#METABOLOMICS WORKBENCH babiodun_20241031_133624 DATATRACK_ID:5329 STUDY_ID:ST003577 ANALYSIS_ID:AN005873 PROJECT_ID:PR002207
VERSION             	1
CREATED_ON             	November 15, 2024, 10:19 am
#PROJECT
PR:PROJECT_TITLE                 	Multi-omics indicate depth-discrete partitioning of nitrogen metabolism in a
PR:PROJECT_TITLE                 	toxic Planktothrix rubescens bloom in the winter water column
PR:PROJECT_TYPE                  	Multiomics characterization of Meads Quarry Water after Harmful Algae Bloom
PR:PROJECT_SUMMARY               	This project attempts to: 1. Identify the dominant cyanobacteria bloom former in
PR:PROJECT_SUMMARY               	a local lake in Knoxville (Meads Quarry) Tennessee using metagenomics. 2. Use a
PR:PROJECT_SUMMARY               	complementary technique of metabolomics to understand the metabolism within this
PR:PROJECT_SUMMARY               	cyanobacteria 3. Identify how different physiochemical parameters affect the
PR:PROJECT_SUMMARY               	metabolism of the cyanobacteria identified across various limniological
PR:PROJECT_SUMMARY               	stratification
PR:INSTITUTE                     	University of Tennessee
PR:DEPARTMENT                    	Microbiology
PR:LABORATORY                    	Steve Wilhelm
PR:LAST_NAME                     	Abiodun
PR:FIRST_NAME                    	Blessing
PR:ADDRESS                       	652 Buehler Dabney Hall, Knoxville
PR:EMAIL                         	babiodun@vols.utk.edu
PR:PHONE                         	8653349978
#STUDY
ST:STUDY_TITLE                   	Multi ‘omics indicate depth-discrete partitioning of nitrogen metabolism in a
ST:STUDY_TITLE                   	toxic Planktothrix rubescens bloom in the winter water column
ST:STUDY_TYPE                    	Metabolomics
ST:STUDY_SUMMARY                 	Limnological sampling efforts have traditionally focused on the summer water
ST:STUDY_SUMMARY                 	column of larger lakes. For example, hundreds of researchers visit prominent
ST:STUDY_SUMMARY                 	systems like the Laurentian Great Lakes to investigate summer cyanobacterial
ST:STUDY_SUMMARY                 	blooms. Concurrently, the recent advancement of meta-omics techniques has led to
ST:STUDY_SUMMARY                 	a surge in molecular studies within these systems. However, these approaches
ST:STUDY_SUMMARY                 	have created several knowledge gaps: smaller, regional lakes remain largely
ST:STUDY_SUMMARY                 	unstudied, the winter ecology of lakes is poorly understood, and meta-omics
ST:STUDY_SUMMARY                 	techniques often fail to make meaningful scientific contributions when used in
ST:STUDY_SUMMARY                 	isolation. To address these gaps, we conducted a combined metagenomic and
ST:STUDY_SUMMARY                 	metabolomic field study focused on a local community concern: the winter pink
ST:STUDY_SUMMARY                 	discoloration of Mead’s Quarry (Knoxville, TN). This case study was designed
ST:STUDY_SUMMARY                 	and executed by seven graduate students at the University of Tennessee-Knoxville
ST:STUDY_SUMMARY                 	enrolled in the course MICR 669: Advanced Techniques in Field Microbiology
ST:STUDY_SUMMARY                 	(Spring 2023). Utilizing a $3,000 budget, the team carried out in-depth
ST:STUDY_SUMMARY                 	meta-omics research. Our metagenomic analyses and phylogenetic confirmations
ST:STUDY_SUMMARY                 	identified a bloom of the pink cyanobacterium Planktothrix rubescens as the
ST:STUDY_SUMMARY                 	primary cause. Physiochemical measurements revealed that the bloom was
ST:STUDY_SUMMARY                 	concentrated in the light-limited (<1 μmol photons m⁻² s⁻¹) metalimnion
ST:STUDY_SUMMARY                 	of the cold (<14°C) water column. Additionally, microcystin-LR levels exceeded
