#METABOLOMICS WORKBENCH adavison_20220513_073938 DATATRACK_ID:3248 STUDY_ID:ST002179 ANALYSIS_ID:AN003569 PROJECT_ID:PR001387
VERSION             	1
CREATED_ON             	May 26, 2022, 5:52 pm
#PROJECT
PR:PROJECT_TITLE                 	Impact of nitisinone on the cerebrospinal fluid metabolome of a murine model of
PR:PROJECT_TITLE                 	alkaptonuria
PR:PROJECT_SUMMARY               	Background: Nitisinone induced hypertyrosinaemia is well documented in
PR:PROJECT_SUMMARY               	Alkaptonuria (AKU), and there is uncertainty over whether it may contribute to a
PR:PROJECT_SUMMARY               	decline in cognitive function and or mood by altering neurotransmitter
PR:PROJECT_SUMMARY               	metabolism. The aim of this work was to evaluate the impact of nitisinone on the
PR:PROJECT_SUMMARY               	cerebrospinal fluid (CSF) metabolome in a murine model of AKU, with a view to
PR:PROJECT_SUMMARY               	providing additional insight into metabolic changes that occur following
PR:PROJECT_SUMMARY               	treatment with nitisinone. Methods: 17 CSF samples were collected from BALB/c
PR:PROJECT_SUMMARY               	Hgd-/-mice (n=8, treated with nitisinone – 4 mg/L and n=9, no treatment).
PR:PROJECT_SUMMARY               	Samples were diluted 1:1 with deionised water and analysed using a 1290 Infinity
PR:PROJECT_SUMMARY               	II liquid chromatography system coupled to a 6550 quadrupole time-of-flight mass
PR:PROJECT_SUMMARY               	spectrometry (Agilent, Cheadle, UK). Raw data were processed using a targeted
PR:PROJECT_SUMMARY               	feature extraction algorithm and an established in-house accurate mass retention
PR:PROJECT_SUMMARY               	time database. Matched entities (±10 ppm theoretical accurate mass and ±0.3
PR:PROJECT_SUMMARY               	minutes retention time window) were filtered based on their frequency and
PR:PROJECT_SUMMARY               	variability. Experimental groups were compared using a moderated t-test with
PR:PROJECT_SUMMARY               	Benjamini-Hochberg false-discovery rate adjustment. Results: Tyrosine,
PR:PROJECT_SUMMARY               	acetyl-tyrosine, γ-glutamyl-tyrosine, p-hydroxyphenylacetic acid and
PR:PROJECT_SUMMARY               	3-(4-hydroxyphenyl)lactic acid were shown to increase in abundance (log2 fold
PR:PROJECT_SUMMARY               	change 2.6-6.9, 3/5 were significant p<0.05) in the mice that received
PR:PROJECT_SUMMARY               	nitisinone. Several other metabolites of interest were matched but no
PR:PROJECT_SUMMARY               	significant differences were observed, including the aromatic amino acids
PR:PROJECT_SUMMARY               	phenylalanine and tryptophan, and monoamine metabolites adrenaline,
PR:PROJECT_SUMMARY               	3-methoxy-4-hydroxyphenylglycol and octopamine. Conclusions: Evaluation of the
PR:PROJECT_SUMMARY               	CSF metabolome of a murine model of AKU showed a significant difference in the
PR:PROJECT_SUMMARY               	abundance of a limited number of metabolites. None of these have been reported
PR:PROJECT_SUMMARY               	in CSF from a murine model of AKU previously. Moreover this study confirms that
PR:PROJECT_SUMMARY               	some monoamine metabolites do not appear to be altered following nitisinone
PR:PROJECT_SUMMARY               	therapy.
PR:INSTITUTE                     	University of Liverpool Institute of Life Course & Medical Sciences
PR:LAST_NAME                     	Davison
PR:FIRST_NAME                    	Andrew
PR:ADDRESS                       	1. Department of Clinical Biochemistry and Metabolic Medicine, Liverpool
PR:ADDRESS                       	Clinical Laboratories, Liverpool University Hospitals NHS Foundation Trust,
PR:ADDRESS                       	Liverpool, UK; 2. Department of Musculoskeletal and Ageing Science, Institute of
PR:ADDRESS                       	Life Course and Medical Sciences, University of Liverpool, Liverpool, UK 3.
PR:ADDRESS                       	School of Exercise Science, Liverpool John Moores University, Liverpool, UK
PR:EMAIL                         	andrew.davison@liverpoolft.nhs.uk
PR:PHONE                         	0151 706 4011
PR:CONTRIBUTORS                  	Davison AS1,2,*, Norman BP2, Sutherland H2,3, Milan AM1,2, Jarvis JC3, Gallagher
PR:CONTRIBUTORS                  	JA2, Ranganath LR1,2
#STUDY
ST:STUDY_TITLE                   	Impact of nitisinone on the cerebrospinal fluid metabolome of a murine model of
ST:STUDY_TITLE                   	alkaptonuria
ST:STUDY_SUMMARY                 	Background: Nitisinone induced hypertyrosinaemia is well documented in
ST:STUDY_SUMMARY                 	Alkaptonuria (AKU), and there is uncertainty over whether it may contribute to a
ST:STUDY_SUMMARY                 	decline in cognitive function and or mood by altering neurotransmitter
ST:STUDY_SUMMARY                 	metabolism. The aim of this work was to evaluate the impact of nitisinone on the
ST:STUDY_SUMMARY                 	cerebrospinal fluid (CSF) metabolome in a murine model of AKU, with a view to
ST:STUDY_SUMMARY                 	providing additional insight into metabolic changes that occur following
ST:STUDY_SUMMARY                 	treatment with nitisinone. Methods: 17 CSF samples were collected from BALB/c
ST:STUDY_SUMMARY                 	Hgd-/-mice (n=8, treated with nitisinone – 4 mg/L and n=9, no treatment).
