#METABOLOMICS WORKBENCH amroilaiwy_20160210_110337 STUDY_ID:ST000348 ANALYSIS_ID:AN000579 PROJECT_ID:PR000279 VERSION 1 CREATED_ON February 23, 2016, 12:26 pm #PROJECT PR:PROJECT_TITLE Non targeted meatbolomic profiling of transgenic- humanized amylin producing PR:PROJECT_TITLE rats PR:PROJECT_TYPE GC-MS non targeted analysis PR:PROJECT_SUMMARY Non targeted metabolomic analysis on samples from rats expressing human amylin. PR:INSTITUTE University of North Carolina PR:LABORATORY Multiple Centers PR:LAST_NAME Ilaiwy PR:FIRST_NAME Amro PR:ADDRESS 111 Mason Farm road, Chapel Hill, North Carolina, 27599-7126, USA PR:EMAIL amroilaiwy@gmail.com PR:PHONE 210-596-0171 PR:FUNDING_SOURCE NIH, Fondation Leducq #STUDY ST:STUDY_TITLE Metabolomic analysis on samples from rats expressing human amylin (brain ST:STUDY_TITLE tissue). ST:STUDY_TYPE Non targeted metabolomics-Brain tissue ST:STUDY_SUMMARY Human amylin proteotoxicity impairs protein biosynthesis, and alters major ST:STUDY_SUMMARY cellular signaling pathways in the heart, brain and liver of humanized diabetic ST:STUDY_SUMMARY rat model in vivo. The 37 amino acid hormone Amylin is co­secreted with insulin ST:STUDY_SUMMARY from ß cells in the pancreas. In pre­diabetic and obese humans, chronic amylin ST:STUDY_SUMMARY hypersecretion parallels the course of disease and is involved in the ST:STUDY_SUMMARY pathophysiology of beta cell destruction in the pancreas. Recent studies in rats ST:STUDY_SUMMARY with transgenic expression of beta cell amylin (HIP), we have discovered that ST:STUDY_SUMMARY human amylin is prone to misfolding and has proteotoxic effects in vivo, ST:STUDY_SUMMARY resulting in the induction of cell death paralleling the pathophysiology of ST:STUDY_SUMMARY neurodegenerative disease. These misfolded proteotoxic amylin proteins are found ST:STUDY_SUMMARY to migrate to both the brain and heart to induce both neurologic deficits and ST:STUDY_SUMMARY cardiac dysfunction. In the present study, we use non­targeted GC­MS ST:STUDY_SUMMARY metabolomics analysis to investigate the metabolic consequences of amyloidogenic ST:STUDY_SUMMARY and cytotoxic amylin oligomers and diabetes in HIP heart, brain, liver, and ST:STUDY_SUMMARY plasma compared to wild type controls at 1 year of age. We identified that HIP ST:STUDY_SUMMARY hearts had 45 significantly altered metabolites by t­test (p<0.05) compared to ST:STUDY_SUMMARY wildtype control hearts (0.1­34.3 fold different,N=8/group). Similarly, we ST:STUDY_SUMMARY identified 30 metabolites significantly different in the HIP brain by t­test ST:STUDY_SUMMARY (p<0.05) compared to wildtype control brains (0.2­25.2 fold different ST:STUDY_SUMMARY (N=~10/group). HIP livers had 58 metabolites significantly altered by t­test ST:STUDY_SUMMARY (p<0.05) compared to wildtype livers (0.01­99.4 fold different,N=~10/group. ST:STUDY_SUMMARY Pathway enrichment analysis identified a systemic alteration in protein ST:STUDY_SUMMARY biosynthesis in the heart and brain of HIP rats compared to wild type controls. ST:STUDY_SUMMARY Alterations in phenylalanine metabolism and aminoacyl­tRNA biosynthesis were ST:STUDY_SUMMARY specifically affected in heart and