#METABOLOMICS WORKBENCH KeiRodrz30_20220208_122818 DATATRACK_ID:3070 STUDY_ID:ST002080 ANALYSIS_ID:AN003394 PROJECT_ID:PR001320 VERSION 1 CREATED_ON February 8, 2022, 1:51 pm #PROJECT PR:PROJECT_TITLE Effects of Ferroptosis on the Metabolome in Cardiac Cells: The Role of PR:PROJECT_TITLE Glutaminolysis PR:PROJECT_TYPE GCMS PR:PROJECT_SUMMARY Ferroptosis is a novel iron-dependent regulated cell death mechanism that PR:PROJECT_SUMMARY affects cell metabolism; however, a detailed metabolomic analysis of ferroptotic PR:PROJECT_SUMMARY cells is not yet available. Here, we elucidated the metabolome of H9c2 PR:PROJECT_SUMMARY cardioblasts by gas chromatography-mass spectrometry during ferroptosis induced PR:PROJECT_SUMMARY by RSL3, a GPX4 inhibitor, in the presence of ferrostatin-1 (a ferroptosis PR:PROJECT_SUMMARY inhibitor), XJB-5-131 (a mitochondrial-targeted ROS scavenger), or TSM-1005-44 PR:PROJECT_SUMMARY (a newly developed cellular ROS scavenger). Results demonstrated that RSL3 PR:PROJECT_SUMMARY decreased the levels of amino acids involved in glutathione synthesis more than PR:PROJECT_SUMMARY two-fold. In contrast, saturated fatty acids levels were markedly increased in PR:PROJECT_SUMMARY RSL3-challenged cells, with no effects on unsaturated fatty acids. RSL3 PR:PROJECT_SUMMARY significantly altered the levels of mitochondrial tricarboxylic acid cycle PR:PROJECT_SUMMARY intermediates; isocitrate and 2-oxoglutarate were found to increase, whereas PR:PROJECT_SUMMARY succinate was significantly decreased in RSL3-challenged cells. Ferrostatin-1, PR:PROJECT_SUMMARY XJB-5-131, and TSM-1005-44 prevented RSL3-induced cell death and conserved the PR:PROJECT_SUMMARY metabolomic profile of the cells. Since 2-oxoglutarate is involved in the PR:PROJECT_SUMMARY regulation of ferroptosis, particularly through glutamine metabolism, we further PR:PROJECT_SUMMARY assessed the role of glutaminolysis in ferroptosis in H9c2 cardioblasts. Genetic PR:PROJECT_SUMMARY silencing of GLS1, which encodes the K-type mitochondrial glutaminase PR:PROJECT_SUMMARY (glutaminase C), protected against ferroptosis in the early stage. In PR:PROJECT_SUMMARY conclusion, our study demonstrates that RSL3-induced ferroptosis impairs the PR:PROJECT_SUMMARY metabolome of H9c2 cardioblasts. PR:INSTITUTE University of Puerto Rico, School of Medicine PR:DEPARTMENT Physiology PR:LABORATORY Cardiovascular Physiology, DR. Sabzali Javadov's Lab PR:LAST_NAME Rodriguez-Graciani PR:FIRST_NAME Keishla M PR:ADDRESS Medical Sciences Campus, Main Building, 6th Floor, Department of Physiology, San PR:ADDRESS Juan, Puerto Rico, 00936-5067, USA PR:EMAIL keishla.rodriguez20@upr.edu PR:PHONE 7877582525x26888 PR:FUNDING_SOURCE This research was funded by the National Institutes of Health (Grants PR:FUNDING_SOURCE SC1GM128210, R25GM061838, U54MD007600, P20GM103475, HDTRA1-16-1-0041, and PR:FUNDING_SOURCE U19-AI1068021). PR:PUBLICATIONS https://www.mdpi.com/1476010, Antioxidants 2022, 11(2), 278; PR:PUBLICATIONS https://doi.org/10.3390/antiox11020278 PR:CONTRIBUTORS Xavier R. Chapa-Dubocq, Esteban J. Ayala-Arroyo, Ivana Chaves-Negrón, Sehwan PR:CONTRIBUTORS Jang, Nataliya Chorna, Taber S. Maskrey, Peter Wipf and Sabzali Javadov #STUDY ST:STUDY_TITLE Effects of Ferroptosis on the Metabolome in Cardiac Cells: The Role of ST:STUDY_TITLE Glutaminolysis ST:STUDY_TYPE GCMS ST:STUDY_SUMMARY Ferroptosis is a novel iron-dependent regulated cell death mechanism that ST:STUDY_SUMMARY affects cell metabolism; however, a detailed metabolomic analysis of ferroptotic ST:STUDY_SUMMARY cells is not yet available. Here, we elucidated the metabolome of H9c2 ST:STUDY_SUMMARY cardioblasts by gas chromatography-mass spectrometry during ferroptosis induced ST:STUDY_SUMMARY by RSL3, a GPX4 inhibitor, in the presence of ferrostatin-1 (a ferroptosis ST:STUDY_SUMMARY inhibitor), XJB-5-131 (a mitochondrial-targeted ROS scavenger), or TSM-1005-44 ST:STUDY_SUMMARY (a newly developed cellular ROS scavenger). Results demonstrated that RSL3 ST:STUDY_SUMMARY decreased the levels of amino acids involved in glutathione synthesis more than ST:STUDY_SUMMARY two-fold. In contrast, saturated fatty acids levels were markedly increased in ST:STUDY_SUMMARY RSL3-challenged cells, with no effects on unsaturated fatty acids. RSL3 ST:STUDY_SUMMARY significantly altered the levels of mitochondrial tricarboxylic acid cycle ST:STUDY_SUMMARY intermediates; isocitrate and 2-oxoglutarate were found to increase, whereas ST:STUDY_SUMMARY succinate was significantly decreased in RSL3-challenged cells. Ferrostatin-1, ST:STUDY_SUMMARY XJB-5-131, and TSM-1005-44 prevented RSL3-induced cell death and conserved the ST:STUDY_SUMMARY metabolomic profile of the cells. Since 2-oxoglutarate is involved in the ST:STUDY_SUMMARY regulation of ferroptosis, particularly through glutamine metabolism, we further ST:STUDY_SUMMARY assessed the role of glutaminolysis in ferroptosis in H9c2 cardioblasts. Genetic ST:STUDY_SUMMARY silencing of GLS1, which encodes the K-type mitochondrial glutaminase ST:STUDY_SUMMARY (glutaminase C), protected against ferroptosis in the early stage. In ST:STUDY_SUMMARY conclusion, our study demonstrates that RSL3-induced ferroptosis impairs the ST:STUDY_SUMMARY metabolome of H9c2 cardioblasts. ST:INSTITUTE University of Puerto Rico, School of Medicine ST:DEPARTMENT Physiology ST:LABORATORY Cardiovascular Physiology, DR. Sabzali Javadov's Lab ST:LAST_NAME Rodriguez-Graciani ST:FIRST_NAME Keishla M ST:ADDRESS Medical Sciences Campus, Main Building, 6th Floor, Department of Physiology, San ST:ADDRESS Juan, Puerto Rico, 00936-5067, USA ST:EMAIL keishla.rodriguez20@upr.edu ST:PHONE 7877582525x26888 ST:NUM_GROUPS 8 ST:PUBLICATIONS https://www.mdpi.com/1476010, Antioxidants 2022, 11(2), 278; ST:PUBLICATIONS https://doi.org/10.3390/antiox11020278 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Rattus norvegicus SU:TAXONOMY_ID 10116 SU:CELL_BIOSOURCE_OR_SUPPLIER American Type Culture Collection (ATCC) SU:CELL_STRAIN_DETAILS H9c2(2-1) is a subclone of the original clonal cell line derived from embryonic SU:CELL_STRAIN_DETAILS BD1X rat heart tissue by B. Kimes and B. Brandt and exhibits many of the SU:CELL_STRAIN_DETAILS properties of skeletal muscle. SU:CELL_PASSAGE_NUMBER 3-10 SU:CELL_COUNTS 80-90% confluency #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 C Control_ 