#METABOLOMICS WORKBENCH amroilaiwy_20160528_113623_mwtab.txt DATATRACK_ID:640 STUDY_ID:ST000431 ANALYSIS_ID:AN000681 PROJECT_ID:PR000334 VERSION 1 CREATED_ON July 29, 2016, 11:14 am #PROJECT PR:PROJECT_TITLE C2C12 stretch cessation models muscle atrophy and anaplerotic changes in PR:PROJECT_TITLE metabolism PR:PROJECT_TYPE GC-MS non targeted qualitative analysis PR:PROJECT_SUMMARY Studies of skeletal muscle disuse either in patients on bed rest or PR:PROJECT_SUMMARY experimentally in animals(immobilization) have demonstrated that decreased PR:PROJECT_SUMMARY protein synthesis is common, with transient parallel increases in protein PR:PROJECT_SUMMARY degradation. Muscle disuse atrophy involves a process of transition from slow to PR:PROJECT_SUMMARY fast myosin fiber types 6 . A shift toward glycolysis, decreased capacity for PR:PROJECT_SUMMARY fat oxidation, and substrate accumulation in atrophied muscles have been PR:PROJECT_SUMMARY reported as has accommodation of the liver with an increased gluconeogenic PR:PROJECT_SUMMARY capacity. Recent studies have modeled skeletal muscle disuse by using cyclic PR:PROJECT_SUMMARY stretch of differentiated myotubes (C2C12), which mimics the loading pattern of PR:PROJECT_SUMMARY mature skeletal muscle, followed by cessation of stretch.We utilized this model PR:PROJECT_SUMMARY to determine the metabolic changes using non-targeted metabolomics analysis of PR:PROJECT_SUMMARY the media. We identified increases in amino acids resulting from protein PR:PROJECT_SUMMARY degradation (largely sarcomere) that occurs with muscle atrophy that are PR:PROJECT_SUMMARY involved in feeding the Kreb’s cycle through anaplerosis. Specifically, we PR:PROJECT_SUMMARY identified increased alanine/proline metabolism (significantly elevated proline, PR:PROJECT_SUMMARY alanine, glutamine, and asparagine) and increased -ketoglutaric acid, the PR:PROJECT_SUMMARY proposed Kreb’s cycle intermediate being fed by the alanine/proline metabolic PR:PROJECT_SUMMARY anaplerotic mechanism. Additionally, several unique pathways not clearly PR:PROJECT_SUMMARY delineated in previous studies of muscle unloading were seen, including: 1) PR:PROJECT_SUMMARY elevated ethanolamine and elevated keto-acids (e.g. 2-ketoleucine and PR:PROJECT_SUMMARY 2-keovaline) represent intermediates in the Ehlrich amino acid degradation PR:PROJECT_SUMMARY pathway, which feeds into a metabolic pathway supplying acetyl-CoA and PR:PROJECT_SUMMARY 2-hydroxybutyrate (also significantly increased); and 2) elevated guanine, an PR:PROJECT_SUMMARY intermediate of purine metabolism, was seen at 12 hours unloading. Given the PR:PROJECT_SUMMARY interest in targeting different aspects of the ubiquitin proteasome system to PR:PROJECT_SUMMARY inhibit protein degradation, this C2C12 system may allow the identification of PR:PROJECT_SUMMARY direct and indirect alterations in metabolism due to anaplerosis or through PR:PROJECT_SUMMARY other yet to be identified mechanisms using a non-targeted metabolomics PR:PROJECT_SUMMARY approach. PR:INSTITUTE University of North Carolina PR:DEPARTMENT McAllister heart Institute, Department of Internal medicine PR:LABORATORY Multiple Centers PR:LAST_NAME Ilaiwy, Willis PR:FIRST_NAME Amro, Monte PR:ADDRESS 111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA PR:EMAIL amroilaiwy@gmail.com, monte_willis@med.unc.edu PR:PHONE 210-596-0171 PR:FUNDING_SOURCE NIH, Fondation Leducq #STUDY ST:STUDY_TITLE C2C12 