plasma. Tyrosine metabolism is affected ST:STUDY_SUMMARY across organs, including decreased tyrosine (heart), phenylalanine (heart, ST:STUDY_SUMMARY liver, brain), and increased fumarate (heart, liver, brain). Increased urea and ST:STUDY_SUMMARY urea cycle were identified in heart and liver. As protein degradation is a major ST:STUDY_SUMMARY up­regulator of the urea cycle in human rat diabetic models, these findings ST:STUDY_SUMMARY suggest a broader connection between amylin, diabetes, protein catabolism, and ST:STUDY_SUMMARY effects on the urea cycle, which may contribute to the increased morbidity and ST:STUDY_SUMMARY mortality in diabetics at a multi­system level beyond the effects on glucose ST:STUDY_SUMMARY metabolism. ST:INSTITUTE Duke University ST:DEPARTMENT Sarah W. Stedman Nutrition and Metabolism Center ST:LABORATORY Metabolomics lab ST:LAST_NAME Ilaiwy ST:FIRST_NAME Amro 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 210-596-0171 ST:STUDY_COMMENTS Brain tissue-Metabolomic analysis was performed at Duke Metabolomics lab #SUBJECT SU:SUBJECT_TYPE Animal SU:SUBJECT_SPECIES Rattus norvegicus #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - 20 Genotype:WT Brain SUBJECT_SAMPLE_FACTORS - 32 Genotype:WT Brain SUBJECT_SAMPLE_FACTORS - 30 Genotype:WT Brain SUBJECT_SAMPLE_FACTORS - 33 Genotype:WT Brain SUBJECT_SAMPLE_FACTORS - 11 Genotype:WT Brain SUBJECT_SAMPLE_FACTORS - 25 Genotype:WT Brain SUBJECT_SAMPLE_FACTORS - 9 Genotype:WT Brain SUBJECT_SAMPLE_FACTORS - SD1 Genotype:WT Brain SUBJECT_SAMPLE_FACTORS - SD4 Genotype:WT Brain SUBJECT_SAMPLE_FACTORS - SD2 Genotype:WT Brain SUBJECT_SAMPLE_FACTORS - 15 Genotype:HIP Brain SUBJECT_SAMPLE_FACTORS - 48 Genotype:HIP Brain SUBJECT_SAMPLE_FACTORS - 34 Genotype:HIP Brain SUBJECT_SAMPLE_FACTORS - 23 Genotype:HIP Brain SUBJECT_SAMPLE_FACTORS - 22 Genotype:HIP Brain SUBJECT_SAMPLE_FACTORS - 18 Genotype:HIP Brain SUBJECT_SAMPLE_FACTORS - 32HIP Genotype:HIP Brain SUBJECT_SAMPLE_FACTORS - 33HIP Genotype:HIP Brain SUBJECT_SAMPLE_FACTORS - 46 Genotype:HIP Brain SUBJECT_SAMPLE_FACTORS - 10 Genotype:HIP Brain #COLLECTION CO:COLLECTION_SUMMARY Brain tissue was harvested amd then flash frozen in a liquid nitrogen cooled CO:COLLECTION_SUMMARY biopress #TREATMENT TR:TREATMENT_SUMMARY Fraction of brain tissue weighed (25–50 mg wet weight), then the finely cut up TR:TREATMENT_SUMMARY tissue quickly added to fresh pre-made buffer (50 % acetyl-nitrile, 50 % water, TR:TREATMENT_SUMMARY 0.3% formic acid) at a standard concentration of 25 mg/475 mcl buffer then fully TR:TREATMENT_SUMMARY homogenized on ice for 10–25 s and placed on dry ice/stored at - 80C #SAMPLEPREP SP:SAMPLEPREP_SUMMARY The samples were crash deprotonized by methanol precipitation and spiked with SP:SAMPLEPREP_SUMMARY D27-deuterated myristic acid (D27-C14:0) as an internal standard for SP:SAMPLEPREP_SUMMARY retention-time locking and dried. The trimethylsilyl-D27-C14:0 standard SP:SAMPLEPREP_SUMMARY retention time (RT) was set at 16.727 min. Reactive carbonyls were stabilized at SP:SAMPLEPREP_SUMMARY 50C 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 50C. #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 20 32 