1 Treatment:Control RAW_FILE_NAME=C1_UNK-0010_7132021_10 SUBJECT_SAMPLE_FACTORS C Control_ 2 Treatment:Control RAW_FILE_NAME=C2_UNK-0030_7132021_30 SUBJECT_SAMPLE_FACTORS C Control_ 3 Treatment:Control RAW_FILE_NAME=C4_UNK-0021_7132021_21 SUBJECT_SAMPLE_FACTORS C Control_ 4 Treatment:Control RAW_FILE_NAME=C5_UNK-0017_7132021_17 SUBJECT_SAMPLE_FACTORS C Control_ 5 Treatment:Control RAW_FILE_NAME=C6_UNK-0041_7132021_41 SUBJECT_SAMPLE_FACTORS R RSL3_1 Treatment:RSL3 RAW_FILE_NAME=R1_UNK-0012_7132021_12 SUBJECT_SAMPLE_FACTORS R RSL3_2 Treatment:RSL3 RAW_FILE_NAME=R2_UNK-0043_7132021_43 SUBJECT_SAMPLE_FACTORS R RSL3_3 Treatment:RSL3 RAW_FILE_NAME=R4_UNK-0037_7132021_37 SUBJECT_SAMPLE_FACTORS R RSL3_4 Treatment:RSL3 RAW_FILE_NAME=R5_UNK-0009_7132021_9 SUBJECT_SAMPLE_FACTORS R RSL3_5 Treatment:RSL3 RAW_FILE_NAME=R6_UNK-0034_7132021_34 SUBJECT_SAMPLE_FACTORS R RSL3_6 Treatment:RSL3 RAW_FILE_NAME=R8_UNK-0040_7132021_40 SUBJECT_SAMPLE_FACTORS F Fer1-1 Treatment:Ferrostatin-1 RAW_FILE_NAME=F1_UNK-0003_7132021_3 SUBJECT_SAMPLE_FACTORS F Fer1-2 Treatment:Ferrostatin-1 RAW_FILE_NAME=F2_UNK-0036_7132021_36 SUBJECT_SAMPLE_FACTORS F Fer1-3 Treatment:Ferrostatin-1 RAW_FILE_NAME=F4_UNK-0046_7132021_46 SUBJECT_SAMPLE_FACTORS F Fer1-4 Treatment:Ferrostatin-1 RAW_FILE_NAME=F5_UNK-0020_7132021_20 SUBJECT_SAMPLE_FACTORS F Fer1-5 Treatment:Ferrostatin-1 RAW_FILE_NAME=F6_UNK-0027_7132021_27 SUBJECT_SAMPLE_FACTORS F Fer1-6 Treatment:Ferrostatin-1 RAW_FILE_NAME=F8_UNK-0011_7132021_11 SUBJECT_SAMPLE_FACTORS X XJB_1 Treatment:XJB-5-131 RAW_FILE_NAME=X1_UNK-0025_7132021_25 SUBJECT_SAMPLE_FACTORS X XJB_2 Treatment:XJB-5-131 RAW_FILE_NAME=X2_UNK-0015_7132021_15 SUBJECT_SAMPLE_FACTORS X XJB_3 Treatment:XJB-5-131 RAW_FILE_NAME=X5_UNK-0019_7132021_19 SUBJECT_SAMPLE_FACTORS X XJB_4 Treatment:XJB-5-131 RAW_FILE_NAME=X6_UNK-0006_7132021_6 SUBJECT_SAMPLE_FACTORS X XJB_5 Treatment:XJB-5-131 RAW_FILE_NAME=X8_UNK-0028_7132021_28 SUBJECT_SAMPLE_FACTORS T TSM_1 Treatment:TSM-1005-44 RAW_FILE_NAME=T1_UNK-0023_7132021_23 SUBJECT_SAMPLE_FACTORS T TSM_3 Treatment:TSM-1005-44 RAW_FILE_NAME=T4_UNK-0032_7132021_32 SUBJECT_SAMPLE_FACTORS T TSM_4 Treatment:TSM-1005-44 RAW_FILE_NAME=T5_UNK-0035_7132021_35 SUBJECT_SAMPLE_FACTORS T TSM_5 Treatment:TSM-1005-44 RAW_FILE_NAME=T6_UNK-0048_7132021_48 SUBJECT_SAMPLE_FACTORS T TSM_6 Treatment:TSM-1005-44 RAW_FILE_NAME=T8_UNK-0008_7132021_8 SUBJECT_SAMPLE_FACTORS RF RF_1 Treatment:RSL3 + Ferrostatin-1 RAW_FILE_NAME=RF1_UNK-0004_7132021_4 SUBJECT_SAMPLE_FACTORS RF RF_2 Treatment:RSL3 + Ferrostatin-1 RAW_FILE_NAME=RF2_UNK-0014_7132021_14 SUBJECT_SAMPLE_FACTORS RF RF_3 Treatment:RSL3 + Ferrostatin-1 RAW_FILE_NAME=RF4_UNK-0024_7132021_24 SUBJECT_SAMPLE_FACTORS RF RF_4 Treatment:RSL3 + Ferrostatin-1 RAW_FILE_NAME=RF5_UNK-0044_7132021_44 SUBJECT_SAMPLE_FACTORS RF RF_5 Treatment:RSL3 + Ferrostatin-1 RAW_FILE_NAME=RF6_UNK-0047_7132021_47 SUBJECT_SAMPLE_FACTORS RF RF_6 Treatment:RSL3 + Ferrostatin-1 RAW_FILE_NAME=RF8_UNK-0029_7132021_29 SUBJECT_SAMPLE_FACTORS RX RX_1 Treatment:RSL3 + XJB-5-131 RAW_FILE_NAME=RX1_UNK-0031_7132021_31 SUBJECT_SAMPLE_FACTORS RX RX_2 Treatment:RSL3 + XJB-5-131 RAW_FILE_NAME=RX2_UNK-0007_7132021_7 SUBJECT_SAMPLE_FACTORS RX RX_3 Treatment:RSL3 + XJB-5-131 RAW_FILE_NAME=RX4_UNK-0016_7132021_16 SUBJECT_SAMPLE_FACTORS RX RX_4 Treatment:RSL3 + XJB-5-131 RAW_FILE_NAME=RX5_UNK-0026_7132021_26 SUBJECT_SAMPLE_FACTORS RX RX_5 Treatment:RSL3 + XJB-5-131 RAW_FILE_NAME=RX6_UNK-0038_7132021_38 SUBJECT_SAMPLE_FACTORS RX RX_6 Treatment:RSL3 + XJB-5-131 RAW_FILE_NAME=RX8_UNK-0013_7132021_13 SUBJECT_SAMPLE_FACTORS RT RT_1 Treatment:RSL3 + TSM-1005-44 RAW_FILE_NAME=RT1_UNK-0042_7132021_42 SUBJECT_SAMPLE_FACTORS RT RT_2 Treatment:RSL3 + TSM-1005-44 RAW_FILE_NAME=RT4_UNK-0022_7132021_22 SUBJECT_SAMPLE_FACTORS RT RT_3 Treatment:RSL3 + TSM-1005-44 RAW_FILE_NAME=RT5_UNK-0039_7132021_39 SUBJECT_SAMPLE_FACTORS RT RT_4 Treatment:RSL3 + TSM-1005-44 RAW_FILE_NAME=RT6_UNK-0001_7152021_1 SUBJECT_SAMPLE_FACTORS RT RT_5 Treatment:RSL3 + TSM-1005-44 RAW_FILE_NAME=RT8_UNK-0045_7132021_45 #COLLECTION CO:COLLECTION_SUMMARY H9c2 cardioblasts were cultured according to the manufacturer’s CO:COLLECTION_SUMMARY recommendations (ATCC, Manassas, VA, USA) with minor modifications. Briefly, the CO:COLLECTION_SUMMARY cells were incubated in DMEM based modified media containing 4 mM L-glutamine, CO:COLLECTION_SUMMARY 4.5 g/L glucose, 1 mM sodium pyruvate, and 1.5 g/L sodium bicarbonate, pH 7.4, CO:COLLECTION_SUMMARY and supplemented with 10% fetal bovine serum and 1% antibiotic solution CO:COLLECTION_SUMMARY (HyClone, Logan, UT, USA) in a CO2 incubator containing 95% air and 5% CO2 at 37 CO:COLLECTION_SUMMARY ◦C. Cells with 80–90% confluence from passages 3–10 were used for CO:COLLECTION_SUMMARY experiments. All chemicals were purchased from Sigma-Aldrich (St. Louis, MO). CO:COLLECTION_SUMMARY H9c2 cardioblasts demonstrate similar to the primary cardiomyocytes hypertrophic CO:COLLECTION_SUMMARY response as well as mitochondrial metabolism and morphology. CO:SAMPLE_TYPE Cultured cells #TREATMENT TR:TREATMENT_SUMMARY Cultured H9c2 cells reaching 80–90% confluence from passages 3–10 were used TR:TREATMENT_SUMMARY for experiments. Ferroptosis was induced by incubating the cells with 0.5 µM TR:TREATMENT_SUMMARY RSL3 for 3 h. In addition, the cells were exposed to RSL3 in the presence of 1 TR:TREATMENT_SUMMARY µM Fer-1, 0.2 µM XJB, and 0.6 µM TSM. Thus, experiments were performed in the TR:TREATMENT_SUMMARY following 8 groups: (i) control (n = 5), (ii) RSL3 (n = 6), (iii) Fer-1 (n = 6), TR:TREATMENT_SUMMARY (iv) XJB (n = 5), (v) TSM (n = 5), (vi) RSL3 + Fer-1 (n = 6), (vii) RSL3 + XJB TR:TREATMENT_SUMMARY (n = 6), and (viii) RSL3 + TSM (n = 5). Fer-1, XJB, or TSM were added to the TR:TREATMENT_SUMMARY culture medium simultaneously with RSL3. At the end of the incubation, the cells TR:TREATMENT_SUMMARY were harvested and processed for analysis by GC-MS. TR:CELL_PCT_CONFLUENCE 80-90% #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Metabolites were extracted as previously described using 1 mL of cold MeOH/H2O SP:SAMPLEPREP_SUMMARY (85:15). Samples were sonicated for 15 s (3×) on ice and centrifuged at 1400 SP:SAMPLEPREP_SUMMARY rpm × 10 min at 4 ◦C (Rotor model: Eppendorf, FA-45-30-11). For protein SP:SAMPLEPREP_SUMMARY quantification, the pellets were dried for 15 min in a rotary vacuum