stretch cessation models muscle atrophy and anaplerotic changes in ST:STUDY_TITLE metabolism ST:STUDY_TYPE GC-MS non-targeted metabolomic profiling ST:STUDY_SUMMARY Studies of skeletal muscle disuse either in patients on bed rest or ST:STUDY_SUMMARY experimentally in animals(immobilization) have demonstrated that decreased ST:STUDY_SUMMARY protein synthesis is common, with transient parallel increases in protein ST:STUDY_SUMMARY degradation. Muscle disuse atrophy involves a process of transition from slow to ST:STUDY_SUMMARY fast myosin fiber types 6 . A shift toward glycolysis, decreased capacity for ST:STUDY_SUMMARY fat oxidation, and substrate accumulation in atrophied muscles have been ST:STUDY_SUMMARY reported as has accommodation of the liver with an increased gluconeogenic ST:STUDY_SUMMARY capacity. Recent studies have modeled skeletal muscle disuse by using cyclic ST:STUDY_SUMMARY stretch of differentiated myotubes (C2C12), which mimics the loading pattern of ST:STUDY_SUMMARY mature skeletal muscle, followed by cessation of stretch.We utilized this model ST:STUDY_SUMMARY to determine the metabolic changes using non-targeted metabolomics analysis of ST:STUDY_SUMMARY the media. We identified increases in amino acids resulting from protein ST:STUDY_SUMMARY degradation (largely sarcomere) that occurs with muscle atrophy that are ST:STUDY_SUMMARY involved in feeding the Kreb’s cycle through anaplerosis. Specifically, we ST:STUDY_SUMMARY identified increased alanine/proline metabolism (significantly elevated proline, ST:STUDY_SUMMARY alanine, glutamine, and asparagine) and increased -ketoglutaric acid, the ST:STUDY_SUMMARY proposed Kreb’s cycle intermediate being fed by the alanine/proline metabolic ST:STUDY_SUMMARY anaplerotic mechanism. Additionally, several unique pathways not clearly ST:STUDY_SUMMARY delineated in previous studies of muscle unloading were seen, including: 1) ST:STUDY_SUMMARY elevated ethanolamine and elevated keto-acids (e.g. 2-ketoleucine and ST:STUDY_SUMMARY 2-keovaline) represent intermediates in the Ehlrich amino acid degradation ST:STUDY_SUMMARY pathway, which feeds into a metabolic pathway supplying acetyl-CoA and ST:STUDY_SUMMARY 2-hydroxybutyrate (also significantly increased); and 2) elevated guanine, an ST:STUDY_SUMMARY intermediate of purine metabolism, was seen at 12 hours unloading. Given the ST:STUDY_SUMMARY interest in targeting different aspects of the ubiquitin proteasome system to ST:STUDY_SUMMARY inhibit protein degradation, this C2C12 system may allow the identification of ST:STUDY_SUMMARY direct and indirect alterations in metabolism due to anaplerosis or through ST:STUDY_SUMMARY other yet to be identified mechanisms using a non-targeted metabolomics ST:STUDY_SUMMARY approach. ST:INSTITUTE University of North Carolina ST:DEPARTMENT McAllister Heart Institute ST:LABORATORY Mutliple Centers ST:LAST_NAME Ilaiwy, WIllis ST:FIRST_NAME Amro, Monte ST:ADDRESS 111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA ST:EMAIL amroilaiwy@gmail.com, monte_willis@med.unc.edu ST:PHONE 919-3607599 #SUBJECT SU:SUBJECT_TYPE Animal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - p0 #1 Group:6 hour stretch SUBJECT_SAMPLE_FACTORS - p0 #2 Group:6 hour stretch SUBJECT_SAMPLE_FACTORS - p0 #3 Group:6 hour stretch SUBJECT_SAMPLE_FACTORS - p1 #1 Group:1 hour reversal SUBJECT_SAMPLE_FACTORS - p1 #2 Group:1 hour reversal SUBJECT_SAMPLE_FACTORS - p1 #3 Group:1 hour reversal