30 33 11 25 9 SD1 SD4 SD2 15 48 34 23 22 18 32HIP 33HIP 46 10 Factors Genotype:WT Brain Genotype:WT Brain Genotype:WT Brain Genotype:WT Brain Genotype:WT Brain Genotype:WT Brain Genotype:WT Brain Genotype:WT Brain Genotype:WT Brain Genotype:WT Brain Genotype:HIP Brain Genotype:HIP Brain Genotype:HIP Brain Genotype:HIP Brain Genotype:HIP Brain Genotype:HIP Brain Genotype:HIP Brain Genotype:HIP Brain Genotype:HIP Brain Genotype:HIP Brain C10 hydrocarbon 12.76 15.56 12.29 11.55 16.45 12.81 12.34 16.41 12.49 11.94 14.75 11.82 13.97 14.79 13.62 12.76 Decamethyltetrasiloxane 14.75 13.12 13.81 13.60 13.22 13.69 13.72 13.71 13.87 14.04 13.98 13.65 13.82 14.25 13.27 13.85 14.27 13.91 2-Hydroxypyridine 16.91 15.52 15.60 16.29 15.98 15.65 14.79 16.03 16.34 15.70 15.92 15.70 15.66 16.17 16.59 16.54 15.98 15.89 16.72 15.43 Phenol 14.62 13.49 13.83 12.76 13.67 Lactic acid 25.10 23.86 23.81 24.06 23.61 23.69 23.62 24.23 24.11 24.26 25.01 24.44 24.76 25.66 24.98 24.51 24.13 24.57 24.80 23.86 Alanine 19.62 18.72 18.90 18.95 18.31 18.21 17.73 19.03 18.85 18.71 19.23 18.19 18.83 19.40 19.01 17.36 18.02 18.71 19.02 17.55 C11 hydrocarbon 14.68 13.11 13.91 14.71 14.34 14.16 14.17 14.41 14.47 14.53 13.82 14.53 14.56 13.88 14.01 14.51 14.19 14.45 14.64 14.65 2-Hydroxybutyric acid 20.81 21.57 21.71 21.53 21.22 21.40 21.59 21.45 21.41 21.47 22.24 21.23 21.40 21.58 21.37 22.58 21.04 21.67 21.62 21.18 Butyro-1,4-lactam 13.51 14.76 15.50 15.64 13.53 14.70 15.03 beta-Hydroxybutyric acid 16.99 15.85 17.65 17.52 17.35 17.24 16.54 17.13 16.92 16.94 18.08 19.59 19.46 19.93 19.39 18.52 17.62 19.24 18.52 17.78 Lactamide 20.54 19.95 20.28 20.33 20.18 19.90 20.46 20.13 19.99 20.20 19.37 20.21 20.14 20.26 20.09 20.11 19.96 19.58 19.99 19.97 2-Aminobutanoic acid 12.92 13.15 12.48 13.00 12.04 13.23 12.19 O-Methylphosphate 19.84 20.14 20.52 20.82 19.77 20.77 21.20 20.94 20.40 21.02 17.54 20.30 20.59 21.43 21.24 20.77 20.84 20.47 20.49 21.22 Malonic acid 17.73 19.21 18.98 18.90 18.90 19.36 19.21 18.85 18.54 18.92 19.01 17.58 18.90 19.38 18.94 19.06 18.68 18.75 18.78 18.62 Valine 16.14 17.08 16.66 16.81 16.13 16.37 16.17 17.11 16.62 16.32 17.05 15.61 17.22 18.24 17.08 17.41 16.68 16.99 17.24 15.75 C12 hydrocarbon 13.00 13.18 13.20 13.63 12.72 13.25 13.42 13.20 12.98 12.88 12.85 12.94 13.46 13.18 13.07 13.31 13.36 Urea 22.66 21.83 21.84 22.05 21.88 21.46 21.96 21.91 21.48 21.70 23.54 22.76 23.21 24.06 23.95 22.32 22.41 22.43 23.69 22.07 Benzoic acid 16.78 16.11 16.24 16.10 16.28 15.99 16.09 16.10 15.96 15.98 16.48 15.97 16.02 16.13 16.06 16.09 15.73 16.09 16.40 16.02 Ethanolamine 16.93 16.65 17.42 17.67 17.38 17.17 17.70 17.99 17.93 17.37 16.18 16.63 16.35 17.49 17.02 16.83 16.60 16.54 17.15 16.70 Phosphoric acid 25.91 25.52 25.73 26.12 25.44 25.81 25.80 25.91 25.69 25.67 25.64 24.43 25.69 26.50 26.08 25.45 25.27 25.25 25.61 25.63 Glycerol 20.93 20.39 19.97 19.83 19.43 20.25 20.41 20.17 20.14 19.55 20.13 19.91 22.44 21.40 19.54 18.92 19.31 20.10 19.75 C13 hydrocarbon 15.75 14.74 15.32 15.44 15.61 15.32 15.60 15.14 15.51 15.15 15.53 15.17 15.16 15.32 15.52 15.10 15.39 15.34 15.36 15.38 Isoleucine/Leucine 16.81 16.61 16.47 17.64 17.79 17.64 17.16 17.50 16.12 16.63 15.11 