#METABOLOMICS WORKBENCH fernandezlab_20210505_151314 DATATRACK_ID:2623 STUDY_ID:ST001778 ANALYSIS_ID:AN002886 PROJECT_ID:000000 VERSION 1 CREATED_ON May 8, 2021, 10:02 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. 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 II) PR:LAST_NAME Huang #STUDY ST:STUDY_TITLE Comparison of High-Resolution Fourier Transform Mass Spectrometry Platforms for ST:STUDY_TITLE Metabolite Annotation 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. 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:8M - neg RAW_FILE_NAME=MeOH_8M_neg_1.mzML SUBJECT_SAMPLE_FACTORS - Model mixture 2 Experimental Condition:8M - neg RAW_FILE_NAME=MeOH_8M_neg_2.mzML SUBJECT_SAMPLE_FACTORS - Model mixture 3 Experimental Condition:8M - pos RAW_FILE_NAME=MeOH_8M_pos_1.mzML SUBJECT_SAMPLE_FACTORS - Model mixture 4 Experimental Condition:8M - pos RAW_FILE_NAME=MeOH_8M_pos_2.mzML SUBJECT_SAMPLE_FACTORS - Model mixture 5 Experimental Condition:8M - pos RAW_FILE_NAME=MeOH_8M_pos_3.mzML SUBJECT_SAMPLE_FACTORS - PD1074 C. elegans mixture 1 Experimental Condition:8M - neg RAW_FILE_NAME=Biomatrix_8M_neg_1.mzML SUBJECT_SAMPLE_FACTORS - PD1074 C. elegans mixture 2 Experimental Condition:8M - neg RAW_FILE_NAME=Biomatrix_8M_neg_2.mzML SUBJECT_SAMPLE_FACTORS - PD1074 C. elegans mixture 3 Experimental Condition:8M - neg RAW_FILE_NAME=Biomatrix_8M_neg_3.mzML SUBJECT_SAMPLE_FACTORS - PD1074 C. elegans mixture 4 Experimental Condition:8M - pos RAW_FILE_NAME=Biomatrix_8M_pos_1.mzML SUBJECT_SAMPLE_FACTORS - PD1074 C. elegans mixture 5 Experimental Condition:8M - pos RAW_FILE_NAME=Biomatrix_8M_pos_2.mzML SUBJECT_SAMPLE_FACTORS - PD1074 C. elegans mixture 6 Experimental Condition:8M - pos RAW_FILE_NAME=Biomatrix_8M_pos_3.mzML #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_TYPE None (Direct infusion) CH:INSTRUMENT_NAME none CH:COLUMN_NAME none #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Bruker Solarix FT-ICR-MS MS:INSTRUMENT_TYPE FT-ICR MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS Samples were analyzed in positive and negative ion modes with direct infusion MS:MS_COMMENTS FT-ICR MS. A spectral size of 8 M was used for data acquisition with an ion MS:MS_COMMENTS accumulation time of 0.025 s and a scan range of 73.71 – 1000.00 m/z. Average MS:MS_COMMENTS scans were set to 200. Raw data files were converted into mzML format using MS:MS_COMMENTS MSConvert. Spectral features were extracted using MZmine 2.51. MS:CAPILLARY_VOLTAGE 5.0kV MS:DRY_GAS_FLOW 4.0 L/min MS:DRY_GAS_TEMP 180 °C MS:NEBULIZER 0.5bar #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Da MS_METABOLITE_DATA_START Samples Model mixture 3 Model mixture 4 Model mixture 5 PD1074 C. elegans mixture 4 PD1074 C. elegans mixture 5 PD1074 C. elegans mixture 6 Factors Experimental Condition:8M - pos Experimental Condition:8M - pos Experimental Condition:8M - pos Experimental Condition:8M - pos Experimental Condition:8M - pos Experimental Condition:8M - pos glycolic acid 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 Alanine 90.05495 90.05495 90.05494 90.05495 90.05495 90.05496 L-Lactate n/a n/a n/a n/a n/a n/a Acetoacetate n/a n/a n/a 103.03899 103.03899 103.039 2-Aminoisobutyric acid 104.07062 104.07062 104.07062 104.07063 104.07063 104.07063 Choline 104.10699 104.10699 104.10699 104.107 104.107 