SUBJECT_SAMPLE_FACTORS - p3 #1 Group:3 hour reversal SUBJECT_SAMPLE_FACTORS - p3 #2 Group:3 hour reversal SUBJECT_SAMPLE_FACTORS - p3 #3 Group:3 hour reversal SUBJECT_SAMPLE_FACTORS - p6 #1 Group:6 hour reversal SUBJECT_SAMPLE_FACTORS - p6 #2 Group:6 hour reversal SUBJECT_SAMPLE_FACTORS - p6 #3 Group:6 hour reversal SUBJECT_SAMPLE_FACTORS - p12 #1 Group:12 hour reversal SUBJECT_SAMPLE_FACTORS - p12 #2 Group:12 hour reversal SUBJECT_SAMPLE_FACTORS - p12 #3 Group:12 hour reversal SUBJECT_SAMPLE_FACTORS - p0 no stretch #1 Group:no stretch-serum free SUBJECT_SAMPLE_FACTORS - p1 no stretch #1 Group:no stretch-serum free SUBJECT_SAMPLE_FACTORS - p3 no stretch #1 Group:no stretch-serum free SUBJECT_SAMPLE_FACTORS - p6 no stretch #1 Group:no stretch-serum free SUBJECT_SAMPLE_FACTORS - serum 6hr no stretch #1 Group:serum-no stretch SUBJECT_SAMPLE_FACTORS - serum 6hr no stretch #2 Group:serum-no stretch SUBJECT_SAMPLE_FACTORS - serum 6hr no stretch #3 Group:serum-no stretch #COLLECTION CO:COLLECTION_SUMMARY Media was collected from all different 6 wells, and stored in -80 C #TREATMENT TR:TREATMENT_SUMMARY N/A #SAMPLEPREP SP:SAMPLEPREP_SUMMARY N/A #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY GC/MS methods follow previous studies using a 6890 N GC connected to a 5975 CH:CHROMATOGRAPHY_SUMMARY Inert single quadrupole MS (Agilent Technologies, Santa Clara, CA) (Bonikos et CH:CHROMATOGRAPHY_SUMMARY al. 1975; Fiehn 2008; Kind et al. 2009). The two wall-coated, open-tubular GC CH:CHROMATOGRAPHY_SUMMARY columns connected in series are both from J&W/Agilent (part 122–5512), DB5-MS, CH:CHROMATOGRAPHY_SUMMARY 15 meters in length, 0.25 mm in diameter, with an 0.25-l m luminal film. CH:CHROMATOGRAPHY_SUMMARY Positive ions generated with conventional electron-ionization at 70 eV are CH:CHROMATOGRAPHY_SUMMARY scanned broadly from 600 to 50 m/z in the detector throughout the 45 min cycle CH:CHROMATOGRAPHY_SUMMARY time. CH:CHROMATOGRAPHY_TYPE GC CH:INSTRUMENT_NAME Agilent 6890N CH:COLUMN_NAME Agilent DB5-MS (15m × 0.25mm, 0.25um) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:MS_COMMENTS - MS:INSTRUMENT_NAME Agilent 5975 MS:INSTRUMENT_TYPE Single quadrupole MS:MS_TYPE EI MS:ION_MODE POSITIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Peak values (Log transformed) MS_METABOLITE_DATA_START Samples p0 #1 p0 #2 p0 #3 p1 #1 p1 #2 p1 #3 p3 #1 p3 #2 p3 #3 p6 #1 p6 #2 p6 #3 p12 #1 p12 #2 p12 #3 p0 no stretch #1 p1 no stretch #1 p3 no stretch #1 p6 no stretch #1 serum 6hr no stretch #1 serum 6hr no stretch #2 serum 6hr no stretch #3 Factors Group:6 hour stretch Group:6 hour stretch Group:6 hour stretch Group:1 hour reversal Group:1 hour reversal Group:1 hour reversal Group:3 hour reversal Group:3 hour reversal Group:3 hour reversal Group:6 hour reversal Group:6 hour reversal Group:6 hour reversal Group:12 hour reversal Group:12 hour reversal Group:12 hour reversal Group:no stretch-serum free Group:no stretch-serum free Group:no stretch-serum free Group:no stretch-serum free Group:serum-no stretch Group:serum-no stretch Group:serum-no stretch Glyoxylic acid 13.08 12.66 13.58 13.08 12.94 13.08 13.34 12.85 12.8 12.96 13 C10 hydrocarbon 13.29 13.57 14.07 14.87 13.76 14.93 15.02 14.7 14.88 15.27 13.54 14.63 14.66 14.87 14.84 14.65 14.82 14.87 15.29 15.04 14.73 13.38 epsilon-Caprolactam 12.61 13.15 12.46 11.74 14.34 12.04 13.88 13.41 12.73 13.09 14.41 13.84 14.46 14.31 13.46 14.07 11.24 14.7 14.37 13.09 2-Hydroxypyridine 