16.70 18.19 17.83 16.97 17.04 16.74 17.23 16.48 Proline 15.59 15.69 15.48 15.42 14.92 15.31 15.16 16.62 15.90 15.02 15.75 13.42 14.78 15.91 15.57 14.58 13.82 15.30 16.07 13.66 Maleic acid 20.66 20.30 18.17 18.20 17.00 17.61 19.74 18.71 17.79 17.51 17.22 16.36 Glycine 18.69 19.10 20.23 20.34 20.10 20.60 21.64 20.94 20.59 20.65 18.46 19.76 19.66 20.59 20.17 19.90 19.80 20.08 20.31 21.79 5-(Hydroxymethyl)furfural 15.33 15.11 15.27 15.43 13.52 14.80 14.05 13.83 13.44 14.29 Succinic acid 16.49 17.32 17.78 16.67 2,3-Dihydroxypyridine 15.21 11.70 13.70 15.21 15.19 15.01 15.68 14.24 12.37 15.13 MSTFA artifact 16.15 15.97 17.06 17.17 16.87 16.96 17.30 17.16 17.31 17.13 16.17 16.01 17.00 17.75 17.62 17.01 16.69 16.82 17.48 17.15 Uracil 18.01 17.68 17.24 17.51 17.12 17.16 17.27 17.70 16.76 17.14 16.46 15.97 15.43 16.81 16.00 15.77 15.49 15.19 15.78 15.82 Fumaric acid 17.86 16.73 17.31 17.32 16.69 16.81 16.86 16.70 16.98 16.88 17.55 17.40 17.65 18.19 17.72 17.40 16.47 17.02 17.23 16.56 Serine 21.48 20.77 20.99 20.90 20.27 19.76 19.50 20.96 20.64 19.69 21.25 18.96 20.27 21.13 20.82 20.78 19.38 20.41 20.79 19.19 6-Methyluracil 14.69 13.01 14.57 12.23 Threonic acid/erythronic acid 16.62 15.97 16.29 17.07 16.10 16.43 16.81 16.56 17.18 15.75 17.03 17.42 17.06 16.90 16.23 15.73 16.49 15.90 Threonine 20.68 19.72 20.00 19.94 19.46 19.54 19.50 20.21 19.50 19.37 19.96 18.11 19.07 20.33 19.54 19.36 18.30 19.04 19.65 18.88 Hydroquinone 13.59 14.45 14.69 13.61 13.70 14.62 13.97 14.34 14.21 14.89 14.49 10.62 13.43 13.32 13.75 14.51 Aspartic acid 18.88 20.56 20.76 20.79 20.46 20.13 20.24 20.56 20.06 19.36 21.10 19.16 20.36 21.45 20.95 20.41 19.80 19.34 19.64 19.94 beta-Alanine 11.86 12.43 13.75 14.05 14.26 15.92 14.28 15.29 14.53 14.44 12.46 12.65 13.84 14.80 15.55 14.60 15.21 15.03 15.35 14.30 Aminomalonic acid 14.05 15.02 16.61 17.02 16.17 17.07 18.01 17.46 17.02 16.95 13.86 15.66 16.18 17.34 16.46 16.69 16.30 16.50 16.41 18.30 Nicotinamide 17.84 17.16 17.51 17.75 17.15 17.42 17.08 17.47 17.11 16.99 17.66 16.89 17.48 18.25 17.73 17.48 17.04 16.86 17.32 16.55 Malic acid 21.36 20.65 21.25 21.17 20.38 20.65 20.41 20.66 20.69 20.39 21.61 20.76 21.30 21.95 21.47 21.08 20.32 21.05 21.05 20.66 C15 hydrocarbon 16.21 15.07 15.75 15.52 15.44 15.94 15.52 15.81 15.28 15.17 15.78 15.20 15.31 15.72 15.43 15.51 15.24 15.10 15.66 15.67 Threitol/erythritol 16.79 15.35 15.67 16.01 15.82 14.85 15.10 15.04 16.19 15.19 18.15 16.64 17.81 18.73 18.01 17.16 17.56 17.02 18.00 16.39 Tris 12.26 12.06 13.90 13.39 Hydroxyprolines 15.28 13.26 14.08 14.54 14.42 13.47 14.37 14.53 14.14 15.57 14.78 15.01 14.26 14.73 12.66 gamma-Aminobutyric acid 21.94 22.55 23.46 23.52 23.21 23.92 22.71 23.67 23.35 23.21 22.93 22.09 23.36 23.87 24.03 23.33 23.39 23.41 23.49 22.17 Erythronic acid/Threonic acid 16.32 14.51 15.49 16.33 15.36 15.48 14.72 15.89 15.71 15.87 13.97 14.96 16.07 15.88 14.59 14.41 14.46 15.53 15.08 7H-Purine/Phenylalanine 15.21 14.93 14.73 13.98 Creatinine/Cysteine 23.01 22.30 22.40 22.47 22.13 21.93 21.96 22.21 22.09 22.22 22.48 21.48 22.48 22.92 22.49 22.15 21.86 21.74 22.27 21.93 2-Hydroxyglutaric acid 14.93 14.48 15.32 Phosphoenolpyruvic acid 12.06 12.76 13.23 14.09 