104.107 Malonic acid n/a n/a n/a n/a n/a n/a L-Serine 106.04987 106.04989 106.04987 106.04988 106.04989 106.0499 L-Glyceric acid n/a n/a n/a n/a n/a n/a Cytosine 112.05055 112.05056 112.05055 112.05058 112.05058 112.05058 Histamine 112.08694 112.08694 112.08694 112.08695 112.08695 112.08696 Uracil 113.03457 113.03458 113.03457 113.03458 113.03458 113.03458 L-Proline (Reagents Plus) 116.07062 116.07063 116.07062 116.07063 116.07063 116.07063 Fumarate dibasic n/a n/a n/a n/a n/a n/a N-Acetylglycine n/a n/a n/a n/a n/a n/a L-Valine 118.08627 118.08627 118.08627 118.08628 118.08628 118.08628 Succinate (dibasic) n/a n/a n/a n/a n/a n/a L-Threonine 120.06553 120.06554 120.06553 120.06554 120.06554 120.06555 D-(-)-Erythrose n/a n/a n/a n/a n/a n/a Nicotinamide 123.05531 123.05531 123.0553 123.05532 123.05532 123.05533 Nicotinic acid 124.03931 124.03932 124.03931 124.03934 124.03934 124.03934 Taurine 126.02193 126.02194 126.02193 n/a 126.02192 126.02196 Pyroglutamic acid 130.04987 130.04987 130.04987 130.04988 130.0499 130.04988 Citraconic acid n/a n/a n/a n/a n/a n/a trans-4-Hydroxy-L-proline 132.06554 132.06555 132.06554 132.06555 132.06555 132.06557 creatine 132.07677 132.07677 132.07675 132.07678 132.07678 132.07678 L-Leucine 132.10191 132.10191 132.10191 132.10193 132.10193 132.10193 Glutaric acid 133.04956 133.04953 133.04948 133.04951 133.04956 133.04953 L-Asparagine 133.06078 133.06078 133.06078 133.06079 133.06081 133.06081 L-Leucic acid 133.08594 133.08595 133.08594 133.08594 133.08595 133.08595 L-Ornithine 133.09715 133.09715 133.09714 133.09717 133.09717 133.09717 L-Aspartic acid n/a n/a n/a n/a n/a n/a DL-Malic acid n/a n/a n/a n/a n/a n/a Adenine 136.06175 136.06175 136.06175 136.06177 136.06177 136.06177 Hypoxanthine 137.04578 137.04579 137.04578 137.04581 137.04581 137.04581 Anthranilic acid 138.05496 138.05498 138.05498 138.05498 138.05498 138.05498 Acetylcholine 146.11757 146.11757 146.11757 146.11757 146.11758 146.11758 alpha-Ketoglutaric acid n/a n/a n/a n/a n/a n/a Adipic acid 147.0652 147.06522 147.06517 147.06522 147.06519 147.06522 L-Glutamine 147.07643 147.07645 147.07643 147.07646 147.07646 147.07646 L-Lysine 147.11281 147.11281 147.11281 147.11282 147.11282 147.11282 L-Glutamic acid n/a n/a n/a 148.06044 148.06046 148.06044 L-Methionine 150.05833 150.05833 150.05833 150.05835 150.05835 150.05836 Xanthine 153.04073 153.04073 153.04071 153.04073 153.04074 153.04074 Cystamine 153.05145 153.05145 153.05145 153.05147 153.05147 153.05147 Histidine 156.07674 156.07675 156.07674 156.07677 156.07677 156.07677 Orotic acid n/a n/a n/a n/a n/a n/a Allantoin 159.05127 159.05127 159.05124 159.05128 159.0513 159.05127 DL-2-Aminoadipic acid 162.07611 162.07613 162.07611 162.07611 162.07613 162.07613 Carnitine 162.11246 162.11247 162.11246 162.11246 162.11246 162.11246 Phenylpyruvic acid 165.05464 165.05464 165.05463 165.05464 165.05466 165.05466 7-methylguanine 166.07233 166.07233 166.07233 166.07234 166.07234 166.07234 L-Phenylalanine 166.08624 166.08624 166.08624 166.08626 166.08626 166.08626 Phospho(enol)pyruvic acid n/a n/a n/a n/a n/a n/a Uric Acid n/a n/a n/a 169.03563 169.03561 169.03566 trans-Aconitic acid n/a n/a n/a n/a n/a n/a Shikimic acid 175.06015 n/a n/a 175.06013 175.06017 175.06013 Arginine 175.11896 175.11896 175.11896 175.11897 175.11897 175.11899 L-citrulline 