820435296.0000	224748264.0000
FAD	531979.0000	284295.0000	333458.0000	409569.0000	401.0000	254826.0000	179189.0000	298578.0000	345017.0000	319502.0000	360070.0000	460155.0000	396876.0000	313414.0000	332169.0000	377653.0000
gamma-Aminobutyric acid	20167529.0000	29881149.0000	29678621.0000	23898727.0000	6732904.0000	25620679.0000	11321754.0000	17689230.0000	22074229.0000	21550245.0000	24225782.0000	17351638.0000	27968630.0000	22014489.0000	19468941.0000	37085009.0000
Glutamic acid	124153182.0000	132860684.0000	82621598.0000	112373272.0000	3775173.0000	104055406.0000	36931053.0000	22018534.0000	89160264.0000	100785274.0000	105323504.0000	115108540.0000	165726612.0000	93302722.0000	69096947.0000	167786700.0000
Glutamine	909410144.0000	565860752.0000	594384512.0000	779517920.0000	28376340.0000	436992640.0000	369696368.0000	397101552.0000	594267888.0000	700350080.0000	715807712.0000	729505408.0000	1011832480.0000	610582784.0000	892982720.0000	679431392.0000
GSH	5816101.0000	2584905.0000	1483289.0000	5723024.0000	443712.0000	2513813.0000	2678761.0000	2957803.0000	3636234.0000	4115172.0000	4498051.0000	6552290.0000	4120606.0000	2367503.0000	5260439.0000	5003955.0000
Guanine	73443900.0000	57108636.0000	41119470.0000	76297026.0000	3652728.0000	99526142.0000	44235419.0000	63559974.0000	73201708.0000	75410842.0000	74041246.0000	105168774.0000	60097422.0000	27844549.0000	75923162.0000	137574724.0000
Guanosine	9183095.0000	7276600.0000	12613059.0000	1728273.0000	319688.0000	6225212.0000	4381079.0000	3339203.0000	5303948.0000	5598515.0000	5964033.0000	3121260.0000	4414674.0000	4432140.0000	2751491.0000	3956718.0000
Hypoxanthine	569208192.0000	500134048.0000	582747536.0000	713636832.0000	16132447.0000	905917792.0000	227610832.0000	385361856.0000	612766528.0000	643997872.0000	644232560.0000	719377744.0000	808337984.0000	520670992.0000	912036480.0000	716546704.0000
Isoleucine	355107216.0000	373987460.0000	269908506.0000	329575708.0000	12340165.0000	675832040.0000	94497860.0000	165794602.0000	331033256.0000	350624604.0000	352811712.0000	328605684.0000	532908440.0000	260638052.0000	254612790.0000	411186964.0000
Kynurenine	2895013.0000	1435869.0000	3943523.0000	29869118.0000	331985.0000	1854061.0000	2974681.0000	3311271.0000	9976287.0000	10382526.0000	10599091.0000	5402855.0000	50195817.0000	7178212.0000	10460752.0000	2540822.0000
Lysine	40391859.0000	56605104.0000	28598747.0000	51096020.0000	2329872.0000	68274026.0000	14902909.0000	9951363.0000	43791750.0000	42196275.0000	40790435.0000	56534236.0000	66563676.0000	37414672.0000	35945769.0000	60663290.0000
Methionine	120885736.0000	130559532.0000	107506612.0000	53109626.0000	3391455.0000	154204008.0000	36379174.0000	31916473.0000	87676684.0000	93012480.0000	95669532.0000	70844260.0000	123514956.0000	93364148.0000	57364412.0000	118388616.0000
N-Acetylglutamic acid	35386024.0000	22978832.0000	24259859.0000	29867410.0000	792673.0000	33487100.0000	9962953.0000	19154891.0000	25600094.0000	28974842.0000	30238336.0000	34623819.0000	40638144.0000	21508104.0000	33671591.0000	43731059.0000
N-Acetyllysine	29942560.0000	51703274.0000	22708996.0000	19970116.0000	750399.0000	57667738.0000	9936309.0000	23578699.0000	27528205.0000	31949191.0000	32079424.0000	28858851.0000	21930102.0000	27484562.0000	31180732.0000	38617067.0000