ST:STUDY_SUMMARY                 	Samples were diluted 1:1 with deionised water and analysed using a 1290 Infinity
ST:STUDY_SUMMARY                 	II liquid chromatography system coupled to a 6550 quadrupole time-of-flight mass
ST:STUDY_SUMMARY                 	spectrometry (Agilent, Cheadle, UK). Raw data were processed using a targeted
ST:STUDY_SUMMARY                 	feature extraction algorithm and an established in-house accurate mass retention
ST:STUDY_SUMMARY                 	time database. Matched entities (±10 ppm theoretical accurate mass and ±0.3
ST:STUDY_SUMMARY                 	minutes retention time window) were filtered based on their frequency and
ST:STUDY_SUMMARY                 	variability. Experimental groups were compared using a moderated t-test with
ST:STUDY_SUMMARY                 	Benjamini-Hochberg false-discovery rate adjustment. Results: Tyrosine,
ST:STUDY_SUMMARY                 	acetyl-tyrosine, γ-glutamyl-tyrosine, p-hydroxyphenylacetic acid and
ST:STUDY_SUMMARY                 	3-(4-hydroxyphenyl)lactic acid were shown to increase in abundance (log2 fold
ST:STUDY_SUMMARY                 	change 2.6-6.9, 3/5 were significant p<0.05) in the mice that received
ST:STUDY_SUMMARY                 	nitisinone. Several other metabolites of interest were matched but no
ST:STUDY_SUMMARY                 	significant differences were observed, including the aromatic amino acids
ST:STUDY_SUMMARY                 	phenylalanine and tryptophan, and monoamine metabolites adrenaline,
ST:STUDY_SUMMARY                 	3-methoxy-4-hydroxyphenylglycol and octopamine. Conclusions: Evaluation of the
ST:STUDY_SUMMARY                 	CSF metabolome of a murine model of AKU showed a significant difference in the
ST:STUDY_SUMMARY                 	abundance of a limited number of metabolites. None of these have been reported
ST:STUDY_SUMMARY                 	in CSF from a murine model of AKU previously. Moreover this study confirms that
ST:STUDY_SUMMARY                 	some monoamine metabolites do not appear to be altered following nitisinone
ST:STUDY_SUMMARY                 	therapy.
ST:INSTITUTE                     	University of Liverpool Institute of Life Course & Medical Sciences
ST:LAST_NAME                     	Davison
ST:FIRST_NAME                    	Andrew
ST:ADDRESS                       	1. Department of Clinical Biochemistry and Metabolic Medicine, Liverpool
ST:ADDRESS                       	Clinical Laboratories, Liverpool University Hospitals NHS Foundation Trust,
ST:ADDRESS                       	Liverpool, UK; 2. Department of Musculoskeletal and Ageing Science, Institute of
ST:ADDRESS                       	Life Course and Medical Sciences, University of Liverpool, Liverpool, UK 3.
ST:ADDRESS                       	School of Exercise Science, Liverpool John Moores University, Liverpool, UK
ST:EMAIL                         	andrew.davison@liverpoolft.nhs.uk
ST:PHONE                         	0151 706 4011
ST:NUM_GROUPS                    	2
ST:TOTAL_SUBJECTS                	17
#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           	-	Hgd81.1 + Nitisinone NEG	Treatment group:Nitisinone	RAW_FILE_NAME=NEG_AKU_+ NTBC_81.1_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd81.2 + Nitisinone NEG	Treatment group:Nitisinone	RAW_FILE_NAME=NEG_AKU_+ NTBC_81.2_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd81.3 + Nitisinone NEG	Treatment group:Nitisinone	RAW_FILE_NAME=NEG_AKU_+ NTBC_81.3_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd82.1 + Nitisinone NEG	Treatment group:Nitisinone	RAW_FILE_NAME=NEG_AKU_+ NTBC_82.1_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd82.2 + Nitisinone NEG	Treatment group:Nitisinone	RAW_FILE_NAME=NEG_AKU_+ NTBC_82.2_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd83.1 + Nitisinone NEG	Treatment group:Nitisinone	RAW_FILE_NAME=NEG_AKU_+ NTBC_83.1_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd83.2 - No treatment NEG	Treatment group:Nitisinone	RAW_FILE_NAME=NEG_AKU_+ NTBC_83.2_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd80.3 - No treatment NEG	Treatment group:No Treatment (control)	RAW_FILE_NAME=NEG_AKU_80.3_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd80.4 - No treatment NEG	Treatment group:No Treatment (control)	RAW_FILE_NAME=NEG_AKU_80.4_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd81.4 - No treatment NEG	Treatment group:No Treatment (control)	RAW_FILE_NAME=NEG_AKU_81.4_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd81.5 - No treatment NEG	Treatment group:No Treatment (control)	RAW_FILE_NAME=NEG_AKU_81.5_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd81.6 - No treatment NEG	Treatment group:No Treatment (control)	RAW_FILE_NAME=NEG_AKU_81.6_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd82.3 - No treatment NEG	Treatment group:No Treatment (control)	RAW_FILE_NAME=NEG_AKU_82.3_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd82.4 - No treatment NEG	Treatment group:No Treatment (control)	RAW_FILE_NAME=NEG_AKU_82.4_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd83.3 - No treatment NEG	Treatment group:No Treatment (control)	RAW_FILE_NAME=NEG_AKU_83.3_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd83.4 - No treatment NEG	Treatment group:No Treatment (control)	RAW_FILE_NAME=NEG_AKU_83.4_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd80.2 + Nitisinone POS	Treatment group:Nitisinone	RAW_FILE_NAME=POS_AKU_+ NTBC_80.2_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd81.1 + Nitisinone POS	Treatment group:Nitisinone	RAW_FILE_NAME=POS_AKU_+ NTBC_81.1_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd81.2 + Nitisinone POS	Treatment group:Nitisinone	RAW_FILE_NAME=POS_AKU_+ NTBC_81.2_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd81.3 + Nitisinone POS	Treatment group:Nitisinone	RAW_FILE_NAME=POS_AKU_+ NTBC_81.3_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd82.1 + Nitisinone POS	Treatment group:Nitisinone	RAW_FILE_NAME=POS_AKU_+ NTBC_82.1_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd82.2 + Nitisinone POS	Treatment group:Nitisinone	RAW_FILE_NAME=POS_AKU_+ NTBC_82.2_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd83.1 + Nitisinone POS	Treatment group:Nitisinone	RAW_FILE_NAME=POS_AKU_+ NTBC_83.1_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd83.2 - No treatment POS	Treatment group:Nitisinone	RAW_FILE_NAME=POS_AKU_+ NTBC_83.2_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd80.3 - No treatment POS	Treatment group:No Treatment (control)	RAW_FILE_NAME=POS_AKU_80.3_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd80.4 - No treatment POS	Treatment group:No Treatment (control)	RAW_FILE_NAME=POS_AKU_80.4_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd81.4 - No treatment POS	Treatment group:No Treatment (control)	RAW_FILE_NAME=POS_AKU_81.4_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd81.5 - No treatment POS	Treatment group:No Treatment (control)	RAW_FILE_NAME=POS_AKU_81.5_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd81.6 - No treatment POS	Treatment group:No Treatment (control)	RAW_FILE_NAME=POS_AKU_81.6_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd82.3 - No treatment POS	Treatment group:No Treatment (control)	RAW_FILE_NAME=POS_AKU_82.3_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd82.4 - No treatment POS	Treatment group:No Treatment (control)	RAW_FILE_NAME=POS_AKU_82.4_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd83.3 - No treatment POS	Treatment group:No Treatment (control)	RAW_FILE_NAME=POS_AKU_83.3_CSF
SUBJECT_SAMPLE_FACTORS           	-	Hgd83.4 - No treatment POS	Treatment group:No Treatment (control)	RAW_FILE_NAME=POS_AKU_83.4_CSF
#COLLECTION
CO:COLLECTION_SUMMARY            	A murine model of AKU was used for all experiments as described previously
CO:COLLECTION_SUMMARY            	(Preston et al., 2014). Seventeen BALB/c Hgd-/- mice (Figure 5 for age and
CO:COLLECTION_SUMMARY            	gender) were included in this study, 8 mice were administered nitisinone (4
CO:COLLECTION_SUMMARY            	mg/L) through their drinking water for 6 days, the remaining 9 mice received no
CO:COLLECTION_SUMMARY            	nitisinone. All animals were housed in air conditioned rooms (with a 12 h
CO:COLLECTION_SUMMARY            	dark/light cycle) at 20 °C and 53 % humidity, with access to food and water ad
CO:COLLECTION_SUMMARY            	libitum at Liverpool John Moores University. All animal experiments complied
CO:COLLECTION_SUMMARY            	with the ARRIVE guidelines and were carried out in accordance with the U.K.
