#METABOLOMICS WORKBENCH fernandezlab_20210505_000411_mwtab.txt DATATRACK_ID:2619 STUDY_ID:ST001777 ANALYSIS_ID:AN002884 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 POSITIVE 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 3.5 kV MS:AUTOMATIC_GAIN_CONTROL 5e4, 1e5, 5e5 #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Da MS_METABOLITE_DATA_START Samples Model mixture 10 Model mixture 11 Model mixture 12 Model mixture 13 Model mixture 14 Model mixture 15 Model mixture 16 Model mixture 17 Model mixture 18 PD1074 C. elegans mixture 10 PD1074 C. elegans mixture 11 PD1074 C. elegans mixture 12 PD1074 C. elegans mixture 13 PD1074 C. elegans mixture 14 PD1074 C. elegans mixture 15 PD1074 C. elegans mixture 16 PD1074 C. elegans mixture 17 PD1074 C. elegans mixture 18 Factors Experimental Condition:AGC 5e4 - R 120K - pos Experimental Condition:AGC 5e4 - R 240K - pos Experimental Condition:AGC 5e4 - R 500K - pos Experimental Condition:AGC 1e5 - R 120K - pos Experimental Condition:AGC 1e5 - R 240K - pos Experimental Condition:AGC 1e5 - R 500K - pos Experimental Condition:AGC 5e5 - R 120K - pos Experimental Condition:AGC 5e5 - R 240K - pos Experimental Condition:AGC 5e5 - R 500K - pos Experimental Condition:AGC 5e4 - R 120K - pos Experimental Condition:AGC 5e4 - R 240K - pos Experimental Condition:AGC 5e4 - R 500K - pos Experimental Condition:AGC 1e5 - R 120K - pos Experimental Condition:AGC 1e5 - R 240K - pos Experimental Condition:AGC 1e5 - R 500K - pos Experimental Condition:AGC 5e5 - R 120K - pos Experimental Condition:AGC 5e5 - R 240K - pos Experimental Condition:AGC 5e5 - R 500K - pos glycolic 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 Pyruvate 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 Alanine 90.05498 90.05499 90.05501 90.05496 90.05498 90.05499 90.05494 90.05496 90.05497 90.05497 90.05499 90.05499 90.05495 90.05497 90.05498 90.05493 90.05494 90.05496 L-Lactate 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 Acetoacetate 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 2-Aminoisobutyric acid 104.07064 104.07065 104.07067 104.07063 104.07064 104.07065 104.07059 104.07061 104.07062 104.07057 104.0706 104.07061 104.07057 104.07058 104.07058 104.0705 104.07051 104.07053 Choline 104.10702 104.10704 104.10705 104.10701 104.10703 104.10703 104.10701 104.10701 104.10702 104.10696 104.10698 104.10699 104.10693 104.10696 104.10696 104.10687 104.10688 104.1069 Malonic 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 L-Serine 106.04989 106.04994 106.04995 106.04989 106.04991 106.04993 106.04986 106.04986 106.0499 106.04985 106.04989 106.04988 106.04983 106.04987 106.04988 106.04985 106.04986 106.04986 L-Glyceric 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 Cytosine 112.0506 112.05061 112.05063 112.05059 112.0506 112.05062 112.05058 112.05059 112.0506 112.0505 112.05053 112.05054 112.05049 112.05051 112.05051 112.05045 112.05046 112.05047 Histamine 112.08698 112.087 112.08701 112.08697 112.08698 112.087 112.08696 112.08697 112.08698 112.08688 112.08691 112.08692 112.08686 112.08689 112.0869 112.08684 112.08685 112.08686 Uracil 113.03461 113.03463 113.03463 113.0346 113.03461 113.03461 113.03457 113.03457 113.03458 113.03452 113.03455 113.03455 113.03452 113.03454 113.03454 113.03449 113.03449 113.03452 L-Proline (Reagents Plus) 116.07066 116.07068 116.07069 116.07066 116.07067 116.07068 116.07064 116.07066 116.07066 116.07058 116.0706 116.07061 116.07056 116.07058 116.07059 116.07056 116.07056 