evaporator, SP:SAMPLEPREP_SUMMARY then resuspended in 60 µL of denaturation buffer and sonicated for 1 min. All SP:SAMPLEPREP_SUMMARY samples were centrifuged for 10 min at the maximum speed and the supernatant was SP:SAMPLEPREP_SUMMARY used for protein quantification using the Bradford method. Supernatants were SP:SAMPLEPREP_SUMMARY collected and evaporated to dryness using a SpeedVac (Savant AS160, Farmingdale, SP:SAMPLEPREP_SUMMARY NY, USA). The metabolite samples were first derivatized through methoxyamination SP:SAMPLEPREP_SUMMARY by adding 30 µL of 20 mg/mL solution of methoxyamine hydrochloride Antioxidants SP:SAMPLEPREP_SUMMARY 2022, 11, 278 4 of 16 in pyridine (Sigma-Aldrich, St. Louis, MO, USA) and SP:SAMPLEPREP_SUMMARY incubated at 37 ◦C for 2 h. Afterward, trimethylsilylation was performed by SP:SAMPLEPREP_SUMMARY adding 30 µL of N-tert-butyldimethylsilyl-Nmethyltrifluoroacetamide (MTBSTFA + SP:SAMPLEPREP_SUMMARY 1% TBDMSCl; Sigma-Aldrich, St. Louis, MO, USA) and incubated for 1 h at 65 ◦C. SP:SAMPLEPREP_SUMMARY The reaction mixture was centrifuged at 1300 rpm × 10 min at RT. Supernatants SP:SAMPLEPREP_SUMMARY were transferred to analytical vials. Then, 20 µL per sample was added to glass SP:SAMPLEPREP_SUMMARY vials with inserts followed by the addition of 1 mM 2-fluobiphenyl SP:SAMPLEPREP_SUMMARY (Sigma-Aldrich, St. Louis, MO, USA) as an internal standard. Samples were SP:SAMPLEPREP_SUMMARY processed by GC-MS-QP2010 (Shimadzu Scientific Instruments, Inc., Columbia, MD) SP:SAMPLEPREP_SUMMARY using analytical conditions as previously described. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE GC CH:INSTRUMENT_NAME Shimadzu GCMS-QP2010 ultra CH:COLUMN_NAME Shimadzu SH-RXI (30M x 0.25mm x 0.25um) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Shimadzu QP2010 Ultra MS:INSTRUMENT_TYPE Single quadrupole MS:MS_TYPE EI MS:ION_MODE POSITIVE MS:MS_COMMENTS Raw chromatography data were obtained and processed in GC-MS Labsolution Postrun MS:MS_COMMENTS Analysis software (Shimadzu Scientific Instruments, Inc., Columbia, MD) for MS:MS_COMMENTS identification of metabolites from their electron impact mass spectra by MS:MS_COMMENTS comparison to the database (NIST14/2014/EPA/NIH) [26]. Peak integration for all MS:MS_COMMENTS metabolites and multiple searches in the mass spectral library database resulted MS:MS_COMMENTS in a final database of 52 metabolic features chosen for this analysis. To assess MS:MS_COMMENTS analytical accuracy and precision, an external quality evaluation was performed MS:MS_COMMENTS using 2-fluorobiphenyl spiked into derivatization blank samples before running MS:MS_COMMENTS on the GC-MS. The quantitative analysis corresponding to the metabolite MS:MS_COMMENTS concentrations in each sample was calculated based on the internal standard in MS:MS_COMMENTS mM. A table in the format of comma-delimited (* csv) was created and uploaded to MS:MS_COMMENTS MetaboAnalyst.ca. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS mM MS_METABOLITE_DATA_START Samples Control_ 1 Control_ 2 Control_ 3 Control_ 4 Control_ 5 RSL3_1 RSL3_2 RSL3_3 RSL3_4 RSL3_5 RSL3_6 Fer1-1 Fer1-2 Fer1-3 Fer1-4 Fer1-5 Fer1-6 XJB_1 XJB_2 XJB_3 XJB_4 XJB_5 TSM_1 TSM_3 TSM_4 TSM_5 TSM_6 RF_1 RF_2 RF_3 RF_4 RF_5 RF_6 RX_1 RX_2 RX_3 RX_4 RX_5 RX_6 RT_1 RT_2 RT_3 RT_4 RT_5 Factors Treatment:Control Treatment:Control Treatment:Control Treatment:Control Treatment:Control Treatment:RSL3 Treatment:RSL3 Treatment:RSL3 Treatment:RSL3 Treatment:RSL3 Treatment:RSL3 Treatment:Ferrostatin-1 Treatment:Ferrostatin-1 Treatment:Ferrostatin-1 Treatment:Ferrostatin-1 Treatment:Ferrostatin-1 Treatment:Ferrostatin-1 Treatment:XJB-5-131 Treatment:XJB-5-131 Treatment:XJB-5-131 Treatment:XJB-5-131 Treatment:XJB-5-131 Treatment:TSM-1005-44 Treatment:TSM-1005-44 Treatment:TSM-1005-44 Treatment:TSM-1005-44 Treatment:TSM-1005-44 Treatment:RSL3 + Ferrostatin-1 Treatment:RSL3 + Ferrostatin-1 Treatment:RSL3 + Ferrostatin-1 Treatment:RSL3 + Ferrostatin-1 Treatment:RSL3 + Ferrostatin-1 Treatment:RSL3 + Ferrostatin-1 Treatment:RSL3 + XJB-5-131 Treatment:RSL3 + XJB-5-131 Treatment:RSL3 + XJB-5-131 Treatment:RSL3 + XJB-5-131 Treatment:RSL3 + XJB-5-131 Treatment:RSL3 + XJB-5-131 Treatment:RSL3 + TSM-1005-44 Treatment:RSL3 + TSM-1005-44 Treatment:RSL3 + TSM-1005-44 Treatment:RSL3 + TSM-1005-44 Treatment:RSL3 + TSM-1005-44 L-Isoleucine 0.023855059 0.015279335 0.014545088 0.022637229 0.021945321 0.050670915 0.014616325 0.003900552 0.01927543 0.032504553 0.016565365 0.016415414 0.014712377 0.017001869 0.016562404 0.013118972 0.01568373 0.029007589 0.014969662 0.017246555 0.012363484 0.025104264 0.029031745 0.017385558 0.02136262 0.018028206 0.014678176 0.016185173 0.028696025 0.039483825 0.011957117 0.027405923 0.014321944 0.030243222 0.011542392 0.007411324 0.009537635 0.023201998 0.00568887 0.018364857 0.017640258 0.0151836 0.008833551 0.01138563 Lactic acid 0.125939804 0.09337406 0.129137651 0.107762424 0.126047214 0.290160595 0.219909143 0.047871635 0.229090977 0.213727032 0.171058054 0.099142715 0.092978155 0.137681627 0.122138602 0.095156206 0.112740426 0.20080092 0.119458102 0.134235546 0.104478705 0.147034117 0.124025908 0.171012849 0.148407457 0.101565986 0.123562074 0.111703872 0.173516058 0.172300966 0.085654391 0.146647882 0.121644252 0.182441848 0.082091554 0.130021092 0.073764815 0.130506573 0.106830095 0.133670191 0.11004941 0.1309109 0.075495975 0.11368298 Glycolic acid 0.003298776 0.002750013 0.00279402 0.00225361 0.002337595 0.008359305 0.00928996 0.005528053 0.008023406 0.004075016 0.007062717 0.002854614 0.002689416 0.002704701 0.002724213 0.002101356 0.003206289 0.005256121 0.003379105 0.002769912 0.00278366 0.003064505 0.003892295 0.004619428 0.003239358 0.002078777 0.003094245 0.003272378 0.004418248 0.00407853 0.001797896 0.002541607 0.003040179 0.004709193 0.002570095 0.007665224 0.001704929 0.002923804 0.00465892 0.003328813 0.00298039 0.002979955 0.001582257 0.002760576 Alanine 0.019916353 0.0124674 0.023820018 0.007436345 0.015180484 0.005457954 0.007740356 0.001858652 0.012240356 0.012090271 0.003304846 0.025523637 0.017383314 0.013572095 0.024339889 0.016856794 0.015847044 0.041730264 0.040934403 0.034221722 0.057073815 0.013994412 0.004819665 0.032199667 0.022640662 0.016491615 0.02545502 0.009007378 0.038142154 0.023005063 0.014487864 0.020140526 0.024724845 0.033358202 0.014606169 