#METABOLOMICS WORKBENCH PJTrim_20161222_162316 DATATRACK_ID:804 STUDY_ID:ST000547 ANALYSIS_ID:AN000834 PROJECT_ID:PR000401 VERSION 1 CREATED_ON February 8, 2017, 1:13 pm #PROJECT PR:PROJECT_TITLE Intergenerational murine gut microbiome variation PR:PROJECT_TYPE untargeted metabolomics PR:PROJECT_SUMMARY Inbred mice are used to investigate many aspects of human physiology, including PR:PROJECT_SUMMARY susceptibility to disease and response to therapies. Despite increasing evidence PR:PROJECT_SUMMARY that the composition and function of the murine intestinal microbiota can PR:PROJECT_SUMMARY substantially influence a broad range of experimental outcomes, relatively PR:PROJECT_SUMMARY little is known about microbiome dynamics within experimental mouse populations. PR:PROJECT_SUMMARY We investigated changes in the intestinal microbiome between C57BL/6J mice PR:PROJECT_SUMMARY spanning six generations (assessed at generations 1, 2, 3 and 6), following PR:PROJECT_SUMMARY their introduction to a stringently controlled facility. Faecal microbiota PR:PROJECT_SUMMARY composition and function were assessed by 16S rRNA gene amplicon sequencing and PR:PROJECT_SUMMARY liquid chromatography mass spectrometry, respectively. Significant divergence of PR:PROJECT_SUMMARY the intestinal microbiota between founder and second generation mice, as well as PR:PROJECT_SUMMARY continuing inter-generational variance, was observed. Bacterial taxa whose PR:PROJECT_SUMMARY relative abundance changed significantly included Akkermansia, Turicibacter and PR:PROJECT_SUMMARY Bifidobacterium (p< 0.05), all of which are recognised as having the potential PR:PROJECT_SUMMARY to substantially influence host physiology. Shifts in microbiota composition PR:PROJECT_SUMMARY were mirrored by corresponding differences in the faecal metabolome (r=0.57, PR:PROJECT_SUMMARY p=0.0001), with notable differences in levels of tryptophan pathway metabolites PR:PROJECT_SUMMARY and amino acids, including glutamine, glutamate and aspartate. The magnitude of PR:PROJECT_SUMMARY these changes in the intestinal microbiota and metabolome characteristics during PR:PROJECT_SUMMARY acclimation were on a scale with those observed between populations housed in PR:PROJECT_SUMMARY separate facilities, which differed in regards to husbandry, barrier conditions PR:PROJECT_SUMMARY and dietary intake. The microbiome variance reported here has major implications PR:PROJECT_SUMMARY for experimental reproducibility, and as a consequence, experimental design and PR:PROJECT_SUMMARY the interpretation of research outcomes across as wide range of contexts. PR:INSTITUTE South Australian Health and Medical Research Institute PR:DEPARTMENT Infection and Immunity Theme PR:LAST_NAME Choo PR:FIRST_NAME Jocelyn PR:ADDRESS SAHMRI, North Terrace, Adelaide, SA 5000, Australia PR:EMAIL Jocelyn.choo@sahmri.com PR:PHONE N/A #STUDY ST:STUDY_TITLE Intergenerational murine gut microbiome variation ST:STUDY_TYPE Intergenerational ST:STUDY_SUMMARY Inbred mice are used to investigate many aspects of human physiology, including ST:STUDY_SUMMARY susceptibility to disease and response to therapies. Despite increasing evidence ST:STUDY_SUMMARY that the composition and function of the murine intestinal microbiota can ST:STUDY_SUMMARY substantially influence a broad range of experimental outcomes, relatively ST:STUDY_SUMMARY little is known about microbiome dynamics within experimental mouse populations. ST:STUDY_SUMMARY We investigated changes in the intestinal microbiome between C57BL/6J mice ST:STUDY_SUMMARY spanning six generations (assessed at generations 1, 2, 3 and 6), following ST:STUDY_SUMMARY their introduction to a stringently controlled facility. Faecal