116.07058 Fumarate dibasic 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 N-Acetylglycine 118.04992 118.04995 118.04994 118.04991 118.04992 118.04993 118.04986 118.04989 n/a 118.04982 118.04983 118.04983 118.0498 118.04981 118.04983 118.04975 118.04976 118.04977 L-Valine 118.08629 118.08631 118.08632 118.08628 118.0863 118.08631 118.08626 118.08627 118.08628 118.0862 118.08622 118.08623 118.08617 118.0862 118.0862 118.08614 118.08615 118.08617 Succinate (dibasic) 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-Threonine 120.06556 120.06558 120.06559 120.06555 120.06556 120.06558 120.0655 120.06552 120.06552 120.06548 120.06549 120.0655 120.06545 120.06547 120.06548 120.06542 120.06542 120.06542 D-(-)-Erythrose 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 Nicotinamide 123.05535 123.05536 123.05537 123.05534 123.05536 123.05537 123.05541 123.05543 123.05541 123.05524 123.05526 123.05527 123.05523 123.05526 123.05526 123.05521 123.05522 123.05529 Nicotinic acid 124.03935 124.03935 124.03937 124.03932 124.03933 124.03934 124.03927 124.03927 124.0393 124.03925 124.03927 124.03928 124.03923 124.03925 124.03923 124.03919 124.03921 124.03923 Taurine 126.02199 126.022 126.02201 126.02197 126.02199 126.022 126.02194 126.02195 126.02195 126.0219 126.02191 126.02192 126.02188 126.0219 126.0219 126.02186 126.02188 126.02186 Pyroglutamic acid 130.04994 130.04995 130.04996 130.04992 130.04994 130.04994 130.04992 130.04993 130.04993 130.04982 130.04985 130.04985 130.04981 130.04983 130.04984 130.04981 130.04981 130.04983 Citraconic 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 trans-4-Hydroxy-L-proline 132.06558 132.0656 132.06561 132.06558 132.06559 132.0656 132.06558 132.0656 132.06562 132.06547 132.06549 132.06549 132.06545 132.06547 132.06548 132.06545 132.06547 132.06548 creatine 132.0768 132.07682 132.07684 132.07678 132.07679 132.07684 132.07674 132.07679 132.07683 132.07668 132.0767 132.07672 132.07665 132.07666 132.0767 132.07664 132.07658 132.07667 L-Leucine 132.10196 132.10197 132.10199 132.10195 132.10196 132.10197 132.10192 132.10193 132.10194 132.10185 132.10186 132.10187 132.10182 132.10184 132.10185 132.10179 132.10179 132.10182 Glutaric acid 133.04959 n/a n/a 133.04957 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-Asparagine 133.0608 133.06083 133.06085 133.06079 133.06081 133.06081 133.06079 133.06079 133.06079 133.06069 133.06073 133.06073 133.06068 133.0607 133.06071 133.06067 133.06068 133.06069 L-Leucic acid 133.08599 133.086 133.08602 133.08597 133.08598 133.08598 133.08595 133.08598 133.08598 133.08587 133.08587 133.08589 133.08585 133.08587 133.08589 133.08583 133.08584 133.08584 L-Ornithine 133.0972 133.09722 133.09723 133.09721 133.09721 133.09721 133.09719 133.0972 133.09719 133.09709 133.09711 133.09711 133.09707 133.0971 133.0971 133.09704 133.09705 133.09706 L-Aspartic acid 134.04484 134.04486 134.04487 134.04482 134.04483 n/a 134.04478 n/a n/a 134.04472 134.04473 134.04474 134.04469 134.04471 134.04471 134.04468 134.04468 134.04469 DL-Malic 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 Adenine 136.06185 136.06186 136.06187 136.06184 136.06185 136.06185 136.06184 136.06185 136.06185 136.06172 136.06174 136.06174 136.06171 136.06173 136.06173 136.06172 136.06173 136.06174 Hypoxanthine 137.04581 137.04582 137.04584 137.04578 137.04579 137.04579 137.04572 137.0457 137.04572 137.04571 137.04572 137.04572 137.04569 137.0457 137.04571 137.04566 137.04567 137.04566 Anthranilic