ST:STUDY_SUMMARY                 	the U.S. EPA limit for recreational exposure (8 μg L⁻¹). Untargeted
ST:STUDY_SUMMARY                 	metabolomics indicated that P. rubescens communities were differentially
ST:STUDY_SUMMARY                 	partitioning nitrogen metabolism strategies by depth (0.5 m vs. 2.0 m). This
ST:STUDY_SUMMARY                 	study highlights the previously underappreciated roles of arginine biosynthesis
ST:STUDY_SUMMARY                 	and pyrimidine metabolism in the ecological success of P. rubescens. We propose
ST:STUDY_SUMMARY                 	a novel ecological hypothesis regarding the proliferation of this harmful
ST:STUDY_SUMMARY                 	cyanobacterial bloom former in the light-limited metalimnion. Overall, our
ST:STUDY_SUMMARY                 	findings suggest that meaningful meta-omics research can be conducted locally
ST:STUDY_SUMMARY                 	with limited funding as part of a course-based curriculum involving at least
ST:STUDY_SUMMARY                 	seven graduate students. Furthermore, we demonstrate that the synergistic use of
ST:STUDY_SUMMARY                 	multiple meta-omics techniques is feasible on a small scale while yielding
ST:STUDY_SUMMARY                 	significant scientific impact.
ST:INSTITUTE                     	University of Tennessee
ST:DEPARTMENT                    	Microbiology
ST:LABORATORY                    	Steve Wilhelm
ST:LAST_NAME                     	Abiodun
ST:FIRST_NAME                    	Blessing
ST:ADDRESS                       	652 Buehler Dabney Hall, Knoxville
ST:EMAIL                         	babiodun@vols.utk.edu
ST:PHONE                         	8653349978
ST:NUM_GROUPS                    	3
ST:TOTAL_SUBJECTS                	12
#SUBJECT
SU:SUBJECT_TYPE                  	Water sample
#FACTORS
#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           	D1	D1_R1	Sample source:Water | Depth:0.5m	RAW_FILE_NAME=D1_R1.mzml
SUBJECT_SAMPLE_FACTORS           	D1	D1_R2	Sample source:Water | Depth:0.5m	RAW_FILE_NAME=D1_R2.mzml
SUBJECT_SAMPLE_FACTORS           	D1	D1_R3	Sample source:Water | Depth:0.5m	RAW_FILE_NAME=D1_R3.mzml
SUBJECT_SAMPLE_FACTORS           	D1	D1_R4	Sample source:Water | Depth:0.5m	RAW_FILE_NAME=D1_R4.mzml
SUBJECT_SAMPLE_FACTORS           	D2	D2_R1	Sample source:Water | Depth:2.0m	RAW_FILE_NAME=D2_R1.mzml
SUBJECT_SAMPLE_FACTORS           	D2	D2_R2	Sample source:Water | Depth:2.0m	RAW_FILE_NAME=D2_R2.mzml
SUBJECT_SAMPLE_FACTORS           	D2	D2_R3	Sample source:Water | Depth:2.0m	RAW_FILE_NAME=D2_R3.mzml
SUBJECT_SAMPLE_FACTORS           	D2	D2_R4	Sample source:Water | Depth:2.0m	RAW_FILE_NAME=D2_R4.mzml
SUBJECT_SAMPLE_FACTORS           	D3	D3_R1	Sample source:Water | Depth:6.0m	RAW_FILE_NAME=D3_R1.mzml
SUBJECT_SAMPLE_FACTORS           	D3	D3_R2	Sample source:Water | Depth:6.0m	RAW_FILE_NAME=D3_R2.mzml
SUBJECT_SAMPLE_FACTORS           	D3	D3_R3	Sample source:Water | Depth:6.0m	RAW_FILE_NAME=D3_R3.mzml
SUBJECT_SAMPLE_FACTORS           	D3	D3_R4	Sample source:Water | Depth:6.0m	RAW_FILE_NAME=D3_R4.mzml
#COLLECTION
CO:COLLECTION_SUMMARY            	Water sampling for metabolomics analysis was conducted using a standardized
CO:COLLECTION_SUMMARY            	filtration method to ensure clean and precise sample collection. We prepared
CO:COLLECTION_SUMMARY            	materials, including a 0.2 µm filter, syringes, swinnex filter holders, and
CO:COLLECTION_SUMMARY            	pre-labeled cryovials. After rinsing the sampling tripod and ensuring the
CO:COLLECTION_SUMMARY            	collection bucket was thoroughly mixed, we collected water samples. Each sample
CO:COLLECTION_SUMMARY            	was filtered by assembling the swinnex with the filter and pushing 120 mL
CO:COLLECTION_SUMMARY            	through the apparatus, recording the total volume filtered until saturation.