12.14 12.27 11.48 14.04 11.86 14.38 11.37 12.73 Pyruvic acid 22.39 22.32 22.99 22.5 22.36 22.7 22.44 22.41 22.54 22.28 22.43 22.57 21.9 21.93 21.81 22.35 22.8 22.22 21.68 21.21 21.34 21.46 Lactic acid 26.03 25.81 26.19 25.84 26.15 26.25 26.35 26.26 26.23 26.48 26.87 26.63 26.84 27.11 26.93 25.98 26.24 26.48 26.41 27.24 27.7 27.63 Glycolic acid 14.16 13.99 14.12 14.15 14.18 14.1 13.96 14.04 14.22 15.27 15.28 14.86 13.72 11.92 15.42 15.78 15.33 Alanine 21.96 21.91 22.43 22.16 22.23 22.57 22.46 22.4 22.42 22.71 23.01 23.05 23.13 23.27 23.19 22.29 22.49 22.65 22.68 22.76 23.3 23.11 C11 hydrocarbon 15.31 15.69 15.41 16.22 15.46 15.83 14.93 15.65 13.58 13.84 15.41 15.6 13.73 15.8 13.75 13.93 13.71 13.83 16.02 15.9 15.61 15.19 2-Hydroxy-2-methylbutyric acid 15.12 15.9 15.63 15.1 15.25 15.48 14.75 15.81 15.36 15.3 2-Ketovaline 16.61 16.27 16.96 16.27 16.19 17.11 16.83 17.35 17.02 17.2 17.52 17.65 18.01 18.31 18.21 16.91 17.28 17.42 17.42 17.88 18.5 18.4 2-Hydroxybutyric acid 23.17 23.19 23.82 23.71 23.61 23.87 23.36 23.33 23.89 23.29 23.69 23.86 23.23 23.3 23.57 23.4 23.6 23.4 22.99 23.18 24.12 23.59 Sulfuric acid 16.57 17.4 12.47 13.85 12.55 14.6 15.39 17 16.18 Dodecamethylpentasiloxane 17.14 16.68 16.83 16.29 16.84 16.92 17.54 16.64 17.05 17.39 17.71 17.71 17.61 17.66 17.55 15.93 16.35 15.87 15.89 17.43 17.92 18.07 Malonic acid 17.24 17.75 17.17 16.45 17.88 16.97 17.85 16.92 17.27 15.99 17.1 17.11 16.5 17.47 16.94 16.45 16.52 17.27 17.07 17.44 17.23 16.58 2-Ketoleucine 19.5 19.23 20.09 19.51 18.94 20.05 19.95 19.84 19.99 20.11 20.36 20.49 20.69 20.94 20.77 19.76 20.03 20.04 20.14 20.28 21 21 Valine 23.48 23.26 24.58 24.05 23.53 24.71 23.28 23.96 24.43 24.08 24.06 24.47 24.56 23.94 24.82 24.42 24.77 24.06 24.3 24.21 24.84 24.75 C12 hydrocarbon 13.57 14.66 13.98 14.57 13.82 14.82 14.93 14.2 14.51 14.42 14.24 14.44 14.18 14.36 13.85 14.36 14.36 14.32 14.58 14.49 15.16 14.64 Benzoic acid 16.69 16.67 16.76 16.84 16.66 16.8 16.86 16.83 17.05 17.03 16.68 16.76 16.83 16.84 16.84 16.64 16.67 16.69 16.33 17.06 17.23 16.91 Ethanolamine 16.38 16.37 16.97 16.69 16.63 16.82 17.05 16.77 17.03 16.9 16.87 17.1 17.7 17.65 17.57 16.97 17.37 17.22 17.5 17.36 17.84 17.68 Phosphoric acid 17.65 17.45 17.46 16.87 16.85 18.11 17.35 17.34 17.2 18.41 17.11 17.68 17.34 17.02 16.97 17.12 16.53 17.99 17.97 17.67 17.59 1,3-Dihydroxyacetone 23.24 23.35 23.62 23.72 23.42 23.9 23.75 23.63 24 23.82 23.71 Leucine 24.08 24.25 24.84 24.13 24.42 25 24.53 24.04 24.69 24.5 24.49 24.69 24.88 24.64 25.03 24.59 25 24.31 24.54 24.46 24.85 24.91 Glycerol 23.24 23.35 21.41 19.76 23.62 25 21.28 23.42 23.9 23.75 20.72 20.6 21.5 22.04 22.78 24.59 20.63 23.71 20.44 20.82 20.37 22.56 Ethylmalonic acid 23.24 23.35 23.62 22.48 23.72 23.42 23.9 23.75 23.63 24 23.82 22.78 23.71 22.56 C13 hydrocarbon 16.42 16.85 16.77 17.18 16.57 17 17.08 16.78 16.97 17.16 16.65 16.67 17.06 16.89 16.78 16.59 16.75 16.83 17.09 17.03 17.02 16.28 Isoleucine 24.28 24.64 24.97 24.35 24.55 25.05 24.86 24.47 24.76 24.35 24.75 24.78 24.96 24.92 24.97 24.63 25.09 24.75 24.68 24.41 24.91 24.98 Proline 16.21 16.91 17.72 17.12 17.22 18.01 17.81 17.55 17.95 18 18.45 18.62 19.29 19.29 19.54 17.83 18.2 18.6 18.36 19.61 20.37 20.34 Glycine 23.6 23.7 24.03 23.72 23.75 24.31 23.98 23.87 24.08 23.7 24.14 24.24 24.39 24.29 24.53 24.03 24.4 24.14 23.95 23.98 24.66 24.65 Succinic acid 16.71 17.08 17.48 17.92 17.11 17.47 17.57 17.59 17.87 17.37 17.39 17.74 16.79 17.64 18.37 18.24 Glyceric acid 