microbiota ST:STUDY_SUMMARY composition and function were assessed by 16S rRNA gene amplicon sequencing and ST:STUDY_SUMMARY liquid chromatography mass spectrometry, respectively. Significant divergence of ST:STUDY_SUMMARY the intestinal microbiota between founder and second generation mice, as well as ST:STUDY_SUMMARY continuing inter-generational variance, was observed. Bacterial taxa whose ST:STUDY_SUMMARY relative abundance changed significantly included Akkermansia, Turicibacter and ST:STUDY_SUMMARY Bifidobacterium (p< 0.05), all of which are recognised as having the potential ST:STUDY_SUMMARY to substantially influence host physiology. Shifts in microbiota composition ST:STUDY_SUMMARY were mirrored by corresponding differences in the faecal metabolome (r=0.57, ST:STUDY_SUMMARY p=0.0001), with notable differences in levels of tryptophan pathway metabolites ST:STUDY_SUMMARY and amino acids, including glutamine, glutamate and aspartate. The magnitude of ST:STUDY_SUMMARY these changes in the intestinal microbiota and metabolome characteristics during ST:STUDY_SUMMARY acclimation were on a scale with those observed between populations housed in ST:STUDY_SUMMARY separate facilities, which differed in regards to husbandry, barrier conditions ST:STUDY_SUMMARY and dietary intake. The microbiome variance reported here has major implications ST:STUDY_SUMMARY for experimental reproducibility, and as a consequence, experimental design and ST:STUDY_SUMMARY the interpretation of research outcomes across as wide range of contexts. ST:INSTITUTE South Australian Health and Medical Research Institute ST:DEPARTMENT Infection and Immunity Theme ST:LAST_NAME Rogers ST:FIRST_NAME Geraint ST:ADDRESS SAHMRI, North Terrace, Adelaide, SA 5000, Australia ST:EMAIL Geraint.rogers@sahmri.com ST:PHONE N/A ST:NUM_GROUPS 4 ST:TOTAL_SUBJECTS 82 #SUBJECT SU:SUBJECT_TYPE Animal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:GENOTYPE_STRAIN C57BL/6 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - SG1 Generation:1st Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG2 Generation:1st Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG3 Generation:1st Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG4 Generation:1st Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG5 Generation:1st Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG6 Generation:1st Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG7 Generation:1st Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG8 Generation:1st Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG9 Generation:1st Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG10 Generation:1st Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG11 Generation:1st Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG12 Generation:1st Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG13 Generation:1st Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG14 Generation:1st Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG15 Generation:1st Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG16 Generation:1st Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG17 Generation:1st Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG18 Generation:1st Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG19 Generation:1st Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG108 Generation:1st Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG109 Generation:1st Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG110 Generation:1st Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG111 Generation:1st Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG112 Generation:1st Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG156 