CO:COLLECTION_SUMMARY            	After filtration, we carefully removed the filter, folded it to preserve the
CO:COLLECTION_SUMMARY            	cellular material, and placed it into a pre-labeled cryovial. The samples were
CO:COLLECTION_SUMMARY            	then stored in liquid nitrogen, and subsequently at -80°C, until extraction.
CO:COLLECTION_SUMMARY            	This process minimized contamination and optimized sample integrity for
CO:COLLECTION_SUMMARY            	downstream metabolomics analysis.
CO:SAMPLE_TYPE                   	Water
CO:STORAGE_CONDITIONS            	-80℃
#TREATMENT
TR:TREATMENT_SUMMARY             	No treatment
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	The flash-frozen filters were placed in empty petri dishes and allowed to thaw
SP:SAMPLEPREP_SUMMARY            	gradually at 4°C. Once thawed, the filters were unfolded with the cell side
SP:SAMPLEPREP_SUMMARY            	facing down, and metabolites were extracted following established protocols (Lu
SP:SAMPLEPREP_SUMMARY            	et al., 2010; Rabinowitz & Kimball, 2007). Specifically, 1.5 mL of an extraction
SP:SAMPLEPREP_SUMMARY            	solvent (40% methanol, 20% water, 40% acetonitrile, and 0.1 M formic acid) was
SP:SAMPLEPREP_SUMMARY            	added to fully immerse each filter. The petri dishes were then chilled at -20°C
SP:SAMPLEPREP_SUMMARY            	for 20 minutes to enhance metabolite release. After chilling, the extraction
SP:SAMPLEPREP_SUMMARY            	solvent containing the released metabolites was transferred to microcentrifuge
SP:SAMPLEPREP_SUMMARY            	tubes. The filters were turned cell-side up, and any remaining cells were
SP:SAMPLEPREP_SUMMARY            	extracted with an additional 400 µL of solvent, which was also transferred to
SP:SAMPLEPREP_SUMMARY            	the tubes. The tubes were vortexed and centrifuged at 15,000 rpm for 5 minutes
SP:SAMPLEPREP_SUMMARY            	at 4°C to separate the metabolite-rich supernatant from the pellet. The
SP:SAMPLEPREP_SUMMARY            	supernatants were carefully transferred to new microcentrifuge tubes and dried
SP:SAMPLEPREP_SUMMARY            	under nitrogen gas. Once dried, the samples were reconstituted in 300 µL of
SP:SAMPLEPREP_SUMMARY            	LC-MS grade water, vortexed, and centrifuged again at 15,000 rpm for 5 minutes
SP:SAMPLEPREP_SUMMARY            	at 4°C to ensure uniformity. The final aliquots were then placed into
SP:SAMPLEPREP_SUMMARY            	autosampler vials for UHPLC-HRMS analysis.
