#METABOLOMICS WORKBENCH Sethjparker_20241101_174808 DATATRACK_ID:5335 STUDY_ID:ST003557 ANALYSIS_ID:AN005847 PROJECT_ID:PR002191
VERSION             	1
CREATED_ON             	November 4, 2024, 12:56 pm
#PROJECT
PR:PROJECT_TITLE                 	N-methyl-arginine is taken up into the liver, brain, and kidney of mice.
PR:PROJECT_TYPE                  	Manuscript
PR:PROJECT_SUMMARY               	N-methyl-arginine was identified as a transported substrate through multiple
PR:PROJECT_SUMMARY               	cationic amino acid transporters, which facilitate the uptake of lysine,
PR:PROJECT_SUMMARY               	arginine, and other structurally related amino acids. In cell culture,
PR:PROJECT_SUMMARY               	N-methyl-arginine inhibits the uptake and metabolism of lysine. In this project,
PR:PROJECT_SUMMARY               	we aimed to determine if N-methyl-arginine administered to mice at 80 mg/kg was
PR:PROJECT_SUMMARY               	sufficient to inhibit tissue lysine uptake and metabolism. Following a single
PR:PROJECT_SUMMARY               	injection, N-methyl-arginine significantly accumulated in mouse plasma and
PR:PROJECT_SUMMARY               	tissues but failed to significantly inhibit the level of lysine metabolites
PR:PROJECT_SUMMARY               	(e.g., saccharopine). These results are in-line with in vitro results suggesting
PR:PROJECT_SUMMARY               	that N-methyl-arginine is a relatively low affinity transport inhibitor.
PR:INSTITUTE                     	University of British Columbia
PR:DEPARTMENT                    	Biochemistry & Molecular Biology
PR:LABORATORY                    	Parker laboratory
PR:LAST_NAME                     	Parker
PR:FIRST_NAME                    	Seth
PR:ADDRESS                       	950 W 28th Ave, Vancouver, British Columbia, V6H 0B3, Canada
PR:EMAIL                         	seth.parker@bcchr.ca
PR:PHONE                         	6048753121
#STUDY
ST:STUDY_TITLE                   	Metabolomics analysis of kidney, brain, liver, and plasma from Aldh7a1-/- mice
ST:STUDY_TITLE                   	administered PBS or N-methyl-arginine (80 mg/kg) via a single intraperitoneal
ST:STUDY_TITLE                   	injection.
ST:STUDY_SUMMARY                 	Metabolite analysis of kidney, brain, liver, and plasma isolated from Aldh7a1-/-
ST:STUDY_SUMMARY                 	mice fed diets consisting of 0.9% w/w lysine and 18 ppm pyridoxine. Tissues were
ST:STUDY_SUMMARY                 	cryo-homogenized using a liquid nitrogen cooled mortar and pestle into a fine
ST:STUDY_SUMMARY                 	powder and approximately 25-50 mg of each tissue was transferred to a
ST:STUDY_SUMMARY                 	pre-weighed homogenization tube to which ~20 2.3 mm zirconia beads and 1 mL of
ST:STUDY_SUMMARY                 	80% ice cold methanol were added. Tissues were homogenized using a bead mill and
ST:STUDY_SUMMARY                 	cleared by centrifugation at 4C, where each of the 3-4 cycles consisted of a 60
ST:STUDY_SUMMARY                 	second homogenization at 6 m/s followed by a 30 second pause. A volume
ST:STUDY_SUMMARY                 	equivalent to 25 mg of tissue extract was transferred to a new tube, and 250 ul
ST:STUDY_SUMMARY                 	of 80% methanol containing 1 ul of stable isotope-labeled internal amino acid
ST:STUDY_SUMMARY                 	standard mix was added. Metabolites were concentrated using a SpeedVac until