acid 138.05502 138.05503 138.05504 138.05501 138.05503 138.05503 138.05502 138.05504 138.05503 138.0549 138.0549 138.0549 138.05488 138.05489 138.0549 138.05486 138.05487 138.05488 Acetylcholine 146.11759 146.1176 146.11761 146.11757 146.11758 146.11759 146.11755 146.11757 146.11758 146.11747 146.11749 146.11749 146.11745 146.11748 146.11748 146.11746 146.11747 146.11748 alpha-Ketoglutaric 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 Adipic 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 L-Glutamine 147.07643 147.07646 147.07646 147.07643 147.07643 147.07644 147.07639 147.0764 147.07642 147.07633 147.07635 147.07637 147.07631 147.07633 147.07633 147.07634 147.07634 147.07635 L-Lysine 147.11283 147.11284 147.11285 147.11281 147.11283 147.11283 147.11278 147.11279 147.1128 147.11271 147.11273 147.11274 147.1127 147.11271 147.11273 147.1127 147.1127 147.11271 L-Glutamic acid 148.06046 148.06046 148.0605 148.06044 148.06046 148.06047 148.06041 148.06041 n/a 148.06034 148.06036 148.06037 148.06033 148.06034 148.06034 148.06033 148.06033 148.06036 L-Methionine 150.05837 150.05838 150.05839 150.05835 150.05837 150.05837 150.05832 150.05834 150.05834 150.05828 150.05829 150.05829 150.05826 150.05828 150.05827 150.05825 150.05825 150.05826 Xanthine 153.04072 153.04074 153.04074 153.04071 153.04073 153.04074 153.04067 153.04069 153.0407 153.04064 153.04065 153.04066 153.04064 153.04065 153.04063 153.04062 153.04063 153.04064 Cystamine 153.05149 153.0515 153.05151 153.05147 153.05148 153.05149 153.05144 153.05146 153.05146 153.05138 153.05141 153.05142 153.05137 153.05139 153.05141 153.05136 153.05137 153.05138 Histidine 156.07678 156.07679 156.07681 156.07677 156.07678 156.07679 156.07674 156.07675 156.07677 156.07671 156.07672 156.07673 156.0767 156.07671 156.07672 156.07669 156.07669 156.0767 Orotic 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 Allantoin 159.05128 159.0513 159.05133 159.05128 159.05129 159.0513 159.05124 159.05128 n/a 159.05122 159.05122 159.05125 159.05118 159.05123 159.05122 159.0512 159.05121 159.05122 DL-2-Aminoadipic acid 162.07611 162.07611 162.07612 162.07611 162.07611 162.07611 162.0761 162.07612 162.0761 162.07602 162.07604 162.07605 162.07602 162.07603 162.07603 162.07605 162.07605 162.07605 Carnitine 162.1125 162.11251 162.11251 162.11249 162.1125 162.1125 162.11249 162.11249 162.11249 162.11241 162.11243 162.11243 162.1124 162.11241 162.11243 162.1124 162.11241 162.11244 Phenylpyruvic acid 165.05466 165.05466 165.05467 165.05465 165.05465 165.05464 165.05464 165.05464 165.05466 165.05457 165.05459 165.05457 165.05455 165.05456 165.05456 165.05455 165.05454 165.05456 7-methylguanine 166.07238 166.07239 166.07239 166.07237 166.07238 166.07238 166.07237 166.07237 166.07239 166.07229 166.0723 166.0723 166.07228 166.0723 166.0723 166.0723 166.0723 166.07231 L-Phenylalanine 166.0863 166.08629 166.0863 166.08629 166.08628 166.08628 166.08627 166.08626 166.08627 166.0862 166.08621 166.08621 166.08618 166.08619 166.08621 166.08618 166.08618 166.0862 Phospho(enol)pyruvic 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 Uric 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 trans-Aconitic 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 Shikimic 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 Arginine 175.11897 175.11898 175.11898 175.11895 175.11895 175.11897 175.11892 175.11893 175.11895 175.1189 175.1189 175.11891 175.11888 175.1189 175.11891 175.11889 175.11889 175.1189 L-citrulline 