Generation:2nd Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG157 Generation:2nd Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG158 Generation:2nd Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG159 Generation:2nd Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG160 Generation:2nd Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG161 Generation:2nd Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG162 Generation:2nd Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG163 Generation:2nd Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG166 Generation:2nd Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG167 Generation:2nd Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG168 Generation:2nd Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG170 Generation:2nd Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG164 Generation:3rd Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG165 Generation:3rd Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG169 Generation:3rd Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG171 Generation:3rd Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG172 Generation:3rd Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG173 Generation:3rd Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG174 Generation:3rd Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG175 Generation:3rd Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG176 Generation:3rd Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG177 Generation:3rd Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG178 Generation:3rd Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG179 Generation:3rd Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG180 Generation:3rd Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG181 Generation:3rd Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG182 Generation:3rd Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG183 Generation:3rd Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG184 Generation:3rd Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG185 Generation:3rd Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG186 Generation:3rd Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG187 Generation:3rd Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG188 Generation:3rd Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG189 Generation:3rd Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG190 Generation:3rd Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG191 Generation:3rd Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG192 Generation:3rd Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG202 Generation:6th Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG203 Generation:6th Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG204 Generation:6th Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG205 Generation:6th Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG206 Generation:6th Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG207 Generation:6th Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG208 Generation:6th Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG214 Generation:6th Gen | Gender:M SUBJECT_SAMPLE_FACTORS - SG221 Generation:6th Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG222 Generation:6th Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG223 Generation:6th Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG224 Generation:6th Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG225 Generation:6th Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG231 Generation:6th Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG232 Generation:6th Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG233 