CO:COLLECTION_SUMMARY            	Animals (Scientific Procedures) Act, 1986 and associated guidelines, EU
CO:COLLECTION_SUMMARY            	Directive 2010/63/EU for animal experiments. All mice were culled by carbon
CO:COLLECTION_SUMMARY            	dioxide asphyxiation and CSF was removed from the cisterna magna following the
CO:COLLECTION_SUMMARY            	puncture technique of Liu et al. (56), with some modifications. The cisterna
CO:COLLECTION_SUMMARY            	magna was exposed by dissecting skin and overlaying muscles. Cauterisation was
CO:COLLECTION_SUMMARY            	used to dry up any bleeding from surrounding tissues. A pulled glass pipette
CO:COLLECTION_SUMMARY            	with an internal diameter of ~0.4 mm was attached to silicone tubing and
CO:COLLECTION_SUMMARY            	connected to a 1 mL syringe with a 19 g needle. The tip of the glass pipette was
CO:COLLECTION_SUMMARY            	used to puncture the membrane and held still just below the membrane by 1
CO:COLLECTION_SUMMARY            	person. Through a double-headed microscope a second person then used the 1 mL
CO:COLLECTION_SUMMARY            	syringe to apply gentle pressure to encourage the CSF to flow up the capillary
CO:COLLECTION_SUMMARY            	tube. Only clear CSF samples (i.e. non blood stained) were collected. Once
CO:COLLECTION_SUMMARY            	collected CSF samples were transferred to clear Eppendorf tubes and stored at
CO:COLLECTION_SUMMARY            	-80 °C until analysis. Samples were not acidified. The same glass pipette was
CO:COLLECTION_SUMMARY            	used to collect CSF from each animal, and was washed with pure water between
CO:COLLECTION_SUMMARY            	each animal.
CO:SAMPLE_TYPE                   	CSF
#TREATMENT
TR:TREATMENT_SUMMARY             	9 mice control 8 mice received 4 mg/L nitisinone for 6 days in drinking water
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Murine CSF samples were prepared by diluting 2 µL CSF with 2 µL deionized
SP:SAMPLEPREP_SUMMARY            	water (DIRECT-Q 3UV Millipore water purification system). CSF samples were
SP:SAMPLEPREP_SUMMARY            	pipetted directly into a 150 µL glass insert with polymer feet, which sat in a
SP:SAMPLEPREP_SUMMARY            	2 mL amber screw vial (Agilent, Cheadle, UK). These vials were used to ensure
SP:SAMPLEPREP_SUMMARY            	maximal recovery of sample. Deionised water was also added directly to the glass
SP:SAMPLEPREP_SUMMARY            	insert, and mixed with the CSF sample using a pipette. The glass vials were
SP:SAMPLEPREP_SUMMARY            	centrifuged at 600 rpm for 10 min to ensure the diluted sample was at the bottom
SP:SAMPLEPREP_SUMMARY            	of the vial for sampling. 1 µL of diluted CSF was analysed in negative and
SP:SAMPLEPREP_SUMMARY            	positive polarities, respectively.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	Liquid chromatography (LC) was performed on an Agilent 1290 Infinity II LC
CH:CHROMATOGRAPHY_SUMMARY        	system. An Atlantis dC18 column (3.0x100mm, 3µm, Waters, UK) was maintained at
CH:CHROMATOGRAPHY_SUMMARY        	60°C with a flow rate of 0.4mL/min. Mobile phases were (A) water and (B)
CH:CHROMATOGRAPHY_SUMMARY        	methanol both containing 5mmol/L ammonium formate and 0.1% formic acid. The
CH:CHROMATOGRAPHY_SUMMARY        	elution gradient started at 5% mobile phase B at 0-1 min increasing linearly to
CH:CHROMATOGRAPHY_SUMMARY        	100% B by 12 min, held at 100% B until 14 min, returning to 95% A for 5 min to
CH:CHROMATOGRAPHY_SUMMARY        	recondition the column. Injection volume was 1µL.