13.50 2-Hydroxybiphenyl 17.73 13.82 18.33 17.21 16.69 17.04 19.20 17.79 18.13 18.02 Benzene-1,2,4-triol/hypotaurine 17.05 17.06 17.52 18.73 18.45 17.94 18.30 17.77 17.88 17.38 18.85 17.22 18.07 18.77 18.68 17.47 17.52 17.53 17.64 17.82 5-Aminovaleric acid 16.76 16.27 17.13 16.08 16.41 16.71 16.57 17.14 16.88 17.73 17.11 16.43 14.55 Glutamic acid 25.04 23.85 24.50 24.60 23.75 23.41 23.21 24.20 24.40 23.64 24.97 23.94 24.65 25.03 24.86 24.40 23.76 24.02 24.49 23.06 Phenylalanine 18.64 18.63 18.10 17.84 17.43 18.10 18.12 18.76 17.91 17.94 17.37 17.02 16.45 18.00 17.24 16.93 16.92 16.73 17.49 16.45 N-Acetylaspartic acid 23.98 23.51 24.10 24.28 23.48 23.36 22.94 24.09 23.72 23.17 24.03 23.10 23.91 24.43 24.29 23.99 23.35 23.34 23.76 23.20 Pyrophosphate 15.03 13.51 15.75 15.44 14.31 16.72 15.62 16.72 15.91 16.16 14.17 15.70 15.40 15.95 16.67 16.37 16.48 16.50 16.30 15.93 Asparagine 16.65 16.38 15.95 15.47 15.41 16.39 16.91 16.07 Taurine 17.79 16.81 16.27 16.27 17.50 17.19 16.56 18.50 16.32 19.45 17.01 16.19 18.90 Aldopentoses 18.73 18.25 18.33 18.21 18.46 18.33 18.22 18.69 18.04 18.08 18.39 18.08 17.94 19.39 18.15 17.82 17.55 17.80 18.01 16.32 Cysteinesulfinic acid 16.56 16.79 17.39 17.66 17.04 18.00 16.57 17.95 17.40 17.19 15.70 16.46 17.85 15.67 2-Aminoadipic acid 17.97 17.50 17.46 17.33 16.28 17.62 17.04 17.88 17.16 17.63 17.34 17.87 17.47 17.45 16.74 17.18 16.87 Pentitols 16.92 16.31 16.32 17.08 16.52 14.69 15.61 13.91 16.17 15.59 17.76 16.91 17.15 17.20 17.69 16.85 16.26 16.37 17.01 15.45 Ornithine 13.10 14.24 16.11 15.18 14.48 14.59 Putrescine 14.59 13.30 13.10 14.95 14.24 13.76 13.88 13.58 14.71 16.09 16.11 15.18 14.48 14.59 13.97 13.92 Dihydroxyacetone phosphate 16.29 15.25 15.15 14.97 14.97 15.42 15.94 15.11 14.17 15.06 14.62 15.47 15.46 15.40 15.06 15.35 Orotic acid 19.23 20.09 19.93 20.42 20.41 19.11 20.13 19.38 19.80 19.44 Glycerol 1-phosphate 17.88 16.60 17.13 17.44 17.96 19.23 17.73 16.38 17.69 17.58 20.09 18.29 19.93 20.42 20.41 19.11 20.13 19.38 19.80 19.44 Glutamine 21.72 21.84 20.87 21.12 21.27 20.23 20.95 19.70 19.43 20.01 21.59 18.87 20.33 20.84 21.17 18.37 19.74 19.62 19.16 20.55 Pentonic acids 12.95 12.08 14.40 13.70 13.56 14.21 15.00 13.54 13.13 13.01 14.37 O-Phosphocolamine 17.87 17.98 20.07 20.33 20.25 20.38 19.35 20.44 20.73 19.62 19.16 19.74 20.15 20.07 21.54 20.62 20.39 20.59 20.87 19.72 Gentisic acid 14.16 13.32 14.35 14.39 13.98 14.74 Hypoxanthine 20.56 20.90 20.35 20.52 19.85 20.02 20.09 20.54 20.17 20.20 19.53 19.75 18.95 20.45 19.17 19.23 18.53 18.89 19.26 18.77 Citric acid/isocitric acid 18.80 18.23 19.13 19.13 18.78 18.07 18.00 19.08 18.83 19.00 18.65 19.25 18.83 19.89 18.90 19.00 18.60 18.16 18.68 18.47 deuterated Myristic Acid (RTL std) 21.97 21.80 22.14 21.77 21.63 21.59 21.73 21.70 21.75 21.45 22.26 21.71 21.92 21.91 22.05 22.01 21.45 21.50 21.59 21.37 Dehydroascorbic acid 21.32 20.86 20.92 21.39 20.52 20.65 19.58 20.83 21.26 20.60 21.38 20.70 21.05 21.89 21.38 21.37 20.51 20.92 21.16 19.49 Myristic acid or Pentadecanol 17.65 17.39 18.01 17.22 17.77 17.85 17.65 17.41 17.42 17.44 18.13 17.35 17.62 17.68 17.38 17.66 17.44 17.31 17.29 17.43 cis-3,5-Dimethoxycinnamic acid 14.16 13.58 13.43 13.63 