0.007120641 0.013942522 0.017292926 0.011129884 0.031020603 0.024222223 0.016493338 0.022994569 0.018048993 Glycine 0.063840525 0.055057513 0.072067617 0.024409483 0.052730001 0.012455841 0.019791921 0.004031489 0.033243655 0.047786735 0.011219615 0.067215485 0.07845006 0.05836014 0.076499691 0.064205831 0.065271092 0.12748852 0.111181359 0.088308854 0.090294642 0.060506428 0.01950663 0.100818504 0.082785148 0.060985084 0.088640106 0.034444617 0.119104367 0.076702189 0.053142508 0.060760672 0.085520447 0.096927682 0.044709419 0.018230028 0.047627833 0.053244145 0.030408211 0.087822205 0.08649512 0.070001044 0.051205763 0.069497966 Uracil 0.003162802 0.001429961 0.002009151 0.002401971 0.001621994 0.003262433 0.003176915 0.001582265 0.002487188 0.001544836 0.00000291 0.002658518 0.001721513 0.001493722 0.00123582 0.001013025 0.001103685 0.002510905 0.002192024 0.001785973 0.001606453 0.000801079 0.00173 0.001141859 0.0012855 0.001030812 0.001241423 0.002421561 0.001955378 0.002287316 0.001186378 0.001141896 0.000996756 0.0024939 0.001715902 0.002234229 0.001307734 0.00073331 0.001432043 0.001668969 0.001238628 0.000994586 0.001063623 0.001031085 beta-Alanine 0.011396068 0.01234891 0.013041611 0.001847765 0.008002482 0.00000215 0.001886448 0.00000304 0.004131464 0.007797701 0.00000291 0.013948524 0.01822585 0.009334721 0.017109656 0.016923626 0.01206034 0.0278 0.02855452 0.0211 0.021619516 0.009560429 0.000996 0.019132842 0.018813057 0.012603783 0.01843297 0.003974625 0.025698802 0.012172658 0.0113064 0.009325586 0.016088631 0.019742929 0.009140778 0.002897013 0.011472803 0.006917111 0.004890424 0.018848649 0.019229682 0.012208137 0.012511541 0.012592513 Urea 0.025203151 0.021177962 0.02105714 0.004486013 0.011758218 0.013045691 0.022690503 0.013805086 0.032874656 0.011604238 0.017193939 0.025987716 0.016250056 0.009802435 0.017986778 0.01552988 0.015578684 0.045296618 0.027834518 0.025615686 0.025055257 0.02017146 0.006068766 0.016815185 0.017004105 0.013560557 0.035363828 0.00661209 0.048122711 0.027918167 0.011511029 0.024877044 0.016415371 0.034104469 0.009460894 0.012440713 0.009558572 0.009997798 0.013172927 0.027555483 0.016481395 0.012539299 0.008635967 0.011491087 Valine 0.020787539 0.014269447 0.022485845 0.004873203 0.015756248 0.012161129 0.016355478 0.005846182 0.024094407 0.018629614 0.014179758 0.024051179 0.015893344 0.011651557 0.023011971 0.020650298 0.014564074 0.03309837 0.030163173 0.027814001 0.037959146 0.023039821 0.003626005 0.024327372 0.020962109 0.018524942 0.021619451 0.007433427 0.038921444 0.031898968 0.013817861 0.02869375 0.018854831 0.033690348 0.012627753 0.008780852 0.013498171 0.020783212 0.011965586 0.027968478 0.020281415 0.014792831 0.015394921 0.013783072 Capric acid 0.004974783 0.003301786 0.00424601 0.003879527 0.004294504 0.010000442 0.014341568 0.009766327 0.01286882 0.00000153 0.011966246 0.004389389 0.004132422 0.000454279 0.004146455 0.003677007 0.004969304 0.006964006 0.005569322 0.004595295 0.004616695 0.00542 0.004479175 0.006026406 0.004506783 0.003411426 0.005339432 0.005319985 0.006811212 0.006178835 0.002975503 0.004434405 0.004891581 0.006900755 0.004111617 0.012046049 0.002766703 0.002961368 0.007084671 0.004586982 0.004960107 0.004863502 0.002669682 0.004365559 Leucine 0.016711224 0.010679271 0.02005207 0.002923072 0.012816949 0.006743363 0.012452284 0.003228068 0.017757923 0.013559507 0.009061017 0.019799853 0.012984735 0.009176078 0.01929537 0.017083137 0.011621984 0.028694287 0.02731802 0.024318887 0.036607297 0.017865028 0.001881491 0.021018692 0.017069881 0.01561812 0.018663946 0.005097496 0.032113724 0.024267202 0.011505864 0.023792654 0.017341417 0.028298857 0.010063972 0.005914626 0.011147035 0.016873182 0.009926102 0.025326094 0.016561839 0.012356541 0.013399795 0.012074539 Nicotinamide N-oxide 0.00592835 0.003754863 0.00467984 0.00024683 0.003576068 0.002911867 0.003612366 0.00079789 0.004745878 0.005106227 0.001406985 0.00539224 0.004611871 0.004170515 0.005393097 0.003703522 0.004126976 0.007739983 0.005581857 0.006159536 0.005158855 0.004553699 0.00000133 0.005818934 0.00598125 0.003415925 0.005289264 0.001221427 0.007635139 0.006748857 0.003062027 0.004283403 0.004952085 0.00649 0.003308456 0.003041291 0.003017948 0.004120671 0.002452072 0.005032907 0.00476337 0.004335304 0.002809818 0.003740921 Isoleucine 0.016457639 0.011137714 0.019215012 0.002642118 0.013302931 0.00620341 0.010198287 0.002179869 0.016964999 0.014577685 0.007854586 0.019302386 0.012644601 0.009529272 0.019312243 0.0171309 0.011644083 0.028588968 0.025856808 0.024346699 0.036555308 0.018807974 0.002241494 0.02068118 0.01776113 0.016215847 0.017956655 0.004821652 0.032815108 0.026307814 0.01147224 0.024670974 0.015800568 0.027361851 0.010102372 0.004273026 0.011715826 0.018021415 0.008109857 0.0247067 0.017139275 0.011897637 0.012915785 0.011135413 5-oxoprolinate 0.045893503 0.034936198 0.031363909 0.098743207 0.051221782 0.11116158 0.014674322 0.00247599 0.023131999 0.07531309 0.020008806 0.017199658 0.028393791 0.053001295 0.037748695 0.026448178 0.029852092 0.052716846 0.014271896 0.033711698 0.015504803 0.053299401 0.140169477 0.040128297 0.060896067 0.043523893 0.021107561 0.057264596 0.058329373 0.085288127 0.024628653 0.058349468 0.03089371 0.05616077 0.026006916 0.011245135 0.025024594 0.081115667 0.005180568 0.031497623 0.040509975 0.035130337 0.023193764 0.020795909 Proline 0.013602531 0.011306159 0.015639253 0.006642107 0.009390786 0.00000215 0.00000308 0.00000304 0.00000346 0.00000153 0.00000291 0.0145 0.012400678 0.011959959 0.016349986 0.016050642 0.010803889 0.0219 0.0231 0.0213 0.028 0.011 0.00000133 0.018106809 0.015420932 0.012047229 0.015881187 0.00000101 0.022798308 0.0152 0.010149154 0.0128 0.015309812 0.0172 0.010690827 0.00000258 0.010430888 0.00000141 0.007222242 0.01542592 0.014463693 0.004649499 0.011562772 0.007276708 DL-Phenylalanine 0.012581463 0.008255818 0.007390117 0.016155214 0.011860658 0.028195791 0.006250167 0.00000304 0.008064901 0.01683354 0.007331471 0.007695679 0.007026093 0.009703644 0.009125882 0.006823166 0.00772665 0.013687601 0.0063041 0.00914 0.007229454 0.013363015 0.017991573 0.008863763 0.013175776 0.009506744 0.007271802 0.01051546 0.014781919 0.023101169 0.005531548 0.013876977 0.007590943 0.014079225 0.006370904 0.003311994 0.00525744 0.015110433 0.002907351 0.007809166 