Generation:6th Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG234 Generation:6th Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG235 Generation:6th Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG236 Generation:6th Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG237 Generation:6th Gen | Gender:F SUBJECT_SAMPLE_FACTORS - SG238 Generation:6th Gen | Gender:F #COLLECTION CO:COLLECTION_SUMMARY Fresh faecal pellets were collected homogenised in PBS, centrifuged. Supernatant CO:COLLECTION_SUMMARY was extracted using SPE. CO:SAMPLE_TYPE Faeces CO:VOLUMEORAMOUNT_COLLECTED one faecal pellet CO:STORAGE_CONDITIONS -80C CO:COLLECTION_VIALS 1.5mL Eppendorff CO:STORAGE_VIALS 1.5mL Eppendorff CO:COLLECTION_TUBE_TEMP 4C #TREATMENT TR:TREATMENT_SUMMARY No treatment #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Faeces were homogenised in PBS , equivelent of 125 µg faecal supernatant was SP:SAMPLEPREP_SUMMARY extracted using solid phase extraction prior to UPLC-MS analysis. SP:PROCESSING_METHOD Homogenisation, SPE extraction SP:PROCESSING_STORAGE_CONDITIONS -80C SP:EXTRACTION_METHOD Faecal pellets were dispersed in 1 mL PBS by vortexing. Suspensions were then SP:EXTRACTION_METHOD centrifuged at 13000 x g for 5 mins and the supernatant collected for analysis. SP:EXTRACT_ENRICHMENT A 50 µL aliquot was placed in a pre-washed (1 mL acetonitrile) and equilibrated SP:EXTRACT_ENRICHMENT (1 mL 0.1% TFA aqueous) Oasis HLB 10 mg SPE cartridge (Waters Corporation, SP:EXTRACT_ENRICHMENT Milford, MA, USA). The sample was washed with 1 mL 0.1% TFA and eluted with 1 mL SP:EXTRACT_ENRICHMENT 0.1% TFA in 70% acetonitrile. SP:EXTRACT_STORAGE The eluent was lyophilized overnight in a RVC 2-33CD plus rotational vacuum SP:EXTRACT_STORAGE concentrator (Martin Christ Gefriertrocknungsanlagen GmbH, Osterode am Harz, SP:EXTRACT_STORAGE Germany) operated at 10 mBar and room temperature. SP:SAMPLE_RESUSPENSION Samples were reconstituted in 50 µL 0.1% FA, vortexed and centrifuged at 16 000 SP:SAMPLE_RESUSPENSION x g for 15 min. The supernatant was transferred into LC-MS-grade glass vials SP:SAMPLE_RESUSPENSION (avoiding the pellet) and stored at 6°C until use. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY RP C18 +ve and -ve Ion CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Waters Acquity CH:COLUMN_NAME Waters Acquity BEH C18 (150 x 2.1mm, 1.7um) CH:COLUMN_NAME (Milford, MA, USA) CH:FLOW_RATE 400 µL/min CH:COLUMN_TEMPERATURE 45C CH:SOLVENT_A 0.1 % Formic Acid CH:SOLVENT_B 99.9% Acetonitrile 0.1% Formic Acid CH:WEAK_WASH_SOLVENT_NAME Water CH:WEAK_WASH_VOLUME 600 µL CH:STRONG_WASH_SOLVENT_NAME Acetonitrile CH:STRONG_WASH_VOLUME 200 µL CH:TARGET_SAMPLE_TEMPERATURE 6C CH:SAMPLE_LOOP_SIZE 20 µL CH:RANDOMIZATION_ORDER Random #ANALYSIS AN:ANALYSIS_TYPE MS AN:LABORATORY_NAME SAHMRI, Mass Spectrometry Core Facility AN:OPERATOR_NAME Paul Trim AN:DETECTOR_TYPE TOF AN:SOFTWARE_VERSION MassLynx 4.1, EzInfo 3, Progenesis AN:DATA_FORMAT .raw #MS MS:INSTRUMENT_NAME Waters Synapt G1 MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS Every 3rd injection was a PBQC sample MS:CAPILLARY_VOLTAGE 2.8 kV MS:COLLISION_ENERGY Trap CE 6, Transfer CE 4 MS:COLLISION_GAS Nitrogen MS:SOURCE_TEMPERATURE 120 C MS:DESOLVATION_GAS_FLOW 900 L/h MS:DESOLVATION_TEMPERATURE 450 C #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Normalised Intensity MS_METABOLITE_DATA_START Samples SG1 SG10 SG108 SG109 SG11 SG110 SG111 SG112 SG12 SG13 SG14 SG15 SG16 SG17 SG18 SG19 SG2 SG3 SG4 SG5 SG6 SG7 SG8 SG9 SG156 SG157 SG158 SG159 SG160 SG161 SG162 SG163 SG166 SG167 SG168 SG170 SG164 SG165 SG169 SG171 SG172 SG173 SG174 SG175 SG176 SG177 SG178 SG179 SG180 SG181 SG182 SG183 SG184 SG185 SG186 SG187 