#METABOLOMICS WORKBENCH montewillis_20171030_110750 DATATRACK_ID:1259 STUDY_ID:ST000924 ANALYSIS_ID:AN001517 VERSION 1 CREATED_ON 02-08-2024 #PROJECT PR:PROJECT_TITLE MuRF1-Related Metabolic Alterations in HL-1 Cardiomyocyte Induced by Cyclic PR:PROJECT_TITLE Stretch PR:PROJECT_TYPE Cardiomyocyte cell culture PR:PROJECT_SUMMARY We collected cell media and performed GC-MS non-targeted metabolomics to PR:PROJECT_SUMMARY identify the role of MuRF1 in the dynamic metabolic changes in cardiomyocytes. PR:INSTITUTE University of North Carolina at Chapel Hill PR:DEPARTMENT Pathology & Laboratory Medicine PR:LABORATORY Willis PR:LAST_NAME Willis PR:FIRST_NAME Monte PR:ADDRESS 111 Mason Farm Road PR:EMAIL monte_willis@med.unc.edu PR:PHONE 9849995431 PR:FUNDING_SOURCE National Institutes of Health R01HL104129 PR:DOI http://dx.doi.org/10.21228/M8Z40Z PR:CONTRIBUTORS Wei Tang, PhD1,*, Jessica E. Rodríguez, PhD2,*, Amro Ilaiwy, MD3,4, James R. PR:CONTRIBUTORS Bain, PhD3,4, Michael J. Muehlbauer, PhD3, Megan T. Quintana, MD5, Alexa PR:CONTRIBUTORS Hartman, BS6, Cam Patterson, MD1,7, Janet Rubin, MD8, Monte S. Willis, MD, PR:CONTRIBUTORS PhD1,2,9 #STUDY ST:STUDY_TITLE MuRF1-Related Metabolic Alterations in HL-1 Cardiomyocyte Induced by Cyclic ST:STUDY_TITLE Stretch ST:STUDY_TYPE Cardiomyocyte cell culture ST:STUDY_SUMMARY We collected cell media and performed GC-MS non-targeted metabolomics to ST:STUDY_SUMMARY identify the role of MuRF1 in the dynamic metabolic changes in cardiomyocytes. ST:INSTITUTE University of North Carolina at Chapel Hill ST:DEPARTMENT Pathology & Laboratory Medicine ST:LABORATORY Willis ST:LAST_NAME Willis ST:FIRST_NAME Monte ST:ADDRESS 111 Mason Farm Road ST:EMAIL monte_willis@med.unc.edu ST:PHONE 9849995431 ST:SUBMIT_DATE 2017-10-30 #SUBJECT SU:SUBJECT_TYPE Cell culture SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:CELL_STRAIN_DETAILS C57BL/6 SU:CELL_PRIMARY_IMMORTALIZED AT1 SU:CELL_COUNTS ~60% confluency 6 well plate #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - 18 type:Ad.shMuRF1 | Stretch Cycle:15 min cyclic stretch SUBJECT_SAMPLE_FACTORS - 30 type:Ad.shMuRF1 | Stretch Cycle:15 min cyclic stretch SUBJECT_SAMPLE_FACTORS - 6 type:Ad.shMuRF1 | Stretch Cycle:15 min cyclic stretch SUBJECT_SAMPLE_FACTORS - 19 type:Ad.shMuRF1 | Stretch Cycle:30 min cyclic stretch SUBJECT_SAMPLE_FACTORS - 31 type:Ad.shMuRF1 | Stretch Cycle:30 min cyclic stretch SUBJECT_SAMPLE_FACTORS - 7 type:Ad.shMuRF1 | Stretch Cycle:30 min cyclic stretch SUBJECT_SAMPLE_FACTORS - 20 type:Ad.shMuRF1 | Stretch Cycle:60 min cyclic stretch SUBJECT_SAMPLE_FACTORS - 32 type:Ad.shMuRF1 | Stretch Cycle:60 min cyclic stretch SUBJECT_SAMPLE_FACTORS - 8 type:Ad.shMuRF1 | Stretch Cycle:60 min cyclic stretch SUBJECT_SAMPLE_FACTORS - 17 type:Ad.shMuRF1 | Stretch Cycle:No stretch SUBJECT_SAMPLE_FACTORS - 29 type:Ad.shMuRF1 | Stretch Cycle:No stretch SUBJECT_SAMPLE_FACTORS - 5 type:Ad.shMuRF1 | Stretch Cycle:No stretch SUBJECT_SAMPLE_FACTORS - 14 type:Ad.shRNA Scramble | Stretch Cycle:15 min cyclic stretch SUBJECT_SAMPLE_FACTORS - 2 type:Ad.shRNA Scramble | Stretch Cycle:15 min cyclic stretch SUBJECT_SAMPLE_FACTORS - 26 type:Ad.shRNA Scramble | Stretch Cycle:15 min cyclic stretch SUBJECT_SAMPLE_FACTORS - 15 type:Ad.shRNA Scramble | Stretch Cycle:30 min cyclic stretch SUBJECT_SAMPLE_FACTORS - 27 type:Ad.shRNA Scramble | Stretch Cycle:30 min cyclic stretch SUBJECT_SAMPLE_FACTORS - 3 type:Ad.shRNA Scramble | Stretch Cycle:30 min cyclic stretch SUBJECT_SAMPLE_FACTORS - 16 type:Ad.shRNA Scramble | Stretch Cycle:60 min cyclic stretch SUBJECT_SAMPLE_FACTORS - 28 type:Ad.shRNA Scramble | Stretch Cycle:60 min cyclic stretch SUBJECT_SAMPLE_FACTORS - 4 type:Ad.shRNA Scramble | Stretch Cycle:60 min cyclic stretch SUBJECT_SAMPLE_FACTORS - 1 type:Ad.shRNA Scramble | Stretch Cycle:No stretch SUBJECT_SAMPLE_FACTORS - 13 type:Ad.shRNA