#METABOLOMICS WORKBENCH biswapriya_20180516_102729 DATATRACK_ID:1404 STUDY_ID:ST000972 ANALYSIS_ID:AN001592 PROJECT_ID:PR000665 VERSION 1 CREATED_ON May 22, 2018, 1:17 pm #PROJECT PR:PROJECT_TITLE High Resolution GC-MS Metabolomics of Non-Human Primate Serum PR:PROJECT_TYPE Method Development in Metabolomics PR:PROJECT_SUMMARY Rationale: Metabolomics analyses using gas chromatography mass spectrometry PR:PROJECT_SUMMARY (GC-MS) - based metabolomics are heavily impeded by the lack of high-resolution PR:PROJECT_SUMMARY mass spectrometers and limited spectral libraries to complement the excellent PR:PROJECT_SUMMARY chromatography that GC platforms offer, a challenge that is being addressed with PR:PROJECT_SUMMARY the implementation of high resolution (HR) platforms such as GC-Orbitrap-MS. PR:PROJECT_SUMMARY Methods: We used serum samples from a non-human primate (NHP), a baboon (Papio PR:PROJECT_SUMMARY hamadryas), with suitable quality controls to quantify the chemical space using PR:PROJECT_SUMMARY an advanced HR MS platform for confident metabolite identification and robust PR:PROJECT_SUMMARY quantification to assess the suitability of the platform for routine clinical PR:PROJECT_SUMMARY metabolomics research. In a comparative approach, we also analyzed the same PR:PROJECT_SUMMARY serum samples using a two-dimensional gas chromatography time-of-flight PR:PROJECT_SUMMARY mass-spectrometer (2D GC-ToF-MS) for metabolite identification and PR:PROJECT_SUMMARY quantification following established standard protocols. Results: Overall, the PR:PROJECT_SUMMARY 2D GC-ToF-MS and GC-Orbitrap-MS analyses enabled identification and PR:PROJECT_SUMMARY quantification of 555 total metabolites from the NHP serum with a spectral PR:PROJECT_SUMMARY similarity score Rsim ≥ 900 and S/R ratio of > 25. A common set of 30 PR:PROJECT_SUMMARY metabolites with HMDB and KEGG IDs were quantified in the serum samples by both PR:PROJECT_SUMMARY platforms where the 2D GC-ToF-MS enabled quantification of a total 384 PR:PROJECT_SUMMARY metabolites (118 HMDB IDs) and the GC-Orbitrap-MS analysis quantification of a PR:PROJECT_SUMMARY total 200 metabolites (47 HMDB IDs). Conclusions: Our study provides insights PR:PROJECT_SUMMARY into the benefits and limitations of the use of a higher mass accuracy PR:PROJECT_SUMMARY instrument for untargeted GC-MS-based metabolomics with multi-dimensional PR:PROJECT_SUMMARY chromatography in future studies addressing clinical conditions or exposome PR:PROJECT_SUMMARY studies. PR:INSTITUTE Wake Forest School of Medicine PR:DEPARTMENT Center for Precision Medicine PR:LABORATORY Michael Olivier Laboratory PR:LAST_NAME Misra PR:FIRST_NAME Biswapriya PR:ADDRESS NRC Building, Room G#43, Medical Center Boulevard, Winston Salem, NC, USA PR:EMAIL bmisra@wakehealth.edu PR:PHONE 3522156040 PR:FUNDING_SOURCE NA PR:PROJECT_COMMENTS NA PR:PUBLICATIONS In process PR:CONTRIBUTORS Biswapriya B. Misra, Ekong Bassey, Andrew C. Bishop, David T. Kusel, Laura A. PR:CONTRIBUTORS Cox, Michael Olivier #STUDY ST:STUDY_TITLE High Resolution GC-MS Metabolomics of Non-Human Primate Serum ST:STUDY_TYPE Non-human Primate Serum ST:STUDY_SUMMARY Rationale: Metabolomics analyses using gas chromatography mass spectrometry ST:STUDY_SUMMARY (GC-MS) - based metabolomics are heavily impeded by the lack of high-resolution ST:STUDY_SUMMARY mass spectrometers and limited spectral libraries to complement the excellent ST:STUDY_SUMMARY chromatography that GC platforms offer, a challenge that is being addressed with ST:STUDY_SUMMARY the implementation of high resolution (HR) platforms such as GC-Orbitrap-MS. ST:STUDY_SUMMARY Methods: We used serum samples from a non-human primate (NHP), a baboon (Papio ST:STUDY_SUMMARY hamadryas), with suitable quality controls to quantify the chemical space using ST:STUDY_SUMMARY an advanced HR MS platform for confident metabolite identification and robust ST:STUDY_SUMMARY quantification to assess the suitability of the platform for routine clinical ST:STUDY_SUMMARY metabolomics research. In a comparative approach, we