#METABOLOMICS WORKBENCH fernandezlab_20210505_000411_mwtab.txt DATATRACK_ID:2619 STUDY_ID:ST001777 ANALYSIS_ID:AN002885 PROJECT_ID:PR001131 VERSION 1 CREATED_ON May 8, 2021, 9:41 am #PROJECT PR:PROJECT_TYPE MS based metabolomics PR:PROJECT_SUMMARY Fourier transform ion cyclotron resonance (FT-ICR) and Orbitrap mass PR:PROJECT_SUMMARY spectrometry (MS) are among the highest-performing analytical platforms in PR:PROJECT_SUMMARY metabolomics. Their high mass measurement accuracy and mass resolving power PR:PROJECT_SUMMARY enable detailed investigation of biological metabolomes. Non-targeted MS PR:PROJECT_SUMMARY experiments, however, yield extremely complex datasets that make metabolite PR:PROJECT_SUMMARY annotation very challenging, if not impossible. High-resolution accurate mass PR:PROJECT_SUMMARY measurements greatly facilitate this process by reducing mass errors and PR:PROJECT_SUMMARY spectral overlaps. When applied together with relative isotopic abundance (RIA) PR:PROJECT_SUMMARY measurements, heuristic rules, and constraints during searches, the number of PR:PROJECT_SUMMARY candidate elemental formula(s) can be significantly reduced. Here, we evaluate PR:PROJECT_SUMMARY the performance of two leading analytical MS platforms, Orbitrap ID-X and 12T PR:PROJECT_SUMMARY solariX FT-ICR mass spectrometers, in terms of mass accuracy and RIA PR:PROJECT_SUMMARY measurements, and how these factors affect the assignment of the correct PR:PROJECT_SUMMARY elemental formulae in metabolite annotation. Quality of the mass measurements PR:PROJECT_SUMMARY was evaluated under various experimental conditions (resolution: 120 K, 240 K, PR:PROJECT_SUMMARY 500 K; automatic gain control: 5e4, 1e5, 5e5) for the Orbitrap MS platform. PR:INSTITUTE Georgia Institute of Technology PR:DEPARTMENT Chemistry PR:LABORATORY Fernández PR:FIRST_NAME Danning PR:ADDRESS 901 Atlantic Dr NE, Atlanta, GA, 30332, USA PR:EMAIL dhuang74@gatech.edu PR:PHONE 404-512-7523 PR:PROJECT_TITLE Comparison of High-Resolution Fourier Transform Mass Spectrometry Platforms for PR:PROJECT_TITLE Metabolite Annotation (part I) PR:LAST_NAME Huang #STUDY ST:STUDY_TITLE Comparison of High-Resolution Fourier Transform Mass Spectrometry Platforms for ST:STUDY_TITLE Metabolite Annotation - C. elegans ST:STUDY_SUMMARY Fourier transform ion cyclotron resonance (FT-ICR) and Orbitrap mass ST:STUDY_SUMMARY spectrometry (MS) are among the highest-performing analytical platforms in ST:STUDY_SUMMARY metabolomics. Their high mass measurement accuracy and mass resolving power ST:STUDY_SUMMARY enable detailed investigation of biological metabolomes. Non-targeted MS ST:STUDY_SUMMARY experiments, however, yield extremely complex datasets that make metabolite ST:STUDY_SUMMARY annotation very challenging, if not impossible. High-resolution accurate mass ST:STUDY_SUMMARY measurements greatly facilitate this process by reducing mass errors and ST:STUDY_SUMMARY spectral overlaps. When applied together with relative isotopic abundance (RIA) ST:STUDY_SUMMARY measurements, heuristic rules, and constraints during searches, the number of ST:STUDY_SUMMARY candidate elemental formula(s) can be significantly reduced. Here, we evaluate ST:STUDY_SUMMARY the performance of two leading analytical MS platforms, Orbitrap ID-X and 12T ST:STUDY_SUMMARY solariX FT-ICR mass spectrometers, in terms of mass accuracy and RIA ST:STUDY_SUMMARY measurements, and how these factors affect the assignment of the correct ST:STUDY_SUMMARY elemental formulae in metabolite annotation. Quality of the mass measurements ST:STUDY_SUMMARY was evaluated under various experimental conditions (resolution: 120 K, 240 K, ST:STUDY_SUMMARY 500 K; automatic gain control: 5e4, 1e5, 5e5) for the Orbitrap MS platform. ST:INSTITUTE Georgia Institute of Technology ST:DEPARTMENT Chemistry ST:LABORATORY Fernández ST:LAST_NAME Huang ST:FIRST_NAME Danning ST:ADDRESS 901 Atlantic Dr NE ST:EMAIL dhuang74@gatech.edu ST:STUDY_TYPE MS based metabolomics ST:PHONE 4045127523 #SUBJECT SU:SUBJECT_TYPE Synthetic sample #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 - Model mixture 1 Experimental Condition:AGC 5e4 - R 120K - neg RAW_FILE_NAME=MeOH_AGC5e4_R120K_neg.raw