13.71 16.25 15.27 MSTFA artifact 13.04 14.54 15.91 15.17 15.86 14.2 15.31 16.15 15.58 15.36 15.42 14.45 15.05 17.39 15.93 15.97 15.58 15.45 12.87 15.92 15.93 Fumaric acid 11.28 11.84 12.48 12.02 12.11 12.35 13.1 12.33 12.5 13.18 13.22 11.41 13.53 13.92 13.84 12.01 13.85 12.57 12.51 14.2 15.32 14.88 Serine 23.26 23.43 23.76 23.22 23.54 23.73 23.64 23.42 23.63 22.76 23.64 23.54 23.72 23.64 23.65 23.55 24.01 23.59 23.31 22.14 22.88 22.78 Threonine 24.83 24.9 25.34 24.83 25.06 25.2 25.14 24.96 25.2 24.78 25.14 25.28 25.34 25.36 25.43 25.04 25.4 25.18 25.08 24.67 25.25 25.26 Hydroquinone 14.78 14.88 15.05 14.98 15.84 14.95 15.03 15.44 15.7 15.16 15.48 15.14 15.41 15.92 15.48 15.6 16.69 16 C14 Hydrocarbon 13.33 14.11 14.12 13.61 13.49 13.18 14.21 13.14 13.52 14.11 beta-Alanine 12.53 12.52 12.19 11.83 9.32 12.65 12.47 12.85 12.57 12.3 12.43 Threose/erythrose 14.21 12.9 12.94 13.14 13.1 13.13 11.07 13.53 12.95 12.98 12.91 12.57 11.64 14.15 Aminomalonic acid 20.71 19.23 20.55 20.24 20.88 20.6 20.63 19.77 19.96 19.74 21.26 20.4 20.19 20.76 20.37 20 20.75 19.8 20.1 19.86 22.15 21.05 Aspartic acid 17.29 15.5 17.25 16.78 17.64 16.92 17.28 15.68 16.05 17.46 18.13 17.05 16.79 18.39 17.74 15.97 17.12 16.47 17.72 16.33 19.97 18.45 Malic acid 15.68 15.52 15.95 15.65 16.26 16.5 16.71 16.2 16.36 16.35 17.12 17.05 17.28 17.56 17.94 16 16.2 16.65 16.28 17.12 18.51 18.41 Nicotinamide 16.1 16.66 16.9 15.54 16.46 17.07 16.92 16.59 16.7 17.33 16.95 17.14 17.58 17.08 17.29 17.02 16.8 13.98 17.51 17.51 C15 hydrocarbon 17.01 17.05 16.98 17.19 17.04 17.4 17.33 14.87 Hydroxyprolines 13.72 11.84 12.99 11.97 11.85 13.14 12.81 13.34 12.68 13.31 12.65 12.98 12.95 13.34 Threitol/erythritol 15 14.85 15.41 15.64 15.97 15.97 15.51 15.79 15.59 15.06 16.23 16.66 Methionine 21.5 21.66 22.42 21.56 21.74 22.18 21.77 21.83 22.23 21.5 22.11 22.21 22.31 22.3 22.35 22.07 22.55 22.07 22.13 21.33 22.18 22.26 Glutamic acid 25.32 25.22 25.61 24.87 25.51 25.6 25.56 25.32 25.56 25.17 25.81 25.67 25.71 25.95 25.99 25.48 25.71 25.55 25.5 25.05 25.87 25.88 Erythronic acid/threonic acid 14.67 15.24 15.05 15.08 15.35 14.95 14.58 14.64 15.6 15.31 15.68 15.69 15.97 14.76 15.39 15.43 15.65 15.33 16.39 16.31 Cysteine 16.39 16.98 17.76 16.81 16.89 17.84 17.27 17.34 18.02 17.31 17.77 18.36 17.89 18.03 18.45 17.88 18.12 18.14 18 17.76 18.18 18.67 alpha Ketoglutaric acid 16.1 16.88 15.84 16.07 16.96 16.45 16.63 17.23 16.39 17 17.52 17.46 17.89 18.21 16.01 16.56 16.72 16.94 17.03 18.31 18.22 2-Isopropylmalic acid 21.24 18.8 20.2 19.72 20.94 19.35 20.21 17.88 17.7 19.76 20.7 18.5 18.69 20.28 18.82 18.7 20.02 18.54 20.02 18.24 22.22 20.17 Phenylalanine 23.9 23.77 24.38 23.59 23.95 24.35 24.09 23.87 24.18 23.69 24.26 24.39 24.38 24.43 24.56 24.22 24.45 24.22 24.15 23.54 24.44 24.4 Succinylacetone 13.18 13.46 13.85 13.3 13.65 13.68 13.45 13.98 12.43 13.81 13.19 14.05 13.69 13.88 13.94 13.49 Asparagine 16.54 17.17 17.87 17.04 17.57 17.32 17.76 17.69 17.54 17.6 18.11 18.1 18.77 18.74 18.9 17.52 17.76 18.04 17.81 18.2 19.06 18.97 Cysteinesulfinic acid 14.94 13.02 13.84 13.54 13.34 13.11 13.21 13.22 13.61 12.64 14.12 11.59 12.53 Pentitols 13.81 14.01 14.12 13.73 13.46 15.2 14.02 14.31 14.58 14.61 15.13 16.41 16.83 16.89 14.61 14.36 15.99 15.77 16.49 17.71 17.55 Putrescine 13.63 13.38 14.28 13.11 14.3 13.87 13.9 13.97 13.87 14.29 14.18 13.61 13.24 13.62 13.61 14.87 16.1 15.73 Glutamine 26.35 26.44 26.73 26.31 26.48 26.61 26.51 26.46 26.62 26.1 26.59 26.72 26.6 26.59 26.66 