also analyzed the same ST:STUDY_SUMMARY serum samples using a two-dimensional gas chromatography time-of-flight ST:STUDY_SUMMARY mass-spectrometer (2D GC-ToF-MS) for metabolite identification and ST:STUDY_SUMMARY quantification following established standard protocols. Results: Overall, the ST:STUDY_SUMMARY 2D GC-ToF-MS and GC-Orbitrap-MS analyses enabled identification and ST:STUDY_SUMMARY quantification of 555 total metabolites from the NHP serum with a spectral ST:STUDY_SUMMARY similarity score Rsim ≥ 900 and S/R ratio of > 25. A common set of 30 ST:STUDY_SUMMARY metabolites with HMDB and KEGG IDs were quantified in the serum samples by both ST:STUDY_SUMMARY platforms where the 2D GC-ToF-MS enabled quantification of a total 384 ST:STUDY_SUMMARY metabolites (118 HMDB IDs) and the GC-Orbitrap-MS analysis quantification of a ST:STUDY_SUMMARY total 200 metabolites (47 HMDB IDs). Conclusions: Our study provides insights ST:STUDY_SUMMARY into the benefits and limitations of the use of a higher mass accuracy ST:STUDY_SUMMARY instrument for untargeted GC-MS-based metabolomics with multi-dimensional ST:STUDY_SUMMARY chromatography in future studies addressing clinical conditions or exposome ST:STUDY_SUMMARY studies. ST:INSTITUTE Wake Forest School of Medicine ST:DEPARTMENT Center for Precision Medicine ST:LABORATORY Michael Olivier Laboratory ST:LAST_NAME Misra ST:FIRST_NAME Biswapriya ST:ADDRESS NRC Building, Medical Center Boulevard ST:EMAIL bmisra@wakehealth.edu ST:PHONE 3522156040 ST:TOTAL_SUBJECTS 1 ST:NUM_MALES 1 ST:STUDY_COMMENTS NA ST:PUBLICATIONS In process #SUBJECT SU:SUBJECT_TYPE Other SU:SUBJECT_SPECIES Papio hamadryas SU:TAXONOMY_ID 9557 SU:AGE_OR_AGE_RANGE 18 SU:GENDER Male #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - S1_1 Platform Type:2D GC-ToF-MS SUBJECT_SAMPLE_FACTORS - S1_2 Platform Type:2D GC-ToF-MS SUBJECT_SAMPLE_FACTORS - S1_3 Platform Type:2D GC-ToF-MS SUBJECT_SAMPLE_FACTORS - S2_1 Platform Type:2D GC-ToF-MS SUBJECT_SAMPLE_FACTORS - S2_2 Platform Type:2D GC-ToF-MS SUBJECT_SAMPLE_FACTORS - S2_3 Platform Type:2D GC-ToF-MS SUBJECT_SAMPLE_FACTORS - S3_1 Platform Type:2D GC-ToF-MS SUBJECT_SAMPLE_FACTORS - S3_2 Platform Type:2D GC-ToF-MS SUBJECT_SAMPLE_FACTORS - S3_3 Platform Type:2D GC-ToF-MS SUBJECT_SAMPLE_FACTORS - S6_1 Platform Type:GC-Orbitrap-MS SUBJECT_SAMPLE_FACTORS - S6_2 Platform Type:GC-Orbitrap-MS SUBJECT_SAMPLE_FACTORS - S6_3 Platform Type:GC-Orbitrap-MS SUBJECT_SAMPLE_FACTORS - S7_1 Platform Type:GC-Orbitrap-MS SUBJECT_SAMPLE_FACTORS - S7_2 Platform Type:GC-Orbitrap-MS SUBJECT_SAMPLE_FACTORS - S7_3 Platform Type:GC-Orbitrap-MS SUBJECT_SAMPLE_FACTORS - S8_1 Platform Type:GC-Orbitrap-MS SUBJECT_SAMPLE_FACTORS - S8_2 Platform Type:GC-Orbitrap-MS SUBJECT_SAMPLE_FACTORS - S8_3 Platform Type:GC-Orbitrap-MS #COLLECTION CO:COLLECTION_SUMMARY All procedures involving animals were reviewed and approved by the Texas CO:COLLECTION_SUMMARY Biomedical Research Institute’s Institutional Animal Care and Use Committee CO:COLLECTION_SUMMARY and conducted in AAALAC approved facilities. For this study, we utilized a CO:COLLECTION_SUMMARY healthy adult male olive baboon (Papio hamadryas) maintained as part of the CO:COLLECTION_SUMMARY baboon colony at the Southwest National Primate Research Center, located on the CO:COLLECTION_SUMMARY campus of the Texas Biomedical Research Institute, San Antonio, Texas. The male CO:COLLECTION_SUMMARY baboon used in this study was 18 yrs. old. The baboon had been raised and CO:COLLECTION_SUMMARY maintained on a standard monkey chow diet (high complex carbohydrates; low fat) CO:COLLECTION_SUMMARY prior to the fasting blood collection. All procedures involving animals were CO:COLLECTION_SUMMARY reviewed and approved by the Texas Biomedical Research Institute’s CO:COLLECTION_SUMMARY Institutional Animal Care and Use Committee (IACUC). Freshly collected serum CO:COLLECTION_SUMMARY samples were stored in aliquots at -80 C until analysis. CO:SAMPLE_TYPE Blood (serum) CO:COLLECTION_LOCATION Southwest National Primate Research Center, San Antonio, Texas, USA