176.10299 176.10299 176.10298 176.10296 176.10296 176.10299 176.10293 176.10295 176.10295 176.1029 176.10292 176.10293 176.1029 176.10292 176.10292 176.10292 176.10292 176.10293 Hippuric acid 180.06552 180.06553 180.06555 180.06552 180.06553 180.0655 180.06553 180.06556 180.06554 180.06546 180.06547 180.06548 180.06546 180.06549 180.06549 180.06549 180.0655 180.06549 4-Hydroxyphenylpyruvic 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 myo-Inositol 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-Tyrosine 182.08119 182.0812 182.08119 182.08117 182.08118 182.08119 182.08114 182.08115 182.08116 182.08112 182.08114 182.08114 182.08112 182.08113 182.08113 182.08114 182.08113 182.08114 Homovanillic 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 D-Sorbitol 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 4-Pyridoxic acid 184.06044 184.06045 184.06045 184.06043 184.06043 184.06044 184.0604 184.06041 184.06043 184.06039 184.06041 184.06041 184.06039 184.06041 184.06041 184.06041 184.06041 184.06041 DL-Normetanephrine 184.09682 184.09685 184.09684 184.09682 184.09681 184.0968 184.09675 184.09675 184.09681 184.09677 184.09679 184.09678 184.09676 184.09677 184.09676 184.09676 184.09677 184.09676 Kynurenic acid 190.04987 190.04988 190.04989 190.04986 190.04986 190.04986 190.04984 190.04986 190.04988 190.04984 190.04985 190.04986 190.04984 190.04985 190.04985 190.04985 190.04985 190.04987 D-Glucuronic 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 L-Tryptophan 205.09719 205.09719 205.0972 205.09718 205.09717 205.09719 205.09717 205.09717 205.09716 205.09718 205.09718 205.09718 205.09717 205.09717 205.09716 205.09716 205.09716 205.09717 Xanthurenic acid 206.04481 206.04481 206.04482 206.04481 206.0448 206.0448 206.04481 206.04481 206.04482 206.0448 206.0448 206.04481 206.0448 206.0448 206.0448 206.04477 206.04477 206.04477 L-Kynurenine 209.09208 209.09208 209.09209 209.09207 209.09207 209.09207 209.09201 209.09201 209.09202 209.09207 209.09207 209.09208 209.09206 209.09206 209.09207 209.092 209.09201 209.09201 undecanedioic 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 5'-Hydroxy-L-tryptophan 221.09213 221.09213 221.09214 221.09213 221.09212 221.09211 221.0921 221.09212 221.09212 221.09208 221.09209 221.09209 221.09206 221.09208 221.09207 221.09205 221.09205 221.09206 3-Nitro-L-Tyrosine 227.06627 227.06628 227.06627 227.06626 227.06626 227.06624 227.06621 227.0662 227.06623 227.06625 227.06626 227.06624 227.06623 227.06625 227.06625 227.06617 227.06619 227.06621 2'-Deoxyuridine 229.08196 229.08198 229.08199 229.08196 229.08195 229.08199 229.082 229.08206 n/a 229.08195 229.08196 229.08194 229.08194 229.08199 229.08191 229.08195 229.08193 n/a 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 233.12853 233.12855 233.12855 233.12853 233.12854 233.12855 233.12855 233.12855 233.12857 233.12849 233.1285 233.12853 233.1285 233.12851 233.12854 233.12851 233.12852 233.12857 L-Cystine 241.03121 241.03119 241.03123 241.03121 241.0312 n/a 241.03122 241.03122 n/a n/a n/a n/a n/a n/a n/a n/a n/a n/a Cytidine 244.09293 244.09295 244.09294 244.09293 244.09292 244.09292 244.09294 244.09295 244.09296 244.09284 244.09284 244.09283 244.09281 244.09283 244.09283 244.0928 244.0928 244.09279 Uridine 245.07696 245.07698 245.07697 245.07699 245.07695 245.07698 245.07701 245.07704 245.07705 245.07684 245.07685 245.07685 245.07684 245.07687 245.07683 245.07687 245.07681 n/a Biotin 245.09556 245.09558 245.09557 245.09557 245.09557 