ST:STUDY_SUMMARY                 	dry. For plasma metabolite analysis, 10 µl of plasma was added to 250 µl of
ST:STUDY_SUMMARY                 	80% methanol containing 1 µl of stable isotope-labeled internal amino acid
ST:STUDY_SUMMARY                 	standard mix, vortexed at 4C for 10 minutes, and centrifuged. 0.9 ml of
ST:STUDY_SUMMARY                 	supernatant was transferred to a new tube and concentrated using a SpeedVac
ST:STUDY_SUMMARY                 	until dry. Metabolites were reconstituted into 25-50 µl of water, vortexed,
ST:STUDY_SUMMARY                 	centrifuged, and transferred to vials for analysis by LCMS. LCMS was performed
ST:STUDY_SUMMARY                 	using a ZIC-pHILIC LC column coupled to a Vanquish LC and a flow gradient
ST:STUDY_SUMMARY                 	consisting of 10 mM ammonium carbonate in water and pure acetonitrile. The LC
ST:STUDY_SUMMARY                 	was coupled to an Exploris 240 mass spectrometer operated in a polarity
ST:STUDY_SUMMARY                 	switching data-dependent Top 5 mode. Full MS scan parameters for both positive
ST:STUDY_SUMMARY                 	and negative mode were set to 67-1000 m/z at a resolution of 120k and ddMS2 were
ST:STUDY_SUMMARY                 	collected at a resolution of 30k.
ST:INSTITUTE                     	University of British Columbia
ST:DEPARTMENT                    	Biochemistry & Molecular Biology
ST:LABORATORY                    	Parker laboratory
ST:LAST_NAME                     	Parker
ST:FIRST_NAME                    	Seth
ST:ADDRESS                       	950 W 28th Ave, Vancouver, British Columbia, V6H 0B3, Canada
ST:EMAIL                         	seth.parker@bcchr.ca
ST:PHONE                         	6048753121
#SUBJECT
SU:SUBJECT_TYPE                  	Mammal
SU:SUBJECT_SPECIES               	Mus musculus
SU:TAXONOMY_ID                   	10090
SU:GENDER                        	Male and female
SU:ANIMAL_FEED                   	0.9% w/w lysine, 18 ppm pyridoxine (standard diets)
#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           	-	1	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Brain	Sex=M; Age (weeks)=8.4; RAW_FILE_NAME(Raw file name)=1.RAW
SUBJECT_SAMPLE_FACTORS           	-	2	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Brain	Sex=M; Age (weeks)=8.4; RAW_FILE_NAME(Raw file name)=2.RAW
SUBJECT_SAMPLE_FACTORS           	-	3	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Brain	Sex=F; Age (weeks)=15.3; RAW_FILE_NAME(Raw file name)=3.RAW
SUBJECT_SAMPLE_FACTORS           	-	4	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Brain	Sex=F; Age (weeks)=15.3; RAW_FILE_NAME(Raw file name)=4.RAW
SUBJECT_SAMPLE_FACTORS           	-	5	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Brain	Sex=M; Age (weeks)=21.4; RAW_FILE_NAME(Raw file name)=5.RAW
SUBJECT_SAMPLE_FACTORS           	-	6	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Brain	Sex=M; Age (weeks)=21.4; RAW_FILE_NAME(Raw file name)=6.RAW
SUBJECT_SAMPLE_FACTORS           	-	7	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Brain	Sex=M; Age (weeks)=32.9; RAW_FILE_NAME(Raw file name)=7.RAW
SUBJECT_SAMPLE_FACTORS           	-	8	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Brain	Sex=M; Age (weeks)=32.9; RAW_FILE_NAME(Raw file name)=8.RAW
SUBJECT_SAMPLE_FACTORS           	-	9	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Brain	Sex=F; Age (weeks)=5.9; RAW_FILE_NAME(Raw file name)=9.RAW