SG188 SG189 SG190 SG191 SG192 SG202 SG203 SG204 SG205 SG206 SG207 SG208 SG214 SG221 SG222 SG223 SG224 SG225 SG231 SG232 SG233 SG234 SG235 SG236 SG237 SG238 Factors Generation:1st Gen | Gender:F Generation:1st Gen | Gender:M Generation:1st Gen | Gender:M Generation:1st Gen | Gender:M Generation:1st Gen | Gender:M Generation:1st Gen | Gender:M Generation:1st Gen | Gender:M Generation:1st Gen | Gender:M Generation:1st Gen | Gender:F Generation:1st Gen | Gender:F Generation:1st Gen | Gender:F Generation:1st Gen | Gender:F Generation:1st Gen | Gender:F Generation:1st Gen | Gender:F Generation:1st Gen | Gender:F Generation:1st Gen | Gender:F Generation:1st Gen | Gender:F Generation:1st Gen | Gender:F Generation:1st Gen | Gender:F Generation:1st Gen | Gender:M Generation:1st Gen | Gender:M Generation:1st Gen | Gender:M Generation:1st Gen | Gender:M Generation:1st Gen | Gender:M Generation:2nd Gen | Gender:F Generation:2nd Gen | Gender:M Generation:2nd Gen | Gender:F Generation:2nd Gen | Gender:M Generation:2nd Gen | Gender:F Generation:2nd Gen | Gender:M Generation:2nd Gen | Gender:F Generation:2nd Gen | Gender:F Generation:2nd Gen | Gender:F Generation:2nd Gen | Gender:F Generation:2nd Gen | Gender:F Generation:2nd Gen | Gender:M Generation:3rd Gen | Gender:M Generation:3rd Gen | Gender:F Generation:3rd Gen | Gender:M Generation:3rd Gen | Gender:F Generation:3rd Gen | Gender:F Generation:3rd Gen | Gender:M Generation:3rd Gen | Gender:F Generation:3rd Gen | Gender:F Generation:3rd Gen | Gender:F Generation:3rd Gen | Gender:F Generation:3rd Gen | Gender:M Generation:3rd Gen | Gender:M Generation:3rd Gen | Gender:M Generation:3rd Gen | Gender:M Generation:3rd Gen | Gender:M Generation:3rd Gen | Gender:F Generation:3rd Gen | Gender:F Generation:3rd Gen | Gender:M Generation:3rd Gen | Gender:M Generation:3rd Gen | Gender:F Generation:3rd Gen | Gender:F Generation:3rd Gen | Gender:M Generation:3rd Gen | Gender:F Generation:3rd Gen | Gender:F Generation:3rd Gen | Gender:M Generation:6th Gen | Gender:M Generation:6th Gen | Gender:M Generation:6th Gen | Gender:M Generation:6th Gen | Gender:M Generation:6th Gen | Gender:M Generation:6th Gen | Gender:M Generation:6th Gen | Gender:M Generation:6th Gen | Gender:M Generation:6th Gen | Gender:F Generation:6th Gen | Gender:F Generation:6th Gen | Gender:F Generation:6th Gen | Gender:F Generation:6th Gen | Gender:F Generation:6th Gen | Gender:F Generation:6th Gen | Gender:F Generation:6th Gen | Gender:F Generation:6th Gen | Gender:F Generation:6th Gen | Gender:F Generation:6th Gen | Gender:F Generation:6th Gen | Gender:F Generation:6th Gen | Gender:F 1.02_152.0579m/z 114.371 938.2469 19.16438 47.17603 551.235 106.7196 187.6733 19.2736 531.669 800.9884 172.7799 111.754 100.8193 186.6514 109.1461 121.8063 44.08902 395.777 289.2316 35.0765 52.70324 146.315 108.6394 42.52484 140.0879 148.7785 249.7724 79.74386 32.74383 35.12083 331.9673 107.5442 212.235 175.0903 53.51262 59.08891 20.91263 43.26562 179.8307 82.04258 75.8469 115.2731 79.99027 87.42145 68.12063 23.967 96.691 47.41916 64.01226 32.60588 55.37533 126.9643 13.84173 25.69627 27.5675 82.43374 21.94208 58.0789 39.81719 57.44122 27.44138 73.59114 48.79221 27.98821 93.78629 1.02_118.0865m/z 104.0546 53.01867 128.1928 25.64869 55.09583 101.811 135.3252 34.90168 30.56724 72.85414 65.91308 56.99779 93.47777 98.03689 70.82063 71.85104 123.8135 105.0421 142.8051 182.3381 155.8902 49.14475 47.84225 155.1485 78.79138 85.59551 54.39988 107.2993 105.9223 18.47892 50.0463 86.08257 108.4541 52.79869 105.7423 135.8593 88.40834 153.3109 154.5343 73.36633 80.88462 53.65744 53.59845 39.42834 69.03901 35.31881 33.9216 46.56536 48.66396 91.37925 54.20597 114.7283 33.57941 87.58337 36.28942 53.8378 