SUBJECT_SAMPLE_FACTORS - Model mixture 2 Experimental Condition:AGC 5e4 - R 240K - neg RAW_FILE_NAME=MeOH_AGC5e4_R240K_neg.raw SUBJECT_SAMPLE_FACTORS - Model mixture 3 Experimental Condition:AGC 5e4 - R 500K - neg RAW_FILE_NAME=MeOH_AGC5e4_R500K_neg.raw SUBJECT_SAMPLE_FACTORS - Model mixture 4 Experimental Condition:AGC 1e5 - R 120K - neg RAW_FILE_NAME=MeOH_AGC1e5_R120K_neg.raw SUBJECT_SAMPLE_FACTORS - Model mixture 5 Experimental Condition:AGC 1e5 - R 240K - neg RAW_FILE_NAME=MeOH_AGC1e5_R240K_neg.raw SUBJECT_SAMPLE_FACTORS - Model mixture 6 Experimental Condition:AGC 1e5 - R 500K - neg RAW_FILE_NAME=MeOH_AGC1e5_R500K_neg.raw SUBJECT_SAMPLE_FACTORS - Model mixture 7 Experimental Condition:AGC 5e5 - R 120K - neg RAW_FILE_NAME=MeOH_AGC5e5_R120K_neg.raw SUBJECT_SAMPLE_FACTORS - Model mixture 8 Experimental Condition:AGC 5e5 - R 240K - neg RAW_FILE_NAME=MeOH_AGC5e5_R240K_neg.raw SUBJECT_SAMPLE_FACTORS - Model mixture 9 Experimental Condition:AGC 5e5 - R 500K - neg RAW_FILE_NAME=MeOH_AGC5e5_R500K_neg.raw SUBJECT_SAMPLE_FACTORS - Model mixture 10 Experimental Condition:AGC 5e4 - R 120K - pos RAW_FILE_NAME=MeOH_AGC5e4_R120K_pos.raw SUBJECT_SAMPLE_FACTORS - Model mixture 11 Experimental Condition:AGC 5e4 - R 240K - pos RAW_FILE_NAME=MeOH_AGC5e4_R240K_pos.raw SUBJECT_SAMPLE_FACTORS - Model mixture 12 Experimental Condition:AGC 5e4 - R 500K - pos RAW_FILE_NAME=MeOH_AGC5e4_R500K_pos.raw SUBJECT_SAMPLE_FACTORS - Model mixture 13 Experimental Condition:AGC 1e5 - R 120K - pos RAW_FILE_NAME=MeOH_AGC1e5_R120K_pos.raw SUBJECT_SAMPLE_FACTORS - Model mixture 14 Experimental Condition:AGC 1e5 - R 240K - pos RAW_FILE_NAME=MeOH_AGC1e5_R240K_pos.raw SUBJECT_SAMPLE_FACTORS - Model mixture 15 Experimental Condition:AGC 1e5 - R 500K - pos RAW_FILE_NAME=MeOH_AGC1e5_R500K_pos.raw SUBJECT_SAMPLE_FACTORS - Model mixture 16 Experimental Condition:AGC 5e5 - R 120K - pos RAW_FILE_NAME=MeOH_AGC5e5_R120K_pos.raw SUBJECT_SAMPLE_FACTORS - Model mixture 17 Experimental Condition:AGC 5e5 - R 240K - pos RAW_FILE_NAME=MeOH_AGC5e5_R240K_pos.raw SUBJECT_SAMPLE_FACTORS - Model mixture 18 Experimental Condition:AGC 5e5 - R 500K - pos RAW_FILE_NAME=MeOH_AGC5e5_R500K_pos.raw SUBJECT_SAMPLE_FACTORS - PD1074 C. elegans mixture 1 Experimental Condition:AGC 5e4 - R 120K - neg RAW_FILE_NAME=Biomatrix_AGC5e4_R120K_neg.raw SUBJECT_SAMPLE_FACTORS - PD1074 C. elegans mixture 2 Experimental Condition:AGC 5e4 - R 240K - neg RAW_FILE_NAME=Biomatrix_AGC5e4_R240K_neg.raw SUBJECT_SAMPLE_FACTORS - PD1074 C. elegans mixture 3 Experimental Condition:AGC 5e4 - R 500K - neg RAW_FILE_NAME=Biomatrix_AGC5e4_R500K_neg.raw SUBJECT_SAMPLE_FACTORS - PD1074 C. elegans mixture 4 Experimental Condition:AGC 1e5 - R 120K - neg RAW_FILE_NAME=Biomatrix_AGC1e5_R120K_neg.raw SUBJECT_SAMPLE_FACTORS - PD1074 C. elegans mixture 5 Experimental Condition:AGC 1e5 - R 240K - neg RAW_FILE_NAME=Biomatrix_AGC1e5_R240K_neg.raw SUBJECT_SAMPLE_FACTORS - PD1074 C. elegans mixture 6 Experimental Condition:AGC 1e5 - R 500K - neg RAW_FILE_NAME=Biomatrix_AGC1e5_R500K_neg.raw SUBJECT_SAMPLE_FACTORS - PD1074 C. elegans mixture 7 Experimental Condition:AGC 5e5 - R 120K - neg RAW_FILE_NAME=Biomatrix_AGC5e5_R120K_neg.raw SUBJECT_SAMPLE_FACTORS - PD1074 C. elegans mixture 8 Experimental Condition:AGC 5e5 - R 240K - neg RAW_FILE_NAME=Biomatrix_AGC5e5_R240K_neg.raw SUBJECT_SAMPLE_FACTORS - PD1074 C. elegans mixture 9 Experimental Condition:AGC 5e5 - R 500K - neg RAW_FILE_NAME=Biomatrix_AGC5e5_R500K_neg.raw SUBJECT_SAMPLE_FACTORS - PD1074 C. elegans mixture 10 Experimental Condition:AGC 5e4 - R 120K - pos RAW_FILE_NAME=Biomatrix_AGC5e4_R120K_pos.raw SUBJECT_SAMPLE_FACTORS - PD1074 C. elegans mixture 11 Experimental Condition:AGC 5e4 - R 240K - pos RAW_FILE_NAME=Biomatrix_AGC5e4_R240K_pos.raw SUBJECT_SAMPLE_FACTORS - PD1074 C. elegans mixture 12 Experimental Condition:AGC 5e4 - R 500K - pos RAW_FILE_NAME=Biomatrix_AGC5e4_R500K_pos.raw SUBJECT_SAMPLE_FACTORS - PD1074 C. elegans mixture 13 Experimental Condition:AGC 1e5 - R 120K - pos RAW_FILE_NAME=Biomatrix_AGC1e5_R120K_pos.raw SUBJECT_SAMPLE_FACTORS - PD1074 C. elegans mixture 14 Experimental Condition:AGC 1e5 - R 240K - pos RAW_FILE_NAME=Biomatrix_AGC1e5_R240K_pos.raw SUBJECT_SAMPLE_FACTORS - PD1074 C. elegans mixture 15 Experimental Condition:AGC 1e5 - R 500K - pos RAW_FILE_NAME=Biomatrix_AGC1e5_R500K_pos.raw SUBJECT_SAMPLE_FACTORS - PD1074 C. elegans mixture 16 Experimental Condition:AGC 5e5 - R 120K - pos RAW_FILE_NAME=Biomatrix_AGC5e5_R120K_pos.raw SUBJECT_SAMPLE_FACTORS - PD1074 C. elegans mixture 17 Experimental Condition:AGC 5e5 - R 240K - pos RAW_FILE_NAME=Biomatrix_AGC5e5_R240K_pos.raw SUBJECT_SAMPLE_FACTORS - PD1074 C. elegans mixture 18 Experimental Condition:AGC 5e5 - R 500K - pos RAW_FILE_NAME=Biomatrix_AGC5e5_R500K_pos.raw #COLLECTION CO:COLLECTION_SUMMARY PD1074 C. elegans samples (~10 mg) were collected and lyophilized. CO:SAMPLE_TYPE Worms CO:STORAGE_CONDITIONS -80℃ #TREATMENT TR:TREATMENT_SUMMARY standard PD1047 C.elegans treatment #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Model Mixture Sample Preparation One hundred and four selected chemical standard SP:SAMPLEPREP_SUMMARY compounds were purchased from Sigma-Aldrich (St. Louis, MO, USA) and used to SP:SAMPLEPREP_SUMMARY prepare stock solutions at a concentration of 0.001 M in LC-MS grade methanol SP:SAMPLEPREP_SUMMARY (Fisher Scientific, Pittsburgh, PA, USA). These standards were selected because SP:SAMPLEPREP_SUMMARY they map to key metabolic pathways, some involved in cancer. If a chemical could SP:SAMPLEPREP_SUMMARY not be completely dissolved in methanol, LC-MS grade water (Fisher Scientific, SP:SAMPLEPREP_SUMMARY Pittsburgh, PA, USA) was added to increase solubility. A pooled sample of 104 SP:SAMPLEPREP_SUMMARY standard compounds was diluted with methanol at a final concentration of 5 μM. SP:SAMPLEPREP_SUMMARY PD1074 C. elegans Bio-mixture Sample Preparation Three 2.0 mm zirconium oxide SP:SAMPLEPREP_SUMMARY beads and ~75 μL volume of 0.5 mm glass beads were added to each lyophilized SP:SAMPLEPREP_SUMMARY PD1074 C. elegans sample (~10 mg). Samples were placed in a TissueLyser II SP:SAMPLEPREP_SUMMARY (QIAGEN, Hilden, Germany) and homogenized at 1,800 rpm, -80 °C for 3 min. One SP:SAMPLEPREP_SUMMARY and a half mL of 80% methanol (in water) was added to each homogenized sample. SP:SAMPLEPREP_SUMMARY Samples were then shaken using an Isotemp high speed shaker (Fisher Scientific, SP:SAMPLEPREP_SUMMARY Pittsburgh, PA, USA) at 1,500 rpm for 30 min, and centrifuged at 22,100 g for 5 SP:SAMPLEPREP_SUMMARY min. The supernatant was collected, dried and stored at -80 °C. Prior to MS SP:SAMPLEPREP_SUMMARY analysis, dried C. elegans matrices were resuspended with 1 mL LC-MS grade SP:SAMPLEPREP_SUMMARY methanol containing the 104 standard compounds at a 5 μM concentration. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY HILIC method CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Thermo Vanquish CH:COLUMN_NAME Waters Acquity BEH Amide (150 x 2.1mm, 1.7um) CH:FLOW_RATE 0.4 ml/min CH:COLUMN_TEMPERATURE 40 CH:SOLVENT_A water/acetonitrile (80:20 v/v), with 10 mM formate and 0.1% formic acid CH:SOLVENT_B acetonitrile with 0.1% formic acid CH:SAMPLE_INJECTION 2μL #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Orbitrap ID-X Tribrid MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS Samples were analyzed in positive and negative ion modes for HILIC MS:MS_COMMENTS chromatography. Spectral features were extracted from the raw data using MZmine MS:MS_COMMENTS 2.51. MS:SPRAY_VOLTAGE -2.5 kV MS:AUTOMATIC_GAIN_CONTROL 5e4, 1e5, 5e5 #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Da MS_METABOLITE_DATA_START Samples Model mixture 1 Model mixture 2 Model mixture 3 Model mixture 4 Model mixture 5 Model mixture 6 Model mixture 7 Model mixture 8 Model mixture 9 PD1074 C. elegans mixture 1 PD1074 C. elegans mixture 2 PD1074 C. elegans mixture 3 PD1074 C. elegans mixture 4 PD1074 C. elegans mixture 5 PD1074 C. elegans mixture 6 PD1074 C. elegans mixture 7 PD1074 C. elegans mixture 8 PD1074 C. elegans mixture 9 Factors Experimental Condition:AGC 5e4 - R 120K - neg Experimental Condition:AGC 5e4 - R 240K - neg Experimental Condition:AGC 5e4 - R 500K - neg Experimental Condition:AGC 1e5 - R 120K - neg Experimental Condition:AGC 1e5 - R 240K - neg Experimental Condition:AGC 1e5 - R 500K - neg Experimental Condition:AGC 5e5 - R 120K - neg