#METABOLOMICS WORKBENCH Sethjparker_20241101_174808 DATATRACK_ID:5335 STUDY_ID:ST003557 ANALYSIS_ID:AN005848 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 NEGATIVE 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 PIPECOLATE 3267190.42 2459096.856 4772595.805 3744580.555 2913888.843 2997165.679 2962459.076 1132528.111 3519034.469 4329782.56 1345416.866 773795.0803 1702220.903 2231057.066 752997.2457 1499956.584 835180.7627 2064595.059 1718924.692 1167865.234 1630229.174 4488683.268 1762488.697 4003763.632 8502440.425 2708743.334 3432169.022 6072395.359 3935664.047 470388.1395 351046.9878 876103.5673 795581.5598 516244.9966 581338.0982 352103.8412 412174.5298 620664.1561 747377.1841 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Mode m/z Retention Time (min) HMDB ID PIPECOLATE Negative 128.0717 9.32 HMDB0000716 METABOLITES_END #END