N-Acetylphenylalanine	29352883.0000	30095463.0000	21903417.0000	22133491.0000	840683.0000	24626563.0000	5981731.0000	5758857.0000	19793690.0000	20500566.0000	20654827.0000	20618631.0000	24442453.0000	28197884.0000	8499562.0000	35328340.0000
N-Acetylputrescine	461927744.0000	889571200.0000	368008880.0000	263836112.0000	5108277.0000	431342416.0000	77809272.0000	259334696.0000	363391392.0000	384113808.0000	385578848.0000	427681296.0000	275881152.0000	359578160.0000	475683120.0000	371546496.0000
N-Acetyltryptophan	7186193.0000	6115523.0000	3407367.0000	5541149.0000	18413.0000	4575710.0000	1085689.0000	1736195.0000	4502034.0000	4815943.0000	4650061.0000	5326438.0000	4926907.0000	4873473.0000	3064543.0000	8468834.0000
NAD	5190279.0000	3236736.0000	2556615.0000	4991874.0000	138108.0000	4048584.0000	1310661.0000	3177422.0000	3040810.0000	3613110.0000	3495239.0000	3052878.0000	2863323.0000	2567012.0000	3021271.0000	3991547.0000
N-Methylglutamate	4497804.0000	6166753.0000	4208349.0000	4677413.0000	36648.0000	5861924.0000	1360112.0000	2489792.0000	3966011.0000	4460406.0000	4079036.0000	5218952.0000	4993934.0000	3808413.0000	4272484.0000	5628141.0000
Ornithine	2170098.0000	2198449.0000	1696682.0000	30525179.0000	336001.0000	13661922.0000	2042588.0000	6989217.0000	7611278.0000	6883034.0000	7269187.0000	1890502.0000	4193948.0000	1457222.0000	1838066.0000	27132099.0000
Phenylalanine	1128397552.0000	1082062928.0000	820388712.0000	815454688.0000	30239393.0000	1487703568.0000	288529020.0000	358191608.0000	845808248.0000	891402544.0000	900586816.0000	694443624.0000	1309379312.0000	684319120.0000	660911520.0000	1097927600.0000
Phosphocholine	2398800652.0000	1501790268.0000	922733800.0000	2170165620.0000	52010663.0000	659313003.0000	590264011.0000	1705833182.0000	1509862193.0000	1682912479.0000	1775985058.0000	1378393151.0000	2733384208.0000	874082364.0000	2663058891.0000	2149433865.0000
Pipecolic acid	136415052.0000	227722118.0000	97665264.0000	129372294.0000	3264232.0000	313825300.0000	39699107.0000	149995051.0000	174223559.0000	184625076.0000	184821118.0000	224928299.0000	195089081.0000	206680616.0000	293316580.0000	222960300.0000
Proline	9449364494.0000	6897737624.0000	6607732384.0000	8268277436.0000	393232048.0000	7191507524.0000	2991539180.0000	5237837443.0000	7946156120.0000	8275109788.0000	8381113274.0000	8575054296.0000	15078910916.0000	10514281898.0000	6990796224.0000	7391484158.0000
S-Adenosylhomocysteine	1694417.0000	1040055.0000	685921.0000	1156964.0000	50792.0000	687897.0000	316312.0000	459207.0000	1181580.0000	1250237.0000	1355457.0000	1125893.0000	1256512.0000	1148566.0000	1847433.0000	4001324.0000
S-Adenosylmethionine	4174833.0000	2765103.0000	2916644.0000	4742687.0000	441487.0000	9128883.0000	4882326.0000	15792211.0000	6925516.0000	7136513.0000	7378918.0000	2666731.0000	8729581.0000	3172527.0000	8534542.0000	4134322.0000
Serine	8809501.0000	4988973.0000	7698412.0000	7965949.0000	528742.0000	15772595.0000	2621084.0000	5204005.0000	7222126.0000	7755986.0000	8467424.0000	4509053.0000	16585458.0000	5561085.0000	5992763.0000	7688183.0000
Thiamine	2504838.0000	2631858.0000	1955154.0000	1809599.0000	38908.0000	991960.0000	956641.0000	429060.0000	2050606.0000	2171082.0000	2195589.0000	3324484.0000	2442800.0000	2598474.0000	2671198.0000	3656370.0000