SP:PROCESSING_STORAGE_CONDITIONS 	4℃
SP:EXTRACT_STORAGE               	4℃
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Thermo Dionex Ultimate 3000
CH:COLUMN_NAME                   	Phenomenex Synergi Hydro-RP (100 x 2mm,2.5um)
CH:SOLVENT_A                     	97% water/3% methanol; 11mM tributylamine; 15 mM acetic acid
CH:SOLVENT_B                     	100% methanol
CH:FLOW_GRADIENT                 	0 min, 0% B; 2.5 min, 0% B; 5 min, 20% B; 7.5 min, 20% B; 13 min, 55% B; 15.5
CH:FLOW_GRADIENT                 	min, 95% B; 18.5 min, 95% B; 19 min, 0% B; 25 min, 0% B
CH:FLOW_RATE                     	0.2 mL/min
CH:COLUMN_TEMPERATURE            	25
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Thermo Q Exactive Orbitrap
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	NEGATIVE
MS:MS_COMMENTS                   	Raw mass spectral files were converted to mzML files using a package from
MS:MS_COMMENTS                   	ProteoWizard, msConverter. All mzML files were imported into an open-source
MS:MS_COMMENTS                   	software, metabolomics analysis and visualization engine (El-MAVEN) where
MS:MS_COMMENTS                   	metabolites were manually identified using an in-house library based on exact
MS:MS_COMMENTS                   	mass (±5 ppm) and retention time (±2 min)
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	Peak area
MS_METABOLITE_DATA_START
Samples	D1_R1	D1_R2	D1_R3	D1_R4	D2_R1	D2_R2	D2_R3	D2_R4	D3_R1	D3_R2	D3_R3	D3_R4
Factors	Sample source:Water | Depth:0.5m	Sample source:Water | Depth:0.5m	Sample source:Water | Depth:0.5m	Sample source:Water | Depth:0.5m	Sample source:Water | Depth:2.0m	Sample source:Water | Depth:2.0m	Sample source:Water | Depth:2.0m	Sample source:Water | Depth:2.0m	Sample source:Water | Depth:6.0m	Sample source:Water | Depth:6.0m	Sample source:Water | Depth:6.0m	Sample source:Water | Depth:6.0m
Creatinine	5.09E+03	1.28E+04	1.32E+06	2.79E+04	6.47E+04	1.04E+06	1.00E+02	2.15E+03	1.00E+02	1.00E+02	1.00E+02	6.17E+03
N-Carbamoyl-L-aspartate	5.46E+04	5.55E+04	1.91E+04	2.82E+04	5.40E+04	1.26E+06	1.38E+05	5.21E+04	1.85E+04	1.00E+04	8.50E+03	1.59E+03
Indole	1.00E+02	1.00E+02	1.00E+02	1.18E+03	1.00E+02	1.47E+05	6.12E+03	6.88E+03	1.00E+02	1.00E+02	8.64E+02	1.00E+02
Glutamate	1.24E+07	8.71E+06	7.51E+06	8.83E+06	1.90E+07	2.57E+08	4.13E+07	3.88E+07	5.76E+06	4.38E+06	3.06E+06	8.85E+05