CO:COLLECTION_FREQUENCY 1 CO:COLLECTION_DURATION NA CO:VOLUMEORAMOUNT_COLLECTED 40 mL CO:STORAGE_CONDITIONS -80℃ CO:COLLECTION_TUBE_TEMP 4 C CO:ADDITIVES None #TREATMENT TR:TREATMENT_SUMMARY None #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Aliquots of serum (30 µL) samples were subjected to sequential solvent SP:SAMPLEPREP_SUMMARY extraction once each with 1 mL of acetonitrile: isopropanol: water (3:3:2) and SP:SAMPLEPREP_SUMMARY 500 µL of acetonitrile: water (1:1) mixtures at 4 C.22 Adonitol and SP:SAMPLEPREP_SUMMARY d4-succinic acid (both 5 µL from 10 mg/ml stock) were added to each aliquots as SP:SAMPLEPREP_SUMMARY two internal standards prior to the extraction. The pooled extracts (~ 1500 µL) SP:SAMPLEPREP_SUMMARY from the two steps were dried under vacuum at 4 C prior to chemical SP:SAMPLEPREP_SUMMARY derivatization. Dummy extractions performed on blank tubes served as extraction SP:SAMPLEPREP_SUMMARY blanks to account for background (extraction) noise and other sources of SP:SAMPLEPREP_SUMMARY contamination. Six (S1, S2, S3, S6, S7, S8) samples were then sequentially SP:SAMPLEPREP_SUMMARY derivatized with methoxyamine hydrochloride (MeOX) and 1% TMCS in SP:SAMPLEPREP_SUMMARY N-methyl-N-trimethylsilyl-trifluoroacetamide (MSTFA) as described SP:SAMPLEPREP_SUMMARY elsewhere.23,24 Steps involved addition of 10 μL of MeOX (20 mg mL-1) in SP:SAMPLEPREP_SUMMARY pyridine incubated under shaking at 55 °C for 60 min followed by SP:SAMPLEPREP_SUMMARY trimethylsilylation at 60 °C for 60 min after adding 90 μL MSTFA. SP:PROCESSING_METHOD Fiehn et al., 2008 SP:PROCESSING_STORAGE_CONDITIONS On ice SP:EXTRACTION_METHOD Fiehn et al., 2008 SP:EXTRACT_ENRICHMENT None SP:EXTRACT_CLEANUP None SP:EXTRACT_STORAGE -80℃ SP:SAMPLE_DERIVATIZATION Methoxyamination + silylation (MSTFA) SP:SAMPLE_SPIKING Adonitol, d4-succinic acid #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY 1µL of derivatized sample into a split/splitless (SSL) injector at 250 °C CH:CHROMATOGRAPHY_SUMMARY using a splitless injection on a Thermo Scientific™ TRACE™ 1310 GC. Helium CH:CHROMATOGRAPHY_SUMMARY carrier gas at a flow rate of 1 mL/min was used for separation on a Thermo CH:CHROMATOGRAPHY_SUMMARY Scientific™ TraceGOLD™ TG-5SILMS 30 m length × 0.25 mm i.d. × 0.25 µm CH:CHROMATOGRAPHY_SUMMARY film thickness column. The initial oven temperature was held at 70 °C for 4 CH:CHROMATOGRAPHY_SUMMARY min, followed by an initial gradient of 20 °C/min ramp rate. The final CH:CHROMATOGRAPHY_SUMMARY temperature was 320 °C and held for 8 min. CH:CHROMATOGRAPHY_TYPE GC CH:INSTRUMENT_NAME Leco Pegasus 4D GC CH:COLUMN_NAME Restek Rtx-5Sil MS (30 x 0.25mm, 0.25um) CH:FLOW_RATE 1 ml/min CH:INJECTION_TEMPERATURE 250 CH:INTERNAL_STANDARD Adonitol, d4-succinic acid CH:SAMPLE_INJECTION 1 uL CH:RUNNING_BUFFER Helium CH:TRANSFERLINE_TEMPERATURE 250 CH:RANDOMIZATION_ORDER Yes CH:CHROMATOGRAPHY_COMMENTS 2D GC #ANALYSIS AN:ANALYSIS_TYPE MS AN:LABORATORY_NAME Michael Olivier Laboratory AN:OPERATOR_NAME Biswapriya Misra AN:DETECTOR_TYPE ToF AN:SOFTWARE_VERSION ChromaToF 4.71.0.0 AN:DATA_FORMAT .cdf #MS MS:MS_COMMENTS - MS:INSTRUMENT_NAME Leco Pegasus 4D GCxGC TOF MS:INSTRUMENT_TYPE GC x GC-TOF MS:MS_TYPE EI MS:ION_MODE POSITIVE MS:FRAGMENT_VOLTAGE -70 eV MS:FRAGMENTATION_METHOD EI MS:HELIUM_FLOW 1 ml/min MS:ION_SOURCE_TEMPERATURE 250 MS:MASS_ACCURACY Unit resolution MS:SCAN_RANGE_MOVERZ 40-600 MS:SCANNING 200 scans/s MS:MS_RESULTS_FILE ST000972_AN001592_Results.txt UNITS:Arbitrary units #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Arbitrary units MS_METABOLITE_DATA_START Samples S1_1 S1_2 S1_3 S2_1 S2_2 S2_3 S3_1 S3_2 S3_3 Factors Platform Type:2D GC-ToF-MS Platform Type:2D GC-ToF-MS Platform Type:2D GC-ToF-MS Platform Type:2D GC-ToF-MS Platform Type:2D GC-ToF-MS Platform Type:2D GC-ToF-MS Platform Type:2D GC-ToF-MS Platform Type:2D GC-ToF-MS Platform Type:2D GC-ToF-MS Butyric acid 614169 661804 724580 566896 132738 684411 1151271 1154392 1187018 Acetic acid 2889622 11057002 1039731 952564 1071585 972112 28711 37148 Cholesterol 1011009 1710691 1676832 1636837 1613140 