Experimental Condition:AGC 5e5 - R 240K - neg Experimental Condition:AGC 5e5 - R 500K - neg Experimental Condition:AGC 5e4 - R 120K - neg Experimental Condition:AGC 5e4 - R 240K - neg Experimental Condition:AGC 5e4 - R 500K - neg Experimental Condition:AGC 1e5 - R 120K - neg Experimental Condition:AGC 1e5 - R 240K - neg Experimental Condition:AGC 1e5 - R 500K - neg Experimental Condition:AGC 5e5 - R 120K - neg Experimental Condition:AGC 5e5 - R 240K - neg Experimental Condition:AGC 5e5 - R 500K - neg glycolic acid 75.00861 75.00863 75.00865 75.00862 75.00864 75.00865 75.00862 75.00864 75.00865 75.00862 75.00865 75.00866 75.00863 75.00865 75.00866 75.00863 75.00867 75.00871 Pyruvate 87.00867 87.00869 87.0087 87.00868 87.0087 87.00872 87.00869 87.00872 87.00873 87.00869 87.00871 87.00871 87.0087 87.00871 87.00872 87.00869 87.00877 87.00881 Alanine 88.04035 88.04036 88.04037 88.04036 88.04038 88.04037 88.04038 88.04044 88.04049 88.04035 88.04037 88.04038 88.04036 88.04038 88.0404 88.04039 88.04034 88.04052 L-Lactate 89.02434 89.02436 89.02437 89.02435 89.02437 89.02438 89.02436 89.02439 89.02441 89.02436 89.02437 89.02438 89.02437 89.02438 89.02439 89.02435 89.02438 89.02436 Acetoacetate 101.02438 101.02437 101.02439 101.02437 101.02437 101.02441 101.02439 101.02441 101.02444 101.02437 101.02442 101.0244 101.02438 101.02441 101.02441 101.02441 101.02438 101.02451 2-Aminoisobutyric acid 102.05601 n/a n/a 102.05602 n/a n/a n/a n/a n/a 102.05603 102.05604 n/a 102.05602 102.05602 n/a 102.05607 n/a n/a Choline n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a Malonic acid 103.00368 103.00365 103.00366 103.00364 103.00365 103.00367 103.00365 103.00367 103.00369 103.00365 103.00368 103.00369 103.00366 103.00368 103.00369 103.00365 103.00369 103.00369 L-Serine 104.03528 104.0353 104.03532 104.03529 104.03531 104.03531 104.03526 104.03539 104.03544 104.0353 104.03533 104.03532 104.0353 104.03531 104.03534 104.03538 104.0353 104.03553 L-Glyceric acid 105.01928 105.0193 105.01932 105.01929 105.01932 105.01933 105.01931 105.01944 105.01948 105.01923 105.01933 105.01934 105.01931 105.01933 105.01934 105.01935 105.0193 105.01955 Cytosine n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a Histamine n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a Uracil 111.01995 111.01997 111.01999 111.01997 111.02 111.02 111.01999 111.02003 111.02005 111.01998 111.02 111.02 111.02 111.02001 111.02002 111.02001 111.02006 111.02019 L-Proline (Reagents Plus) n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a Fumarate dibasic 115.00363 115.00365 115.00366 115.00364 115.00365 115.00366 115.00365 115.00366 115.00365 115.00364 115.00365 115.00366 115.00364 115.00364 115.00365 115.0036 115.00366 115.00368 N-Acetylglycine 116.03523 116.03525 116.03526 116.03522 116.03522 116.03523 116.03525 116.0353 116.03505 116.03523 116.03524 116.03524 116.03521 116.03522 116.03524 116.03517 116.03521 116.03515 L-Valine n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a Succinate (dibasic) 117.01923 117.01924 117.01923 117.01923 117.01927 117.01924 117.01934 117.01921 117.01919 117.01922 117.01921 117.01926 117.01923 117.01927 117.0192 117.01943 117.01931 117.019 L-Threonine 118.05088 118.05089 118.0509 118.05085 118.05087 118.05087 118.05068 118.05087 118.05092 118.05087 118.05091 118.0509 118.05085 118.05087 118.05088 118.05084 118.05074 118.05099 D-(-)-Erythrose 119.03495 119.03491 n/a 119.0349 119.03493 n/a 119.03491 n/a n/a 119.03493 119.03493 119.035 119.03493 119.03495 n/a 119.03493 119.035 n/a Nicotinamide n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a Nicotinic acid 122.0247 122.02471 122.02472 122.0247 122.02471 122.02472 122.02464 122.02467 122.0247 122.0247 122.02472 122.02474 122.0247 122.02472 122.02473 122.02465 122.02469 122.02462 Taurine 124.00731 124.00734 124.00734 124.00732 124.00734 124.00733 124.00731 124.00741 124.00745 124.00734 124.00736 124.00736 124.00733 124.00735 124.00736 124.00733 124.00726 124.00751 Pyroglutamic acid 128.03526 128.03528 128.03529 128.03526 128.03526 128.03526 128.03512 128.03511 128.03518 128.03527 128.03528 128.03527 128.03525 128.03525 128.03523 128.03531 128.03531 128.03526 Citraconic acid 129.01929 