Glutamine	3.17E+05	1.87E+05	2.61E+05	2.02E+05	1.83E+05	1.33E+06	2.76E+05	2.84E+05	1.33E+05	1.66E+05	6.57E+04	2.88E+04
Glutathione	2.26E+05	4.15E+05	9.74E+04	1.00E+02	7.88E+04	1.17E+05	8.46E+03	2.88E+05	1.00E+02	1.00E+02	1.00E+02	1.00E+02
Glutathione disulfide	2.04E+06	1.64E+06	8.85E+05	1.08E+06	1.59E+06	2.97E+07	3.55E+06	4.23E+06	2.70E+05	2.53E+05	1.53E+05	1.00E+02
Homoserine/Threonine	6.70E+05	7.48E+05	6.30E+05	6.09E+05	7.81E+05	4.93E+06	1.14E+06	1.79E+06	6.63E+05	5.80E+05	4.93E+05	5.38E+05
Methionine	1.00E+02	3.76E+03	3.86E+03	8.92E+03	3.53E+04	6.35E+05	3.27E+04	9.42E+04	4.29E+03	7.00E+03	1.00E+02	1.00E+02
Ophthalmate	2.10E+04	1.58E+04	9.76E+03	1.36E+04	1.54E+04	6.18E+05	6.68E+04	7.72E+04	2.42E+03	1.87E+03	1.09E+03	1.00E+02
phosphothreonine	2.59E+04	3.57E+04	5.11E+03	1.10E+04	9.32E+03	4.84E+05	2.23E+04	2.76E+04	1.00E+02	6.93E+02	1.00E+02	1.00E+02
Aspartate	3.77E+06	4.86E+06	2.52E+06	2.87E+06	5.13E+06	9.45E+07	9.46E+06	8.37E+06	2.10E+06	2.04E+06	1.19E+06	1.24E+06
Lysine	5.50E+04	6.14E+04	5.55E+04	5.98E+04	9.33E+04	9.95E+05	1.40E+05	2.78E+05	2.91E+04	5.70E+04	1.43E+04	5.07E+03
Ornithine	1.20E+05	2.07E+05	1.94E+05	7.59E+04	1.11E+05	1.59E+06	3.54E+05	7.15E+05	1.11E+05	1.66E+05	4.97E+04	1.36E+05
Phenylalanine	2.79E+05	2.47E+05	2.99E+05	2.44E+05	3.89E+05	3.13E+06	5.71E+05	8.36E+05	2.44E+05	2.61E+05	1.51E+05	1.67E+05
Tyrosine	4.19E+05	3.19E+05	2.96E+05	2.99E+05	4.22E+05	3.80E+06	5.62E+05	8.47E+05	2.09E+05	2.07E+05	1.42E+05	1.43E+05
Valine/betaine	8.00E+05	9.58E+05	8.84E+05	7.63E+05	1.14E+06	7.54E+06	1.68E+06	2.03E+06	9.71E+05	7.82E+05	7.19E+05	7.44E+05
Alanine/Sarcosine	2.57E+05	3.07E+05	2.59E+05	1.98E+05	4.10E+05	4.38E+06	9.50E+05	1.03E+06	2.59E+05	2.37E+05	1.83E+05	1.67E+05
Citrulline	9.01E+04	1.25E+05	7.16E+04	7.24E+04	1.12E+05	1.78E+06	2.25E+05	3.33E+05	8.20E+04	6.92E+04	7.03E+03	1.62E+04
Leucine/Isoleucine	2.29E+06	1.06E+06	1.15E+06	9.56E+05	2.26E+06	2.98E+07	3.84E+06	6.94E+06	1.22E+06	1.11E+06	1.29E+06	1.05E+06
N-Acetylglutamate	1.60E+06	1.20E+06	8.63E+05	1.02E+06	2.67E+06	1.57E+07	5.16E+06	3.66E+06	9.49E+05	8.71E+05	7.42E+05	3.65E+05
Allantoate	1.70E+04	1.59E+04	1.33E+04	1.42E+04	2.64E+04	6.27E+05	7.12E+04	9.42E+04	1.82E+03	1.00E+02	1.00E+02	1.00E+02
Tryptophan	3.75E+04	7.21E+04	1.77E+05	1.25E+05	6.18E+05	1.22E+07	1.27E+06	1.69E+06	1.10E+05	1.16E+05	9.46E+04	4.27E+03