13.77 13.75 Fructose or similar ketohexose 15.44 15.88 14.98 15.35 14.88 14.02 14.94 16.78 15.60 15.81 19.09 20.81 20.05 21.16 19.83 20.23 20.82 19.38 19.86 19.14 Tyramine 12.68 12.69 12.68 13.62 13.82 13.67 Glucose and other aldohexoses 18.98 20.61 18.90 17.45 17.30 18.67 18.28 18.55 18.51 18.05 21.00 23.35 21.91 23.77 20.74 23.28 22.91 21.97 21.56 22.43 Sedoheptulose or similar sugar 15.31 15.50 15.41 15.68 15.67 15.47 15.94 15.68 15.32 14.99 15.84 14.72 15.25 15.36 15.47 14.25 14.67 14.64 15.17 14.78 p-Hydroxyphenyllactic acid 13.48 13.31 12.70 C19 hydrocarbon 15.55 15.16 15.14 15.61 15.00 14.71 14.49 15.56 15.40 15.61 15.42 14.87 15.65 14.60 Lysine 17.27 18.10 17.76 18.11 18.42 18.30 18.96 17.81 17.40 17.80 17.78 17.84 17.15 18.28 18.01 16.88 17.80 17.35 16.79 18.36 Hexitols 16.84 16.33 16.99 16.50 15.85 16.11 17.35 16.84 16.55 19.99 19.48 20.13 20.13 20.16 19.42 19.61 19.22 19.15 18.23 Tyrosine 16.73 17.56 16.16 15.96 16.03 15.99 17.01 15.85 14.92 15.73 16.88 15.66 14.21 15.54 14.63 11.63 14.92 14.53 14.57 14.75 Ascorbic acid 16.72 16.89 17.21 18.52 18.21 17.56 12.21 17.34 15.13 18.27 17.47 17.01 17.91 19.22 19.33 18.22 18.26 17.19 18.48 16.15 Pentadecanoic acid 15.76 14.88 15.24 15.29 15.53 14.57 15.20 14.57 15.27 15.29 14.93 14.27 14.94 14.96 14.45 14.19 Erythrose-4-phosphate 12.72 13.41 12.54 12.24 12.49 Ononitol or similar methoxyinositol 15.25 15.59 14.36 15.30 15.53 13.65 17.19 15.54 17.07 17.29 Gly-Pro 11.62 14.86 15.22 11.76 14.36 15.56 14.01 Hexadecanol 14.31 14.72 14.93 14.56 13.82 13.83 14.85 14.59 13.60 13.96 15.52 13.47 15.21 15.34 14.45 13.40 14.09 14.53 13.99 16.06 Pantothenic acid 12.32 11.81 11.98 15.24 14.75 14.46 14.44 13.68 17.32 15.17 14.61 15.18 14.63 14.82 13.75 Xanthine 18.04 18.42 17.70 17.77 18.29 17.44 17.97 17.45 16.98 17.97 16.40 17.39 15.03 17.84 13.62 15.81 15.70 15.08 16.32 16.20 Palmitic acid 23.62 23.36 23.17 23.11 22.99 22.73 22.83 22.88 22.86 22.75 23.73 23.09 23.07 23.19 22.93 22.77 22.51 22.62 22.61 22.47 Dopamine/5-Hydroxylysine 13.81 15.45 10.67 14.76 Myoinositol 25.36 25.16 25.39 25.70 25.12 25.75 25.71 25.22 25.70 25.61 25.58 25.66 25.95 26.59 26.03 26.06 25.88 25.93 25.74 25.68 Ketopentose-5-phosphate 16.78 16.10 15.74 16.82 15.67 15.98 15.16 16.09 16.89 14.92 17.47 17.11 16.77 18.26 17.01 17.37 16.72 16.94 16.16 16.31 C21 Hydrocarbon 15.21 14.90 14.34 15.77 15.53 15.07 14.67 Heptadecanoic acid/Octadecanol 19.03 18.50 18.15 18.61 19.11 17.85 18.18 17.62 17.89 17.82 19.88 17.84 18.07 18.36 18.20 17.59 17.46 17.55 17.76 17.66 C22 hydrocarbon 13.85 13.59 14.80 14.12 13.17 13.65 13.29 Spermidine 13.33 15.74 17.10 17.89 18.69 18.77 19.41 17.61 17.21 19.34 16.71 16.87 16.97 19.00 18.27 16.58 18.20 17.69 17.70 19.20 Linoleic acid 15.38 15.37 13.68 15.08 14.43 14.15 17.35 15.92 16.34 Tryptophan 14.36 11.67 13.96 14.88 13.10 12.71 Oleic acid 22.59 22.53 22.06 22.03 22.25 21.59 22.13 21.69 21.30 21.46 22.63 21.65 21.45 21.87 21.52 21.12 20.97 20.92 21.04 21.73 Stearic acid 24.07 23.89 23.90 23.86 23.56 23.43 23.60 23.48 23.43 23.62 24.17 23.61 23.75 23.89 23.62 23.33 23.42 23.47 23.33 23.16 Fructose-6-phosphate 16.15 18.03 15.84 14.09 16.00 17.18 17.22 17.82 17.91 18.11 21.16 19.35 18.64 