176.10297 176.10298 176.10297 176.10298 176.103 176.103 Hippuric acid 180.06554 180.06557 180.06554 180.06557 180.06558 180.06558 4-Hydroxyphenylpyruvic acid 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 L-Tyrosine 182.08116 182.08116 182.08116 182.08118 182.08118 182.08119 Homovanillic acid 183.06522 183.06519 183.0652 183.0652 183.0652 183.06522 D-Sorbitol 183.08632 183.08635 183.08635 183.08636 183.08635 183.08635 4-Pyridoxic acid 184.06042 184.06044 184.06042 184.06044 184.06044 184.06044 DL-Normetanephrine 184.09682 184.09683 184.09682 184.09685 184.09685 184.09685 Kynurenic acid 190.0499 190.0499 190.0499 190.04993 190.04991 190.04993 D-Glucuronic acid n/a n/a n/a n/a n/a n/a L-Tryptophan 205.09715 205.09717 205.09715 205.09718 205.09718 205.09718 Xanthurenic acid 206.04477 206.04477 206.04475 206.04478 206.0448 206.04478 L-Kynurenine 209.09206 209.09206 209.09206 209.09209 209.09209 209.09209 undecanedioic acid 217.14345 217.14345 217.14345 217.14348 217.14348 217.14348 5'-Hydroxy-L-tryptophan 221.09207 221.09207 221.09206 221.0921 221.0921 221.0921 3-Nitro-L-Tyrosine 227.06621 227.06622 227.06622 227.06624 227.06624 227.06624 2'-Deoxyuridine 229.08186 229.08188 229.08188 229.08188 229.08189 229.08189 Myristic acid 229.21619 229.2162 229.21619 229.2162 229.2162 229.2162 Melatonin 233.12843 233.12843 233.12842 233.12846 233.12846 233.12846 L-Cystine 241.03111 241.03113 241.03114 n/a n/a n/a Cytidine 244.0928 244.0928 244.09279 244.09283 244.09283 244.09283 Uridine 245.0768 245.07681 245.07681 245.07683 245.07681 245.07684 Biotin 245.09546 245.09547 245.09546 245.09549 245.0955 245.0955 Pyridoxal 5' phosphate 248.03183 248.03181 248.03183 248.03185 248.03186 248.03186 α-D-Glucose 1-phosphate n/a n/a n/a n/a n/a n/a O-phospho-L-tyrosine 262.04755 262.04755 262.04755 262.04761 262.04761 262.04761 Adenosine 268.10406 268.10406 268.10406 268.1041 268.1041 268.1041 Inosine 269.08807 269.08807 269.08807 269.0881 269.0881 269.0881 Heptadecanoic acid 271.26318 271.26321 271.26318 271.26321 271.26321 271.26324 Guanosine 284.09897 284.099 284.09897 284.099 284.099 284.099 Xanthosine 285.08298 285.08298 285.08298 285.08301 285.08301 285.08304 2'-Deoxycytidine 5'-monophosphate 308.06427 308.06427 308.06424 308.0643 308.0643 308.0643 L-Glutathione Reduced 308.09116 308.09119 308.09116 308.09119 308.09119 308.09119 Cytidine 5'-Monophosphate 324.0592 324.0592 324.0592 324.05923 324.05923 324.05923 D-Fructose 1,6-biphosphate n/a n/a n/a n/a n/a n/a Sucrose n/a n/a n/a n/a n/a n/a Gibberellic acid n/a n/a n/a n/a n/a n/a Adenosine 5'-Monophosphate 348.0704 348.0704 348.0704 348.07043 348.07047 348.07047 Inosine 5'-monophosphate n/a n/a n/a n/a n/a n/a Chenodeoxycholate 393.30002 393.29993 393.30002 393.29999 393.30005 393.29993 Glycochenodeoxycholate 450.32147 450.3215 450.32147 450.32153 450.32153 450.3215 Glycocholic acid 466.31635 466.31641 466.31638 466.31641 466.31641 466.31641 Adenosine 5'-triphosphate n/a n/a n/a n/a n/a n/a L-Glutathione Oxidized n/a n/a n/a n/a n/a n/a MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Elemental Formula Theoretical Mass (Pos) glycolic acid C2H4O3 77.02332 Pyruvate C3H4O3 89.02332 Alanine C3H7NO2 90.05495 L-Lactate C3H6O3 91.039 Acetoacetate C4H6O3 103.03897 2-Aminoisobutyric acid C4H9NO2 104.07061 Choline C5H14NO 104.10699 Malonic acid C3H4O4 