Scramble | Stretch Cycle:No stretch SUBJECT_SAMPLE_FACTORS - 25 type:Ad.shRNA Scramble | Stretch Cycle:No stretch #COLLECTION CO:COLLECTION_SUMMARY Media harvested at 14, 30, and 60 min post stretch (1Hz, 15%) CO:COLLECTION_PROTOCOL_ID None CO:COLLECTION_PROTOCOL_COMMENTS Non-targeted metabolomics analysis. After HL-1 cells were cultured for 16 h in CO:COLLECTION_PROTOCOL_COMMENTS specialized 6well BioFlex culture plates in Claycomb medium, medium was changed CO:COLLECTION_PROTOCOL_COMMENTS to DMEM supplemented 1% penicillin-streptomycin and 10% serum. Transient CO:COLLECTION_PROTOCOL_COMMENTS knockdown of MuRF1 was carried out using recombinant Ad.shRNA MuRF1 at MOI 30. CO:COLLECTION_PROTOCOL_COMMENTS After transduction for 48 h, HL-1 cells were used for mechanical stretch at 15% CO:COLLECTION_PROTOCOL_COMMENTS strain and for different times (15, 30, and 60 min) in a computer-regulated CO:COLLECTION_PROTOCOL_COMMENTS Flexcell FX-5000 Compression System (Flexcell International Corporation, CO:COLLECTION_PROTOCOL_COMMENTS Hillsborough, NC). Non-stretch cells (0 min) were used as the control. After CO:COLLECTION_PROTOCOL_COMMENTS HL-1 cells were stretched, HL-1 cells (at 0 and 60 min) and DMEM medium (at 0, CO:COLLECTION_PROTOCOL_COMMENTS 15, 30, and 60 min) were collected for GC/MS measurement of metabolites, and CO:COLLECTION_PROTOCOL_COMMENTS samples were placed on dry ice/stored at -80C. Samples were then analyzed by CO:COLLECTION_PROTOCOL_COMMENTS GC/MS, as previously described [24]. The raw, transformed, and sorted data used CO:COLLECTION_PROTOCOL_COMMENTS for each of the three comparisons in the metabolomics analyses can be found in CO:COLLECTION_PROTOCOL_COMMENTS Supplemental Table 1. Up to 1 missing value per group was imputed using lowest CO:COLLECTION_PROTOCOL_COMMENTS value in the same group, with groups missing 2 or more excluded from the CO:COLLECTION_PROTOCOL_COMMENTS analysis. The data obtained in this study will be accessible at the NIH Common CO:COLLECTION_PROTOCOL_COMMENTS Fund’s Data Repository and Coordinating Center (supported by NIH grant, CO:COLLECTION_PROTOCOL_COMMENTS U01-DK097430) website, http://www.metabolomicsworkbench.org. CO:SAMPLE_TYPE Media CO:COLLECTION_METHOD Pipette CO:COLLECTION_LOCATION Tissue culture. CO:COLLECTION_FREQUENCY q 15-30 minutes CO:COLLECTION_DURATION 1 hour CO:VOLUMEORAMOUNT_COLLECTED 10 ul CO:STORAGE_CONDITIONS -80C CO:COLLECTION_VIALS Cryovials CO:STORAGE_VIALS Cryovials CO:COLLECTION_TUBE_TEMP Ice #TREATMENT TR:TREATMENT_SUMMARY Cell culture, adenovirus transduction, and MuRF1 knock-down. The TR:TREATMENT_SUMMARY cardiomyocyte-derived HL-1 cell line was maintained as previously described TR:TREATMENT_SUMMARY [11-13]. Briefly, cells were split and cultured to ~90% confluency in 6-well TR:TREATMENT_SUMMARY BioFlex culture plates (Flexcell International Corporation, Hillsborough, NC) TR:TREATMENT_SUMMARY coated with 0.02% gelatin (wt/vol) and 0.5% fibronectin (vol/vol) for ∼30 min TR:TREATMENT_SUMMARY and grown in complete Claycomb media containing 10% serum (Cat. #51800C, TR:TREATMENT_SUMMARY Sigma-Aldrich; St. Louis, MO), as previously described [13]. Media was changed TR:TREATMENT_SUMMARY to serum-free Dulbecco’s modified Eagle’s medium (DMEM) supplemented 1% TR:TREATMENT_SUMMARY penicillin-streptomycin. HL-1 cells were transduced with 30 MOI adenovirus TR:TREATMENT_SUMMARY expressing shRNA MuRF1 (Ad.shRNA MuRF1) or shRNA scramble (Ad.shRNA Scr) control TR:TREATMENT_SUMMARY (custom made by Vector Biolabs, Philadelphia, PA) to transiently knock down TR:TREATMENT_SUMMARY MuRF1, as previously described [13]. TR:TREATMENT Ad.shRNA MuRF1 (or Ad.shRNA Scramble) treatment TR:TREATMENT_COMPOUND Adenovirus TR:TREATMENT_ROUTE Media TR:TREATMENT_DOSE 30 MOI TR:TREATMENT_DOSEVOLUME <10 ul TR:TREATMENT_DOSEDURATION 48 hours TR:TREATMENT_VEHICLE DMEM TR:CELL_MEDIA DMEM / No serum TR:CELL_ENVIR_COND Stretch TR:CELL_PCT_CONFLUENCE ~60% TR:CELL_MEDIA_LASTCHANGED At start of stretch period. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY The samples were “crash” deprotonized by methanol precipitation and spiked SP:SAMPLEPREP_SUMMARY with D27-deuterated myristic acid (D27-C14:0) as an internal standard for SP:SAMPLEPREP_SUMMARY retention-time locking and dried. The trimethylsilyl (TMS)-D27-C14:0 standard SP:SAMPLEPREP_SUMMARY retention time (RT) was set at ~16.727 min. Reactive carbonyls were stabilized SP:SAMPLEPREP_SUMMARY at 50 °C with methoxyamine hydrochloride in dry pyridine. Metabolites were made SP:SAMPLEPREP_SUMMARY volatile with TMS groups using N-methyl-N-(trimethylsilyl) trifluoroacetamide or SP:SAMPLEPREP_SUMMARY MSTFA with catalytic trimethylchlorosilane at 50 °C. GC/MS methods generally SP:SAMPLEPREP_SUMMARY follow those of Roessner et al. (2000), Fiehn et al. (2008), and Kind et al. SP:SAMPLEPREP_SUMMARY (2009), and used a 6890 N GC connected to a 5975 Inert single-quadrupole MS SP:SAMPLEPREP_SUMMARY (Agilent Technologies, Santa Clara, CA). The two wall-coated, open-tubular SP:SAMPLEPREP_SUMMARY (WCOT) GC columns connected in series are both from J&W/Agilent (part SP:SAMPLEPREP_SUMMARY 122–5512), DB5-MS, 15 meters in length, 0.25 mm in diameter, with an 0.25-μm SP:SAMPLEPREP_SUMMARY luminal film. Positive ions generated with conventional electron-ionization (EI) SP:SAMPLEPREP_SUMMARY at 70 eV are scanned broadly from 600 to 50 m/z in the detector throughout the SP:SAMPLEPREP_SUMMARY 45 min cycle time. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Metabolites were made volatile with trimethylsilyl (TMS) groups using N-methyl- CH:CHROMATOGRAPHY_SUMMARY N-(trimethylsilyl) trifluoroacetamide or MSTFA with catalytic CH:CHROMATOGRAPHY_SUMMARY trimethylchlorosilane at 50 C. GC/MS methods generally follow those of Roessner CH:CHROMATOGRAPHY_SUMMARY et al. (Roessner et al. 2000), Fiehn et al. (2008), and Kind et al. (2009), and CH:CHROMATOGRAPHY_SUMMARY used a 6890N GC connected to a 5,975 inert single-quadrupole MS (Agilent CH:CHROMATOGRAPHY_SUMMARY Technologies, Santa Clara, CA). The two wall-coated, open-tubular (WCOT) GC CH:CHROMATOGRAPHY_SUMMARY columns connected in series are both from J&W/Agilent (part 122-5512), DB5- MS, CH:CHROMATOGRAPHY_SUMMARY 15 m in length, 0.25 mm in diameter, with an 0.25-lm luminal film. Positive ions CH:CHROMATOGRAPHY_SUMMARY generated with conventional electron-ionization (EI) at 70 eV are scanned CH:CHROMATOGRAPHY_SUMMARY broadly from 600 to 50 m/z in the detector throughout the 45 min cycle time. CH:INSTRUMENT_NAME Agilent 6890N CH:COLUMN_NAME GCs connected in series are both from J&W/Agilent (part 122-5512) CH:CHROMATOGRAPHY_TYPE GC #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Agilent 5975 MS:INSTRUMENT_TYPE Single quadrupole MS:MS_TYPE ESI MS:ION_MODE POSITIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Peak values (Log transformed) MS_METABOLITE_DATA_START Samples 18 30 6 19 31 7 20 32 8 17 29 5 14 2 26 15 27 3 16 28 4 1 13 25 Factors type:Ad.shMuRF1 | Stretch Cycle:15 min cyclic stretch type:Ad.shMuRF1 | Stretch Cycle:15 min cyclic stretch type:Ad.shMuRF1 | Stretch Cycle:15 min cyclic stretch type:Ad.shMuRF1 | Stretch Cycle:30 min cyclic stretch type:Ad.shMuRF1 | Stretch Cycle:30 min cyclic stretch type:Ad.shMuRF1 | Stretch Cycle:30 min cyclic stretch type:Ad.shMuRF1 | Stretch Cycle:60 min cyclic stretch type:Ad.shMuRF1 | Stretch Cycle:60 min cyclic stretch type:Ad.shMuRF1 | Stretch Cycle:60 min cyclic stretch type:Ad.shMuRF1 | Stretch Cycle:No stretch type:Ad.shMuRF1 | Stretch Cycle:No stretch type:Ad.shMuRF1 | Stretch Cycle:No stretch type:Ad.shRNA Scramble | Stretch Cycle:15 min cyclic stretch type:Ad.shRNA Scramble | Stretch Cycle:15 min cyclic stretch