1625919 1985389 1103624 2737914 Pipecolic acid 20788 19236 90375 60944 37140 34371 30420 26074 Ethanol 79095 93402 11913 34949 2733.8 16660 97325 18141 13212 D-Glucose 869336 63693 348350 3716354 6667283 6701342 1090528 Glycerol 1605184 6906127 8548479 2037477 7185064 2499252 2318.2 11048885 D-Galactose 195215 6056.8 19058 13993 3871.7 6809.2 7650 L-Tyrosine 51274 48884 53998 33552 38209 51412 87372 L-Phenylalanine 24266 35062 51037 50044 56769 52290 68811 51410 51549 L-Proline 720238 653250 697368 2167181 1672147 12477 2192473 1434060 1290554 L-Threonine 2591886 2849667 2901038 3900295 3704968 3629549 4590806 4933638 4267054 D-Mannose 1817673 5885421 7825.7 3157631 2851067 3990551 3635727 1150688 9571425 L-Aspartic acid 2148 2524.6 14394 58080 60378 50592 65512 58191 55118 Phenylacetic acid 87392 98459 18592 11925 26415 37781 40404 26915 myo-Inositol 3702529 3886817 3738657 3753812 3944842 3710460 4938069 4530395 4315292 Palmitic acid 1004605 3905809 5340097 3197963 3062853 3586072 2724810 2716872 3172045 Propionic acid 2810125 738055 11377821 911910 3957301 10976746 1050171 6867255 34136433 Pyridoxine 2443423 276897 1218567 1231021 1240947 3630736 3448188 3193249 Urea 11198881 36344697 10742612 21312242 20506071 43119005 20495674 49672100 16159624 3-Hexenedioic acid 4997.8 24622 24221 5614.8 11884 N-Alpha-acetyllysine 21052 69718 4773.1 178011 25203 Adipic acid 2205 1170.5 1960.2 3758.4 1808.8 2136.2 1827.8 2026.2 Capric acid 30108 25942 13320 17865 15950 15723 17730 30045 Caproic acid 663198 1367826 1409116 17978 10242 5537.8 45303 1336927 Elaidic acid 1620989 683176 691106 607319 621445 584884 751069 691945 727997 Citraconic acid 38661 28488 52068 57527 46831 46259 53652 64984 Dodecanoic acid 146405 59967 51053 33583 39905 40393 60784 58852 68977 Glutaric acid 4439.8 35636 4457 2328.9 2813.4 4965.4 Heptanoic acid 31773 29248 29364 20266 18132 22228 34910 34371 33955 Isocaproic acid 16409 1242.9 1456.1 12985 53129 9822.7 5137.3 Malonic acid 28651 19703 37870 2362.8 10474 27215 2741228 5325.9 29765 Pseudouridine 5980.6 6440.4 7914.2 11974 10370 9930.5 Azelaic acid 10262 13555 13513 66444 7730.1 8359.7 14438 16345 17123 Pyrrolidonecarboxylic acid 290840 137238 142493 139866 145789 199990 184818 183171 Myristic acid 3858.2 5775.3 1760.1 1533.9 3161.3 4427 7716.2 3628.5 Pentadecanoic acid 79196 48143 48260 34097 19836 45082 52577 61301 Stearic acid 50401 47979 48897 46101 49916 41927 61931 67127 59334 Pelargonic acid 300925 114696 119910 103871 100544 97131 153734 122790 132736 Ribonic acid 67036 70962 51362 542576 772659 626239 723031 131273 591679 L-Valine 2484629 5851378 3084136 255091 4065316 8025858 4241 4454039 4581361 Trimethylamine 2897643 3182055 4068571 9036 18952 3925166 1267240 3832695 4427866 L-Tryptophan 281822 292864 364014 98855 322011 360531 313471 105211 320949 Arachidonic acid 24910 53902 55474 50608 29742 53615 62356 62512 67206 2-Hydroxyphenethylamine 697667 680402 13882 711570 15957 5758.3 Aminomalonic acid 165300 188613 207714 109118 288347 306621 364260 367248 381356 Methyl propenyl ketone 2227055 282412 2070074 93044 1075351 1025152 1417295 368965 1877706 Purine 9163.1 58594 44031 91025 67363 86227 6864.9 Alpha-Linolenic acid 14199 39930 9291.8 6174 13176 9648.5 7877.5 19566 Putrescine 2850.9 3719.1 11545 1993.1 6532.5 Gamma-Tocopherol 18048 17074 16226 17243 16403 21604 21052 21158 Benzoic acid 2554319 3259762 3733075 1148045 2696619 1822245 2636649 3263961 3835589 2-Pyrrolidinone 7719.6 9832.7 17491 5356.7 7548.3 Phthalic acid 195939 201025 212176 251255 247304 239786 309362 287007 275253 Phosphoric acid 15862 61632 18312 46081 51412 Docosahexaenoic acid 5514.6 7017.9 8507.6 6901.8 7919.5 6713.1 7495.4 7637.7 Heptadecanoic acid 20895 56212 42079 7840.7 45849 Cadaverine 3792.7 10317 16953 16380 20080 7039.4 87500 Oxalic acid 3912236 37570260 13738698 5152022 4578014 4374286 61584877 36446124 3758781 Terephthalic acid 5000.3 5940.7 7028.9 3413.4 3728.6 3069.7 2943.9 2678 1,5-Anhydrosorbitol 291767 304847 306027 332771 200658 323767 440400 397432 405953 Erythritol 63824 17888 15011 