129.0193 129.0193 129.01928 129.01928 129.01927 129.01926 129.01927 129.0193 129.01932 129.0193 129.01926 129.01935 129.01936 129.01934 129.01941 129.01943 129.01921 trans-4-Hydroxy-L-proline 130.05093 130.05094 130.05095 130.05095 130.05095 130.05098 130.051 130.05107 130.05115 130.05094 130.05096 130.05097 130.05094 130.05097 130.05099 130.05105 130.05097 130.05124 creatine 130.06216 130.06217 130.06219 130.06218 130.06219 130.06219 130.06222 130.06231 130.06238 130.06217 130.06219 130.06219 130.06218 130.06219 130.06222 130.06227 130.06216 130.06245 L-Leucine 130.0873 n/a n/a 130.08728 n/a n/a n/a n/a n/a 130.08731 130.08733 n/a 130.0873 130.08734 n/a 130.0873 n/a n/a Glutaric acid 131.03493 131.03491 131.03492 131.03491 131.03487 131.0349 131.035 131.03477 131.03486 131.03492 131.03492 131.03491 131.03492 131.03489 131.03488 131.03493 131.03474 131.03474 L-Asparagine 131.0462 131.04622 131.04621 131.04622 131.04622 131.04625 131.04616 131.04634 131.0464 131.04622 131.04622 131.04623 131.04622 131.04624 131.04625 131.04633 131.04622 131.04652 L-Leucic acid 131.0713 131.07131 131.07132 131.07129 131.07131 131.07131 131.07117 131.07118 131.07125 131.07131 131.07132 131.07132 131.07129 131.0713 131.0713 131.07113 131.0714 131.0713 L-Ornithine 131.08257 131.08259 131.0826 131.0826 131.08261 131.08264 131.08256 131.08271 131.08279 131.08258 131.0826 131.08261 131.0826 131.08263 131.08264 131.08269 131.08259 131.0829 L-Aspartic acid 132.03022 132.03023 n/a 132.03024 n/a n/a n/a n/a n/a 132.03027 132.03024 132.03027 132.03024 132.03026 132.03028 132.03036 n/a n/a DL-Malic acid 133.01421 133.01422 133.01424 133.01421 133.01422 133.01425 133.01413 n/a n/a 133.01422 133.01424 133.01423 133.01425 133.01427 133.01425 133.01432 133.01437 133.0145 Adenine 134.04717 134.04718 134.04719 134.04717 134.04718 134.04718 134.04714 134.04712 134.04713 134.04718 134.0472 134.0472 134.04719 134.0472 134.0472 134.04717 134.04721 134.04715 Hypoxanthine 135.03114 135.03115 135.03116 135.0311 135.03112 135.03112 135.03093 135.03092 135.03088 135.03114 135.03116 135.03113 135.03114 135.03108 135.0311 135.03088 135.03104 135.03088 Anthranilic acid 136.04033 136.0403 136.04036 136.04029 136.04031 136.04031 136.04026 136.0403 136.04031 136.04033 136.04036 136.04036 136.04032 136.04034 136.04034 136.0403 136.04027 136.04037 Acetylcholine n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a alpha-Ketoglutaric acid 145.01417 145.01419 145.0142 145.01416 145.01418 145.01419 145.01411 145.01413 145.01416 145.01417 145.01419 145.0142 145.01416 145.01418 145.01419 145.0141 145.0141 145.0141 Adipic acid 145.05056 145.05057 145.05056 145.05055 145.05055 145.05054 145.05044 145.0505 145.05049 145.05057 145.05058 145.05058 145.05055 145.05056 145.05055 145.05051 145.05048 145.05043 L-Glutamine 145.0618 145.06181 145.06183 145.06178 145.0618 145.06181 145.06169 145.06185 145.06194 145.0618 145.06183 145.06182 145.06178 145.06179 145.06182 145.06186 145.0617 145.06201 L-Lysine n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a L-Glutamic acid 146.04579 146.0458 146.04581 146.04578 146.04579 146.0458 146.04567 146.04583 146.04593 146.0458 146.04581 146.04582 146.04577 146.04578 146.04579 146.04582 146.04569 146.04601 L-Methionine 148.04369 148.04373 148.04372 148.04368 148.04371 148.04371 148.04363 148.04378 148.04385 148.04371 148.04372 148.04374 148.04369 148.04371 148.04371 148.04373 148.0436 148.04393 Xanthine 151.02608 151.0261 151.02611 151.02608 151.02609 151.0261 151.02598 151.02607 151.02611 151.02609 151.02611 151.02608 151.02608 151.02607 151.02606 151.02602 151.02608 151.02579 Cystamine n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a Histidine 154.06215 154.06217 154.06217 154.06214 154.06215 154.06217 154.06205 154.06221 154.0623 154.06216 154.06217 154.06218 154.06214 154.06217 154.06217 154.06226 154.06205 154.06239 Orotic acid 155.00977 155.00979 155.0098 155.00977 155.00979 155.00979 155.00966 155.00964 155.00958 155.00978 155.00979 155.00979 155.00974 155.00975 155.00974 155.00966 155.00951 155.00981 Allantoin 157.03665 157.03667 157.03667 157.03665 157.03665 157.03665 157.03649 157.03648 157.03665 157.03665 157.03666 157.03666 157.03661 