Arginine	2.36E+05	3.08E+05	2.36E+05	2.54E+05	4.08E+05	5.69E+06	1.08E+06	1.06E+06	1.31E+05	1.72E+05	7.65E+04	4.30E+04
Glycine	1.62E+06	1.77E+06	1.54E+06	1.53E+06	1.68E+06	3.74E+06	2.01E+06	2.44E+06	1.70E+06	1.54E+06	1.49E+06	1.46E+06
Taurine	7.97E+03	1.15E+04	3.75E+05	9.78E+03	2.23E+04	8.99E+04	1.88E+04	6.79E+04	6.77E+04	1.51E+04	4.66E+03	2.37E+04
Trehalose/Sucrose	5.58E+05	4.44E+05	3.34E+05	3.98E+05	6.92E+05	6.72E+06	1.69E+06	1.26E+06	2.05E+05	1.90E+05	9.27E+04	4.81E+04
Fructose 1_6-bisphosphate	2.29E+05	2.14E+05	4.15E+04	1.46E+05	7.23E+04	3.36E+06	4.70E+05	4.29E+05	4.58E+04	4.39E+04	3.99E+04	6.56E+03
Glucose phosphate	1.52E+06	1.81E+06	6.12E+05	5.55E+05	1.14E+06	1.63E+07	3.13E+06	2.61E+06	3.14E+05	1.71E+05	2.02E+05	2.55E+04
Glycerone phosphate	7.77E+04	2.87E+04	4.44E+03	8.14E+04	7.22E+04	1.81E+06	2.46E+05	1.52E+05	1.46E+03	1.76E+03	1.25E+03	1.00E+02
Phosphoenolpyruvate	1.61E+05	8.11E+04	3.03E+04	2.76E+04	1.43E+05	1.80E+06	2.63E+05	1.70E+05	6.63E+03	7.18E+03	1.00E+02	1.00E+02
Pyruvate	6.50E+06	6.69E+06	6.17E+06	6.24E+06	8.56E+06	5.38E+07	1.24E+07	1.07E+07	7.44E+06	6.72E+06	6.55E+06	6.59E+06
Lactate	1.35E+08	1.35E+08	1.28E+08	1.12E+08	1.29E+08	5.07E+08	1.65E+08	2.31E+08	1.35E+08	1.33E+08	1.13E+08	1.29E+08
3-Phosphoglycerate	1.57E+06	1.66E+06	4.19E+05	4.91E+05	1.28E+06	3.08E+07	3.21E+06	2.73E+06	5.24E+04	4.10E+04	1.09E+04	3.11E+03
aminocaproic acid	2.37E+06	1.11E+06	1.21E+06	1.02E+06	2.32E+06	3.00E+07	3.89E+06	7.02E+06	1.27E+06	1.17E+06	1.33E+06	1.11E+06
2-Isopropylmalate	1.60E+06	1.79E+06	1.71E+06	1.45E+06	1.55E+06	4.97E+06	2.16E+06	2.34E+06	1.50E+06	1.46E+06	1.53E+06	1.44E+06
Cytidine	1.00E+02	3.11E+03	3.25E+03	3.18E+03	2.78E+04	8.45E+05	8.61E+04	1.00E+05	8.16E+03	4.27E+03	1.00E+02	1.00E+02
dCMP	7.48E+02	1.00E+02	1.76E+03	1.00E+02	2.92E+03	6.69E+04	7.65E+03	1.64E+03	1.00E+02	8.92E+02	8.31E+02	8.08E+02
1-Methyladenosine	1.51E+03	1.00E+02	1.00E+02	1.00E+02	1.85E+03	8.23E+04	5.97E+03	1.63E+03	1.00E+02	1.00E+02	1.00E+02	1.62E+03
dTMP	1.95E+03	1.41E+03	2.27E+03	1.07E+03	1.13E+04	2.90E+05	2.94E+04	3.17E+04	2.36E+03	1.76E+03	1.00E+02	1.00E+02
FAD	9.61E+03	4.68E+03	2.20E+03	1.73E+03	4.41E+03	2.37E+05	1.52E+04	1.82E+04	1.00E+02	1.00E+02	1.00E+02	1.00E+02
GDP	2.88E+04	3.03E+04	1.50E+03	1.31E+03	1.00E+02	3.54E+05	1.72E+03	8.24E+03	9.92E+02	1.00E+02	1.20E+03	1.00E+02