Threonine	52387600.0000	48853191.0000	41415875.0000	50608604.0000	1495792.0000	74594102.0000	15123868.0000	20076541.0000	41965204.0000	45930140.0000	48009878.0000	52866294.0000	67373030.0000	49014903.0000	30517128.0000	48613406.0000
Trigonelline	424401739.0000	336519357.0000	362066227.0000	346649429.0000	6272576.0000	249936934.0000	115386431.0000	194368938.0000	303955348.0000	325837560.0000	331527595.0000	449514246.0000	454169736.0000	392165669.0000	338308685.0000	409883676.0000
Trimethyllysine	463645144.0000	474809904.0000	119966454.0000	669202224.0000	11463394.0000	507080680.0000	165609396.0000	422191928.0000	484116832.0000	502712800.0000	517905288.0000	372064656.0000	996872640.0000	230703524.0000	1118959520.0000	869565040.0000
Tryptophan	138552960.0000	166326488.0000	87033860.0000	137119172.0000	5235063.0000	164762084.0000	39610931.0000	71744916.0000	124818368.0000	133186448.0000	135546192.0000	156258424.0000	177272072.0000	98359704.0000	98239028.0000	178988704.0000
Tyramine	6726405.0000	5632478.0000	6028086.0000	3691143.0000	81124.0000	4064284.0000	1692604.0000	3099077.0000	4820113.0000	5372029.0000	5611482.0000	6781467.0000	6875834.0000	6773369.0000	4576946.0000	9801474.0000
Tyrosine	153248328.0000	196606744.0000	96364020.0000	112479148.0000	3388688.0000	62092930.0000	32478366.0000	37598528.0000	106308060.0000	113740920.0000	114716144.0000	108808708.0000	165095916.0000	90156472.0000	95442806.0000	167807900.0000
Valine	140610923.0000	131653057.0000	116399276.0000	121610571.0000	7780353.0000	258184062.0000	41952827.0000	55895765.0000	124089607.0000	129990324.0000	133530958.0000	127923057.0000	206958678.0000	117343729.0000	91333395.0000	136812102.0000
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	pubchem_id	inchi_key	kegg_id	other_id	other_id_type	ri	ri_type	moverz_quant	
1-Methylhistidine			C01152						
3-Methylhistidine			C01152						
3-Ureidopropionic acid			C02642						
4-Guanidinobutyric acid			C01035						
4-Hydroxyproline			C01157						
5-Aminovaleric acid			C00431						
Acetylcarnitine			C02571						
Acetylcholine			C01996						
Adenine			C00147						
Alanine			C00041						
AMP			C00020						
Arginine			C00062						
beta-Alanine			C00099						
Betaine			C00719						
Carnitine			C00318						
Carnosine			C00386						
CDP-choline			D03528						
Choline			C00114						
Citrulline			C00327						
Dimethylarginine			C03626						
FAD			C00016						
gamma-Aminobutyric acid			C00334						
Glutamic acid			C00025						
Glutamine			C00064						
GSH			C00051						
Guanine			C00242						
Guanosine			C00387						
Hypoxanthine			C00262						
Isoleucine			C00407						
Kynurenine			C01718						
Lysine			C00047						
Methionine			C01733						
N-Acetylglutamic acid			C00624						
N-Acetyllysine			C12989						
N-Acetylphenylalanine			C03519						
N-Acetylputrescine			C02714						
N-Acetyltryptophan			C03137						
NAD			C00003						
N-Methylglutamate			C01046						
Ornithine			C00077						
Phenylalanine			C02057						
Phosphocholine			C00588						
Pipecolic acid			C00408						
Proline			C00148						
S-Adenosylhomocysteine			C00021						
S-Adenosylmethionine			C00019						
Serine			C00065						
Thiamine			D08580						
Threonine			C00188						
Trigonelline			C01004						
Trimethyllysine			C03793						
Tryptophan			C00806						
Tyramine			C00483						
Tyrosine			C00082						
Valine			C00183						
METABOLITES_END
#END