26.57 26.85 26.56 26.55 26.14 26.47 26.74 Gentisic acid 14.83 14.52 13.6 14.93 15.75 13.97 15.32 14.94 15.61 15.38 Unoximated aldohexose 12.28 14.78 13.26 Hypoxanthine 14.69 14.03 13.56 14.29 14.07 14.86 15.13 15.42 12.46 14.85 14.51 Citric acid/isocitric acid 19.35 19.53 18.89 19.5 20.2 20.08 20.48 20.05 19.47 19.71 21.16 20.9 21.65 21.69 21.69 19.84 19.97 20.38 20.28 22.22 23.04 23.08 Ornithine 20.57 18.71 19.42 18.61 19.12 19.38 19.4 19.1 19.53 19.15 19.63 19.79 19.58 20.01 19.93 19.06 19.46 19.7 19.51 19.6 20.25 20.49 Arginine/Citrulline 18.05 18.06 18.83 17.74 18.42 19.12 18.51 18.16 18.97 18.54 18.83 19.03 18.63 19.01 19.82 18.65 19.3 18.87 18.95 16.27 18.53 18.67 deuterated Myristic Acid (RTL std) 22.05 22.23 22.39 22.04 22.27 22.42 21.99 22.37 22.39 22.26 22.24 22.38 22.34 22.14 22.49 22.38 22.17 22.38 22.29 22.1 22.35 22.55 Myristic acid or Pentadecanol 14.96 16.33 14.75 16.25 13.31 14.22 16.52 14.98 14.93 16.07 16.24 14.19 14.66 15.07 15.01 14.38 14.67 15.12 14.38 16.08 Methionine sulfoxide 15.68 15.84 13.52 14.41 17.22 15 16.71 16.14 14.72 14.82 14.63 14.36 15 14.56 16.28 15.16 14.75 14.9 15.28 15.36 15.25 Fructose or similar ketohexose 22.71 22.52 22.13 19.55 22.15 21.92 21.95 21.43 21.75 21.59 21.91 21.23 22 21.9 22.09 21.06 21.87 21.51 22.08 20.84 21.3 21.89 Gluconic acid or similar sugar acid 22.93 22.58 23.24 22.38 22.97 23.29 23.14 23.08 23.06 22.87 23.3 23.11 23 23.28 23.53 22.78 23.33 23.09 23.15 20.84 21.3 22.74 Pyridoxine 18.68 18.96 20.53 18.56 18.86 20.74 18.78 19.51 16.82 19.09 19.02 22.15 19.69 19.23 20.31 19.71 19.56 19.88 18.83 21.42 18.97 19.35 Glucose and other aldohexoses 2 27.68 27.72 27.97 27.58 27.57 27.93 27.16 27.84 28.18 27.74 27.62 27.95 27.9 27.7 27.87 27.98 27.94 27.98 27.96 27.78 26.93 27.79 Glucose and other aldohexoses 1 27.68 27.72 27.97 27.58 27.57 27.93 26.96 27.84 28.18 27.74 27.62 27.95 27.9 27.7 27.87 27.98 27.94 27.98 27.96 27.78 27.58 27.79 Dehydroascorbic acid� 25.56 23.69 25.41 23.55 25.21 24.88 24.67 25.35 25.31 25.86 25.5 25.59 25.6 24.82 25.43 26.94 25.91 24.87 25.4 23.1 22.7 27 Hexitols 26.83 26.87 27.02 26.65 27.07 27.34 26.96 26.96 27.19 26.95 27.05 27.15 27.07 27.02 27.17 26.94 27.17 27.01 27.16 26.97 26.93 27.34 Histidine 20.4 20.78 21.02 20.69 20.78 21.04 20.86 20.72 20.84 20.6 21 20.96 21.08 21.08 21.28 21.05 21.11 20.97 20.63 20.58 21.18 21.23 Lysine 25.16 25.19 25.55 25.2 25.38 25.51 25.39 25.25 25.42 25.13 25.39 25.49 25.52 25.68 25.75 25.41 25.66 25.51 25.52 25.01 25.49 25.52 Tyrosine 24.36 24.4 24.78 24.28 24.54 24.67 24.6 24.39 24.56 24.19 24.75 24.74 24.78 24.88 25.03 24.6 24.89 24.63 24.66 24.14 24.71 24.87 Sedoheptulose or similar sugar 23.47 23.55 23.9 23.52 23.65 23.77 23.79 23.58 23.65 23.36 23.79 23.78 23.92 23.88 24.01 23.83 24.03 23.74 23.73 23.14 23.57 23.78 Glucose and other aldohexoses 3 26.61 26.39 26.1 26.2 26.84 25.94 26.64 25.46 24.14 25.93 27.2 25.45 25.97 26.8 25.76 25.85 26.51 25.1 26.21 25.99 27.38 26.65 Pantothenic acid 19.83 19.58 19.48 19.34 20.1 19.75 19.88 19.01 19.13 19.38 20.01 19.42 19.44 19.97 19.61 19.91 19.96 19.32 19.9 19.13 19.98 20.09 Palmitic acid 20.66 21.8 20.93 22.07 20.89 21.91 22.01 20.94 21.96 21.86 20.87 20.78 22.12 21.79 22.05 21.6 21.72 21.73 22.06 21.99 21.94 20.65 Myoinositol 22.15 22.12 22.27 21.87 22.23 22.34 22.28 22.04 22.07 21.79 22.49 22.38 22.18 22.27 22.41 22.25 22.47 22.22 22.27 21.84 22.42 22.52 Guanine 12.06 12.91 12.42 14.2 14.65 15.12 13.8 13.47 13.75 14.84 C21 Hydrocarbon 