SUBJECT_SAMPLE_FACTORS           	-	10	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Brain	Sex=F; Age (weeks)=5.9; RAW_FILE_NAME(Raw file name)=10.RAW
SUBJECT_SAMPLE_FACTORS           	-	11	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Liver	Sex=M; Age (weeks)=8.4; RAW_FILE_NAME(Raw file name)=11.RAW
SUBJECT_SAMPLE_FACTORS           	-	12	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Liver	Sex=M; Age (weeks)=8.4; RAW_FILE_NAME(Raw file name)=12.RAW
SUBJECT_SAMPLE_FACTORS           	-	13	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Liver	Sex=F; Age (weeks)=15.3; RAW_FILE_NAME(Raw file name)=13.RAW
SUBJECT_SAMPLE_FACTORS           	-	14	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Liver	Sex=F; Age (weeks)=15.3; RAW_FILE_NAME(Raw file name)=14.RAW
SUBJECT_SAMPLE_FACTORS           	-	15	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Liver	Sex=M; Age (weeks)=21.4; RAW_FILE_NAME(Raw file name)=15.RAW
SUBJECT_SAMPLE_FACTORS           	-	16	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Liver	Sex=M; Age (weeks)=21.4; RAW_FILE_NAME(Raw file name)=16.RAW
SUBJECT_SAMPLE_FACTORS           	-	17	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Liver	Sex=M; Age (weeks)=32.9; RAW_FILE_NAME(Raw file name)=17.RAW
SUBJECT_SAMPLE_FACTORS           	-	18	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Liver	Sex=M; Age (weeks)=32.9; RAW_FILE_NAME(Raw file name)=18.RAW
SUBJECT_SAMPLE_FACTORS           	-	19	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Liver	Sex=F; Age (weeks)=5.9; RAW_FILE_NAME(Raw file name)=19.RAW
SUBJECT_SAMPLE_FACTORS           	-	20	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Liver	Sex=F; Age (weeks)=5.9; RAW_FILE_NAME(Raw file name)=20.RAW
SUBJECT_SAMPLE_FACTORS           	-	21	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Kidney	Sex=M; Age (weeks)=8.4; RAW_FILE_NAME(Raw file name)=21.RAW
SUBJECT_SAMPLE_FACTORS           	-	22	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Kidney	Sex=M; Age (weeks)=8.4; RAW_FILE_NAME(Raw file name)=22.RAW
SUBJECT_SAMPLE_FACTORS           	-	23	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Kidney	Sex=F; Age (weeks)=15.3; RAW_FILE_NAME(Raw file name)=23.RAW
SUBJECT_SAMPLE_FACTORS           	-	24	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Kidney	Sex=F; Age (weeks)=15.3; RAW_FILE_NAME(Raw file name)=24.RAW
SUBJECT_SAMPLE_FACTORS           	-	25	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Kidney	Sex=M; Age (weeks)=21.4; RAW_FILE_NAME(Raw file name)=25.RAW
SUBJECT_SAMPLE_FACTORS           	-	26	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Kidney	Sex=M; Age (weeks)=21.4; RAW_FILE_NAME(Raw file name)=26.RAW
SUBJECT_SAMPLE_FACTORS           	-	27	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Kidney	Sex=M; Age (weeks)=32.9; RAW_FILE_NAME(Raw file name)=27.RAW
SUBJECT_SAMPLE_FACTORS           	-	28	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Kidney	Sex=M; Age (weeks)=32.9; RAW_FILE_NAME(Raw file name)=28.RAW
SUBJECT_SAMPLE_FACTORS           	-	29	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Kidney	Sex=F; Age (weeks)=5.9; RAW_FILE_NAME(Raw file name)=29.RAW
SUBJECT_SAMPLE_FACTORS           	-	31	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Plasma	Sex=M; Age (weeks)=8.4; RAW_FILE_NAME(Raw file name)=31.RAW
SUBJECT_SAMPLE_FACTORS           	-	32	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Plasma	Sex=M; Age (weeks)=8.4; RAW_FILE_NAME(Raw file name)=32.RAW