52.18955 27.10742 36.75533 78.39011 17.11444 60.31252 27.18005 111.5487 41.23442 42.96499 68.44189 30.34314 15.3995 91.94395 25.36252 93.14756 22.70814 66.34 22.10225 45.69817 44.99276 99.4674 1.02_208.9870m/z 26.53009 724.521 453.1123 455.5188 409.6145 107.4868 422.9178 1.02_113.9647m/z 211.3249 262.653 209.1113 69.92908 40.47269 174.2845 225.7838 342.1104 141.7938 420.0607 174.2175 220.4041 170.7116 154.6263 386.0761 301.6964 198.3405 228.9601 247.8257 201.5913 279.456 316.6597 258.8817 252.1992 222.0228 171.635 286.9355 205.8027 205.3045 179.4678 339.8715 323.7186 114.24 210.3348 245.064 223.9573 255.4279 251.976 355.3759 248.1745 264.6791 317.2563 414.9135 389.5786 307.8418 358.222 261.6912 164.769 190.5553 159.4096 238.9512 195.8465 168.9765 224.5414 128.309 58.2225 226.0885 155.8682 29.04316 51.7127 146.0215 167.0168 188.6165 131.3217 259.7167 209.2146 255.4868 79.12913 107.503 258.5283 90.80498 159.6267 209.2006 308.3462 300.6935 357.5686 274.9512 199.5597 242.1158 375.0834 249.1737 214.8198 1.02_72.9373m/z 129.4754 192.1391 77.36697 59.68151 127.782 183.235 138.9776 76.43225 273.6617 68.32684 183.1232 139.8579 11.30104 251.8446 215.7413 139.1305 122.0127 95.923 104.7323 181.1025 173.8903 120.7079 200.3643 143.1236 144.3989 176.238 170.7261 158.5884 145.3918 170.767 146.0496 113.2503 125.9061 195.3577 94.32536 154.1335 190.6156 230.8714 175.5878 172.6657 251.1638 256.5587 190.692 191.4795 165.3723 209.6505 56.35912 152.3822 110.9364 144.6846 131.2048 100.4476 150.369 103.2056 35.39674 92.75234 106.2511 38.34062 24.03032 78.39613 113.449 105.4924 76.4151 166.8333 104.1682 140.7527 54.98378 54.61782 158.448 49.75482 74.87979 166.4409 187.2644 181.5543 231.1429 148.225 141.2674 135.5237 214.1137 146.7224 113.2593 1.03_136.0628m/z 861.4928 155.6222 56.41208 29.85955 81.35409 59.10154 94.85072 29.60427 158.8754 102.1158 75.9669 88.25296 194.2643 140.0536 126.4634 161.1363 240.871 110.2157 159.4525 245.315 42.5198 145.8993 66.74849 32.01075 74.61047 25.36745 23.63271 28.17493 184.908 162.7185 25.72078 80.16975 194.5386 143.0997 127.7762 180.6539 207.8304 144.1776 1014.778 586.7827 1195.234 336.101 267.1579 124.935 163.9228 116.3464 91.56992 77.24152 103.3757 22.21616 232.3272 390.2036 534.3421 108.3304 126.2768 410.4912 456.9356 661.8828 336.3494 130.213 449.4371 404.7766 43.8025 356.7088 34.10973 122.4561 152.4667 106.4936 1811.318 630.212 803.5685 1077.79 1312.869 883.1026 1414.851 184.2849 1.09_224.1289m/z 178.1867 212.5523 272.0264 227.5084 260.34 197.1132 312.0083 363.4088 267.5734 175.9287 235.1113 208.8139 132.5452 131.5921 269.096 335.5101 245.261 232.424 298.6562 242.1016 209.7849 314.0096 175.765 256.9908 217.652 168.889 256.7567 196.3351 166.851 181.0616 201.3927 174.1242 104.0344 32.1619 161.9409 157.972 241.7178 241.0459 285.9633 250.3371 246.8286 297.8103 197.2206 290.3192 220.9982 238.2528 241.8552 145.0769 227.9145 181.3135 212.1072 195.3353 279.8338 216.4057 248.7678 74.80776 286.2929 133.1947 38.74273 70.88522 270.254 173.5733 248.8187 172.6783 238.254 183.093 140.5293 124.4068 111.0563 230.1428 191.5532 186.289 186.0479 254.476 254.4702 317.3268 182.9783 143.0049 246.8324 264.7368 172.3832 286.226 1.09_160.1343m/z 30.7592 63.17917 34.77467 109.527 479.4818 91.77969 45.81693 31.14597 21.85103 142.4877 244.0372 124.9371 105.4494 196.8651 119.8682 110.443 223.7255 134.9998 315.0942 184.9251 207.7804 236.5879 125.5388 193.0055 92.33724 97.54806 161.5412 131.7445 167.7095 258.8095 113.4051 277.3689 103.4107 160.6067 100.9825 84.84252 96.13889 93.62777 123.0088 145.9203 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2914.047 2123.484 1398.232 2594.654 1837.126 2019.938 1806.717 2475.379 1626.151 2597.971 2017.071 1965.81 2067.579 2267.635 