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Agilent Infinity II
CH:COLUMN_NAME                   	Atlantis dC18 column (3.0x100mm, 3µm, Waters, UK)
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Agilent 6550 QTOF
MS:INSTRUMENT_TYPE               	Triple quadrupole
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	NEGATIVE
MS:MS_COMMENTS                   	Quadrupole time-of-flight mass spectrometry (QTOF-MS) conditions An Agilent 6550
MS:MS_COMMENTS                   	QTOF-MS equipped with a dual jet stream electrospray ionisation source was
MS:MS_COMMENTS                   	operated in 2GHz mode, over the mass range of 50-1700, in negative and positive
MS:MS_COMMENTS                   	polarities. A reference mass correction solution was continually infused at a
MS:MS_COMMENTS                   	flow rate of 0.5 mL/min via an external isocratic pump (Agilent, UK) for
MS:MS_COMMENTS                   	constant mass correction (see Preparation of reference mass correction
MS:MS_COMMENTS                   	solution). Capillary and fragmentor voltages were 4000 V and 380 V,
MS:MS_COMMENTS                   	respectively. Desolvation gas temperature was 200 °C with flow rate at 15
MS:MS_COMMENTS                   	L/min. The sheath gas temperature was 300 °C with flow rate at 12 L/min and
MS:MS_COMMENTS                   	nebulizer pressure was 40 psi and nozzle voltage 1000 V. Data acquisition rate
MS:MS_COMMENTS                   	was 3 spectra/s. Preparation of reference mass correction solution Reference
MS:MS_COMMENTS                   	mass correction solution was prepared in 95:5 methanol:water containing 5 mmol/L
MS:MS_COMMENTS                   	purine (CAS No. 120-73-0), 100 mmol/L trifluoroacetic acid ammonium salt (TFA,
MS:MS_COMMENTS                   	CAS No. 3336-58-1) and 2.5 mmol/L hexakis(1H, 1H,
MS:MS_COMMENTS                   	3H-tetrafluoropropoxy)phosphazine (HP-0921, CAS No. 58943-98-9) (Agilent,
MS:MS_COMMENTS                   	Cheadle, UK). Reference ions monitored were: purine (m/z 121.0509) and HP-0921
MS:MS_COMMENTS                   	(m/z 922.0098) (positive polarity) and TFA (m/z 112.9856), purine (m/z 119.0363)
MS:MS_COMMENTS                   	and HP-0921 (HP-0921 + formate adduct: m/z 966.0007) (negative polarity). Data
MS:MS_COMMENTS                   	acquisition and handling parameters Data were acquired using Acquisition (Build
MS:MS_COMMENTS                   	06.00, Agilent, Cheadle, UK). Quality checks and processing of raw data files
MS:MS_COMMENTS                   	(Agilent ‘.d’ files) were performed with Qualitative Analysis software
MS:MS_COMMENTS                   	(Build 07.00, Agilent, Cheadle, UK). Extracted ion chromatograms of reference
MS:MS_COMMENTS                   	masses were performed to check mass accuracy remained <5 ppm throughout the run
MS:MS_COMMENTS                   	and that the reference ion signal did not drop out during the chromatographic
MS:MS_COMMENTS                   	run. In addition, to check chromatographic reproducibility binary pump pressure
MS:MS_COMMENTS                   	curves for injections across each analytical sequence were overlaid. Mass
MS:MS_COMMENTS                   	accuracy and chromatographic reproducibility were acceptable for all experiments
MS:MS_COMMENTS                   	performed. Acquired profiling sample data were mined for signals against an
MS:MS_COMMENTS                   	established in-house AMRT database of compounds (contains theoretical accurate
MS:MS_COMMENTS                   	mass, measured retention time, and empirical formula for 469 intermediary
MS:MS_COMMENTS                   	metabolites, MW 72-785) using ‘targeted feature extraction’ with Profinder
MS:MS_COMMENTS                   	software (Build 08.00, Agilent, Cheadle, UK). Targeted feature extraction uses
MS:MS_COMMENTS                   	the molecular formulae from the AMRT database to extract and group spectral
MS:MS_COMMENTS                   	signals (i.e. adducts, isotopes and multimers) that correspond to individual
MS:MS_COMMENTS                   	database compounds. Feature extraction employed a window of theoretical accurate
MS:MS_COMMENTS                   	mass ±10 ppm and database retention time ±0.3 min. Allowed species were: H+,
MS:MS_COMMENTS                   	Na+ and NH4+ (positive polarity) and H- and CHO2- (negative polarity). Dimers
MS:MS_COMMENTS                   	were allowed for both polarities. Charge state range was 1-2. Data files were
MS:MS_COMMENTS                   	then exported from Profinder as ‘.CEF’ files as a whole batch for each
MS:MS_COMMENTS                   	profiling experiment and imported to Mass Profiler Professional (MPP) software
MS:MS_COMMENTS                   	for statistical analysis (Build 14.5, Agilent, Cheadle, UK).
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	area
MS_METABOLITE_DATA_START
Samples	Hgd81.1 + Nitisinone NEG	Hgd81.2 + Nitisinone NEG	Hgd81.3 + Nitisinone NEG	Hgd82.1 + Nitisinone NEG	Hgd82.2 + Nitisinone NEG	Hgd83.1 + Nitisinone NEG	Hgd83.2 - No treatment NEG	Hgd80.3 - No treatment NEG	Hgd80.4 - No treatment NEG	Hgd81.4 - No treatment NEG	Hgd81.5 - No treatment NEG	Hgd81.6 - No treatment NEG	Hgd82.3 - No treatment NEG	Hgd82.4 - No treatment NEG	Hgd83.3 - No treatment NEG	Hgd83.4 - No treatment NEG