157.03661 157.0366 157.03659 157.03655 157.0363 DL-2-Aminoadipic acid 160.0615 160.0615 160.06152 160.06149 160.06151 160.06152 160.06138 160.06162 160.06166 160.06149 160.06152 160.0615 160.06149 160.0615 160.0615 160.06156 160.06142 160.06172 Carnitine n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a Phenylpyruvic acid 163.04003 163.04004 163.04005 163.04003 163.04004 163.04005 163.04003 163.04006 163.04007 163.04004 163.04006 163.04005 163.04004 163.04005 163.04006 163.04002 163.04002 163.04007 7-methylguanine 164.05772 164.05774 164.05773 164.05773 164.05774 164.05774 164.05765 164.05768 164.05777 164.05773 164.05775 164.05774 164.05772 164.05772 164.05772 164.05768 164.05768 164.05754 L-Phenylalanine 164.07163 164.07165 164.07167 164.07163 164.07163 164.07167 164.07161 164.07162 164.07169 164.07164 164.07165 164.07166 164.07162 164.07163 164.07165 164.07157 164.07148 164.07141 Phospho(enol)pyruvic acid 166.97507 n/a n/a 166.97507 n/a n/a n/a n/a n/a 166.97506 166.97507 166.97508 166.97505 166.97507 166.97508 166.97517 n/a n/a Uric Acid 167.02101 167.02102 167.02103 167.021 167.02102 167.02104 167.02103 167.02094 167.02117 167.02102 167.02103 167.02101 167.021 167.02101 167.02103 167.0209 167.02089 167.0213 trans-Aconitic acid 173.0091 173.00911 173.00912 173.00911 173.00913 173.00913 173.00912 173.00913 173.00912 173.00912 173.00913 173.00914 173.00912 173.00914 173.00913 173.00912 173.00918 173.0091 Shikimic acid 173.04549 173.04549 173.0455 173.04548 173.04548 173.0455 173.04525 173.04543 173.04539 173.04549 173.0455 173.04551 173.04548 173.04549 173.04548 173.04541 173.0454 173.0454 Arginine 173.10435 173.10436 173.10437 173.10435 173.10435 173.10439 173.10425 173.10441 173.10453 173.10435 173.10435 173.10437 173.10433 173.10434 173.10435 173.10446 173.10422 173.10461 L-citrulline 174.08835 174.08838 174.08837 174.08836 174.08837 174.0884 174.08826 174.08846 174.08853 174.08836 174.08837 174.08838 174.08834 174.08836 174.08837 174.08842 174.08827 174.08867 Hippuric acid 178.05091 178.0509 178.05092 178.05089 178.05087 178.05088 178.05079 178.05076 178.05084 178.0509 178.05089 178.05089 178.05087 178.05086 178.05082 178.05086 178.05084 178.05075 4-Hydroxyphenylpyruvic acid 179.03494 179.03499 179.03501 179.03499 179.03501 179.03503 179.03494 179.03508 179.0351 179.03497 179.03501 179.03502 179.03505 179.035 179.03503 179.03509 179.03515 179.03514 myo-Inositol 179.0561 179.05614 179.0561 179.05609 179.05608 n/a 179.05598 n/a n/a 179.05609 179.0561 179.05613 179.05609 179.05611 n/a 179.05619 179.056 179.05639 L-Tyrosine 180.0666 180.0666 n/a 180.06658 180.06661 n/a 180.06652 n/a n/a 180.0666 180.06658 180.0666 180.06659 180.06659 180.0666 180.06656 180.06651 180.06693 Homovanillic acid 181.05048 181.05055 n/a 181.05061 181.0506 n/a 181.05061 181.05058 n/a 181.05062 181.05051 181.05055 181.05066 181.05043 181.05042 181.05055 181.05041 n/a D-Sorbitol 181.07171 181.07174 181.07173 181.07171 181.07173 181.07173 181.07163 181.07166 181.07185 181.07171 181.07173 181.07172 181.07169 181.0717 181.07169 181.07158 181.07151 181.07191 4-Pyridoxic acid 182.04582 182.04581 182.04582 182.04581 182.04582 182.04582 182.0458 182.0458 182.04556 182.04581 182.04582 182.04582 182.04579 182.04581 182.04581 182.04572 182.04571 182.0457 DL-Normetanephrine n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a Kynurenic acid 188.03531 188.03533 188.03534 188.03533 188.03533 188.03534 188.03527 188.0353 188.03538 188.0353 188.03532 188.03531 188.03529 188.03529 188.03528 188.03519 188.03517 188.03526 D-Glucuronic acid 193.03536 193.03536 193.03538 193.03535 193.03536 193.03537 193.03524 193.03544 193.0355 193.03536 193.03536 193.03537 193.03534 193.03535 193.03537 193.03543 193.03527 193.03568 L-Tryptophan 203.0826 n/a n/a n/a n/a n/a 203.08253 203.08258 203.08257 203.08258 n/a n/a 203.08256 203.08255 203.08253 203.08252 203.08255 203.08265 Xanthurenic acid 204.03019 204.03019 204.03019 204.03019 204.03019 204.03019 204.03011 204.03009 204.03007 204.03018 204.03018 204.03018 204.03016 204.03017 204.03016 204.03006 204.03001 204.03 L-Kynurenine 207.07749 207.07749 207.07749 207.0775 207.07749 207.07751 207.07745 207.07751 n/a 207.0775 207.07749 