17.86 19.96 18.17 19.84 18.03 18.30 Glucose-6-phosphate or similar 17.67 19.51 17.79 15.81 16.87 18.72 18.87 19.85 19.80 13.93 19.58 22.46 20.51 20.17 19.71 21.61 20.05 21.51 19.79 20.19 1-Eicosanol 14.23 11.79 14.27 Arachidonic acid (also Eicosapentaenoic acid) 21.56 21.40 20.94 21.23 20.99 20.40 20.88 20.97 20.97 20.00 21.58 20.95 20.55 20.88 20.77 20.76 19.83 20.44 20.45 19.79 Purine riboside 15.92 16.38 16.46 17.66 17.40 16.84 17.55 17.52 17.54 17.30 15.95 15.63 16.87 18.83 17.34 16.76 17.22 17.24 17.48 18.06 Myoinositol-2-phosphate 16.25 16.23 16.64 17.06 16.18 17.64 18.76 17.36 17.39 17.50 17.05 17.85 17.69 18.29 17.66 17.79 17.52 17.55 17.59 18.26 Arachidic acid 16.03 16.16 16.17 16.17 17.30 16.15 17.24 15.39 15.50 16.29 17.23 15.83 16.22 16.77 16.28 15.50 16.10 16.03 15.28 17.62 Uridine/pseudouridine 15.96 15.33 15.77 15.47 14.32 15.40 15.82 16.34 17.67 15.60 17.26 15.91 16.39 14.21 15.46 16.13 15.01 O-Benzyltyrosine 14.12 14.16 14.26 13.35 14.34 14.30 14.33 14.42 14.30 14.23 14.75 beta-Monopalmitin 18.19 17.58 17.38 17.70 17.75 17.35 17.44 17.09 17.41 17.21 18.86 16.94 17.25 18.17 17.07 17.22 17.23 17.26 17.17 17.34 Docosahexaenoic acid 21.85 21.23 20.81 20.94 20.87 19.92 19.98 20.69 20.40 20.39 21.93 20.85 20.73 21.18 20.80 20.54 20.43 20.06 20.38 19.43 Inosine 23.24 22.78 22.83 23.11 22.63 22.62 22.45 22.71 22.96 22.68 23.13 23.09 22.71 23.67 22.70 22.66 22.59 22.70 23.03 22.44 alpha-Monopalmitin 21.51 21.30 21.42 21.52 21.69 21.53 21.29 21.57 21.51 21.52 21.87 21.53 21.52 21.57 21.50 21.46 21.40 21.45 21.56 21.51 cis-13-Docosenoic acid 14.77 15.16 15.66 14.86 16.37 14.25 15.39 16.79 16.30 15.23 14.28 16.19 N-Acetylneuraminic acid 17.04 17.19 16.67 16.95 17.41 16.85 17.09 16.87 16.75 16.10 17.84 16.93 16.58 17.56 17.50 16.51 17.11 16.54 16.66 17.10 Sucrose and similar disaccharides 14.21 14.24 Adenosine 18.93 16.22 20.88 21.20 14.77 17.12 17.94 15.68 18.99 16.72 20.06 19.88 19.66 20.63 19.31 18.01 20.04 15.85 Docosanoic acid 14.48 15.40 14.89 14.12 14.35 15.27 15.80 14.34 13.59 14.89 14.67 14.34 14.47 15.78 15.09 13.95 14.51 14.36 16.12 Disaccharide 13.99 16.71 13.48 14.12 13.63 12.67 13.67 13.76 15.67 14.69 16.80 13.44 14.36 14.21 12.76 Tricosanoic acid 13.96 14.75 14.77 14.50 14.61 11.79 12.90 13.09 beta-Monostearin 19.67 19.32 19.42 19.52 19.86 19.57 19.22 19.55 19.60 19.47 20.08 19.52 19.44 19.43 19.50 19.43 19.55 19.29 19.56 19.55 Guanosine 16.10 15.44 17.07 13.65 17.25 13.64 13.47 15.25 16.42 15.45 18.40 16.31 16.50 13.16 16.41 16.92 alpha-Monostearin 24.04 23.77 24.17 24.19 24.38 24.23 24.01 24.29 24.21 24.24 24.09 24.23 24.19 24.13 24.20 24.20 24.22 24.11 24.29 24.30 cis-15-Tetracosenoic acid 16.35 15.53 16.48 16.49 16.36 16.86 18.13 16.06 14.46 17.17 14.78 14.91 14.33 17.50 16.27 13.32 15.29 14.72 14.41 17.26 Uridine 5'-monophosphate 14.88 15.99 15.68 16.67 Melbiose or similar disaccharide 17.21 17.62 17.35 17.32 18.59 16.80 17.37 17.15 18.25 16.18 16.47 17.25 16.40 16.28 16.37 16.92 Tetracosanoic acid 16.69 16.11 16.24 16.48 15.64 16.45 16.92 16.15 15.66 16.24 16.48 16.02 16.57 17.40 17.23 15.75 16.27 15.85 15.80 17.30 Adenosine-5'-monophosphate 15.11 14.02 17.55 15.24 18.41 16.81 16.49 5-Cholesten-3-beta-7-alpha-diol 