Factors	Treatment group:Nitisinone	Treatment group:Nitisinone	Treatment group:Nitisinone	Treatment group:Nitisinone	Treatment group:Nitisinone	Treatment group:Nitisinone	Treatment group:Nitisinone	Treatment group:No Treatment (control)	Treatment group:No Treatment (control)	Treatment group:No Treatment (control)	Treatment group:No Treatment (control)	Treatment group:No Treatment (control)	Treatment group:No Treatment (control)	Treatment group:No Treatment (control)	Treatment group:No Treatment (control)	Treatment group:No Treatment (control)
Inosine	1128769	600575	750496	1462769	1081718	1146996	1220644	939141	925632	1077156	1152140	1134742	1056061	1044233	1287162	1216464
Adipic acid	9009	1	43272	20022	11036	17297	105525	11153	9282	11231	16915	10864	32357	31038	11722	23134
Sarcosine	217770	147203	282444	994577	496851	251892	302044	244631	5745	210810	285484	376666	175021	407808	299528	233228
Ethylmalonic acid	19773	19126	21867	48514	63018	17879	43690	15494	28961	14809	31800	27553	19485	44167	29256	32284
D-(-)-Lyxose	278483	163289	181334	201718	245345	187892	206847	235213	199127	244772	322068	224985	216133	197387	244992	346072
Hippuric acid	2412	1	1	5434	3354	1	3028	3474	3182	600	15233	4499	1144	4175	1	2190
Uridine monophosphate (UMP)	1	1	1	30086	7092	1	1	10320	121338	1	3706	1	1	11672	1	1725
Methyl jasmonate	8881	13159	10510	8229	12124	11495	10983	10348	8196	10797	10048	14307	13706	4872	11810	9912
Dulcitol	55303	27062	29867	46732	41499	45039	35229	54540	44443	40408	148242	47346	32609	41290	43610	32190
Galactaric acid	48382	1	1	69057	1	1	55759	33817	114492	55835	77637	52185	441202	76819	74688	1
meso-Tartaric acid	1	21326	37919	114442	27532	1	1	97576	227109	32080	37255	48418	39909	112178	1	1
Hypoxanthine	891435	469625	717384	1058502	976339	800484	815486	711909	684307	600007	847670	727415	667770	856116	737045	779124
Phosphocreatine	17913	12358	1	22265	1	15307	18793	25432	25971	19550	20785	18714	37834	21835	21115	25110
Thymidine	7050	1	5108	7456	7577	7567	7607	5855	1	5865	6233	6457	7059	7469	6903	7760
Orotic acid	130834	78807	79097	66028	78837	79447	106516	120671	98824	131426	110715	101884	130432	104003	105175	144899
Uric acid	260623	166502	234730	218442	245572	241133	253019	236867	202750	198674	261913	239876	222290	152171	270200	293044
Gluconic acid	1390079	672667	920394	1456643	1473088	1241182	1178866	1638406	1205748	1225294	1435025	1225167	1474564	1568685	1499215	1314853
Aceturic acid	11347	1	1	1	1	1	1	1	1	9742	1	1	13135	12098	10595	1
?-Linolenic Acid	1	1727	1	1548	1407	1	2391	1	3032	1	1	1	1989	7178	1	1
L-Tryptophan	30486	16706	26212	26666	26745	24586	38620	30869	33063	24125	40285	32731	14159	29862	32632	43392
Cytidine	16632	11513	14980	18469	19805	17800	18307	15140	12932	15297	16479	17222	14221	18985	16382	15694
L-Threonine	48560	117283	95799	43820	63163	62046	66332	63431	55995	54300	63861	64302	33153	52984	78782	87400
4-Hydroxyphenyllactic acid	1376458	536068	604074	1298520	811041	865084	978715	30188	32308	29815	34761	38660	42736	36405	44618	55018
Malic acid	825379	559524	1060503	3585395	2028170	949236	1855575	888399	979005	834057	1398644	2045739	1107212	2247099	1339087	1526760
L-Isoleucine	25892	18253	10342	47403	34682	29152	37252	31283	28531	26649	31988	31053	24939	14800	35529	36085
N-Acetyl-L-glutamic acid	1	1	1	1	1	1	1	1	1	1	1	1	1	23759	1	1
D-Mannitol	55303	27062	29867	46732	41499	45039	35229	54540	44443	40408	148242	47346	32609	41290	43610	32190
L-Aspartic Acid	114303	44461	79175	99559	101651	89570	94463	92501	1	78718	113746	78106	92289	116478	92698	135695
Myristic acid	1	2910	1	2618	1	2458	3000	1	1907	1	1	1	2671	5225	1	1
phosphoenol pyruvate	1	1	1	246210	42275	1	26911	31493	45026	29480	25286	19327	1	89739	35380	40275
Oxoglutaric acid	11050	10611	20916	37807	26686	1	9974	218196	61542	58659	15379	17825	17888	19536	60255	14341
Citric acid	2039994	2446245	2471211	3749292	4156968	2197859	3406313	2426531	2756584	2440273	3659776	3753129	5294389	4533789	2938319	3364858
Succinic acid	261864	204846	348569	700447	811825	319805	674079	199920	319347	203442	417370	406116	235224	711046	437307	402747
Methyl N-(a-methylbutyryl)glycine	34583	32491	288107	84954	59610	84977	595783	49514	39168	55642	74959	61644	828908	154006	62365	72866
D-Aspartic acid	114635	44633	79627	100265	102256	89703	94894	92550	1	78718	114250	78649	92511	117101	92897	136217
12-Hydroxydodecanoic acid	1	6698	15217	5546	7639	7047	25442	5000	1	1	1	4608	13909	10171	1	4799
Pantothenic Acid	450261	269316	335589	314520	320484	560943	582807	505320	552106	532948	609399	481084	264297	311180	647807	665806
Dehydroascorbic acid	53897	18043	36011	1	33845	42492	49049	40601	20216	22724	61738	36802	46718	12208	47105	54602
Glutamine	1688220	1453556	1616356	1447027	1819199	1555354	1639762	1795624	1694370	1492094	1696140	1605798	1485114	1702206	1810762	1652460
Arabinose	278482	163289	181334	201718	245345	187892	206850	235213	199127	244772	322068	224985	216133	197387	244992	346072