207.07752 207.07747 207.07749 207.07748 207.07748 207.07741 207.07778 undecanedioic acid 215.12884 215.12885 215.12884 215.12884 215.12884 215.1288 215.12879 215.12875 215.12898 215.12882 215.12884 215.12884 215.12882 215.12882 215.12875 215.12874 215.1288 215.12888 5'-Hydroxy-L-tryptophan 219.0775 219.0775 219.0775 219.0775 219.0775 219.0775 219.07742 219.07765 219.07772 219.0775 219.07748 219.0775 219.07749 219.07749 219.07746 219.07746 219.07735 219.07785 3-Nitro-L-Tyrosine 225.05169 225.0517 225.05168 225.05169 225.05169 225.05168 225.05161 225.05163 225.0516 225.05169 225.05168 225.05167 225.05166 225.05165 225.05165 225.05158 225.05154 225.05145 2'-Deoxyuridine 227.06736 227.06736 227.06736 227.06736 227.06735 227.06735 227.06728 227.0673 227.0673 227.06735 227.06736 227.06734 227.06735 227.06735 227.06734 227.06722 227.0673 227.06726 Myristic acid n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a Melatonin 231.11401 n/a n/a 231.11385 n/a n/a n/a n/a n/a 231.11391 n/a n/a 231.11386 n/a n/a n/a n/a n/a L-Cystine n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a Cytidine 242.07828 242.07829 242.07826 242.07828 242.0783 242.07829 242.07822 242.07822 242.07837 242.07827 242.07826 242.07826 242.07826 242.07826 242.07826 242.07817 242.07809 242.07783 Uridine 243.06226 243.06224 243.06225 243.06225 243.06225 243.06224 243.06217 243.06213 243.06195 243.06225 243.06225 243.06223 243.06224 243.06223 243.06222 243.06214 243.06212 243.06202 Biotin 243.0809 243.0809 243.08088 243.0809 243.08088 243.08086 243.08079 243.08074 243.08069 243.08089 243.08088 243.08087 243.08088 243.08087 243.08085 243.08074 243.08076 243.08072 Pyridoxal 5' phosphate 246.01733 246.01732 246.01733 246.01734 246.01732 246.0173 246.01727 n/a n/a 246.01733 246.01733 246.01732 246.01733 246.01731 246.01728 246.01731 246.01715 246.01766 α-D-Glucose 1-phosphate 259.02253 259.02251 259.02243 259.0225 259.02248 259.02237 259.02227 259.02246 259.02259 259.02252 259.02248 259.02246 259.02248 259.0225 259.02245 259.02266 259.02217 259.02263 O-phospho-L-tyrosine 260.03304 260.03301 260.0329 260.03305 260.03302 260.03291 260.03284 n/a n/a 260.03304 260.03301 260.03304 260.03301 260.03299 260.0329 260.03316 260.03256 260.0331 Adenosine 266.08959 266.08958 266.08962 266.0896 266.0896 266.08961 266.08964 266.08961 266.08948 266.08958 266.08957 266.0896 266.08959 266.08958 266.0896 266.08959 266.08945 266.08962 Inosine 267.07366 267.07365 267.07364 267.0737 267.07367 267.07366 267.07367 267.07373 267.07385 267.07362 267.07362 267.0736 267.07363 267.07359 267.07356 267.07354 267.07334 267.07382 Heptadecanoic acid n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a Guanosine 282.08442 282.08442 282.08441 282.08445 282.08442 282.08444 282.08435 282.08404 282.08411 282.08443 282.08438 282.08441 282.0844 282.08436 282.08429 282.08429 282.08411 282.08461 Xanthosine 283.06845 283.06842 283.0684 283.06844 283.06842 283.0684 283.06832 283.0683 283.06824 283.0684 283.06839 283.06835 283.06838 283.06836 283.06832 283.0682 283.06811 283.06811 2'-Deoxycytidine 5'-monophosphate 306.04972 306.04965 306.04961 306.04972 306.04965 306.04946 306.04951 306.04924 n/a 306.04969 306.04967 306.04965 306.04968 306.0496 306.04951 306.04979 306.04919 306.04966 L-Glutathione Reduced 306.0766 306.07655 306.07647 306.07658 306.07657 306.07642 306.07656 306.07626 n/a 306.07657 306.07653 306.07656 306.07657 306.0765 306.07673 306.0766 306.07564 n/a Cytidine 5'-Monophosphate 322.04456 322.04453 322.04435 322.04458 322.0445 322.04424 322.04427 322.04454 322.04395 322.04456 322.04451 322.04444 322.04451 322.04442 322.0443 322.04466 322.0437 322.04449 D-Fructose 1,6-biphosphate n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a Sucrose 341.10885 341.10886 341.10877 341.10886 341.10875 n/a 341.1086 341.10858 341.10837 341.10884 341.10883 341.10886 341.10887 341.10875 341.1088 341.10882 341.10801 341.1091 Gibberellic acid 345.1343 345.13428 345.13424 345.13431 345.13428 345.13424 345.13409 345.1341 345.13398 345.13429 345.13427 345.13422 345.1343 345.13426 345.13419 345.13404 345.13397 345.13382 Adenosine 5'-Monophosphate 346.05576 346.05572 346.05559 346.05581 346.05571 346.05527 346.05543 346.05492 