SUBJECT_SAMPLE_FACTORS           	-	33	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Plasma	Sex=F; Age (weeks)=15.3; RAW_FILE_NAME(Raw file name)=33.RAW
SUBJECT_SAMPLE_FACTORS           	-	34	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Plasma	Sex=F; Age (weeks)=15.3; RAW_FILE_NAME(Raw file name)=34.RAW
SUBJECT_SAMPLE_FACTORS           	-	35	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Plasma	Sex=M; Age (weeks)=21.4; RAW_FILE_NAME(Raw file name)=35.RAW
SUBJECT_SAMPLE_FACTORS           	-	36	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Plasma	Sex=M; Age (weeks)=21.4; RAW_FILE_NAME(Raw file name)=36.RAW
SUBJECT_SAMPLE_FACTORS           	-	37	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Plasma	Sex=M; Age (weeks)=32.9; RAW_FILE_NAME(Raw file name)=37.RAW
SUBJECT_SAMPLE_FACTORS           	-	38	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Plasma	Sex=M; Age (weeks)=32.9; RAW_FILE_NAME(Raw file name)=38.RAW
SUBJECT_SAMPLE_FACTORS           	-	39	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Plasma	Sex=F; Age (weeks)=5.9; RAW_FILE_NAME(Raw file name)=39.RAW
SUBJECT_SAMPLE_FACTORS           	-	40	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Plasma	Sex=F; Age (weeks)=5.9; RAW_FILE_NAME(Raw file name)=40.RAW
#COLLECTION
CO:COLLECTION_SUMMARY            	Liver, brain, kidney, or plasma samples collected from mice treated with PBS or
CO:COLLECTION_SUMMARY            	N-methyl-arginine (80 mg/kg) as a single intraperitoneal injection. Tissues
CO:COLLECTION_SUMMARY            	collected 7-hours after injection.
CO:SAMPLE_TYPE                   	Tissue or Plasma
#TREATMENT
TR:TREATMENT_SUMMARY             	Mice were administered a single intraperitoneal injection containing either
TR:TREATMENT_SUMMARY             	saline or N-methyl-arginine (80 mg/kg). 7-hours after the injection, the mice
TR:TREATMENT_SUMMARY             	were euthanized by cardiac puncture, and tissues were isolated for analysis by
TR:TREATMENT_SUMMARY             	LCMS.
TR:TREATMENT_COMPOUND            	N-methyl-arginine
TR:TREATMENT_ROUTE               	Intraperitoneal
TR:TREATMENT_DOSE                	80 mg/kg
TR:TREATMENT_VEHICLE             	PBS
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Dried samples were reconstituted in 50 µL of HPLC-grade water. Samples were
SP:SAMPLEPREP_SUMMARY            	vortexed for ~10 minutes, then centrifuged at 21,000 x g for 15 min at 4°C. 40
SP:SAMPLEPREP_SUMMARY            	microliters were transferred to LC vials containing glass inserts for analysis.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_TYPE           	HILIC
CH:INSTRUMENT_NAME               	Thermo Vanquish
CH:COLUMN_NAME                   	Merck SeQuant ZIC-pHILIC (150 x 2.1mm,5um)
CH:SOLVENT_A                     	100% water, 10 mM Ammonium Carbonate, pH 9.0
CH:SOLVENT_B                     	100% acetonitrile
CH:FLOW_GRADIENT                 	80-20%B (0-30 min), 20-20%B (30-40 minute), and 20-80%B (40-40.5 minute); the LC
CH:FLOW_GRADIENT                 	column was re-equilibrated using 80-80%B from 40.5-52 minute before subsequent
CH:FLOW_GRADIENT                 	injections
CH:FLOW_RATE                     	100 uL/min
CH:COLUMN_TEMPERATURE            	25
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Thermo Exploris 240
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	POSITIVE