24202 7377.6 Propal 9424.7 63935 5372.2 4721.5 16495 9287.7 Carbamic acid 15677 17997 15154 11651 5702.2 22506 Arabinofuranose 8051.4 2363 1572.7 3182.7 5019.4 Palmitelaidic acid 57613 53783 66242 44310 52061 73571 71035 67419 1-Hexanol 44181 24463 19735 15687 16616 38295 19995 Ethylamine 113637 13200 33714 35186 195021 175980 18044 Docosanol 25922 23916 23587 20915 29980 28553 28995 Prenol 56897 53548 63332 92313 80908 51082 3-Methyl-3-buten-1-ol 833158 956407 2354703 2271499 2313713 1149712 3729927 3205396 1011699 Acrylic acid 5484.5 78103 3474.1 14372 4238.9 13014 D-Pinitol 8247.6 16458 5383.2 7776.8 19586 7937.4 5297.7 9188.5 9368.4 1,3-Diaminopropane 2950.9 1856.9 23502 4912.1 138052 Ascorbic acid 32381 5520.4 19570 37273 18414 15045 Dihydrouracil 5672262 6566617 7462174 8328611 9043427 8992535 10026834 9457497 5342543 Citric acid 1654294 1726743 1873866 2198985 2172100 2229630 2898129 2888848 2824172 Glyoxylic acid 4494.2 8941.2 10530 17881 4829.8 18968 11638 41898 14746 Glycine 4468.6 3518.3 3781.9 36208 82746 2806655 422598 Guanosine 10128 12636 15067 17414 29393 23115 17963 L-Glutamic acid 102901 109004 121862 951974 151708 143693 123262 161783 185844 Ethanolamine 2375.8 49532 29103 3317.4 2569.8 158513 L-Alanine 2244093 2668839 1914277 3249521 4178757 2508538 D-Maltose 106075 95214 95859 136581 137457 126416 123611 114239 115765 L-Asparagine 3471.3 9370.3 19647 45261 6762.8 15362 12084 3316.2 15823 L-Isoleucine 836475 1086264 1410894 1399084 1386208 1781830 1721041 1677577 L-Lysine 501584 78668 770485 315223 302416 4234120 634028 683974 Alpha-Lactose 11641 9434.7 17189 17962 9109.2 8350.6 14340 11049 L-Serine 541028 622745 667446 5184346 5369232 883009 1155784 1091063 1076890 L-Cystine 3346.7 7299.4 53229 113624 121554 96164 126237 145751 14604 Indoleacetic acid 9265.8 10921 9834.2 8962.3 13184 11011 Ornithine 192502 356845 228387 292893 54703 247716 N-Acetyl-D-glucosamine 1807.8 1547.7 1573 2015.9 1761.6 32660 2563.2 2001.7 1954 Taurine 73615 86646 99979 1007577 1058698 661572 1839074 1509109 1054634 Succinic acid 24572 36571 49366 36414 29411 15155 37177 13424 29190 Sucrose 180254 178607 184358 148624 147750 137203 169148 178163 219036 Pyroglutamic acid 2645474 2011470 4033467 4438826 4296716 4865772 5013290 4199311 Sarcosine 96358 13684 44939 14328 26705 D-Ribose 149879 153375 160339 144077 144718 145731 186030 179727 169015 Uridine 109563 119841 131083 127601 126341 110000 171128 120596 158214 Allantoin 5262.8 4565.9 6866.3 4257.4 6686.5 9571.5 8284.1 7336.8 L-Arginine 6464.4 5787.4 9046.2 16461 21520 17722 16848 18921 15472 Creatinine 15194 3341.8 4043.9 7150.6 25852 D-Phenyllactic acid 2077.7 1984 15609 8797.6 40059 30930 L-Glutamine 103963 121988 217793 319666 375551 1087061 1693142 538179 1525212 D-Fructose 88972 88661 89920 152863 139350 155487 227185 218609 172721 L-Leucine 1457684 1562424 1757934 2352980 2380639 2224855 2290415 1716265 1283426 L-Methionine 6151.3 27671 10431 37613 10751 Hydroxypropionic acid 3674.7 6560 4943.8 5714.8 8890.8 4-Hydroxyproline 40622 23936 7362.7 125761 113902 97965 135921 41262 35646 Coumarin 8328.4 19007 3194.2 10585 13563 22028 1596.5 14021 14190 D-Alanine 93046 128528 18332 27344 13810 26238 12363 25183 D-Ribose 5-phosphate 82785 85292 95522 110906 98935 127631 127566 117986 Phenylglyoxylic acid 6629.4 8049.2 6845.3 4269.8 3472.3 3638.1 2411.4 D-Xylulose 8707.1 7886.7 2875.6 46917 11664 12044 Alpha-Tocopherol 62633 13997 13546 26961 26727 28848 Salicylic acid 2683.5 4216.3 3591.9 3449.6 2581.8 Methionine sulfoxide 29759 50254 48589 63697 57102 49283 Oleamide 11382 29233 13758 31347 20136 N-Methyl-a-aminoisobutyric acid 44794 21369 23163 16098 14400 1558.9 39918 11300 Aniline 7790.3 4872.7 5831 5515.7 6887.6 94931 6232.4 5570.8 4973.4 1-Hexadecanol 19268 112031 10575 21342 7206.2 10851 6181.7 L-Iditol 60837 14134 90122 36936 35314 14124 3997908 3809621 3289198 Tetradecanol 286800 2236.3 10174 2869.4 5352.1 9592.7 761982 Trimethyl sulfonium 5705.8 7344.6 4799.7 4025.5 5645.2 4323.8 1-Undecanol 72746 412491 56952 101806 6007.7 48839 20217 Carbachol 1388.6 