0.009027144 0.008163136 0.004901185 0.005538638 Sarcosine 0.007072394 0.004541216 0.00000021 0.005803044 0.005762509 0.00000215 0.00000308 0.00000304 0.00000346 0.00000153 0.00000291 0.00577 0.00593621 0.00000103 0.000000986 0.000000867 0.00000135 0.00000147 0.00753 0.00000104 0.000000884 0.00000147 0.00000133 0.008979589 0.00000123 0.00000093 0.007153757 0.00000101 0.00934 0.0092 0.00392 0.00601 0.00701 0.00999 0.000000936 0.00000258 0.000000704 0.00000141 0.00000188 0.00657 0.007135597 0.00622 0.00000071 0.00652 Glycerol 0.02232121 0.020662053 0.02616469 0.022147071 0.029562954 0.027941609 0.071093649 0.040483878 0.029116543 0.049421423 0.065764857 0.009679918 0.02973163 0.034791653 0.027441633 0.01905677 0.017262208 0.030163193 0.022498852 0.024270017 0.017111327 0.036431692 0.035889968 0.04572767 0.037226097 0.032503159 0.014912235 0.017420233 0.036650573 0.039296022 0.021388097 0.035957547 0.027860714 0.032300144 0.017354618 0.051010733 0.018245126 0.038746269 0.023293096 0.026968546 0.038584557 0.023403707 0.019327622 0.024017758 Oxoglutaric Acid 0.002882513 0.00162 0.00173 0.00101 0.0014 0.003084288 0.003747063 0.00234334 0.003497851 0.002140029 0.003109714 0.002173809 0.002095069 0.001493222 0.001701986 0.00156301 0.001860643 0.003642472 0.003284981 0.002279353 0.001733062 0.001870925 0.001401388 0.00284829 0.00187116 0.00130578 0.002283662 0.001881103 0.002672374 0.002207466 0.001465366 0.001435732 0.002308715 0.00281496 0.001343381 0.003476132 0.001180338 0.000474301 0.00233908 0.002938834 0.002214279 0.001722588 0.001242434 0.000240042 Taurine 0.000760029 0.00046333 0.001412973 0.000000167 0.001661404 0.00000215 0.00000308 0.00000304 0.00000346 0.00000153 0.00000291 0.000000866 0.000000919 0.00000103 0.000000986 0.000000867 0.00000135 0.00000147 0.000877 0.00000104 0.000000884 0.00000147 0.00000133 0.00000137 0.00232 0.000644 0.00000108 0.000492 0.00000156 0.00000122 0.000000796 0.00000116 0.00000124 0.00000125 0.000000936 0.00000258 0.000000704 0.00000141 0.00000188 0.00000127 0.00000139 0.0000011 0.00000071 0.00000107 Phosphoric acid 0.120988609 0.110822402 0.121381593 0.110787999 0.115535169 0.19792258 0.280560144 0.110703578 0.257993372 0.1859379 0.092859362 0.095160039 0.126614777 0.138561211 0.115005526 0.100777983 0.117673988 0.199742474 0.144806778 0.126529069 0.099288257 0.121315814 0.126247951 0.179946135 0.147225375 0.097193405 0.134143783 0.127705151 0.174239464 0.151632008 0.083712175 0.117005709 0.145816785 0.17062203 0.093839295 0.26055228 0.075031509 0.134659461 0.15264418 0.128652968 0.145008784 0.14364549 0.08073705 0.133929356 5-oxoproline 0.128676005 0.114496532 0.132799642 0.097051473 0.134340909 0.120354168 0.123912674 0.020974438 0.166135972 0.165946419 0.10049738 0.101865802 0.131449744 0.135904954 0.134623135 0.109818909 0.123829456 0.213141601 0.159874379 0.14776077 0.12477698 0.148172892 0.018731651 0.189112675 0.15919145 0.120097393 0.14175552 0.098402005 0.204106908 0.166226946 0.09405281 0.159648884 0.151423709 0.192686025 0.094565007 0.073494433 0.08520344 0.17913479 0.083633707 0.149727645 0.159185346 0.14982088 0.09164245 0.13984058 Methionine 0.006343864 0.004494439 0.007259292 0.00196191 0.007066836 0.003697826 0.003519369 0.002403511 0.005073187 0.004911741 0.002977017 0.009511137 0.006143692 0.004045949 0.006328642 0.007014562 0.005393159 0.010925642 0.008867865 0.009962322 0.013540752 0.006866014 0.001330042 0.009834899 0.007262064 0.006540129 0.008992496 0.002940643 0.011900896 0.010295324 0.005144874 0.011299559 0.00830638 0.012635213 0.0034444 0.001538108 0.004326041 0.007106848 0.002896473 0.010913529 0.006585876 0.006406498 0.004919554 0.006282225 N-Acetyl-aspartic acid 0.00538637 0.004490372 0.005218436 0.004511335 0.007060524 0.014375862 0.010196006 0.002212768 0.010488599 0.005632394 0.004856534 0.006369072 0.005444553 0.004296781 0.004454831 0.004499706 0.00599905 0.008540732 0.00556286 0.006489715 0.007142175 0.007580771 0.00282326 0.008425582 0.006319152 0.005183141 0.007183433 0.005082644 0.010551086 0.01042511 0.003882215 0.008908972 0.006820775 0.010485747 0.00406169 0.005785717 0.003507314 0.006852683 0.006381183 0.007680689 0.007282493 0.006682544 0.003707909 0.00578137 N-Acetyl-glutamic acid 0.164825465 0.150494275 0.157296139 0.159160428 0.162843668 0.121306564 0.068791993 0.007481262 0.144300591 0.249107276 0.026006664 0.109653338 0.15568902 0.192086865 0.158561826 0.128420037 0.148219192 0.246578513 0.158089069 0.167794257 0.116941882 0.171785679 0.147597108 0.209491617 0.217547059 0.138026856 0.15587407 0.168889297 0.226877735 0.215328942 0.111837596 0.153532847 0.1702987 0.211755358 0.118002631 0.125397193 0.104732736 0.188558645 0.061824711 0.153453968 0.183921661 0.175656088 0.1106552 0.145072624 Serine 0.106100208 0.093773153 0.077564976 0.140212347 0.11068827 0.055884191 0.053029393 0.005224395 0.078200931 0.176033219 0.014407168 0.065131363 0.07057596 0.105889777 0.095477212 0.075237315 0.099712294 0.142513248 0.141215068 0.123652075 0.101503522 0.108323602 0.124496107 0.161424493 0.146467124 0.09986049 0.120622941 0.114578619 0.172235675 0.120984531 0.084794751 0.140087266 0.122032985 0.13840832 0.063385045 0.070459847 0.094368488 0.098954348 0.055170577 0.137234973 0.151980058 0.139989932 0.074815483 0.111467555 Fumaric Acid 0.012821704 0.002954312 0.005169022 0.002690813 0.001759206 0.007824747 0.010208502 0.00595156 0.009106159 0.004903642 0.008278559 0.005575336 0.005010674 0.004325836 0.004907991 0.004004585 0.005186278 0.008241042 0.006949508 0.006055347 0.005625736 0.004851302 0.002914683 0.00689217 0.005550162 0.003720844 0.006510021 0.003707353 0.007839345 0.005829301 0.003119936 0.004466819 0.005597978 0.006837304 0.00411267 0.00846178 0.003078843 0.004875647 0.006011514 0.005141262 0.005594584 0.00557942 0.003245768 0.004632054 Threonine 0.037431741 0.025076351 0.035401782 0.009632452 0.045178799 0.010507435 0.019198344 0.00000304 0.019110298 0.022294361 0.00000291 0.033140131 0.043426901 0.028222558 0.040509457 0.03406041 0.02884009 0.071840105 0.055267766 0.0531 0.046240585 0.032122888 0.00517 0.068702061 0.043096471 0.042565889 0.039344576 0.018595731 0.052441585 0.051301546 0.035164956 0.058753946 0.050955354 0.065390293 0.017380012 0.013937495 0.02561982 0.041438779 0.018042876 0.06399282 0.045736707 0.035306096 0.033289193 0.050546546 Azelaic acid 0.005049156 0.005968772 