Palmitic acid	34909	35665	34944	44299	47212	41272	39666	40482	41344	36536	37368	38499	36586	51786	38053	37278
Itaconic acid	16775	17403	18824	24675	25646	26541	26347	17191	15951	16895	11471	29083	26193	17821	21518	20265
DL-a-Hydroxybutyric acid	24167	75119	27120	68574	99855	27156	48896	23890	52315	36367	58498	50304	52586	100495	27376	39148
Adenosine5'-monophosphate	1	5824	45308	396358	30994	12303	7965	26690	95369	23480	17710	4715	18060	169935	7567	11444
D-Mannose 6-phosphate	8969	1	35419	114088	40176	1	27439	1	41603	13533	29840	1	1	44126	10400	1
Dihydroxyacetone phosphate	1	29206	22413	191209	21096	1	23609	1	1	1	4013	1	1	53487	1	1
a-N-Acetylglucosamine	1	1	1	8477	1	1	1	1	8285	8124	1	1	1	8202	1	1
N-Acetyl-DL-serine	267193	109833	150007	101758	119872	215604	207582	186052	140090	176672	183125	166533	188559	183457	222128	247464
Linoleic acid	14059	19087	16661	42002	15802	15811	23173	22093	33321	15774	14762	20432	26726	70706	15682	16625
L-Anserine	23065	9103	9280	25784	13020	16788	21182	21669	18627	16612	21861	16405	20501	18328	22659	22332
D-Fructose 6-phosphate	8969	1	35419	114088	40176	1	27439	1	41603	13533	29840	1	1	44126	10400	1
Maleic acid	26932	33022	30552	24647	54659	61853	48042	36833	27130	31288	1	48684	38852	37116	31762	52326
Pimelic acid	1	1	18312	1	1	6219	36522	1	1	1	5040	1	15589	11247	1	7799
Suberic acid	16537	13107	96177	27733	22775	24653	168704	19923	17174	23032	29458	19621	69454	49335	24327	25998
tartaric acid	1	21326	37919	114442	27532	1	1	97576	227109	32080	37255	48418	39909	112178	1	1
D-Ribose	278479	163289	181334	201718	245345	187892	206866	235213	199127	244772	322068	224985	216133	197387	244992	346072
?-D-Glucose 1-phosphate	8969	1	35419	114088	40176	1	27439	1	41603	13533	29840	1	1	44126	10400	1
Xanthine	515121	273122	462125	591375	568278	459478	486384	409252	383343	340912	465477	466259	446330	508223	430102	353640
Glyceryl phosphate	265991	241780	395647	495986	461139	273197	286019	400200	365513	263601	347804	376373	197980	481055	324324	301046
L-Norleucine	25849	18253	10342	47403	34682	29152	37252	31102	28552	26649	32009	31082	24954	14800	35507	36122
L-Serine	26769	60674	49098	26684	37131	32851	29564	29476	24545	33000	31137	37984	19663	30071	42374	43469
Xanthosine	6821	1	1	6878	6115	5969	1	4520	1	3227	4512	3259	1	8864	2405	3736
D-(+)-Xylose	278483	163289	181334	201718	245345	187892	206847	235213	199127	244772	322068	224985	216133	197387	244992	346072
N-Acetyl-?-D-mannosamine	1	1	1	8477	1	1	1	1	8285	8124	1	1	1	8202	1	1
DL-Homoserine	48560	117283	95799	43820	63163	62046	66332	63431	55995	54300	63861	64302	33153	52984	78782	87400
D-(+)-Malic acid	803962	533285	1030614	3495055	1979574	927578	1802824	870857	974210	805193	1368476	1972900	1076367	2200004	1297429	1483643
L-Phenylalanine	69288	35148	43927	82162	66328	57555	71332	63719	60992	48777	75370	67295	50312	70100	62896	76246
3-hydroxy-3-methyl-Glutaric acid	21702	12301	19098	21721	31997	23921	26413	21315	26070	13292	27946	39954	30935	22679	31467	18824
L-Methionine	49274	27827	43066	51075	46460	49064	51330	56934	50853	33160	52568	48958	34571	44007	50786	51083
?-D-Glucose	394687	425412	285885	199750	66344	52674	26196	23025	180623	249120	229114	30677	195056	31615	27162	28018
Nicotinamide adenine dinucleotide (NAD)	1	1	2493	2273	1	1	1	1	1	1	1	1	1	11755	1	1
Elaidic Acid	53739	61404	52523	90076	55377	55596	67188	65363	76034	60907	53296	61659	60182	112295	56588	57842
N-Acetyl-L-alanine	20352	11111	13453	22539	19069	16985	20914	16604	16445	16714	20535	19519	20102	21077	19894	18621
cis-9-palmitoleic acid	5747	10898	9344	16623	6601	9606	34882	6988	9842	7983	3897	7980	11279	39558	10665	9455
trans-Aconitate	226505	151786	190795	231769	401135	203920	234898	247130	309868	198857	322855	457119	253854	363646	305449	255774
Citramalic acid	44627	6957	18579	62722	65274	12274	21415	76314	51444	12967	49801	30311	67405	12040	17359	73762
4-Hydroxybenzaldehyde	1	1	1	12621	7171	5541	9988	1	1	5060	1	1	1	14714	1	1
?-Alanine	217770	147203	282444	994577	496849	251892	302044	244631	240431	210810	285484	376666	175021	407808	299488	233228
Glyoxylic acid	62670	1	74766	90064	82327	83204	90348	77549	75664	98154	1	87415	84347	133381	1	76553
Inosine 5'-monophosphate (IMP)	1	1	1	406858	1	1	1	1	1	1	1	1	13549	24727	1	1
Taurine	194490	250695	371728	1546039	688593	260088	333158	184756	223268	217557	300648	375004	160310	372510	308488	236042
D-Glucuronic acid	118644	62984	90324	131243	125282	103792	99224	174578	121976	192105	132867	107278	122458	171074	127769	115361
L-Asparagine	1	7766	9648	1	1	6892	1	1	1	1	1	1	1	6815	1	8595
D-(+)-GALACTURONIC ACID	118644	62984	90324	131243	125282	103792	99224	174578	121976	192105	132867	107278	122458	171074	127769	115361
Urocanic acid	376708	311405	263657	172839	329815	369087	381179	197665	163466	323606	385710	330231	318293	209005	429552	455551
D-Glucose 6-phosphate	8969	1	35419	114088	40176	1	27439	1	41603	13533	29840	1	1	44126	10400	1
Retinoic acid	27649	33858	36904	26932	29124	28424	42014	26929	29583	27271	27320	26764	28312	34435	27159	30553
Allantoin	41924	38990	58030	52013	50770	46315	37962	74725	61883	61628	47138	44588	46072	67122	48610	41878
L-Lysine	23126	22649	20571	19611	17418	23171	20338	25572	26806	26777	19345	22067	27243	16559	12984	32910