17735 18029 1470.3 18314 9767 24622 23078 Propylthiouracil 123845 3273.1 8140.1 28937 13498 14708 7620.3 6766.3 5975.2 Levamisole 2833.4 1673 2318.1 2443 1985.1 Ketamine 59105 67131 73329 92893 88982 86976 129970 117715 109199 Paliperidone 11090 2932 1797.4 5784 5419.5 3927.8 1723.3 Dimethylurea 57003 102478 78401 107344 114428 129407 167187 81564 150820 Allyl propiote 18806 67086 22644 177977 235018 241611 Butyl propiote 2862.7 1334.9 63632 4193.9 1578.5 6498.4 6162.7 Aesculetin 9887.5 31902 3933.6 25592 3313.7 8469.3 1245.5 6182.8 25582 1-Butylamine 2804.3 6039.4 56560 31630 8540.7 9096.2 N-Dodecane 1770298 177228 187227 244875 5600704 205675 1584120 Undecane 14750281 10090850 16305469 5951789 5056881 5621258 7267024 4324525 3483930 N-Decane 261826 385854 21003 4713.5 158469 279012 D-Malic acid 6309.1 16801 38438 6484.5 64612 Octadecane 2556107 2625023 3429667 3147763 99322 27190 694203 12532 1475419 Ethyl benzoate 719733 694357 709254 851331 828765 801105 938346 855845 782899 Methyl benzoate 417420 555648 709255 318108 482237 604773 409057 540715 691721 Tetradecal 369678 340494 458100 377823 352030 413815 461151 432299 408876 Tridecane 4138133 1770153 3945810 3411671 7829905 4969238 3290164 4424556 9160511 Nocosane 13625 8015.6 28664 15270 48491 3295.8 3175.5 Lumichrome 2862734 3244777 3736803 4405346 4248337 4267111 3776340 9051777 4925496 1-phthylamine 2343.5 5507.8 2888.7 75233 3495.3 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name retention index quantified m/z PubChem ID KEGG ID Groups Derivatized MSI Matching Levels HMDB ChEBI METLIN Butyric acid 264 C00246 Butanoic acid Yes Level 2 HMDB0000039 30772 107 Acetic acid 176 C00033 Acetic acid Yes Level 2 HMDB0000042 15366 3206 Cholesterol 11025495 C00187 Cholesterol Yes Level 2 HMDB0000067 1307929 163 Pipecolic acid 849 C00408 2-Piperidinecarboxylic acid Yes Level 2 HMDB0000070 17964 50 Ethanol 702 C00469 Ethanol Yes Level 2 HMDB0000108 16236 3203 D-Glucose 5793 C00031 D-Glucose Yes Level 2 HMDB0000122 4167 133 Glycerol 753 C00116 Glycerol Yes Level 2 HMDB0000131 17522 105 D-Galactose 439357 C00984 d-Galactose Yes Level 2 HMDB0000143 28061 134 L-Tyrosine 6057 C00082 L-Tyrosine Yes Level 2 HMDB0000158 17895 34 L-Phenylalanine 6140 C00079 L-Phenylalanine Yes Level 2 HMDB0000159 17295 28 L-Proline 145742 C00148 L-Proline Yes Level 2 HMDB0000162 17203 29 L-Threonine 6288 C00188 L-Threonine Yes Level 2 HMDB0000167 16857 32 D-Mannose 18950 C00159 d-Mannose Yes Level 2 HMDB0000169 4208 L-Aspartic acid 5960 C00049 L-Aspartic acid Yes Level 2 HMDB0000191 17053 5206 Phenylacetic acid 999 C07086 Benzeneacetic acid Yes Level 2 HMDB0000209 30745 129 myo-Inositol C00137 Myo-Inositol Yes Level 2 HMDB0000211 17268 5221 Palmitic acid 985 C00249 Hexadecanoic acid Yes Level 2 HMDB0000220 15756 187 Propionic acid 1032 C00163 Propanoic acid Yes Level 2 HMDB0000237 30768 3215 Pyridoxine 1054 C00314 Pyridoxine Yes Level 2 HMDB0000239 16709 5245 Urea 1176 C00086 Urea Yes Level 2 HMDB0000294 16199 6 3-Hexenedioic acid 5351896 trans-3-Hexenedioic acid Yes Level 2 HMDB0000393 86952 5382 N-Alpha-acetyllysine 192590 C12989 N-alpha-Acetyl-L-Lysine Yes Level 2 HMDB0000446 35704 5435 Adipic acid 196 C06104 Hexanedioic acid Yes Level 2 HMDB0000448 30832 115 Capric acid 2969 C01571 Decanoic acid Yes Level 2 HMDB0000511 30813 336 Caproic acid 8892 C01585 Hexanoic acid Yes Level 2 HMDB0000535 30776 5520 Elaidic acid 445639 C00712 9-Octadecenoic acid Yes Level 2 HMDB0000573 27997 3406 Citraconic acid 643798 C02226 Methylmaleic acid Yes Level 2 HMDB0000634 17626 5607 Dodecanoic acid 3893 C02679 Dodecanoic acid Yes Level 2 HMDB0000638 30805 5611 Glutaric acid 743 C00489 Pentanedioic acid Yes Level 2 HMDB0000661 17859 3254 Heptanoic acid 8094 C17714 Heptanoic acid Yes Level 2 HMDB0000666 45571 5636 Isocaproic acid 12587 4-Methylvaleric acid Yes Level 2 HMDB0000689 74903 4191 Malonic acid 867 C00383 Malonic acid Yes Level 2 HMDB0000691 30794 3237 Pseudouridine 15047 C02067 Pseudouridine Yes Level 2 HMDB0000767 17802 5734 Azelaic acid 2266 C08261 Azelaic acid Yes Level 2 HMDB0000784 