0.007291422 0.005172823 0.00586692 0.013863295 0.017501754 0.010536147 0.016855325 0.007948808 0.014165882 0.00705836 0.006570059 0.006529387 0.007699167 0.005462106 0.0083758 0.012178068 0.009071533 0.007928983 0.007699097 0.007128824 0.005962524 0.009161322 0.008090877 0.00577943 0.0084 0.007558938 0.010738549 0.008546454 0.005034893 0.006598378 0.008104291 0.009342423 0.00596526 0.014431648 0.004415268 0.007383049 0.009952508 0.007538175 0.007271652 0.007563992 0.004160091 0.00000107 (2R)-Pyrrolidine 0.044177701 0.033288555 0.028768675 0.027218273 0.045378638 0.01086137 0.011822166 0.001126125 0.018193279 0.035568506 0.002077666 0.036870916 0.044309376 0.03439649 0.041788223 0.039779912 0.031837044 0.071785475 0.055982437 0.05069964 0.037893094 0.02744187 0.019313915 0.042113077 0.047760461 0.032431146 0.037627917 0.038463614 0.055290289 0.057029973 0.032647787 0.03142564 0.04638505 0.055410051 0.023268143 0.018116977 0.029596978 0.024130753 0.016257286 0.046347682 0.043661715 0.036778712 0.026651896 0.029132456 Myristic acid 0.008282244 0.005556793 0.006582844 0.005376153 0.00570148 0.011921147 0.013983247 0.009248585 0.014712132 0.007889027 0.01172599 0.007585885 0.006393904 0.00616334 0.006901225 0.004929387 0.007055656 0.01230644 0.008361248 0.007237151 0.007089993 0.006358322 0.005770744 0.009877226 0.008623478 0.004671261 0.008033163 0.007807508 0.009793734 0.008610932 0.004457288 0.006987243 0.007969687 0.010436441 0.004896303 0.014807145 0.003872962 0.006823164 0.008127854 0.008107714 0.006589326 0.007305413 0.003692001 0.005891791 Phenylalanine 0.018012637 0.010350195 0.015673607 0.002737505 0.014847184 0.00744277 0.009964326 0.001575289 0.014019983 0.011325276 0.005689059 0.021367879 0.015568074 0.008394259 0.017578495 0.015089254 0.011906697 0.031718192 0.025343563 0.021955349 0.027398634 0.014669159 0.000479472 0.022164724 0.01578767 0.013396246 0.019461407 0.005688488 0.027282557 0.023039823 0.01185404 0.023264364 0.017095096 0.029354071 0.00818918 0.003777387 0.009764319 0.013577647 0.00826486 0.024492847 0.016119043 0.014480512 0.010810139 0.013114481 Malic acid 0.042037953 0.022989728 0.039704651 0.031590398 0.03129084 0.043375153 0.030471572 0.003441549 0.041412073 0.048729406 0.008332921 0.033224098 0.030001279 0.037767816 0.038503798 0.027470293 0.037031193 0.061635161 0.035551108 0.04592742 0.034360791 0.036620102 0.023457658 0.056536858 0.049694661 0.027168737 0.044196099 0.042093558 0.039180865 0.050294473 0.028504826 0.028501988 0.049736645 0.04954884 0.02022185 0.040701906 0.024266464 0.018492559 0.033914487 0.032788851 0.030984314 0.042342274 0.02654173 0.036870741 Aspartic acid 0.053808295 0.039830528 0.046801476 0.006077402 0.03575348 0.000733283 0.006811066 0.000967959 0.017324213 0.016353057 0.00000291 0.052610318 0.056168395 0.028803576 0.051343484 0.040444866 0.037238784 0.097097403 0.092878616 0.069626094 0.062674825 0.030005771 0.000654 0.066544408 0.051288724 0.033346342 0.067125068 0.019589793 0.07276699 0.047206686 0.038443684 0.037675151 0.059598858 0.067103574 0.022781583 0.013777207 0.030810477 0.02182452 0.020204527 0.065395953 0.056011788 0.04648922 0.035878026 0.047654733 Cysteine 0.003687739 0.002232804 0.003954751 0.000219751 0.00375566 0.000797818 0.000738919 0.00000304 0.000918542 0.002497561 0.00102027 0.005370221 0.003653945 0.001953341 0.00365955 0.003746028 0.002687393 0.00798446 0.006767251 0.00505 0.006283105 0.003979657 0.00000133 0.004745373 0.003685005 0.003107129 0.00553648 0.000944735 0.00595745 0.00413 0.003153986 0.005078013 0.003821748 0.00824 0.001430185 0.00000258 0.002449284 0.001448096 0.000758099 0.006658221 0.003030085 0.002641867 0.002575897 0.002892686 Hypoxanthine 0.012603478 0.010041046 0.011666077 0.000332884 0.003218064 0.00000215 0.003403765 0.00093381 0.004730943 0.004529443 0.001169039 0.009600892 0.01301742 0.008450655 0.010650342 0.006624884 0.003793452 0.0209 0.013464818 0.009313022 0.008932992 0.00381422 0.00000133 0.024967696 0.009041399 0.005386574 0.01243633 0.006859686 0.015403523 0.011918083 0.005314014 0.004523014 0.019980455 0.0157 0.006684094 0.001601287 0.003794584 0.00452235 0.00765299 0.011833709 0.013628058 0.016395616 0.004751999 0.009983919 Palmitoleic Acid 0.005400579 0.003195201 0.003937695 0.001604624 0.003947455 0.015013897 0.005600217 0.003030533 0.00554165 0.002533149 0.002844892 0.008759197 0.004629218 0.006370899 0.005002939 0.003769871 0.003107945 0.015986332 0.00692722 0.006021826 0.005494643 0.002360456 0.002735323 0.006781339 0.005054622 0.003133099 0.005872247 0.004287805 0.006091438 0.004839494 0.003454929 0.004228772 0.004915619 0.007105273 0.001959889 0.003261194 0.002639353 0.002624007 0.002300676 0.006270224 0.003890881 0.00416894 0.002956481 0.001532651 Succinic Acid 0.011322986 0.007248162 0.008603387 0.007739112 0.011380083 0.004721837 0.004751181 0.000586165 0.004770014 0.007522711 0.002342106 0.016885666 0.013711137 0.011226468 0.016662507 0.012551824 0.014729481 0.030588878 0.019015645 0.018303302 0.017104004 0.016783733 0.017707611 0.01644609 0.01624334 0.012363518 0.016225333 0.0115771 0.021176851 0.026089002 0.012213874 0.016032963 0.015631506 0.027311745 0.009730898 0.01939676 0.009995153 0.010567267 0.019695718 0.021838579 0.014679718 0.012097897 0.010970896 0.013504049 Glutamic acid 0.130366424 0.115397463 0.13554732 0.044106736 0.125406239 0.02145544 0.077446225 0.007401358 0.143112462 0.112458888 0.010874607 0.109154969 0.143207795 0.133127669 0.133984853 0.114879118 0.123096857 0.227565078 0.173558938 0.151856775 0.123799692 0.125099944 0.001144323 0.201401295 0.158820937 0.110543603 0.152567309 0.099701165 0.193373084 0.158491354 0.099201842 0.13249267 0.160878297 0.188151207 0.08718369 0.102521395 0.087642976 0.129357926 0.113200361 0.150252302 0.155168828 0.152593939 0.093998276 0.144562246 Palmitic Acid 0.074509329 0.062894936 0.069335595 0.054702858 0.068666401 0.14010547 0.192415421 0.128173854 0.172064068 0.098658382 0.164565139 0.059308791 0.067941414 0.072968626 0.076270542 0.054877171 0.076115572 0.114865971 0.092675412 0.073181084 0.065311474 0.079432061 0.057614827 0.102814791 0.089125406 0.055271078 0.080056523 0.077772486 0.105385111 0.08998844 0.051292431 0.075168012 0.084615221 0.100012095 0.055517194 0.174147519 0.040875091 0.07827497 0.103800242 0.074170357 0.081161864 0.074227622 0.042058765 0.07398654 Asparagine 