Guanosine	1.65E+04	3.38E+04	6.00E+04	3.67E+04	8.83E+04	1.14E+06	1.75E+05	1.71E+05	1.54E+04	3.32E+04	1.10E+04	1.00E+02
Adenosine	3.07E+04	1.04E+04	2.14E+04	3.83E+04	7.80E+04	1.01E+06	1.90E+05	9.14E+04	2.95E+03	1.06E+04	3.74E+03	1.00E+02
Inosine	7.48E+06	5.28E+06	3.84E+06	5.24E+06	9.49E+06	9.02E+06	1.64E+07	7.44E+06	2.03E+06	1.73E+06	1.20E+06	5.67E+04
Thymidine	1.00E+02	1.00E+02	4.79E+03	6.63E+03	1.97E+04	5.07E+05	5.55E+04	6.45E+04	1.34E+04	3.50E+03	1.68E+03	1.00E+02
UDP	6.02E+04	3.35E+04	1.00E+02	3.17E+03	1.22E+03	3.65E+05	7.08E+03	1.59E+04	2.86E+03	1.00E+02	1.00E+02	1.00E+02
UDP-glucose	6.43E+04	2.84E+04	1.20E+04	4.13E+04	2.09E+04	6.48E+05	1.87E+05	1.01E+04	6.46E+03	3.54E+03	1.03E+03	1.00E+02
UDP-glucuronate	1.02E+05	9.33E+04	1.27E+04	1.56E+04	1.45E+03	1.78E+05	2.03E+04	3.37E+03	6.89E+03	1.51E+03	2.09E+03	1.00E+02
UMP	3.09E+05	4.49E+05	1.94E+05	2.18E+05	1.65E+05	3.61E+06	3.88E+05	5.43E+05	5.00E+04	2.48E+04	3.13E+04	1.00E+02
Uridine	9.43E+04	7.22E+04	1.40E+05	1.75E+05	3.59E+05	5.75E+06	7.91E+05	8.15E+05	1.47E+05	1.10E+05	9.00E+04	1.00E+02
Xanthosine	1.00E+02	1.00E+02	1.39E+03	2.57E+03	9.45E+03	3.90E+05	4.73E+04	5.81E+04	6.48E+03	1.00E+02	1.00E+02	1.00E+02
Xanthosine 5'-phosphate	2.14E+04	2.06E+04	4.02E+04	3.29E+04	7.63E+04	2.11E+06	1.73E+05	3.51E+05	1.65E+04	1.58E+04	4.08E+03	1.38E+03
FMN	4.81E+03	5.99E+03	2.76E+03	1.00E+02	5.59E+03	3.61E+05	2.60E+04	1.47E+04	1.00E+02	1.00E+02	1.00E+02	1.00E+02
ADP	4.03E+05	5.73E+05	1.43E+05	1.86E+05	2.44E+05	8.24E+06	6.05E+05	1.06E+06	9.03E+04	6.78E+04	7.69E+04	5.00E+04
CMP	5.90E+04	8.33E+04	2.66E+04	3.23E+04	8.62E+04	1.61E+06	2.16E+05	3.19E+05	8.44E+03	6.47E+03	4.03E+03	1.00E+02
GMP	1.04E+05	1.38E+05	1.75E+04	2.14E+04	8.96E+03	4.73E+05	4.85E+04	2.64E+04	2.63E+03	1.00E+02	1.00E+03	1.00E+02
IMP	5.27E+04	4.92E+04	9.04E+03	2.25E+04	2.57E+04	1.25E+06	1.11E+05	1.86E+05	1.00E+02	9.04E+02	1.03E+03	1.00E+02
NAD+	4.39E+04	3.81E+04	3.71E+03	1.13E+04	7.93E+03	5.29E+05	2.91E+04	7.64E+04	1.00E+02	1.00E+02	1.00E+02	1.00E+02
NADP+	1.21E+05	1.76E+05	1.46E+04	3.81E+04	7.24E+04	2.12E+06	8.51E+04	3.28E+05	2.40E+03	1.00E+02	3.33E+03	1.00E+02
S-Adenosyl-L-homocysteine	2.93E+03	2.44E+03	1.87E+03	2.99E+03	5.00E+03	3.25E+05	1.51E+04	3.84E+04	1.00E+02	1.00E+02	1.00E+02	1.00E+02
UDP-N-acetylglucosamine	3.88E+05	3.14E+05	1.02E+05	1.55E+05	9.24E+04	1.63E+06	2.24E+05	2.49E+05	2.61E+04	1.22E+04	1.29E+04	1.00E+02