type:Ad.shRNA Scramble | Stretch Cycle:15 min cyclic stretch type:Ad.shRNA Scramble | Stretch Cycle:30 min cyclic stretch type:Ad.shRNA Scramble | Stretch Cycle:30 min cyclic stretch type:Ad.shRNA Scramble | Stretch Cycle:30 min cyclic stretch type:Ad.shRNA Scramble | Stretch Cycle:60 min cyclic stretch type:Ad.shRNA Scramble | Stretch Cycle:60 min cyclic stretch type:Ad.shRNA Scramble | Stretch Cycle:60 min cyclic stretch type:Ad.shRNA Scramble | Stretch Cycle:No stretch type:Ad.shRNA Scramble | Stretch Cycle:No stretch type:Ad.shRNA Scramble | Stretch Cycle:No stretch 2-Deoxyglucose 16.8400 16.6600 16.1900 17.2400 16.8600 16.9300 16.6600 16.9900 16.8400 16.6500 17.4400 16.3400 16.5100 16.1300 17.2400 16.7000 16.8500 16.3900 16.5800 16.5600 16.3300 16.9600 16.8400 16.8300 2-Ethylhexanoic acid 12.9200 12.4900 13.3500 13.3000 12.8300 12.7200 13.0300 13.1900 12.7000 14.2500 13.4200 12.7400 12.5900 2-Hydroxybutyric acid 14.5000 15.0800 15.0700 15.0700 15.2900 14.5800 15.2500 15.1800 15.1200 14.8000 15.7400 15.0700 14.6700 14.9200 14.4100 14.9900 12.2900 14.8300 2-Hydroxypyridine 15.3200 15.3500 15.1800 15.4600 15.4100 15.2800 15.1800 15.1900 15.3300 15.2700 15.6200 14.9600 15.4000 15.3400 15.6500 15.1700 15.6400 15.3300 15.0200 15.2300 15.4700 14.8500 15.1500 15.4100 2-Hydroxyvaleric acid 12.1800 13.0100 12.8900 13.1000 12.5800 12.6200 12.7000 12.8800 13.2700 12.5900 14.9500 12.3200 12.7500 12.3300 13.1700 12.3200 13.4700 12.1200 12.2500 12.8800 12.9700 12.3300 13.4300 2-Ketoleucine/ketoisoleucine 19.8100 19.5600 19.2600 19.8900 19.7500 19.7200 19.6000 19.5700 19.6500 19.5600 19.5400 18.6000 19.6000 19.3900 20.1300 19.4800 19.8600 19.3100 19.1700 19.2900 19.3600 18.9800 19.3200 19.2600 2-Ketovaline 17.4000 17.3900 17.1300 17.7800 17.4500 17.2900 17.4900 17.3900 17.6500 17.4100 17.7000 16.3500 17.5100 17.2600 17.8100 17.2900 17.6400 16.9500 16.9300 17.1000 17.0800 16.8800 16.8800 17.0100 3-Indolelactic acid/tryptophan 12.8900 12.8800 12.4300 13.0400 13.2000 12.1800 12.8000 12.9500 12.8400 13.0400 14.1400 12.1400 12.6000 11.5600 13.2000 11.5700 12.0900 12.3700 12.3000 12.3700 3-Phenyllactic acid 13.1100 13.8500 13.2200 13.7400 12.6300 14.0200 13.5400 15.2100 13.2700 12.4600 14.0400 12.5200 12.8600 14.2600 4-Hydroxyphenylpyruvic acid 14.6300 14.2000 15.5300 13.8400 15.3000 16.3000 15.2200 15.7200 14.5300 15.0100 16.1300 14.8900 14.8500 14.9200 14.0800 5-(Hydroxymethyl)furfural 15.1200 15.1300 14.6500 14.6000 15.0200 15.1400 14.7200 15.2300 14.6600 15.0400 14.6100 15.5900 5-Hydroxytryptophan 12.4500 13.1200 13.2900 6-Aminohexanoic acid/delta-Aminolevulinic acid 13.3800 13.0900 13.9400 13.3400 13.2800 13.9600 13.8900 14.3800 13.6000 13.2000 14.3500 13.5300 13.7800 13.5800 14.4100 14.5700 14.1200 12.5300 13.5000 Alanine 22.6300 22.4100 22.4300 22.7400 22.5800 22.3400 22.1500 22.5500 22.4400 22.5700 22.8100 22.2700 22.5400 22.1600 22.7600 22.3600 22.6900 22.3200 22.5100 22.4700 22.3900 22.3600 22.4100 22.5700 Aldopentoses 18.7300 18.6300 18.2200 18.5300 18.3400 16.7200 18.2200 18.3200 18.9100 17.1600 19.0100 18.8100 18.8000 18.2800 18.4000 18.4500 18.1400 18.4700 Allantoin 18.4500 17.6400 18.0200 18.0100 18.2500 17.7000 17.6600 17.8100 18.1400 18.0500 18.1700 17.8500 18.0200 17.9600 17.9800 18.1800 17.9600 18.1300 18.2600 18.0500 18.2300 18.2300 17.7500 17.8600 alpha Ketoglutaric acid 19.6100 19.5900 19.0900 19.9400 19.6900 19.2300 19.2500 19.2800 19.5200 19.6800 20.0800 18.5800 19.6200 18.9700 20.2900 19.2700 19.9800 19.1200 19.3100 19.5900 19.2200 19.2400 19.1600 20.0300 alpha-Monopalmitin 19.6600 19.8100 19.2000 20.0300 20.3900 19.6200 20.0400 19.7900 19.8400 19.8700 20.3800 19.2000 19.9800 19.7200 20.6300 19.5000 20.2100 19.7700 19.5700 19.7500 19.8100 19.6900 19.5400 19.4200 alpha-Monostearin 22.8200 23.0100 22.5200 23.0600 23.6400 22.6500 23.1300 22.8700 22.8900 22.8900 23.4100 22.4500 22.9700 22.8700 