346.05487 346.05578 346.05574 346.05577 346.05575 346.0557 346.05565 346.05576 346.05487 346.05563 Inosine 5'-monophosphate 347.03981 347.03977 347.03949 347.0398 347.03965 347.03928 347.03953 347.03882 347.03897 347.0398 347.03975 347.0397 347.03979 347.0397 347.03944 347.03994 347.03886 347.03934 Chenodeoxycholate 391.28521 391.28524 391.2851 391.28528 391.2853 n/a 391.2853 n/a n/a 391.2852 391.28519 391.28526 391.2852 391.28516 n/a 391.28506 391.28389 n/a Glycochenodeoxycholate 448.30683 448.30677 448.30668 448.30682 448.30672 448.30657 448.3063 448.3061 448.30572 448.30679 448.30673 448.30666 448.3068 448.30673 448.30653 448.30615 448.30591 448.30606 Glycocholic acid 464.30158 464.30149 464.30146 464.30158 464.30151 464.30133 464.30113 464.3009 464.30091 464.30155 464.3015 464.30143 464.30155 464.30148 464.30138 464.30096 464.3007 464.30076 Adenosine 5'-triphosphate n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a L-Glutathione Oxidized 611.14433 611.14412 n/a 611.14417 n/a n/a n/a n/a n/a 611.14436 611.14417 n/a 611.14414 n/a n/a n/a n/a n/a MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Retention Time (min) Elemental Formula glycolic acid 2.27 C2H4O3 Pyruvate 2.38 C3H4O3 Alanine 7.62 C3H7NO2 L-Lactate 1.92 C3H6O3 Acetoacetate 1.37 C4H6O3 2-Aminoisobutyric acid 7.31 C4H9NO2 Choline 4.22 C5H14NO Malonic acid 2.99 C3H4O4 L-Serine 8.3 C3H7NO3 L-Glyceric acid 6.68 C3H6O4 Cytosine 5.33 C4H5N3O Histamine 8.29 C5H9N3 Uracil 3.9 C4H4N2O2 L-Proline (Reagents Plus) 6.9 C5H9NO2 Fumarate dibasic 2.33 C4H4O4 N-Acetylglycine 2.3 C4H7NO3 L-Valine 6.31 C5H11NO2 Succinate (dibasic) 1.81 C4H6O4 L-Threonine 7.93 C4H9NO3 D-(-)-Erythrose 2.02 C4H8O4 Nicotinamide 1.7 C6H6N2O Nicotinic acid 1.67 C6H5NO2 Taurine 6.95 C2H7NO3S Pyroglutamic acid 3.22 C5H7NO3 Citraconic acid 1.42 C5H6O4 trans-4-Hydroxy-L-proline 7.74 C5H9NO3 creatine 7.63 C4H9N3O2 L-Leucine 6.11 C6H13NO2 Glutaric acid 1.63 C5H8O4 L-Asparagine 8.37 C4H8N2O3 L-Leucic acid 1.56 C6H12O3 L-Ornithine 9.03 C5H12N2O2 L-Aspartic acid 8.68 C4H7NO4 DL-Malic acid 8.25 C4H6O5 Adenine 4.01 C5H5N5 Hypoxanthine 3.96 C5H4N4O Anthranilic acid 1.05 C7H7NO2 Acetylcholine 3.04 C7H16NO2 alpha-Ketoglutaric acid 5.74 C5H6O5 Adipic acid 1.57 C6H10O4 L-Glutamine 8.25 C5H10N2O3 L-Lysine 9 C6H14N2O2 L-Glutamic acid 8.18 C5H9NO4 L-Methionine 6.63 C5H11NO2S Xanthine 4.63 C5H4N4O2 Cystamine 8.63 C4H12N2S2 Histidine 8.99 C6H9N3O2 Orotic acid 6.38 C5H4N2O4 Allantoin 4.54 C4H6N4O3 DL-2-Aminoadipic acid 7.9 C6H11NO4 Carnitine 6.5 C7H15NO3 Phenylpyruvic acid 7.05 C9H8O3 7-methylguanine 4.61 C6H7N5O L-Phenylalanine 5.46 C9H11NO2 Phospho(enol)pyruvic acid 8.32 C3H5O6 Uric Acid 6.79 C5H4N4O3 trans-Aconitic acid 3.71 C6H6O6 Shikimic acid 3.9 C7H10O5 Arginine 8.94 C6H14N4O2 L-citrulline 8.45 C6H13N3O3 Hippuric acid 1.53 C9H9NO3 4-Hydroxyphenylpyruvic acid 1.01 C9H8O4 myo-Inositol 8.45 C6H12O6 L-Tyrosine 7.05 C9H11NO3 Homovanillic acid 1.54 C9H10O4 D-Sorbitol 6.62 C6H14O6 4-Pyridoxic acid 2.97 C8H9NO4 DL-Normetanephrine 5.46 C9H13NO3 Kynurenic acid 4.9 C10H7NO3 D-Glucuronic acid 8.55 C6H10O7 L-Tryptophan 6.22 C11H12N2O2 Xanthurenic acid 5.21 C10H7NO4 L-Kynurenine 6.24 C10H12N2O3 undecanedioic acid 1.39 C11H20O4 5'-Hydroxy-L-tryptophan 7.05 C11H12N2O3 3-Nitro-L-Tyrosine 6.18 C9H10N2O5 2'-Deoxyuridine 2.9 C9H12N2O5 Myristic acid 1.05 C14H28O2 Melatonin 1.05 C13H16N2O2 L-Cystine 9.34 C6H12N2O4S Cytidine 5.88 C9H13N3O5 Uridine 3.9 C9H12N2O6 Biotin 2.58 C10H16N2O3S Pyridoxal 5' phosphate 7.02 C8H10NO6P α-D-Glucose 1-phosphate 9.05 C6H13O9P O-phospho-L-tyrosine 9.12 C9H12NO6P Adenosine 4.14 C10H13N5O4 Inosine 5.02 C10H12N4O5 Heptadecanoic acid 1.04 C17H34O2 Guanosine 6.26 C10H13N5O5 Xanthosine 5.52 C10H12N4O6 2'-Deoxycytidine 5'-monophosphate 8.77 C9H14N3O7P L-Glutathione Reduced 8.42 C10H17N3O6S Cytidine 5'-Monophosphate 8.99 C9H14N3O8P D-Fructose 1,6-biphosphate 7.47 C6H14O12P2 Sucrose 8.11 C12H22O11 Gibberellic acid 1.7 C19H22O6 Adenosine 5'-Monophosphate 8.59 C10H14N5O7P Inosine 5'-monophosphate 8.75 C10H13N4O8P Chenodeoxycholate 1.49 C24H40O4 Glycochenodeoxycholate 2.49 C26H43NO5 Glycocholic acid 3.81 C26H43NO6 Adenosine 5'-triphosphate 9.1 C10H16N5O13P3 L-Glutathione Oxidized 9.4 C20H32N6O12S2 METABOLITES_END #END