0.002478965 0.001737498 0.001787708 0.000000167 0.001854091 0.00000215 0.00000308 0.00000304 0.001790598 0.00000153 0.00000291 0.002981421 0.003538032 0.001315468 0.002688572 0.00238779 0.000720982 0.00558 0.00387 0.00342 0.003632298 0.00000147 0.00000133 0.005312901 0.002108509 0.002167583 0.002263773 0.00062642 0.003812822 0.00124 0.001922525 0.002836507 0.004327502 0.00409 0.000369601 0.00000258 0.001130547 0.001158164 0.001547609 0.004180133 0.002922175 0.003767956 0.001367528 0.00281 Lysine 0.007978151 0.002439664 0.007411387 0.000000167 0.01069009 0.00000215 0.00000308 0.00000304 0.00000346 0.00000153 0.00000291 0.008595977 0.009942097 0.007419826 0.008738474 0.005469809 0.00534368 0.00000147 0.0146 0.00814 0.0105 0.0123 0.00000133 0.008739069 0.007553605 0.004831058 0.009159315 0.003185596 0.014373667 0.008402179 0.008573682 0.008314998 0.008529585 0.0133 0.00651348 0.010289642 0.002517756 0.005260466 0.006575068 0.00947 0.012217942 0.010668734 0.004723595 0.00698705 Glutamine 0.007901031 0.005292178 0.00985931 0.000000167 0.007188397 0.00000215 0.008898593 0.001091752 0.000481363 0.00000153 0.00000291 0.015241304 0.017479088 0.004037214 0.012934806 0.010256724 0.00896105 0.0292 0.038494621 0.020502748 0.024324611 0.00417 0.00000133 0.035285606 0.006850334 0.008689067 0.022594694 0.001935604 0.016676782 0.005583804 0.012614786 0.017511282 0.020351837 0.0222 0.002032055 0.003473447 0.00537707 0.003721572 0.010634556 0.026427324 0.012700942 0.015031139 0.009720459 0.01960162 Oleic Acid 0.027919734 0.016362848 0.019987063 0.014222238 0.028637066 0.062872068 0.054003917 0.021900388 0.03348799 0.020757224 0.023528808 0.027773201 0.01854003 0.01806398 0.019493255 0.015628309 0.018357279 0.052708388 0.025349156 0.017728467 0.017862837 0.00000147 0.015224847 0.02768897 0.023040786 0.017722833 0.019369514 0.017645365 0.029676904 0.040761669 0.015637756 0.042079898 0.020718404 0.051611439 0.010072084 0.042876705 0.008747897 0.017022567 0.022268943 0.040392012 0.016320077 0.0167533 0.008390129 0.0167 Stearic acid 0.095315884 0.083183212 0.089543537 0.074922887 0.089295945 0.196752973 0.304801485 0.188164323 0.235823905 0.138334349 0.248729374 0.075460294 0.086984431 0.094369901 0.092949323 0.075050462 0.095035174 0.146631826 0.115229471 0.09357279 0.081779221 0.109435591 0.064418442 0.122202933 0.106936529 0.075000779 0.101233188 0.098795847 0.140594942 0.117202377 0.062440164 0.10344721 0.102374058 0.12960998 0.074285204 0.232850717 0.053367519 0.104287776 0.138023931 0.09836817 0.101335915 0.099936058 0.055179339 0.0949045 Citric acid 0.056152853 0.042749471 0.043146043 0.014053492 0.047981615 0.03353904 0.086260165 0.023634518 0.083116046 0.070928212 0.046372523 0.042111256 0.04891529 0.039705893 0.043676863 0.034172975 0.039563252 0.072852675 0.062875666 0.050298441 0.043398977 0.053950742 0.00000133 0.0381055 0.046443503 0.037828522 0.044307383 0.02993164 0.075624014 0.078721353 0.03127913 0.052734279 0.04538096 0.082 0.03425967 0.052683728 0.029569012 0.03419704 0.037873783 0.051079591 0.049578104 0.038143718 0.026435348 0.031810435 Tyrosine 0.030935585 0.019017757 0.02626668 0.002961756 0.0273143 0.014084846 0.02808103 0.005343837 0.028396489 0.017310368 0.011427078 0.03381467 0.027916623 0.015010427 0.029140958 0.024988391 0.022606871 0.05622667 0.043825445 0.035035562 0.039859037 0.025485261 0.00000133 0.040673505 0.025714057 0.023789648 0.033094738 0.008580446 0.045161204 0.034060386 0.020590238 0.041370537 0.03146477 0.0531 0.011761257 0.013558875 0.015031716 0.021948778 0.018978979 0.041984389 0.028288502 0.026324742 0.017103657 0.026466748 Pantothenic acid 0.012722606 0.010484868 0.011388821 0.006716299 0.017068893 0.012750239 0.02479739 0.00000304 0.014724217 0.01769584 0.012861225 0.008515419 0.013184065 0.009046916 0.011285844 0.011683973 0.012088358 0.029902221 0.02259979 0.0162 0.014757854 0.018251386 0.009389134 0.024798601 0.02254491 0.016778461 0.015032802 0.004720348 0.025820038 0.02224886 0.010775602 0.022742592 0.018520485 0.029713284 0.008154264 0.015924445 0.009279949 0.013582187 0.017831138 0.025834896 0.020634057 0.013527133 0.011100048 0.018130699 Isocitric Acid 0.041492961 0.028811482 0.029896187 0.011581434 0.081847199 0.174130143 0.165227325 0.021622106 0.048529324 0.059677482 0.066107327 0.035350002 0.038483613 0.018916372 0.023177613 0.024877778 0.033016426 0.100146547 0.041218235 0.02160536 0.025810359 0.069835763 0.039762964 0.017553059 0.031630978 0.049096002 0.021956094 0.104952227 0.149 0.036445687 0.032834064 0.116187313 0.028634014 0.14287616 0.000000936 0.022556725 0.010293779 0.021147711 0.032197112 0.104560319 0.029137414 0.015420366 0.01187768 0.010123469 Erythronic Acid 0.015629851 0.008026883 0.007440381 0.003973736 0.029974036 0.07317196 0.061243365 0.005316342 0.014250463 0.013009311 0.012519271 0.014271136 0.012911884 0.005096956 0.006252021 0.007490274 0.010941801 0.037561761 0.013794576 0.005792807 0.007900039 0.017212007 0.012354806 0.005044517 0.00697867 0.013254354 0.007635921 0.036168603 0.0514 0.010637583 0.01031943 0.044533948 0.008989006 0.052993858 0.000000936 0.005446684 0.002711336 0.004935047 0.011456356 0.039330607 0.007604371 0.004567034 0.002979058 0.003442801 Doconexent 0.005264463 0.002224213 0.00228244 0.002071983 0.003997353 0.005821268 0.00655643 0.001977954 0.004702206 0.00281631 0.001798357 0.005043663 0.003607299 0.002424632 0.002090606 0.001994351 0.003905367 0.007849101 0.003841821 0.003136095 0.003177819 0.002674335 0.00212007 0.003640135 0.002914862 0.002585722 0.00419 0.004071735 0.005784481 0.004402171 0.006568714 0.00490501 0.003674837 0.007279915 0.001561501 0.004764867 0.001367373 0.002714627 0.003119998 0.005257441 0.002150621 0.002641187 0.001266928 0.00000107 Cholesterol 0.092273888 0.03418243 0.014223492 0.031637829 0.092256547 0.16050364 0.21052374 0.060500925 0.047496294 0.006620348 0.014389302 0.074066686 0.071873712 0.024545282 0.02738179 0.02543229 0.065836806 0.135726526 0.083119227 0.028395252 0.035570025 0.036442798 0.067201304 0.051734549 0.029812383 0.027831903 0.061405474 0.081617162 0.003579931 0.118985659 0.034873924 0.073137125 0.066745041 0.100143334 0.007435461 0.101531957 0.010927928 0.004092978 0.071332456 0.08211544 0.044083873 0.062954521 0.009023298 0.075302247 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Characteristic Fragment Ions Retention