23.5600 22.6800 23.4100 23.0800 22.9400 22.7000 22.8900 22.8000 22.7800 22.6300 Aminomalonic acid 19.6600 18.0700 18.3800 19.6500 19.3800 19.1500 18.1200 18.9000 18.4500 18.9000 18.7700 18.1700 19.6000 18.8800 19.4600 19.8900 19.0900 18.9700 18.8100 18.6700 18.9300 18.6000 19.2900 18.5200 Arginine 16.5600 15.4700 15.4200 16.8400 17.0600 15.6600 15.4500 15.3900 15.6700 16.2200 17.0600 14.1600 15.6200 15.1500 15.9900 15.9200 15.6200 15.9400 15.9400 15.6300 14.8100 15.0400 14.9100 15.5600 Asparagine 19.1400 18.3100 18.0100 19.1300 18.7500 18.4000 17.8200 18.1400 18.4300 18.8600 18.2500 17.8900 18.9600 18.4000 19.1600 18.8300 18.8100 18.5900 18.7500 18.2700 18.6400 18.3600 18.6600 18.6000 Aspartic Acid 16.3600 15.5600 16.6200 16.1400 15.9400 15.5500 15.8500 15.9400 16.1800 16.3400 15.2400 16.2400 15.6800 16.6600 16.0700 16.0100 15.9000 15.8400 15.9900 15.7300 15.6400 15.7300 15.8400 Benzoic acid 15.7800 16.0800 15.6200 16.6400 16.2700 16.3600 15.7500 15.9800 16.5500 15.7000 16.1500 14.4200 16.0800 15.8100 17.1000 16.0900 16.6800 15.8500 15.7300 15.0800 15.5500 15.3300 16.0600 16.2400 beta-Alanine 14.6900 14.3800 13.5700 15.4500 15.2000 15.1100 14.7100 15.3200 15.0200 15.0100 14.2500 13.3200 14.9500 14.5200 15.3900 15.1700 14.9000 14.6200 14.4300 15.0800 14.1300 13.7700 14.8100 14.3800 beta-Monopalmitin 13.9200 14.6300 14.5300 13.1800 14.9600 13.7600 beta-Monostearin 17.7000 17.7100 17.5500 18.0400 18.5800 17.6800 17.8500 17.8000 17.8600 17.8200 18.3000 17.1700 18.0700 17.7900 18.6100 17.8000 18.2500 18.0600 17.5100 17.6000 17.8700 17.5900 17.7100 17.4100 Cholesterol 17.5200 17.5700 18.0300 17.6700 18.0700 17.7700 17.7800 17.8300 17.9300 18.1600 18.6300 17.9700 18.1100 18.0800 18.4000 18.4600 18.2800 17.7400 17.3300 18.1800 18.5400 18.5400 18.0400 Citric acid/isocitric acid 19.5500 19.7200 19.4600 19.6400 19.3700 19.2000 19.3400 19.5100 19.6800 19.3700 19.9700 19.0100 19.6400 19.2800 20.0400 19.0100 19.6100 18.9300 19.2600 19.4500 19.3700 19.1200 18.7300 19.5100 Cysteine/Creatinine (co-elution) 17.1100 17.0900 16.5100 17.4500 17.2100 16.7500 16.8200 17.3200 16.9600 17.2200 16.9500 16.3100 17.1200 16.4600 17.5800 16.6300 17.3200 16.6600 17.1600 16.9600 16.7200 16.7100 16.8400 16.9800 Cysteinesulfinic acid 14.7300 13.3700 11.9200 12.1500 Cystine 21.0100 20.4000 20.4200 20.8200 20.9300 20.3900 20.1900 20.5600 20.4300 20.6200 20.8500 20.1500 20.8700 20.6900 21.0900 20.6500 20.8500 20.6000 20.3600 20.2200 20.6700 20.4300 20.4300 20.2900 Erythronic acid/Threonic acid 18.8600 18.4800 18.1500 18.9900 18.8000 18.6300 18.3600 18.5000 18.5900 18.8800 18.9400 17.7500 18.6000 18.1000 19.1300 18.5200 18.9200 18.1800 18.3100 18.4900 18.4000 18.1400 18.4100 18.6100 Ethanolamine 13.4400 13.4100 13.4900 13.0800 13.6600 13.5800 13.0200 13.6200 13.7100 13.9200 13.0000 12.9900 13.3800 13.0700 14.2100 14.0900 13.6600 13.4400 13.5300 13.3700 13.7100 13.4200 Fructose or similar ketohexose 24.3800 24.3200 23.9700 24.5300 24.4300 24.1200 24.1500 24.1700 24.1600 24.2400 24.5800 23.7600 24.3200 23.9900 24.7700 24.0900 24.4900 24.0200 24.1000 24.1600 24.0100 23.9700 24.0300 24.3600 Fumaric acid 15.9600 15.8300 15.1700 16.0500 15.8800 15.5900 15.8400 15.5500 16.0300 16.0100 16.5000 14.8500 15.7400 15.5500 16.6200 15.6900 16.2500 15.3000 15.5300 15.5200 15.6500 15.5100 15.6500 16.0300 gamma-Aminobutyric acid 17.5700 17.8200 17.1400 18.1700 17.5600 17.6000 17.6300 17.2200 18.0500 17.8600 18.7000 16.2500 17.7400 17.6600 19.1800 17.4900 18.2300 17.1500 16.7000 16.9100 17.3100 16.7300 17.5100 17.9200 Gluconic acid or similar sugar acid 17.7000 17.5500 16.9700 17.7900 17.6500 17.3800 17.4800 17.3900 17.4000 17.6800 18.0300 16.8200 17.5000 17.2100 17.9600 17.5800 17.6100 17.2700 17.4000 17.2300 17.3500 17.2700 17.3800 17.3000 Glucose and other aldohexoses 27.2200 27.3500 27.6200 27.2800 27.8500 27.6800 27.7100 28.0700 27.9200 