245.09556 245.09561 245.09562 245.09563 245.09549 245.0955 245.0955 245.09548 245.09549 245.0955 245.09549 245.09549 245.09549 Pyridoxal 5' phosphate 248.03198 248.03198 248.03198 248.03197 248.03196 248.03196 248.03196 248.03197 248.03199 248.03187 248.03188 248.03188 248.03186 248.03187 248.03188 248.03185 248.03184 248.03184 α-D-Glucose 1-phosphate 261.03718 261.03719 261.03718 261.03718 261.03717 261.03717 261.03724 261.03725 261.03723 261.03708 261.03707 261.03704 261.03707 261.03706 261.03707 261.03701 261.037 261.03705 O-phospho-L-tyrosine 262.0477 262.04769 262.0477 262.04768 262.04767 262.04767 262.04773 262.04775 262.04774 262.04758 262.04757 262.04755 262.04758 262.04755 262.04755 262.04755 262.04754 262.04754 Adenosine 268.10426 268.10425 268.10423 268.10427 268.10426 268.10423 268.10429 268.10428 268.10428 268.10414 268.10411 268.1041 268.10413 268.10411 268.1041 268.10409 268.10408 268.10412 Inosine 269.08828 269.08827 269.08827 269.08829 269.08828 269.08827 269.08834 269.08833 269.08833 269.08815 269.08812 269.08814 269.08813 269.08813 269.08814 269.08809 269.08808 269.08808 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 284.09914 284.09912 284.09912 284.09916 284.09914 284.09913 284.09917 284.09914 284.09914 284.09905 284.09903 284.09902 284.09903 284.09903 284.09903 284.09892 284.09893 284.099 Xanthosine 285.08317 285.08315 285.08314 285.0832 285.08317 285.08318 285.08323 285.0832 285.08321 285.08307 285.08306 285.08303 285.08304 285.08305 285.08309 285.08298 285.08298 285.08297 2'-Deoxycytidine 5'-monophosphate 308.06438 308.06437 308.06435 308.06439 308.06437 308.06435 308.06438 308.06438 308.06437 308.06429 308.06428 308.0643 308.06429 308.0643 308.06429 308.06424 308.06424 308.06421 L-Glutathione Reduced 308.09125 308.09124 308.09123 308.09125 308.09123 308.09124 308.09123 308.09124 308.09125 308.09116 308.09119 308.09123 308.09115 308.09117 308.09117 308.09112 308.09114 308.09117 Cytidine 5'-Monophosphate 324.05928 324.05927 324.05926 324.05927 324.05927 324.05924 324.05926 324.05925 324.05928 324.05923 324.05922 324.05923 324.05923 324.05924 324.05925 324.05918 324.05919 324.05917 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 343.12368 n/a n/a n/a n/a n/a n/a n/a n/a 343.12377 343.12357 n/a 343.12361 343.12375 n/a n/a n/a n/a Gibberellic 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 Adenosine 5'-Monophosphate 348.0705 348.07049 348.07046 348.0705 348.07048 348.07045 348.07044 348.07046 348.07045 348.07067 348.07057 348.07056 348.07062 348.07056 348.07054 348.07051 348.07045 348.07038 Inosine 5'-monophosphate 349.05451 349.0545 349.0545 349.05452 349.0545 349.05448 349.05447 349.05448 349.0545 349.05465 349.05459 349.05458 349.05461 349.05457 349.0546 349.05451 349.05451 349.05456 Chenodeoxycholate 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 Glycochenodeoxycholate 450.3215 450.32153 450.32152 450.32153 450.32152 450.32153 450.3212 450.32118 450.32123 450.32163 450.32164 450.32165 450.32164 450.32169 450.32169 450.32163 450.32157 450.3216 Glycocholic acid 466.31645 466.31648 466.31641 466.3165 466.31646 466.3164 466.31597 466.31598 466.31611 466.31656 466.31659 466.31653 466.31655 466.3166 466.31652 466.31648 466.31645 466.31616 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 613.15934 613.15935 613.15935 613.15933 613.15929 613.15942 613.15858 613.1584 n/a 613.15944 613.15952 613.15965 613.1595 613.15946 n/a 613.15951 613.15982 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