time HMDB PubChem KEGG Formula Exact mass Super class Main class Sub class L-Isoleucine 86, 170, 188 8.737 HMDB0000172 6306 C00407 C6H13NO2 131 Organic acids Amino acids and peptides Amino acids Lactic acid 189, 233, 261 10.048 HMDB0000190 61503 C00256 C3H6O3 90.0317 Organic acids Short-chain acids Short-chain acids Glycolic acid 189, 247 10.282 HMDB0000115 757 C03547 C2H4O3 76.016 Fatty Acyls Fatty acids Hydroxy FA Alanine 158, 232, 260 10.754 HMDB0000161 5950 C00041 C3H7NO2 89.0477 Organic acids Amino acids and peptides Amino acids Glycine 218, 246 11.071 HMDB0000123 750 C00037 C2H5NO2 75.032 Organic acids Amino acids and peptides Amino acids Uracil 75,  179,  195 11.632 HMDB0000300 1174 C00106 C4H4N2O2 112.0273 Nucleic acids Pyrimidines Pyrimidines beta-Alanine 144, 218, 260 12.236 HMDB0000056 239 C00099 C3H7NO2 89.0477 Organic acids Amino acids and peptides Amino acids Urea 73, 147, 231 12.371 HMDB0000294 2447 C00086 CH4N2O 60.0324 Organic acids Carboximidic acids Isoureas Valine 186, 260, 288 12.471 HMDB0000883 6287 C00183 C5H11NO2 117.079 Organic acids Amino acids and peptides Amino acids Capric acid 75, 229 12.83 HMDB0000511 2969 C01571 C10H20O2 172.1463 Fatty Acyls Fatty acids Saturated FA Leucine 200, 274, 302 13.04 HMDB0000687 6106 C00123 C6H13NO2 131.0946 Organic acids Amino acids and peptides Amino acids Nicotinamide N-oxide 75,  136,  180 13.209 HMDB0001406 936 C00153 C6H6N2O 122.048 Organoheterocyclic compounds Pyridinecarboxylic acids Nicotinamides Isoleucine 200, 274, 302 13.48 HMDB0000172 6306 C00407 C6H13NO2 131.0946 Organic acids Amino acids and peptides Amino acids 5-oxoprolinate 84, 158, 186 13.578 HMDB0000805 499 C02237 C5H7NO3 129.0426 Organoheterocyclic compounds Pyrroline carboxylic acids Pyrroline carboxylic acids Proline 184, 258, 286 13.991 HMDB0000162 145742 C00148 C5H9NO2 115.0633 Organic acids Amino acids and peptides Amino acids DL-Phenylalanine 120, 130, 146 14.32 HMDB0250791 994 C02265 NA 165.079 Organic acids Amino acids and peptides Amino acids Sarcosine 158, 232, 260 14.873 HMDB0000271 1088 C00213 C3H7NO2 89.0477 Organic acids Amino acids and peptides Amino acids Glycerol 245, 289, 377 15.43 HMDB00131 753 C00116 C3H8O3 92.0473 Organic Oxygen Organooxygen Sugar alcohol Oxoglutaric Acid 157, 215, 317 19.22 HMDB0000208 51 C00026 C5H6O5 146.0215 Organic acids TCA acids TCA acids Taurine 88, 158, 296 15.844 HMDB0000251 1123 C00245 C2H7NO3S 125.0147 Organic acids Sulfonic acids Sulfonic acids Phosphoric acid 124, 383, 384 16.013 HMDB0001429 1004 C00009 H3PO4 97.9769 Non-metal compounds Non-metal oxanionic compounds Non-metal phosphates 5-oxoproline 198, 272, 300 16.349 HMDB0000267 7405 C01879 C5H7NO3 129.0426 Organoheterocyclic compounds Pyrroline carboxylic acids Pyrroline carboxylic acids Methionine 218, 292, 320 16.551 HMDB0000696 6137 C00073 C5H11NO2S 149.0511 Organic acids Amino acids and peptides Amino acids N-Acetyl-aspartic acid 73, 225, 346 16.601 HMDB0000812 65065 C01042 C6H9NO5 175.0481 Organic acids Amino acids and peptides Amino acids N-Acetyl-glutamic acid 186, 216, 318 16.8 HMDB0001138 185 C00624 C7H11NO5 189.0637 Organic acids Amino acids and peptides Amino acids Serine 288, 362, 390 16.823 HMDB0000187 5951 C00065 C3H7NO3 105.0426 Organic acids Amino acids and peptides Amino acids Fumaric Acid 147, 287, 329 17.02 HMDB00134 444972 C00122 C4H4O4 116.011 Organic acids TCA acids TCA acids Threonine 303, 376, 404 17.212 HMDB0000167 6288 C00188 C4H9NO3 119.0582 Organic acids Amino acids and peptides Amino acids Azelaic acid 129, 359, 401 17.57 HMDB00784 2266 C08261 C9H16O4 188.1049 Fatty Acyls Fatty Acids Saturated FA (2R)-Pyrrolidine 184, 214, 330 17.487 NA 91742919 NA C18H37NO4Si2 387.7 Organoheterocyclic compounds Pyrroline carboxylic acids Pyrroline carboxylic acids Myristic acid 75, 129, 285 17.892 HMDB0000806 11005 C06424 C14H28O2 228.2089 Fatty Acyls Fatty acids Saturated FA Phenylalanine 234, 302, 336 18.02 HMDB0000159 6140 C00079 C9H11NO2 165.079 Organic acids Amino acids and peptides Amino acids Malic acid 115, 287, 419 18.233 HMDB0000744 525 C03668 C4H6O5 134.0215 Organic acids TCA acids TCA acids Aspartic acid 302, 316, 418 18.933 HMDB0000191 5960 C00049 C4H7NO4 133.0375 Organic acids Amino acids and peptides Amino acids Cysteine 302, 378, 406 19.315 HMDB0000574 5862 C00097 C3H7NO2S 121.0198 Organic acids Amino acids and peptides Amino acids Hypoxanthine 73,  193,  307 19.554 HMDB0000157 790 C00262 C5H4N4O 136.0385 Nucleic acids Purines Hypoxanthines Palmitoleic Acid 75, 311, 353 20.051 HMDB0003229 5312427 C08362 C16H30O2 254.2246 Fatty Acyls Fatty acids Saturated FA Succinic Acid 73,147, 289, 331 14.028 HMDB0000254 1110 C00042 C4H6O4 118.0266 Organic acids TCA acids TCA acids Glutamic acid 272, 330, 432 20.026 HMDB0000148 33032 C00025 C5H9NO4 147.0532 Organic acids Amino acids and peptides Amino acids Palmitic Acid 75, 117, 129, 313 20.12 HMDB0000220 985 C00249 C16H32O2 256.2402 Fatty Acyls Fatty acids Saturated FA Asparagine 302, 400, 417 20.361 HMDB0000168 6267 C00152 C4H8N2O3 132.0535 Organic acids Amino acids and peptides Amino acids Lysine 198, 272, 300 21.144 HMDB0000182 5962 C00047 C6H14N2O2 146.1055 Organic acids Amino acids and peptides Amino acids Glutamine 198, 271, 431 21.596 HMDB0000641 5961 C00064 C5H10N2O3 146.0691 Organic acids Amino acids and peptides Amino acids Oleic Acid 75, 129, 339 21.946 HMDB0000207 445639 C00712 C18H34O2 282.2559 Fatty Acyls Fatty acids Unsaturated FA Stearic acid 75, 341 22.176 HMDB0000827 5281 C01530 C18H36O2 284.2715 Fatty Acyls Fatty acids Saturated FA Citric acid 431, 459, 591 23.494 HMDB0000094 311 C00158 C6H8O7 192.027 Organic acids TCA acids TCA acids Tyrosine 302, 364, 466 23.7 HMDB0000158 6057 C00082 C9H11NO3 181.0739 Organic acids Amino acids and peptides Amino acids Pantothenic acid 75, 87, 504 24.138 HMDB0000210 6613 C00864 C9H17NO5 219.11 Organic Oxygen Organooxygen Secondary alcohols Isocitric Acid 73, 147, 202, 345 24.435 HMDB0000193 1198 C00311 C6H8O7 192.027 Organic acids TCA acids TCA acids Erythronic Acid 261, 344, 476 24.56 HMDB0000613 2781043 NA C4H8O5 136.0372 Carbohydrates Monosaccharides Sugar acids Doconexent 75, 209, 385, 427 25.482 HMDB0002183 445580 C06429 C22H32O2 328.2402 Fatty Acyls Fatty acids Unsaturated FA Cholesterol 75, 367, 443 35.95 HMDB0000067 5997 C00187 C27H46O 386.3549 Sterol Lipids Sterols Cholesterols METABOLITES_END #END