14.58 14.83 15.88 14.15 14.21 15.01 15.02 15.12 Methyl stearate 17.48 17.09 18.78 18.68 17.88 16.82 18.35 19 17.09 18.19 17.89 16.74 18.04 16.6 Tetracosamethylundecasiloxane 14.8 14.72 13.72 12.42 13.5 12.12 Heptadecanoic acid/Octadecanol 13.64 13.89 15.13 15.24 13.25 15.21 15.04 C22 hydrocarbon 13.92 14.6 14.51 14.48 13.72 14.4 14.07 13.21 14.3 14.12 14.47 13.92 13.93 14.23 14.41 14.07 13.76 Tryptamine/Norepinephrine 13.61 15.14 15.08 14.76 14.94 14.79 14.76 14.7 15.82 14.65 14.25 15.74 15.16 15.15 16.19 14.75 14.53 15.49 15.74 14.09 15.09 Tryptophan 20.43 20.52 21.25 20.15 20.6 21.42 20.67 20.5 21.07 20.15 21.2 21.36 20.92 21.18 21.93 21.01 21.59 21.09 21.15 20.46 21.82 21.87 Stearic acid 21.46 22.16 21.63 22.16 21.77 22.25 22.23 21.94 22.11 22.31 21.68 21.76 22.24 22.15 22.35 21.95 22.06 22.12 22.27 22.32 22.16 21.51 Hexacosemethyldodecasiloxane 13.16 13 12.51 14.23 13.96 13.15 12.33 13.27 13.68 14.73 Cystine 22.62 22.51 22.52 22.19 22.56 22.75 22.77 22.35 22.21 21.72 23.13 22.8 22.81 22.8 22.94 22.67 23.04 22.44 22.49 22.4 22.87 22.97 Hexose-6-phosphate 18.89 19.13 17.13 18.74 18.49 16.49 18.9 18.24 17.13 17.11 19.34 19.79 18.24 19.36 5-Hydroxytryptophan 14.6 14.34 14.73 14.44 15.25 15.87 17.06 17.33 17.46 15.1 15.18 15.74 16.61 18.61 17.95 beta-Monopalmitin 17.26 16.81 15.86 16.63 16.82 16.64 16.76 16.65 16.23 16.39 18.19 16.21 16.75 16.7 16.81 16.77 16.72 16.43 14.04 16.85 18.88 17.19 alpha-Monopalmitin 21.8 21.85 21.55 21.97 21.8 22.11 22.08 21.72 22.06 21.87 22.08 22.05 21.91 22.22 22.38 22.24 22.25 22 21.77 22.1 22 22.07 Sucrose and similar disaccharides 13.67 13.37 13.28 11.93 12.12 12.9 13.76 13.71 12.06 13.57 13.93 13.02 Lactose or similar disaccharide 18.3 18.19 18.88 17.93 18.25 18.55 18.57 18.47 18.83 17.92 18.53 19.19 19.06 18.63 19.31 18.42 18.81 18.96 18.48 beta-Monostearin 19.56 19.52 19.08 19.96 20.16 19.57 19.79 19.11 19.7 19.81 19.74 19.52 19.65 19.98 20.05 19.62 20.22 19.79 19.77 19.94 19.99 19.93 Maltose or similar disaccharide 16.42 18.39 18.61 17.42 18.27 18.91 17.58 18.64 18.58 18.29 17.61 18.49 17.8 19.28 18.37 18.55 19.43 17.29 18.5 17.29 18.41 18.21 alpha-Monostearin 24.83 24.92 24.67 25.16 25 25.24 25.21 24.79 25.2 25.04 25.05 25.1 25.08 25.28 25.4 25.35 25.39 25.23 24.93 25.14 24.97 25.08 Isomaltose or similar disaccharide 19.67 19.5 19.73 19.12 19.52 19.78 19.86 19.42 19.49 19.02 20.09 19.96 19.86 19.85 20.07 19.72 20.11 19.71 19.73 19.24 20.21 20.34 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name CAS Retention time Reverse SimPeak Glyoxylic acid 563-96-2 5.916 73.5 7 C10 hydrocarbon 124-18-5 6.185 81.2 8 epsilon-Caprolactam 105-60-2 6.2574 82.5 9 2-Hydroxypyridine 142-08-5 6.4525 85.2 13 Pyruvic acid 127-17-3 6.5922 99 14 Lactic acid 79-33-4 6.7791 99.8 16 Glycolic acid 79-14-1 7.0133 88.1 20 Alanine 56-41-7 7.4056 99 23 C11 hydrocarbon 1120-21-4 7.4866 75.1 24 2-Hydroxy-2-methylbutyric acid 3739-30-8 7.8051 68.9 28 2-Ketovaline 759-05-7 7.8521 94.9 29 2-Hydroxybutyric acid 565-70-8 7.9649 74 32 Sulfuric acid 7664-93-9 8.3068 86.9 35 Dodecamethylpentasiloxane 8.3661 0 36 Malonic acid 141-82-2 8.9088 84.9 46 2-Ketoleucine 816-66-0 9.0128 99.3 49 Valine 72-18-4 9.0754 99.2 50 C12 hydrocarbon 112-40-3 9.2413 74.3 53 Benzoic acid 65-85-0 9.5983 98.4 60 Ethanolamine 141-43-5 9.8107 90.5 64 Phosphoric acid 7664-38-2 9.8485 91.8 65 1,3-Dihydroxyacetone 96-26-4 9.8665 65.6 66 Leucine 61-90-5 9.8713 97.9 67 Glycerol 56-81-5 9.8815 93.4 69 Ethylmalonic acid 601-75-2 