MS:MS_COMMENTS                   	The LC was coupled to a Thermo Scientific Exploris 240 mass spectrometer
MS:MS_COMMENTS                   	operating in heated electrospray ionization mode (HESI) for analysis. The
MS:MS_COMMENTS                   	following parameters were set for HESI: spray voltage 3.4 kV (positive) and 2 kV
MS:MS_COMMENTS                   	(negative), static spray voltage, sheath gas 25, aux gas 5, sweep gas 0.5, ion
MS:MS_COMMENTS                   	transfer tube temperature 320oC, and vaporizer temperature 75oC. The global
MS:MS_COMMENTS                   	parameters included an expected peak width of 20 seconds, mild trapping, and a
MS:MS_COMMENTS                   	default charge state of 1. A 40-min polarity switching data-dependent Top 5
MS:MS_COMMENTS                   	method was used for positive mode and a data-dependent Top 3 method was used for
MS:MS_COMMENTS                   	negative mode. Full MS scan parameters for both positive and negative modes were
MS:MS_COMMENTS                   	set as follows: scan range 67-1000 m/z collected in profile mode, Orbitrap
MS:MS_COMMENTS                   	resolution 120,000, RF lens 70%, AGC target of 300%, and maximum injection time
MS:MS_COMMENTS                   	set to automatic. ddMS2 for positive mode were collected in centroid mode at an
MS:MS_COMMENTS                   	Orbitrap resolution of 30,000, isolation window of 1.5 m/z, an AGC target set to
MS:MS_COMMENTS                   	standard, a maximum injection time set to automatic, and a normalized collision
MS:MS_COMMENTS                   	energy set to 10%, 30%, and 80%. ddMS2 for negative mode were collected in
MS:MS_COMMENTS                   	centroid mode at an Orbitrap resolution of 30,000, isolation window of 2 m/z, an
MS:MS_COMMENTS                   	AGC target set to standard, a maximum injection time set to automatic, and a
MS:MS_COMMENTS                   	normalized collision energy set to 30%. For both positive and negative ddMS2, we
MS:MS_COMMENTS                   	applied an intensity threshold of 5e4 and a dynamic exclusion of 5 ppm for 10
MS:MS_COMMENTS                   	seconds, excluding isotopes. A targeted selected ion monitoring (tSIM) scan was
MS:MS_COMMENTS                   	also included for pipecolate and P6C/P2C, and the retention time ranges were
MS:MS_COMMENTS                   	based on elution of authentic standards (pipecolate) or from positive samples
MS:MS_COMMENTS                   	(Aldh7a1-deficient tissues). The tSIM scan for pipecolate was collected from
MS:MS_COMMENTS                   	8-12 minutes in negative mode at an isolation window of 4 m/z (for metabolomics)
MS:MS_COMMENTS                   	or 18 m/z (for isotope-tracing experiments, to include m/z shifts), an Orbitrap
MS:MS_COMMENTS                   	resolution of 120,000, a RF lens at 70%, an automatic maximum injection time,
MS:MS_COMMENTS                   	and collected in profile mode. The tSIM scan for P6C/P2C was collected from 6-10
MS:MS_COMMENTS                   	minutes in positive mode at an isolation window of 4 m/z (for metabolomics) or
MS:MS_COMMENTS                   	18 m/z (for isotope-tracing experiments, to include m/z shifts), an Orbitrap
MS:MS_COMMENTS                   	resolution of 120,000, a RF lens at 70%, an automatic maximum injection time,
MS:MS_COMMENTS                   	and collected in profile mode.