48131 5750 Pyrrolidonecarboxylic acid 499 C02237 2-Pyrrolidone-5-carboxylic acid Yes Level 2 HMDB0000805 16010 5769 Myristic acid 11005 C06424 Tetradecanoic acid Yes Level 2 HMDB0000806 28875 196 Pentadecanoic acid 13849 C16537 n-Pentadecanoic acid Yes Level 2 HMDB0000826 5789 Stearic acid 5281 C01530 Octadecanoic acid Yes Level 2 HMDB0000827 28842 189 Pelargonic acid 8158 C01601 Nonoic acid Yes Level 2 HMDB0000847 29019 5810 Ribonic acid 5460677 C01685 Ribonic acid Yes Level 2 HMDB0000867 21077 3290 L-Valine 6287 C00183 L-Valine Yes Level 2 HMDB0000883 16414 5842 Trimethylamine 1146 C00565 N-Trimethylamine Yes Level 2 HMDB0000906 18139 3772 L-Tryptophan 6305 C00078 L-Tryptophan Yes Level 2 HMDB0000929 16828 5879 Arachidonic acid 444899 C00219 Arachidonic acid Yes Level 2 HMDB0001043 15843 193 2-Hydroxyphenethylamine 1000 C02735 Phenylethanolamine Yes Level 2 HMDB0001065 16343 59 Aminomalonic acid 100714 C00872 Aminomalonic acid Yes Level 2 HMDB0001147 17475 Methyl propenyl ketone 637920 3-penten-2-one Yes Level 2 HMDB0001184 Purine 1044 C15587 7H-purine Yes Level 2 HMDB0001366 17258 6193 Alpha-Linolenic acid 5280934 C06427 Alpha-Linolenic acid Yes Level 2 HMDB0001388 27432 6208 Putrescine 1045 C00134 1,4-Butanediamine Yes Level 2 HMDB0001414 17148 3226 Gamma-Tocopherol 14986 Gamma-Tocopherol Yes Level 2 HMDB0001492 298402 6276 Benzoic acid 243 C00180 Benzoic acid Yes Level 2 HMDB0001870 30746 1297 2-Pyrrolidinone 12025 C11118 2-pyrrolidinone Yes Level 2 HMDB0002039 36592 6452 Phthalic acid 1017 C01606 1,2-Benzenedicarboxylic acid Yes Level 2 HMDB0002107 29069 6489 Phosphoric acid 1004 C00009 Phosphoric acid Yes Level 2 HMDB0002142 26078 Docosahexaenoic acid 445580 C06429 Docosahexaenoic acid Yes Level 2 HMDB0002183 28125 3457 Heptadecanoic acid 10465 Heptadecanoic acid Yes Level 2 HMDB0002259 32365 6578 Cadaverine 273 C01672 Cadaverine Yes Level 2 HMDB0002322 18127 3236 Oxalic acid 971 C00209 Ethanedioic acid Yes Level 2 HMDB0002329 16995 113 Terephthalic acid 7489 C06337 1,4-Benzenedicarboxylic acid Yes Level 2 HMDB0002428 15702 6676 1,5-Anhydrosorbitol 64960 C07326 1,5-Anhydro-D-sorbitol Yes Level 2 HMDB0002712 16070 3775 Erythritol 222285 C00503 Erythritol Yes Level 2 HMDB0002994 17113 140 Propal 527 C00479 Propal Yes Level 2 HMDB0003366 17153 6906 Carbamic acid 277 C01563 Carbamate Yes Level 2 HMDB0003551 28616 6950 Arabinofuranose 440921 C06115 Arabinofuranose Yes Level 2 HMDB0012325 6178 Palmitelaidic acid 5282745 Palmitelaidic acid Yes Level 2 HMDB0012328 59265 1-Hexanol 8103 C00854 Cyclohexanol Yes Level 2 HMDB0012971 113441 Ethylamine 6341 C00797 Ethylamine Yes Level 2 HMDB0013231 15862 Docosanol 12620 1-Docosanol Yes Level 2 HMDB0014770 31000 Prenol 11173 C01390 3-Methyl-2-buten-1-ol Yes Level 2 HMDB0030124 16019 3-Methyl-3-buten-1-ol 12988 3-Methyl-3-buten-1-ol Yes Level 2 HMDB0030126 62898 Acrylic acid 6581 C00511 2-Propenoic acid Yes Level 2 HMDB0031647 18308 D-Pinitol C03844 D-Pinitol Yes Level 2 HMDB0034219 28548 1,3-Diaminopropane 428 C00986 1,3-Diaminopropane No Level 3 HMDB0000002 15725 5081 Ascorbic acid 54670067 C00072 L-ASCORBIC ACID No Level 3 HMDB0000044 17208 3753 Dihydrouracil 649 C00429 Dihydrouracil No Level 3 HMDB0000076 15901 285 Citric acid 311 C00158 Citrate No Level 3 HMDB0000094 30769 124 Glyoxylic acid 760 C00048 Glyoxylic acid No Level 3 HMDB0000119 16891 3213 Glycine 750 C00037 Glycine No Level 3 HMDB0000123 15428 20 Guanosine 6802 C00387 Guanosine No Level 3 HMDB0000133 16750 87 L-Glutamic acid 33032 C00025 L-Glutamic acid No Level 3 HMDB0000148 16015 5174 Ethanolamine 700 C00189 Ethanolamine No Level 3 HMDB0000149 16000 3207 L-Alanine 5950 C00041 L-Alanine No Level 3 HMDB0000161 16977 D-Maltose 10991489 C00208 Maltose No Level 3 HMDB0000163 47937 413 L-Asparagine 6267 C00152 L-Asparagine No Level 3 HMDB0000168 17196 14 L-Isoleucine 6306 C00407 L-Isoleucine No Level 3 HMDB0000172 17191 5193 L-Lysine 5962 C00047 L-Lysine No Level 3 HMDB0000182 18019 5200 Alpha-Lactose 84571 C00243 Alpha-Lactose No Level 3 HMDB0000186 36219 267 L-Serine 5951 C00065 L-Serine No Level 3 