9.8799 69.9 70 C13 hydrocarbon 629-50-5 10.0037 77.7 72 Isoleucine 443-79-8 10.1788 100 74 Proline 147-85-3 10.2512 96 75 Glycine 56-40-6 10.3698 99 77 Succinic acid 110-15-6 10.4499 75.6 79 Glyceric acid 473-81-4 10.663 71.7 83 MSTFA artifact 24589-78-4 10.7331 86.4 84 Fumaric acid 17013-01-3 10.961 86.3 87 Serine 56-45-1 11.0755 99 88 Threonine 72-19-5 11.4268 99 92 Hydroquinone 123-31-9 11.8527 51 95 C14 Hydrocarbon 629-59-4 11.9049 73.3 96 beta-Alanine 107-95-9 12.0056 75.6 97 Threose/erythrose 95-43-2 12.2856 73.5 98 Aminomalonic acid 1068-84-4 12.5131 65.3 101 Aspartic acid 16504-56-6 12.5295 71.6 102 Malic acid 617-48-1 12.7361 87.3 105 Nicotinamide 98-92-0 12.7709 92.5 106 C15 hydrocarbon 629-62-9 12.8566 82.1 107 Hydroxyprolines 4298-08-2 12.9591 75.8 110 Threitol/erythritol 149-32-6 12.9845 81.5 111 Methionine 63-68-3 13.1669 86.8 112 Glutamic acid 56-86-0 13.199 99.7 113 Erythronic acid/threonic acid 13752-84-6 13.4125 75.4 115 Cysteine 52-90-4 13.5897 93.1 119 alpha Ketoglutaric acid 328-50-7 13.8052 82.2 121 2-Isopropylmalic acid 16048-89-8 14.1449 77 125 Phenylalanine 63-91-2 14.4664 97 128 Succinylacetone 51568-18-4 14.6441 78.8 131 Asparagine 70-47-3 14.9279 93.3 136 Cysteinesulfinic acid 1115-65-7 15.0536 75.6 140 Pentitols 488-82-4 15.5475 81.4 149 Putrescine 110-60-1 15.7483 81 151 Glutamine 56-85-9 16.1276 98 154 Gentisic acid 490-79-9 16.2475 75 157 Unoximated aldohexose 138-59-0 16.4398 55.7 159 Hypoxanthine 68-94-0 16.4736 85.3 160 Citric acid/isocitric acid 5949-29-1 16.5612 98.2 162 Ornithine 70-26-8 16.5768 92.1 163 Arginine/Citrulline 74-79-3 16.6427 96.1 164 deuterated Myristic Acid (RTL std) 60658-41-5 16.7219 99 165 Myristic acid or Pentadecanol 629-76-5 16.9412 94 169 Methionine sulfoxide 3226-65-1 17.001 81.4 171 Fructose or similar ketohexose 87-81-0 17.1235 98.8 173 Gluconic acid or similar sugar acid 1198-69-2 17.2245 83.9 176 Pyridoxine 65-23-6 17.39 93 183 Glucose and other aldohexoses 2 2595-98-4 17.5606 98.7 184 Glucose and other aldohexoses 1 50-99-7 17.5983 99.3 187 Dehydroascorbic acid� 490-83-5 17.5577 73.3 188 Hexitols 87-78-5 17.5814 90.7 189 Histidine 5934-29-2 17.6883 81.1 191 Lysine 56-87-1 17.7219 99 192 Tyrosine 60-18-4 17.8678 99 195 Sedoheptulose or similar sugar 469-90-9 18.0049 75.7 200 Glucose and other aldohexoses 3 50-99-7 18.1978 97.4 206 Pantothenic acid 137-08-6 18.3531 94.6 208 Palmitic acid 57-10-3 18.908 99.9 217 Myoinositol 87-89-8 19.3215 97 221 Guanine 73-40-5 19.6127 85 225 C21 Hydrocarbon 629-94-7 19.6866 66.9 227 Methyl stearate 112-61-8 19.7165 87.8 228 Tetracosamethylundecasiloxane 107-53-9 19.7579 62.7 229 Heptadecanoic acid/Octadecanol 506-12-7 19.8307 81.3 230 C22 hydrocarbon 629-97-0 20.0663 66.5 232 Tryptamine/Norepinephrine 61-54-1 20.235 73 233 Tryptophan 73-22-3 20.4606 100 236 Stearic acid 57-11-4 20.7175 99.1 240 Hexacosemethyldodecasiloxane 2471-08-1 21.0244 64.8 246 Cystine 56-89-3 21.1313 99.4 247 Hexose-6-phosphate 3672-15-9 21.3133 78.7 248 5-Hydroxytryptophan 4350-09-8 22.1245 69.7 255 beta-Monopalmitin 23470-00-0 23.2523 96.1 264 alpha-Monopalmitin 542-44-9 23.5166 99.8 266 Sucrose and similar disaccharides 57-50-1 23.8844 75.2 268 Lactose or similar disaccharide 63-42-3 24.6518 81.6 273 beta-Monostearin 31566-31-1 24.6937 96.6 275 Maltose or similar disaccharide 69-79-4 24.8427 89.5 278 alpha-Monostearin 123-94-4 24.9608 100 279 Isomaltose or similar disaccharide 499-40-1 25.6047 98.2 282 METABOLITES_END #END