105.01824 L-Serine C3H7NO3 106.04987 L-Glyceric acid C3H6O4 107.03389 Cytosine C4H5N3O 112.05054 Histamine C5H9N3 112.08692 Uracil C4H4N2O2 113.03455 L-Proline (Reagents Plus) C5H9NO2 116.07061 Fumarate dibasic C4H4O4 117.01824 N-Acetylglycine C4H7NO3 118.04987 L-Valine C5H11NO2 118.08626 Succinate (dibasic) C4H6O4 119.03389 L-Threonine C4H9NO3 120.06552 D-(-)-Erythrose C4H8O4 121.04954 Nicotinamide C6H6N2O 123.05529 Nicotinic acid C6H5NO2 124.0393 Taurine C2H7NO3S 126.02194 Pyroglutamic acid C5H7NO3 130.04987 Citraconic acid C5H6O4 131.03389 trans-4-Hydroxy-L-proline C5H9NO3 132.06552 creatine C4H9N3O2 132.07675 L-Leucine C6H13NO2 132.10191 Glutaric acid C5H8O4 133.04954 L-Asparagine C4H8N2O3 133.06077 L-Leucic acid C6H12O3 133.08592 L-Ornithine C5H12N2O2 133.09715 L-Aspartic acid C4H7NO4 134.04478 DL-Malic acid C4H6O5 135.0288 Adenine C5H5N5 136.06177 Hypoxanthine C5H4N4O 137.04579 Anthranilic acid C7H7NO2 138.05495 Acetylcholine C7H16NO2 146.11756 alpha-Ketoglutaric acid C5H6O5 147.0288 Adipic acid C6H10O4 147.06519 L-Glutamine C5H10N2O3 147.0764 L-Lysine C6H14N2O2 147.1128 L-Glutamic acid C5H9NO4 148.06043 L-Methionine C5H11NO2S 150.05833 Xanthine C5H4N4O2 153.0407 Cystamine C4H12N2S2 153.05147 Histidine C6H9N3O2 156.07675 Orotic acid C5H4N2O4 157.02438 Allantoin C4H6N4O3 159.05127 DL-2-Aminoadipic acid C6H11NO4 162.07608 Carnitine C7H15NO3 162.11247 Phenylpyruvic acid C9H8O3 165.05462 7-methylguanine C6H7N5O 166.07234 L-Phenylalanine C9H11NO2 166.08626 Phospho(enol)pyruvic acid C3H5O6 168.98965 Uric Acid C5H4N4O3 169.03562 trans-Aconitic acid C6H6O6 175.02371 Shikimic acid C7H10O5 175.0601 Arginine C6H14N4O2 175.11895 L-citrulline C6H13N3O3 176.10297 Hippuric acid C9H9NO3 180.06552 4-Hydroxyphenylpyruvic acid C9H8O4 181.04954 myo-Inositol C6H12O6 181.07066 L-Tyrosine C9H11NO3 182.08117 Homovanillic acid C9H10O4 183.06519 D-Sorbitol C6H14O6 183.08631 4-Pyridoxic acid C8H9NO4 184.06043 DL-Normetanephrine C9H13NO3 184.09682 Kynurenic acid C10H7NO3 190.04987 D-Glucuronic acid C6H10O7 195.04993 L-Tryptophan C11H12N2O2 205.09715 Xanthurenic acid C10H7NO4 206.04478 L-Kynurenine C10H12N2O3 209.09207 undecanedioic acid C11H20O4 217.14344 5'-Hydroxy-L-tryptophan C11H12N2O3 221.09207 3-Nitro-L-Tyrosine C9H10N2O5 227.06625 2'-Deoxyuridine C9H12N2O5 229.0819 Myristic acid C14H28O2 229.21621 Melatonin C13H16N2O2 233.12845 L-Cystine C6H12N2O4S 241.03112 Cytidine C9H13N3O5 244.0928 Uridine C9H12N2O6 245.07681 Biotin C10H16N2O3S 245.09544 Pyridoxal 5' phosphate C8H10NO6P 248.03185 α-D-Glucose 1-phosphate C6H13O9P 261.03699 O-phospho-L-tyrosine C9H12NO6P 262.0475 Adenosine C10H13N5O4 268.10403 Inosine C10H12N4O5 269.08805 Heptadecanoic acid C17H34O2 271.26316 Guanosine C10H13N5O5 284.09895 Xanthosine C10H12N4O6 285.08296 2'-Deoxycytidine 5'-monophosphate C9H14N3O7P 308.06421 L-Glutathione Reduced C10H17N3O6S 308.09108 Cytidine 5'-Monophosphate C9H14N3O8P 324.05913 D-Fructose 1,6-biphosphate C6H14O12P2 341.00333 Sucrose C12H22O11 343.12349 Gibberellic acid C19H22O6 347.14891 Adenosine 5'-Monophosphate C10H14N5O7P 348.07036 Inosine 5'-monophosphate C10H13N4O8P 349.05438 Chenodeoxycholate C24H40O4 393.29994 Glycochenodeoxycholate C26H43NO5 450.3214 Glycocholic acid C26H43NO6 466.31631 Adenosine 5'-triphosphate C10H16N5O13P3 508.00302 L-Glutathione Oxidized C20H32N6O12S2 613.15924 METABOLITES_END #END