14.41 13.73 14.22 13.84 13.79 15.10 14.22 14.37 alpha-Tocopherol 15.25 15.54 15.03 14.82 14.97 15.10 15.46 14.88 14.88 Cholesterol 27.53 27.53 27.54 27.59 27.73 27.67 28.09 27.42 27.20 27.69 27.39 27.16 27.23 27.68 27.73 27.18 27.38 27.11 27.14 28.01 5-alpha-Coprostanol or similar oxysterol 17.98 17.80 18.17 18.01 17.51 18.08 18.01 17.60 Campesterol 18.58 17.92 18.30 17.95 17.64 19.14 17.74 18.78 17.89 18.75 15.17 16.73 17.00 16.68 16.37 14.36 15.61 beta-Sitosterol 16.75 16.33 16.91 16.73 18.29 16.91 17.69 15.54 17.23 14.16 14.95 14.09 12.84 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name C10 hydrocarbon Decamethyltetrasiloxane 2-Hydroxypyridine Phenol Lactic acid Alanine C11 hydrocarbon 2-Hydroxybutyric acid Butyro-1,4-lactam beta-Hydroxybutyric acid Lactamide 2-Aminobutanoic acid O-Methylphosphate Malonic acid Valine C12 hydrocarbon Urea Benzoic acid Ethanolamine Phosphoric acid Glycerol C13 hydrocarbon Isoleucine/Leucine Proline Maleic acid Glycine 5-(Hydroxymethyl)furfural Succinic acid 2,3-Dihydroxypyridine MSTFA artifact Uracil Fumaric acid Serine 6-Methyluracil Threonic acid/erythronic acid Threonine Hydroquinone Aspartic acid beta-Alanine Aminomalonic acid Nicotinamide Malic acid C15 hydrocarbon Threitol/erythritol Tris Hydroxyprolines gamma-Aminobutyric acid Erythronic acid/Threonic acid 7H-Purine/Phenylalanine Creatinine/Cysteine 2-Hydroxyglutaric acid Phosphoenolpyruvic acid 2-Hydroxybiphenyl Benzene-1,2,4-triol/hypotaurine 5-Aminovaleric acid Glutamic acid Phenylalanine N-Acetylaspartic acid Pyrophosphate Asparagine Taurine Aldopentoses Cysteinesulfinic acid 2-Aminoadipic acid Pentitols Ornithine Putrescine Dihydroxyacetone phosphate Orotic acid Glycerol 1-phosphate Glutamine Pentonic acids O-Phosphocolamine Gentisic acid Hypoxanthine Citric acid/isocitric acid deuterated Myristic Acid (RTL std) Dehydroascorbic acid Myristic acid or Pentadecanol cis-3,5-Dimethoxycinnamic acid Fructose or similar ketohexose Tyramine Glucose and other aldohexoses Sedoheptulose or similar sugar p-Hydroxyphenyllactic acid C19 hydrocarbon Lysine Hexitols Tyrosine Ascorbic acid Pentadecanoic acid Erythrose-4-phosphate Ononitol or similar methoxyinositol Gly-Pro Hexadecanol Pantothenic acid Xanthine Palmitic acid Dopamine/5-Hydroxylysine Myoinositol Ketopentose-5-phosphate C21 Hydrocarbon Heptadecanoic acid/Octadecanol C22 hydrocarbon Spermidine Linoleic acid Tryptophan Oleic acid Stearic acid Fructose-6-phosphate Glucose-6-phosphate or similar 1-Eicosanol Arachidonic acid (also Eicosapentaenoic acid) Purine riboside Myoinositol-2-phosphate Arachidic acid Uridine/pseudouridine O-Benzyltyrosine beta-Monopalmitin Docosahexaenoic acid Inosine alpha-Monopalmitin cis-13-Docosenoic acid N-Acetylneuraminic acid Sucrose and similar disaccharides Adenosine Docosanoic acid Disaccharide Tricosanoic acid beta-Monostearin Guanosine alpha-Monostearin cis-15-Tetracosenoic acid Uridine 5'-monophosphate Melbiose or similar disaccharide Tetracosanoic acid Adenosine-5'-monophosphate 5-Cholesten-3-beta-7-alpha-diol alpha-Tocopherol Cholesterol 5-alpha-Coprostanol or similar oxysterol Campesterol beta-Sitosterol METABOLITES_END #END