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	Ion counts
MS_METABOLITE_DATA_START
Samples	1	3	5	7	9	2	4	6	8	10	11	13	15	17	19	12	14	16	18	20	21	23	25	27	29	22	24	26	28	31	33	35	37	39	32	34	36	38	40
Factors	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Brain	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Brain	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Brain	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Brain	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Brain	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Brain	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Brain	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Brain	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Brain	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Brain	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Liver	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Liver	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Liver	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Liver	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Liver	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Liver	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Liver	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Liver	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Liver	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Liver	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Kidney	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Kidney	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Kidney	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Kidney	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Kidney	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Kidney	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Kidney	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Kidney	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Kidney	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Plasma	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Plasma	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Plasma	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Plasma	Genotype:Aldh7a1-/- | Treatment:PBS | Sample source:Plasma	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Plasma	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Plasma	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Plasma	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Plasma	Genotype:Aldh7a1-/- | Treatment:N-methyl-arginine (80 mg/kg) | Sample source:Plasma
LYSINE	7546210.27	2259226.905	6024695.419	7944601.07	9387464.877	6841485.916	5809087.472	694764.2985	7234934.124	7347104.09	13460609.69	10898705.72	7810508.255	6417777.88	19834829.81	10477405.22	15821591.19	18913657.11	12482848.4	6495606.079	13457375.41	13049268.9	8199074.389	11355735.6	5613912.612	15466547.8	12227643.36	2744713.702	8505054.952	7700105.181	2219457.644	3507230.763	2287961.064	99016.1827	3438476.982	102660.2022	4271052.756	1832563.356	3760624.191
SACCHAROPINE	925216.7201	952805.0365	1201715.897	1057473.894	1327349.514	1770386.555	784474.0558	542497.2663	781032.3051	1857121.866	2839519.681	1266117.429	1322223.303	654868.9517	2237486.319	3343912.207	4426710.243	5038074.3	2181537.841	3048595.825	2543169.974	2137491.014	2284319.338	2680219.524	1795280.565	3363221.744	1645679.221	3584547.492	2903134.373	98764.70034	19903.81588	21358.70493	13370.18181	30880.36224	25006.59916	18290.81814	21571.09852	19081.81633	22241.02637
PIPERIDEINE 6-CARBOXYLATE (P6C)	4376418.24	5316705.228	3291530.834	4433116.009	7286092.584	2408849.097	4607338.423	1541657.901	6433322.324	4563928.176	3607206.578	2425546.035	1389685.536	2160681.849	4633378.401	3825004.951	2832931.229	3896055.485	3855844.161	3873090.042	1630229.174	4488683.268	1762488.697	4003763.632	8502440.425	2708743.334	3432169.022	6072395.359	3935664.047	7306099.458	3837529.707	6670191.214	4589707.285	8814347.843	8417972.804	7883629.639	2941763.299	5180551.551	4274569.006
AMINOADIPATE	515977.3967	790312.1645	651313.2598	733737.8086	1070590.669	625812.4851	475280.5337	165790.118	733981.509	654939.0753	2093103.703	1680927.713	660961.7698	681368.6827	6245335.488	4043977.901	3051283.146	5338636.848	1546020.807	5147725.871	3558999.811	5629440.102	2483683.121	2436884.46	7781106.635	8856573.451	3624758.259	2346584.834	2989986.758	358955.5148	332062.1302	405685.1764	215369.5889	667832.9733	647797.8824	286331.6005	337886.5262	283378.7694	439503.6365
NG-METHYL-ARGININE	251827.5662	172270.7422	205892.8916	254848.5867	326780.6727	371231.0304	268826.5391	284185.7235	441177.6328	883400.9922	1371606.966	1145957.909	684140.1755	751846.2347	2074979.969	3444418.176	2086465.688	2411545.828	2890552.629	7084131.18	1331575.427	1439355.702	874599.6526	1551695.998	1482600.48	2731629.474	1715630.564	1483329.774	2206892.236	334270.5886	278164.8449	262560.9507	157854.7304	680615.0368	1113365.621	467282.3679	709699.0189	972886.3619	2684356.854
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	Mode	m/z	Retention Time (min)	HMDB ID
LYSINE	Positive	147.113353	26.05	HMDB0000182
SACCHAROPINE	Positive	277.13901	13.74	HMDB0000279
PIPERIDEINE 6-CARBOXYLATE (P6C)	Positive	128.07061	8.43	HMDB0012130
AMINOADIPATE	Positive	162.0761	12.8	HMDB0000510
NG-METHYL-ARGININE	Positive	189.1346	26.1	HMDB0000182
METABOLITES_END
#END