27.3400 27.0800 27.7000 27.6800 27.7200 27.6400 27.6900 27.7500 27.7000 27.6400 27.7100 27.5800 27.9200 27.6000 27.4200 Glutamic acid 24.8000 24.4900 24.0600 25.0100 24.8900 24.4900 24.3900 24.5400 24.4500 24.5600 24.7400 23.6100 24.6200 24.2100 25.1000 24.6500 24.7900 24.1300 24.2900 24.3400 24.1300 23.9800 24.3200 24.3900 Glutamine 23.2500 22.0700 21.1000 23.0000 23.2000 22.5500 21.7900 22.9000 21.1900 22.7200 20.0200 21.2300 22.5400 22.4400 22.2200 23.1000 21.5900 22.3200 22.2400 21.4000 21.7300 21.0600 22.4200 20.7700 Glyceric acid 15.8600 15.0900 15.2500 16.1800 16.0600 15.8900 15.7600 15.7400 15.9000 15.9400 15.8600 15.0800 15.3100 15.2500 16.2800 15.1900 16.0300 15.2500 15.3800 15.5600 15.3400 15.2700 15.5300 15.8900 Glycerol 23.0900 20.6900 21.5900 22.4500 22.2800 20.8000 21.8700 20.9400 21.8900 22.1700 21.7100 21.3900 22.1900 21.7900 22.6100 22.0700 22.2800 20.6200 20.6100 21.7900 20.4900 20.7600 21.9500 22.1600 Glycerol 1-phosphate 13.8500 14.0100 13.5200 14.6000 14.2500 13.6300 13.4700 14.1800 13.8800 12.7700 14.5900 11.2400 14.3900 13.6200 14.6300 13.9100 14.4000 13.8000 13.9600 13.7800 13.5000 12.5900 14.0400 14.2900 Glycine 23.4200 23.3600 23.0500 23.6100 23.6100 23.4200 23.2800 23.3000 23.4800 23.4700 23.8800 22.7800 23.4300 23.0900 23.9100 23.3300 23.6800 23.0600 23.1900 23.3700 23.0700 23.2000 23.2200 23.5600 Glycolic acid 15.6700 15.1000 15.4000 15.6500 15.5000 15.3800 15.6700 16.1000 15.5900 16.0700 15.3700 15.9500 15.4000 15.0400 15.1200 15.2200 15.3400 Hexitols 22.7100 22.5700 22.1600 22.7600 22.6800 22.4600 22.4600 22.4900 22.4200 22.5900 22.9900 21.9400 22.4600 22.3300 23.0200 22.3500 22.6600 22.3300 22.3100 22.3800 22.0800 22.2200 22.1800 22.6200 Hexuronic acid 13.9800 16.0000 15.7200 15.7900 15.3200 Histidine 20.1700 20.0600 19.6800 20.1300 19.8100 19.8400 19.6100 20.0400 19.9700 19.9900 18.9000 19.6400 20.0500 19.8700 20.3700 20.1400 20.3100 19.8000 19.9600 19.8800 20.1200 19.9100 19.9100 20.2300 Hydroxyprolines 17.3900 17.3900 17.0000 17.2100 17.4700 17.1900 17.0800 17.4700 17.0100 17.5900 17.2500 17.3600 17.2900 16.8400 Isoleucine 23.0200 23.0200 22.7000 23.0500 23.1700 22.9000 22.5900 23.0700 23.0500 23.0300 22.8000 22.3100 23.0200 22.6500 23.4600 23.1000 23.1100 22.7700 22.6600 22.5800 22.6800 22.5300 22.9700 22.7200 Isomaltose or similar disaccharide 16.3900 16.0900 16.1000 15.9100 16.4700 16.0400 16.0800 16.3800 16.0500 16.1200 16.3800 15.8300 16.3200 16.5100 16.8700 16.1900 16.6900 16.2900 15.9000 15.9700 16.4600 16.2800 15.9900 15.9900 Ketopentoses 18.7300 17.1400 17.9700 18.5300 18.3400 16.7200 17.3400 18.3200 18.0900 14.9200 17.1600 17.7000 17.6300 14.3700 17.1900 16.9300 16.6800 15.3900 18.4700 Lactic acid 27.4200 27.4600 27.4300 27.5200 27.4900 27.3900 27.3900 27.3700 27.6100 27.5100 27.7800 27.1100 27.5000 27.3100 27.7900 27.2000 27.6500 27.0900 27.2100 27.3000 27.3300 27.2800 27.3100 27.4700 Leucine 23.0900 22.6700 22.1400 22.8100 22.7000 22.8600 22.1900 22.4800 22.4200 22.5800 22.3500 21.7200 22.9200 22.1200 23.2600 22.9000 23.0000 22.4300 22.6500 22.6500 22.1200 22.5400 22.4100 22.9800 Lysine 24.5200 24.3300 24.0700 24.4800 24.3800 24.2000 24.1000 24.2600 24.1500 24.2800 24.6600 23.8300 24.3600 24.0800 24.6400 24.2600 24.4600 24.0800 24.0600 24.1100 24.1500 24.0500 24.0400 24.3400 Malic acid 18.5600 18.5600 18.2400 19.0200 18.5700 18.4700 18.2600 18.4500 18.6900 18.7700 19.2400 17.8800 18.7000 18.1400 19.3600 18.4300 19.0200 18.0900 18.2900 18.5600 18.1800 18.3100 18.4200 18.9100 Maltose or similar disaccharide 12.8400 14.6300 14.5700 14.5400 14.8300 13.0800 14.5800 14.6900 14.8000 14.7200 14.5200 14.3700 14.7400 14.6500 15.3400 14.7700 15.1700 14.4900 14.5600 14.5100 15.0500 14.2700 Methionine 20.8500 20.6000 20.1200 20.9600 20.8400 20.2800 20.1500 20.5400 20.3200 20.8100 20.5300 19.4800 20.6100 20.0600 20.9000 20.5100 20.7800 20.1400 20.3600 20.3800 