HMDB0000187 17115 5203 L-Cystine 67678 C00491 L-Cystine No Level 3 HMDB0000192 16283 5207 Indoleacetic acid 802 C00954 BETA-INDOLEACETIC ACID No Level 3 HMDB0000197 16411 5211 Ornithine 6262 C00077 L-Ornithine No Level 3 HMDB0000214 15729 27 N-Acetyl-D-glucosamine 439174 C00140 N-Acetyl-D-glucosamine No Level 3 HMDB0000215 506227 3356 Taurine 1123 C00245 Taurine No Level 3 HMDB0000251 15891 31 Succinic acid 1110 C00042 Succite No Level 3 HMDB0000254 15741 114 Sucrose 5988 C00089 Sucrose No Level 3 HMDB0000258 17992 137 Pyroglutamic acid 7405 C01879 L-5-Oxoproline No Level 3 HMDB0000267 18183 3251 Sarcosine 1088 C00213 Sarcosine No Level 3 HMDB0000271 15611 51 D-Ribose 5779 C00121 D-Ribose No Level 3 HMDB0000283 16988 313 Uridine 6029 C00299 Uridine No Level 3 HMDB0000296 16704 90 Allantoin 204 C01551 Allantoin No Level 3 HMDB0000462 15676 89 L-Arginine 6322 C00062 L-Arginine No Level 3 HMDB0000517 16467 5502 Creatinine 588 C00791 Creatinine No Level 3 HMDB0000562 16737 8 D-Phenyllactic acid C05607 D-3-Phenyllactic acid No Level 3 HMDB0000563 8100 5547 L-Glutamine 5961 C00064 L-Glutamine No Level 3 HMDB0000641 18050 5614 D-Fructose 439709 C02336 D-Fructose No Level 3 HMDB0000660 28645 135 L-Leucine 6106 C00123 L-LEUCINE No Level 3 HMDB0000687 15603 24 L-Methionine 6137 C00073 L-Methionine No Level 3 HMDB0000696 16643 5664 Hydroxypropionic acid 68152 C01013 3-hydroxypropionic acid No Level 3 HMDB0000700 33404 5668 4-Hydroxyproline 5810 C01157 trans-4-Hydroxy-L-proline No Level 3 HMDB0000725 18095 257 Coumarin 323 C05851 Coumarin No Level 3 HMDB0001218 28794 3525 D-Alanine 71080 C00133 D-Alanine No Level 3 HMDB0001310 15570 D-Ribose 5-phosphate 439167 C00117 D-Ribose 5-phosphate No Level 3 HMDB0001548 52742 6315 Phenylglyoxylic acid 11915 C02137 Benzoylformate No Level 3 HMDB0001587 18280 D-Xylulose 5289590 C00310 D-Xylulose No Level 3 HMDB0001644 17140 6333 Alpha-Tocopherol 14985 C02477 alpha-Tocopherol No Level 3 HMDB0001893 18145 6376 Salicylic acid 338 C00805 Salicylic acid No Level 3 HMDB0001895 16914 616 Methionine sulfoxide 847 Methionine sulfoxide No Level 3 HMDB0002005 49033 6428 Oleamide 5283387 C19670 OLEYL AMIDE No Level 3 HMDB0002117 116314 N-Methyl-a-aminoisobutyric acid 6951124 N-Methyl-a-aminoisobutyric acid No Level 3 HMDB0002141 134261 6508 Aniline 6115 C00292 Aniline No Level 3 HMDB0003012 17296 489 1-Hexadecanol 2682 C00823 hexadecanol No Level 3 HMDB0003424 16125 6927 L-Iditol 5460044 C01507 L-Iditol No Level 3 HMDB0011632 18202 Tetradecanol 8209 N-tetradecanol No Level 3 HMDB0011638 133255 Trimethyl sulfonium 1147 C01008 Trimethylsulfonium No Level 3 HMDB0012295 17434 1-Undecanol 8184 N-undecanol No Level 3 HMDB0013113 133122 Carbachol 5831 Carbachol No Level 3 HMDB0014555 3385 Propylthiouracil 657298 C07569 Propylthiouracil No Level 3 HMDB0014690 8502 Levamisole 26879 C07070 Levamisole No Level 3 HMDB0014986 484948 Ketamine 3821 C07525 Ketamine No Level 3 HMDB0015352 6121 Paliperidone 115237 9-OH-Risperidone No Level 3 HMDB0015396 568656 Dimethylurea 7293 C16364 Dimethylurea No Level 3 HMDB0029198 80472 Allyl propiote 61319 Propanoic acid, 2-propenyl ester No Level 3 HMDB0029487 Butyl propiote 11529 BUTYL PROPIOTE No Level 3 HMDB0030052 Aesculetin 5281416 C09263 Aesculetin No Level 3 HMDB0030819 490095 1-Butylamine 8007 1-Butamine No Level 3 HMDB0031321 43799 N-Dodecane 8182 C08374 DODECANE No Level 3 HMDB0031444 28817 Undecane 14257 Undecane No Level 3 HMDB0031445 46342 N-Decane 15600 Decane No Level 3 HMDB0031450 32894 D-Malic acid 92824 C00497 D-Malic acid No Level 3 HMDB0031518 30796 Octadecane 11635 N-OCTADECANE No Level 3 HMDB0033721 32926 Ethyl benzoate 7165 C01839 ETHYL BENZOATE No Level 3 HMDB0033967 32807 Methyl benzoate 7150 Benzoic acid, methyl ester No Level 3 HMDB0033968 72775 Tetradecal 31291 Tetradecal No Level 3 HMDB0034283 84067 Tridecane 12388 C13834 Tridecane No Level 3 HMDB0034284 35998 Nocosane 12409 C08384 Nocosane No Level 3 HMDB0034288 7613 Lumichrome C01727 Lumichrome No Level 3 METPA0216 1-phthylamine C14790 1-phthylamine No Level 3 METPA1135 METABOLITES_END #END