Allothreonine	48560	117283	95799	43820	63163	62046	66332	63431	55995	54300	63861	64302	33153	52984	78782	87400
Citrulline	10971	8183	9961	12245	10089	10483	13429	13102	12108	11845	14161	15230	1	13065	13840	38570
D-Tryptophan	30486	16706	26212	26666	26745	24586	38620	30869	33063	24125	40285	32731	14159	29862	32632	43392
Isocitrate	1273843	466382	684960	176292	1004184	962509	1098103	826469	817903	930663	1735094	872804	1267581	4399441	1147744	1591957
Glycerol 2-phosphate	265991	241780	395647	495986	461139	273197	286019	400200	365513	263601	347804	376373	197980	481055	324324	301046
3-Hydroxybenzaldehyde	1	1	1	12621	7169	5541	9988	1	1	5060	1	1	1	14714	1	1
D-Sorbitol	55303	27062	29867	46732	41499	45039	35229	54540	44443	40408	148242	47346	32609	41290	43610	32190
?-D-Galactose 1-phosphate	8969	1	35419	114088	40176	1	27439	1	41603	13533	29840	1	1	44126	10400	1
Uridine	543288	298498	401553	629180	519181	560797	565114	407336	385598	421635	479753	440789	458179	509094	462782	500018
2-hydroxy-butanoic acid	24167	75119	27120	68574	99794	27155	48896	23890	52315	36367	58498	50304	52586	100570	27376	39177
L-Norvaline	10052	6257	1	14132	11076	8181	10652	10916	1	7899	11645	10374	7996	10694	12122	12024
L-Leucine	25862	18253	10342	47403	34682	29152	37252	31107	28548	26649	32004	31076	24951	14800	35515	36114
Lauric acid	1	1	1	1	1	1	2240	1	1	1	1	1	1	1516	1	1
L-Arginine	23747	18352	20518	23524	24526	23225	28969	28619	29030	25920	32885	30850	27638	30997	28829	34027
Fumaric acid	26932	33022	30552	24647	54659	61853	48042	36833	27144	31288	1	48684	38852	37116	31762	52326
Stearic acid	16607	17329	16673	22043	41516	18105	20340	20345	23894	18521	19043	16731	16896	25601	19541	19006
L-Tyrosine	786327	439203	521239	771238	638155	648572	808583	70103	75230	52644	85508	77829	62135	84383	106267	104767
(4Z,7Z,10Z,13Z,16Z,19Z)-4,7,10,13,1 6,19-Docosahexaenoic acid	1961	1	1	77214	3410	1802	1408	14093	31980	7374	1963	12102	4205	42372	1943	1191
N-acetylaspartate	350364	224128	452949	263134	315184	491038	737232	228688	747485	311765	402382	917572	132306	292135	502995	403900
3-Methylglutaric acid	9009	1	43272	20497	11036	17297	105525	11153	9282	11231	16915	10864	32357	31038	11722	23134
Pyridoxal (Vitamin B6)	13568	1	7430	1	6059	15287	12185	9538	11241	11280	12208	12249	12249	10656	15230	9066
N-Acetylneuraminic Acid	1589609	1325414	1561745	1695347	1216490	1470361	1325846	1882922	1770237	1742253	1465367	1492001	1513459	1658889	1535410	1356779
D-(-)-Arabinose	278483	163289	181334	201718	245345	187892	206850	235213	199127	244772	322068	224985	216133	197387	244992	346072
Malonic acid	93926	62826	77042	57132	75838	89558	96383	88775	70995	93374	105914	144004	111805	79206	124228	125665
2-Methylglutaric acid	9009	1	43272	20497	11036	17297	105525	11153	9282	11231	16915	10864	32357	31038	11722	23134
Adenine	228682	121071	169742	108467	146409	284558	286644	171133	153601	133866	175851	180113	169274	180367	306238	208266
Pyroglutamic acid	498716	353848	333422	717282	469822	361549	458484	541186	422830	421049	595883	638384	445500	542838	640164	857606
Uracil	154499	71139	102309	124731	154816	131300	120897	123079	84890	101380	136212	106810	33813	94038	112612	111689
L-Valine	10052	6257	1	14132	11076	8181	10652	10916	1	7899	11645	10374	7996	10694	12122	12024
N-ALPHA-ACETYL-L-ASPARAGINE	48715	12955	24533	13692	23663	31060	28527	27991	20208	25837	29334	18646	35170	27583	30558	45534
L-Histidine	20001	16163	1	14508	1	15375	18081	13711	1	14642	14867	1	16310	1	17940	21019
Acetyl-L-tyrosine	4251	1	1	4043	2609	3569	4342	1	1	1	1	1	1	1	1	1
Guanosine	37039	24135	11759	11384	18974	39053	46198	20091	11075	39468	38564	46700	29335	1	49793	40580
NTBC HT-1	25255	14303	24763	37774	13604	14287	18481	1	1	1	1	1	1	1	13511	1
2,5-Dihydroxybenzaldehyde	3881	1569	1	1	1	1	6601	1	1	3815	4940	1	1	1	1	1
3-(2-Hydroxyphenyl)propionic acid	1	3484	1	1	5703	1	5940	1	1	1	1	1	1	1	1	1
4-Pyridoxic acid	1	4073	1	1	6881	6583	7297	1	1	1	7319	8552	1	1	1	1
Deoxyuridine monophosphate (dUMP)	1	1	1	1	1	1	14468	1	1	1	1	1	1	1	14193	1
O-Phosphorylethanolamine	1	1	12569	1	15125	1	1	9354	16998	1	1	1	6300	1	7999	1
Glucuronolactone	1	1	1	1	1	1	1	1	1	130315	1	1	1	1	1	1
Quinic acid	1	1	1	1	1	1	1	1	1	1	31959	1	1	1	1	32017
Methylglyoxal	1	1	1	1	1	1	1	1	1	1	43922	1	1	1	1	1
Glutamyl-Tyrosine	1	1	1	1	1	1	1	1	1	1	1	1	1	1	2400	1
?,?-Trehalose	1	1	1	1	1	1	1	1	1	1	1	1	13131	1	1	1
Phenyl acetate	1	1	1524	1	1	3102	1	1	1	1	1	1	1	1	1	1
L-Ornithine	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	11604
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	Mass	Retention Time	CAS Number
Inosine	268.0816	2.9830005	58-63-9
Adipic acid	146.058	4.6969995	124-04-9
Sarcosine	89.0481	1.367	107-97-1
Ethylmalonic acid	132.0422	3.9659998	601-75-2
D-(-)-Lyxose	150.0531	1.5060003	1114-34-7
Hippuric acid	179.0588	5.6160007	495-69-2
Uridine monophosphate (UMP)	324.0359	1.71	58-97-9
Methyl jasmonate	224.1413	10.360998	39924-52-2
Dulcitol	182.0792	1.351	608-66-2
Galactaric acid	210.0381	1.356	526-99-8
meso-Tartaric acid	150.0169	1.4540001	147-73-9
Hypoxanthine	136.0389	2.1690001	68-94-0
Phosphocreatine	211.0373	1.389	67-07-2
Thymidine	242.0904	3.9710004	50-89-5
Orotic acid	156.0172	1.7539998	65-86-1