20.1800 19.9100 20.2600 20.5000 Methionine sulfoxide 15.4000 14.9800 14.3200 15.7700 15.0500 16.1500 15.1900 15.8600 15.5700 15.0600 16.6700 16.0300 15.0100 16.2400 15.1600 14.0400 15.2100 14.5500 15.2100 14.6700 16.4800 15.8600 15.3600 Methylglutamic acid 14.1000 13.6900 12.8100 14.1000 13.9700 13.5000 11.9800 13.8500 14.0700 14.2100 13.0700 13.5600 13.6300 12.7600 13.3300 13.4700 Myoinositol 23.1400 22.9000 22.6400 23.1100 23.1300 22.8300 22.8300 22.9400 22.7400 22.9600 23.3200 22.4500 23.1100 22.9100 23.4900 23.0000 23.1600 22.8700 22.8100 22.8400 22.8700 22.8400 22.8400 22.9600 Myristic acid or Pentadecanol 15.5400 15.8200 15.3900 16.2600 16.2100 15.2800 15.1800 16.5000 15.8100 16.6800 14.9800 15.8600 14.9100 15.9000 15.5700 15.8400 15.5400 15.5100 15.6500 15.5600 15.3700 15.2700 15.9800 Nicotinamide 15.5200 15.3200 14.9300 16.2000 15.6200 15.2100 15.5200 15.1900 15.4600 14.4700 16.1400 13.0400 16.2400 15.4700 15.9300 14.9200 15.1700 14.6100 15.3900 15.4400 15.5100 N-Methylalanine/2-Aminobutanoic acid/N-Ethylglycine 13.3600 13.3800 13.1400 13.0500 13.0100 12.9000 Norepinephrine/Methyltryptamine 13.3000 13.0100 12.8000 12.6100 13.2000 12.0900 12.9300 12.6100 12.7000 Ornithine 22.2700 22.3400 21.6100 22.4000 22.2700 21.8800 21.5800 21.9800 21.6700 22.0100 21.8600 21.4100 22.1400 21.7500 22.3600 22.0800 21.9900 21.8400 21.7700 21.8200 21.7100 21.6700 21.6700 21.7900 Palmitic acid 19.8500 19.2400 19.5200 19.7500 19.4400 19.8400 19.9100 19.5300 19.7600 19.9500 19.9600 19.2200 19.5400 19.5800 20.2900 19.8800 20.0900 19.4300 19.4800 19.1800 19.3400 19.6200 19.7600 20.1000 Pantothenic acid 18.9900 18.8100 18.3700 19.0300 19.0000 18.7000 18.7000 18.6900 18.7300 18.8000 19.2800 18.2200 19.0400 18.6300 19.3100 18.7800 18.9000 18.7000 18.4000 18.4800 18.4900 18.4700 18.5100 18.8600 Pentitols 18.3100 18.3700 18.1700 18.4800 18.3700 18.2600 18.2200 18.0900 18.4500 18.2800 18.8300 17.8900 18.1100 17.7800 18.5600 18.0200 18.5700 17.7700 18.1800 18.3200 18.1100 18.1100 17.8000 18.5100 Pentonic acids 14.5100 14.6400 14.3800 14.1100 14.3500 13.4300 15.3900 13.6000 16.0200 14.2500 15.0000 Phenylalanine 23.1500 22.8100 22.4800 23.3300 23.1000 22.9100 22.5700 22.9000 22.7800 22.9700 23.1800 22.1200 22.9100 22.5100 23.3000 22.8700 23.0900 22.4900 22.7100 22.7200 22.4800 22.5300 22.6500 22.8600 Phosphoric acid 19.2100 20.3800 19.8600 20.9300 20.2900 20.0800 19.8700 20.0000 20.2600 20.7000 22.0700 19.9300 20.5900 19.5400 20.8400 19.4800 20.8100 19.6600 20.0200 20.0600 19.7600 20.2800 20.2400 21.6100 Proline 21.4100 21.2200 21.2600 21.4800 21.2300 21.1600 20.9000 21.2200 21.5200 21.4300 21.7100 20.9100 21.2300 20.9200 21.7200 21.0700 21.6700 20.8600 21.0200 21.3100 20.9800 21.2900 21.1700 21.6400 Pseudouridine 15.3800 14.9000 14.2700 14.9700 15.7400 14.6500 15.4100 15.5600 14.7300 14.4200 15.8200 13.8900 14.9100 14.9400 15.8900 15.5400 15.1800 14.2700 14.7100 14.6100 14.8900 14.1400 14.6100 Putrescine 12.8400 13.3800 13.0900 13.9400 13.3400 13.2800 13.9600 13.8900 14.3800 13.6000 13.2000 14.3500 13.5300 13.7800 13.5800 14.4100 14.5700 14.1200 12.5300 13.5000 Pyridoxine 18.9100 19.5300 18.7200 18.9400 18.9000 20.2500 15.8300 19.2000 19.2600 19.1700 17.6100 19.0900 18.8700 20.3300 18.6600 19.9500 19.2600 18.8000 19.0200 18.8200 18.8800 18.6200 20.0700 Pyruvic acid 20.6400 20.0200 19.8300 20.5900 20.5000 20.1200 20.0000 20.2700 20.1700 20.0500 19.4800 19.4600 20.6400 20.4300 20.5300 20.4400 20.5400 20.3000 20.0200 20.1000 20.1000 19.6700 20.1900 19.5200 Serine 23.1800 22.9300 22.5600 23.4100 23.1500 22.8500 22.4900 22.9900 22.7400 23.0900 22.9500 22.3100 22.9700 22.4900 23.3900 22.9000 22.9900 22.5700 22.6600 22.7800 22.5300 22.5400 22.5300 22.8000 Serotonin 11.7300 12.5600 12.5700 12.6200 12.2200 Stearic acid 19.9800 19.9300 19.9200 20.3800 20.2200 20.0700 20.1000 19.6900 20.5700 20.0500 20.6900 19.5800 20.3200 