AMP/dGMP	1.37E+06	1.48E+06	6.15E+05	7.33E+05	7.92E+05	1.60E+07	2.40E+06	2.77E+06	1.66E+05	9.81E+04	7.67E+04	1.00E+02
Guanine	3.83E+04	1.16E+05	7.06E+04	1.23E+04	2.25E+03	2.27E+05	9.20E+03	5.48E+04	1.42E+04	5.98E+03	4.30E+03	1.00E+02
Orotate	9.49E+04	1.14E+05	9.61E+04	8.60E+04	1.30E+05	2.17E+06	3.25E+05	1.42E+05	8.37E+04	4.15E+04	4.80E+04	6.04E+04
ATP	4.88E+04	5.12E+04	3.45E+04	2.89E+04	1.03E+04	1.08E+06	6.96E+04	1.87E+05	2.41E+04	1.26E+04	3.34E+04	1.62E+03
Xanthine	5.06E+04	9.64E+04	2.32E+05	1.99E+05	5.61E+05	1.01E+07	1.37E+06	1.96E+06	2.12E+05	1.21E+05	5.14E+04	1.83E+04
CDP	1.00E+02	1.06E+03	1.00E+02	1.00E+02	1.26E+03	3.61E+05	1.16E+04	2.05E+04	1.43E+03	1.00E+02	1.00E+02	1.00E+02
Allantoin	1.99E+03	2.29E+03	2.12E+04	1.00E+02	2.56E+03	4.69E+04	9.90E+03	7.03E+03	1.00E+02	2.21E+03	1.00E+02	2.16E+03
Uric acid	4.37E+04	5.24E+04	9.46E+05	8.05E+04	2.34E+05	2.60E+06	2.13E+05	2.62E+05	2.83E+03	3.61E+04	7.11E+03	1.00E+02
Uracil	1.84E+05	2.13E+05	2.01E+05	1.58E+05	3.96E+05	3.42E+06	5.83E+05	8.12E+05	2.96E+05	1.99E+05	1.52E+05	2.04E+05
2-Dehydro-D-gluconate	6.41E+04	4.90E+04	2.04E+05	6.97E+04	7.63E+04	4.02E+05	4.31E+04	6.91E+04	6.59E+04	4.36E+04	3.70E+04	6.36E+04
D-Gluconate	7.13E+05	6.58E+05	1.65E+06	7.42E+05	7.16E+05	4.75E+06	7.53E+05	1.38E+06	7.06E+05	7.19E+05	5.91E+05	6.82E+05
alpha-Ketoglutarate	2.80E+06	2.46E+06	3.04E+06	3.14E+06	6.40E+06	9.37E+07	1.17E+07	1.39E+07	3.37E+06	3.38E+06	2.81E+06	2.05E+06
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name
Creatinine
N-Carbamoyl-L-aspartate
Indole
Glutamate
Glutamine
Glutathione
Glutathione disulfide
Homoserine/Threonine
Methionine
Ophthalmate
phosphothreonine
Aspartate
Lysine
Ornithine
Phenylalanine
Tyrosine
Valine/betaine
Alanine/Sarcosine
Citrulline
Leucine/Isoleucine
N-Acetylglutamate
Allantoate
Tryptophan
Arginine
Glycine
Taurine
Trehalose/Sucrose
Fructose 1_6-bisphosphate
Glucose phosphate
Glycerone phosphate
Phosphoenolpyruvate
Pyruvate
Lactate
3-Phosphoglycerate
aminocaproic acid
2-Isopropylmalate
Cytidine
dCMP
1-Methyladenosine
dTMP
FAD
GDP
Guanosine
Adenosine
Inosine
Thymidine
UDP
UDP-glucose
UDP-glucuronate
UMP
Uridine
Xanthosine
Xanthosine 5'-phosphate
FMN
ADP
CMP
GMP
IMP
NAD+
NADP+
S-Adenosyl-L-homocysteine
UDP-N-acetylglucosamine
AMP/dGMP
Guanine
Orotate
ATP
Xanthine
CDP
Allantoin
Uric acid
Uracil
2-Dehydro-D-gluconate
D-Gluconate
alpha-Ketoglutarate
METABOLITES_END
#END