Uric acid	168.0287	2.004	69-93-2
Gluconic acid	196.0592	1.353	526-95-4
Aceturic acid	117.0428	1.699	543-24-8
?-Linolenic Acid	278.2244	13.231001	506-26-3
L-Tryptophan	204.09	4.61	73-22-3
Cytidine	243.0858	1.531	65-46-3
L-Threonine	119.0581	1.3150003	72-19-5
4-Hydroxyphenyllactic acid	182.0585	4.8720007	6482-98-0
Malic acid	134.0219	1.6189996	6915-15-7
L-Isoleucine	131.0951	2.657	73-32-5
N-Acetyl-L-glutamic acid	189.0643	2.274	1188-37-0
D-Mannitol	182.0792	1.351	69-65-8
L-Aspartic Acid	133.0375	1.545	56-84-8
Myristic acid	228.2093	13.173002	544-63-8
phosphoenol pyruvate	167.983	1.5760001	138-08-9
Oxoglutaric acid	146.022	1.7859998	328-50-7
Citric acid	192.0279	1.8859998	77-92-9
Succinic acid	118.0266	2.326	110-15-6
Methyl N-(a-methylbutyryl)glycine	188.1053	8.199999	123-99-9
D-Aspartic acid	133.0375	1.545	1783-96-6
12-Hydroxydodecanoic acid	216.173	10.632001	505-95-3
Pantothenic Acid	219.1115	4.354	137-08-6
Dehydroascorbic acid	174.0167	1.6079999	490-83-5
Glutamine	146.0695	1.3	56-85-9
Arabinose	150.0531	1.5060003	147-81-9
Palmitic acid	256.2402	13.606998	57-10-3
Itaconic acid	130.0269	3.2890005	97-65-4
DL-a-Hydroxybutyric acid	104.0474	3.1549997	600-15-7
Adenosine5'-monophosphate	347.0635	1.7999997	149022-20-8
D-Mannose 6-phosphate	260.0264	1.3529999	3672-15-9
Dihydroxyacetone phosphate	169.9981	1.3599999	57-04-5
a-N-Acetylglucosamine	221.0901	1.469	7512-17-6
N-Acetyl-DL-serine	147.0536	1.691	94-14-3
Linoleic acid	280.2404	13.445002	60-33-3
L-Anserine	240.1221	1.286	10030-52-1
D-Fructose 6-phosphate	260.0264	1.3529999	643-13-0
Maleic acid	116.0112	2.1790004	110-16-7
Pimelic acid	160.074	5.986	111-16-0
Suberic acid	174.0893	7.18	505-48-6
tartaric acid	150.0169	1.4540001	87-69-4
D-Ribose	150.0531	1.5060003	50-69-1
?-D-Glucose 1-phosphate	260.0264	1.3529999	59-56-3
Xanthine	152.034	2.3800006	69-89-6
Glyceryl phosphate	172.0143	1.3689997	57-03-4
L-Norleucine	131.0951	2.657	327-57-1
L-Serine	105.0424	1.2889998	56-45-1
Xanthosine	284.0761	3.539	146-80-5
D-(+)-Xylose	150.0531	1.5060003	58-86-6
N-Acetyl-?-D-mannosamine	221.0901	1.469	7772-94-3
DL-Homoserine	119.0581	1.3150003	1927-25-9
D-(+)-Malic acid	134.0219	1.6189996	636-61-3
L-Phenylalanine	165.0794	3.813	63-91-2
3-hydroxy-3-methyl-Glutaric acid	162.0534	2.78	503-49-1
L-Methionine	149.0513	1.8570002	63-68-3
?-D-Glucose	180.0634	1.332	492-62-6
Nicotinamide adenine dinucleotide (NAD)	663.1075	1.861	53-84-9
Elaidic Acid	282.256	13.690998	112-79-8
N-Acetyl-L-alanine	131.0584	2.6139998	97-69-8
cis-9-palmitoleic acid	254.2249	13.328999	373-49-9
trans-Aconitate	174.0167	2.292	4023-65-8
Citramalic acid	148.0377	2.4090002	2306-22-1
4-Hydroxybenzaldehyde	122.0371	5.944	123-08-0
?-Alanine	89.0481	1.368	107-95-9
Glyoxylic acid	74.0007	1.457	298-12-4
Inosine 5'-monophosphate (IMP)	348.0472	1.919	131-99-7
Taurine	125.0148	1.293	107-35-7
D-Glucuronic acid	194.043	1.34	03-12-6556
L-Asparagine	132.053	1.289	70-47-3
D-(+)-GALACTURONIC ACID	194.043	1.34	91510-62-2
Urocanic acid	138.0421	1.413	104-98-3
D-Glucose 6-phosphate	260.0264	1.3529999	56-73-5
Retinoic acid	300.209	13.020003	302-79-4
Allantoin	158.0441	1.3929998	97-59-6
L-Lysine	146.1053	1.1099999	56-87-1
Allothreonine	119.0581	1.3150003	144-98-9
Citrulline	175.0961	1.3190001	372-75-8
D-Tryptophan	204.09	4.61	153-94-6
Isocitrate	192.0279	1.6079999	320-77-4
Glycerol 2-phosphate	172.0143	1.3689997	17181-54-3
3-Hydroxybenzaldehyde	122.0371	5.944	100-83-4
D-Sorbitol	182.0792	1.351	50-70-4
?-D-Galactose 1-phosphate	260.0264	1.3529999	2255-14-3
Uridine	244.0703	2.2700002	58-96-8
2-hydroxy-butanoic acid	104.0474	3.1549997	600-15-7
L-Norvaline	117.0794	1.6730002	6600-40-4
L-Leucine	131.0951	2.657	61-90-5
Lauric acid	200.1774	12.625	143-07-7
L-Arginine	174.1115	1.2679999	74-79-3
Fumaric acid	116.0112	2.1790004	110-17-8
Stearic acid	284.2713	13.988999	57-11-4
L-Tyrosine	181.0743	2.2229998	60-18-4
(4Z,7Z,10Z,13Z,16Z,19Z)-4,7,10,13,1 6,19-Docosahexaenoic acid	328.2402	13.384002	6217-54-5
N-acetylaspartate	175.0484	1.8700004	997-55-7
3-Methylglutaric acid	146.058	4.6969995	626-51-7
Pyridoxal (Vitamin B6)	167.0586	1.743	66-72-8
N-Acetylneuraminic Acid	309.107	1.4570001	131-48-6
D-(-)-Arabinose	150.0531	1.5060003	10323-20-3
Malonic acid	104.011	1.691	141-82-2
2-Methylglutaric acid	146.058	4.6969995	617-62-9
Adenine	135.0546	1.7339997	73-24-5
Pyroglutamic acid	129.0431	2.114	98-79-3
Uracil	112.0274	1.8559995	66-22-8
L-Valine	117.0794	1.6730002	72-18-4
N-ALPHA-ACETYL-L-ASPARAGINE	174.0645	1.6329999	4033-40-3
L-Histidine	155.0691	1.2869998	71-00-1
Acetyl-L-tyrosine	223.085	5.041	537-55-3
Guanosine	283.092	3.0289996	118-00-3
NTBC HT-1	329.0511	10.38	104206-65-7
2,5-Dihydroxybenzaldehyde	138.0322	5.407	1194-98-5
3-(2-Hydroxyphenyl)propionic acid	166.0629	6.913	495-78-3
4-Pyridoxic acid	183.0534	3.268	82-82-6
Deoxyuridine monophosphate (dUMP)	308.0422	1.933	964-26-1
O-Phosphorylethanolamine	141.0193	1.288	1071-23-4
Glucuronolactone	176.0326	1.519	32449-92-6
Quinic acid	192.0637	1.492	77-95-2
Methylglyoxal	72.0214	1.936	78-98-8
Glutamyl-Tyrosine	310.1196	3.893	9046-27-9
?,?-Trehalose	342.1077	1.3	99-20-7
Phenyl acetate	136.0524	4.87	122-79-2
L-Ornithine	132.0896	1.319	3184-13-2
METABOLITES_END
#END