19.9100 20.6700 20.1300 20.3500 20.2900 19.6300 19.9300 20.1700 20.0300 20.0100 20.1500 Succinic acid 20.3700 20.0200 19.7100 20.6300 20.3900 20.3200 20.0700 20.0900 20.3300 20.2800 20.5600 19.4100 20.2200 19.8600 20.9300 20.2200 20.4700 19.7300 19.7600 19.9100 19.7400 19.8300 19.9700 20.1000 Threitol/Erythritol 21.0300 20.9300 20.4100 21.2700 21.0500 20.9400 20.8500 20.8300 20.9700 21.0300 21.3300 20.1200 21.0100 20.5000 21.3700 20.9500 21.0900 20.5300 20.6400 20.7100 20.5000 20.5400 20.7100 20.9600 Threonine 23.7900 23.4300 23.1400 23.9900 23.7900 23.3800 23.0100 23.5500 23.4700 23.8300 23.7300 22.8200 23.4300 22.9700 23.9800 23.3800 23.7100 22.9800 23.2700 23.4100 23.1500 23.1200 23.2700 23.6700 Thymine 13.7300 14.4100 15.1500 12.1000 13.8000 Tryptophan 20.5400 20.2300 19.8400 20.3600 20.5700 19.9800 19.7600 20.4100 19.9500 20.1600 20.5500 19.6300 20.3300 20.0800 20.7300 20.2900 20.4400 19.9800 20.0300 20.0500 20.0000 19.9400 20.0600 20.0900 Tyrosine 23.7300 23.4800 23.2700 23.6500 23.6000 23.4900 23.2500 23.4100 23.4100 23.5900 23.8200 23.0200 23.5500 23.2300 23.8800 23.4500 23.6300 23.3900 23.3400 23.3900 23.3000 23.3200 23.3000 23.6300 Uracil 16.0100 15.4500 15.1900 16.1600 15.9400 15.7200 15.0400 15.6000 15.8200 15.3600 15.7200 9.5400 15.5400 14.9900 16.3700 15.6500 16.1200 15.1800 15.0700 14.8300 14.9700 14.4600 14.2000 14.9500 Urea 22.5200 22.3000 22.0300 22.7300 22.5200 22.4300 22.3400 22.3400 22.4700 22.5000 22.7900 21.5000 22.4800 22.0900 22.9000 22.4600 22.6800 21.9000 22.0600 22.2200 22.1200 21.9900 22.2700 22.3700 Uric acid 16.9700 16.6700 16.3300 16.9400 17.0700 16.3800 16.3700 16.6700 16.5900 16.7300 17.1500 15.7000 16.8800 16.3800 17.1800 16.6300 16.9500 16.4500 16.5300 16.5000 16.4900 15.6800 16.3100 16.6000 Valine 22.7900 22.4800 22.1300 22.7000 22.7000 22.5000 21.8100 22.4100 22.3200 22.4900 22.6100 21.8200 22.5900 22.1300 22.8500 22.5300 22.6100 22.1900 22.3000 22.2600 22.1500 22.2800 22.4000 22.3600 Xanthine 16.5800 16.3700 15.5100 16.8000 16.9100 16.2200 15.7200 16.4300 15.9300 16.2700 16.8000 14.8600 16.6800 16.2000 17.0200 16.7900 16.7800 15.7300 15.9900 16.1100 16.2800 15.2500 15.5100 16.1100 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name pubchem_id inchi_key kegg_id other_id other_id_type ri ri_type moverz_quant 2-Deoxyglucose 2-Ethylhexanoic acid 2-Hydroxybutyric acid 2-Hydroxypyridine 2-Hydroxyvaleric acid 2-Ketoleucine/ketoisoleucine 2-Ketovaline 3-Indolelactic acid/tryptophan 3-Phenyllactic acid 4-Hydroxyphenylpyruvic acid 5-(Hydroxymethyl)furfural 5-Hydroxytryptophan 6-Aminohexanoic acid/delta-Aminolevulinic acid Alanine Aldopentoses Allantoin alpha Ketoglutaric acid alpha-Monopalmitin alpha-Monostearin Aminomalonic acid Arginine Asparagine Aspartic Acid Benzoic acid beta-Alanine beta-Monopalmitin beta-Monostearin Cholesterol Citric acid/isocitric acid Cysteine/Creatinine (co-elution) Cysteinesulfinic acid Cystine Erythronic acid/Threonic acid Ethanolamine Fructose or similar ketohexose Fumaric acid gamma-Aminobutyric acid Gluconic acid or similar sugar acid Glucose and other aldohexoses Glutamic acid Glutamine Glyceric acid Glycerol Glycerol 1-phosphate Glycine Glycolic acid Hexitols Hexuronic acid Histidine Hydroxyprolines Isoleucine Isomaltose or similar disaccharide Ketopentoses Lactic acid Leucine Lysine Malic acid Maltose or similar disaccharide Methionine Methionine sulfoxide Methylglutamic acid Myoinositol Myristic acid or Pentadecanol Nicotinamide N-Methylalanine/2-Aminobutanoic acid/N-Ethylglycine Norepinephrine/Methyltryptamine Ornithine Palmitic acid Pantothenic acid Pentitols Pentonic acids Phenylalanine Phosphoric acid Proline Pseudouridine Putrescine Pyridoxine Pyruvic acid Serine Serotonin Stearic acid Succinic acid Threitol/Erythritol Threonine Thymine Tryptophan Tyrosine Uracil Urea Uric acid Valine Xanthine METABOLITES_END #END