1533.343 1842.092 2257.267 1859.77 1897.744 2735.604 2032.868 2550.088 2866.865 2236.169 2481.672 1990.711 2403.542 2219.733 2541.039 10.14_149.1573m/z 83.04818 117.9999 209.4179 136.6712 107.551 89.60443 130.831 155.5301 90.6141 120.433 138.8971 152.4787 112.8426 123.1104 160.0562 111.2726 77.29476 51.02411 93.53782 77.14885 85.39383 145.1594 110.3921 154.8786 80.88183 131.866 120.366 119.5768 107.974 90.94678 90.45205 70.81055 95.52682 94.16299 95.20745 117.6447 72.99337 151.9495 163.5695 107.8544 108.7765 124.9452 116.4252 120.8374 104.0035 138.3517 91.03912 90.28761 113.5233 93.49947 96.33559 105.2078 122.1889 134.7652 87.6181 59.72999 111.6525 65.08738 97.35256 109.0179 124.3797 77.64065 113.1001 59.38126 90.72722 97.09165 101.9258 80.7661 86.56983 122.9409 96.38579 79.07612 126.3715 92.82256 117.8849 109.7118 88.79953 103.5929 105.6353 119.207 104.3639 121.9373 10.14_121.0293m/z 252.3376 331.8544 437.7222 362.7664 307.5365 254.737 313.7629 490.042 222.1853 421.7421 375.3997 414.7104 354.7645 244.3537 416.7426 412.6456 245.9983 164.1776 313.1112 260.8193 250.9879 425.3433 305.1264 407.0114 280.0047 372.3067 355.5113 321.2406 333.3156 266.5324 257.1487 228.0078 288.9406 290.9289 356.2954 347.4004 282.9289 384.7892 377.9868 409.7121 341.0418 408.8177 300.9653 416.9955 314.1553 361.1327 321.3202 236.5872 376.6356 269.7027 312.8286 271.7109 399.621 336.5849 277.3505 196.1496 353.5365 243.6355 237.9497 296.9648 314.8036 208.3967 307.6139 268.2632 264.2636 334.4486 320.5443 226.2243 262.2747 346.9541 262.8861 243.7082 365.8664 264.9141 374.6898 392.1129 282.8949 326.0455 277.0558 327.5737 332.7689 367.81 10.14_93.0329m/z 111.324 180.9335 232.0201 207.1987 162.9855 131.8358 154.7053 208.017 179.2747 240.1883 165.7111 169.495 177.4601 187.6274 231.5007 215.452 94.00577 122.4268 163.8313 150.8937 151.8209 216.4626 176.8179 231.9976 142.0944 175.8671 197.2008 222.3087 143.5908 147.8948 129.8745 136.8679 134.9071 114.7995 176.131 208.7513 137.0105 203.869 205.2237 209.1808 185.8661 211.9587 196.028 167.7466 191.893 183.9118 133.9511 128.2927 158.9126 122.8869 157.9736 150.4018 225.6817 173.1642 159.867 79.38843 206.4812 146.035 135.2179 101.8561 191.6801 100.4601 171.3591 128.0158 134.4281 144.9899 173.9245 124.586 147.6847 181.4232 119.6359 136.6256 165.3124 112.5963 238.5634 164.6535 156.0379 151.8263 141.3663 214.4754 182.5317 153.4349 10.14_65.0377m/z 90.52606 136.7911 128.563 107.0127 98.76987 76.75793 94.71835 118.7114 111.4589 133.4497 101.1105 74.93092 40.10981 110.2117 127.5467 89.44176 61.42147 45.27953 85.72133 45.37736 78.91277 85.8271 106.0774 140.6738 78.60439 79.37856 117.6867 101.0492 120.0033 101.2766 84.9333 85.55338 112.7094 89.47153 100.5925 114.8673 84.66094 151.5519 140.4657 94.50643 93.95037 92.90724 127.144 96.81455 86.26462 122.88 72.56902 89.8559 91.15125 97.798 86.97375 96.8047 99.63383 94.59569 81.21838 64.88712 115.3028 78.25963 53.06285 99.65546 139.1646 103.5945 76.20514 41.28623 75.28665 111.1444 92.28584 66.5301 79.35048 86.20376 85.1074 86.75069 144.4394 108.8315 107.7722 110.5424 81.11032 117.0069 69.37535 108.1423 97.96339 76.40189 10.21_105.0341m/z 81.79678 65.64767 116.3709 178.6137 177.3381 143.679 150.796 142.219 15.7257 121.4559 140.785 62.87037 103.1809 34.0689 119.1717 13.01247 115.7326 93.72767 58.99571 115.1418 181.8333 122.0905 66.66654 16.66444 100.9456 155.1978 69.60104 191.0906 107.2925 72.76902 176.6128 95.50873 44.70323 106.6336 98.99109 54.25974 99.53576 90.92872 155.6789 73.30064 101.8952 126.5229 71.96235 166.3235 138.5196 153.0012 45.56627 138.1186 118.4977 10.33_215.1263m/z 88.95578 89.83383 85.59935 201.8775 113.5283 109.6193 128.0083 108.1607 73.95709 69.08714 38.03259 70.56462 181.942 29.4654 57.9821 97.47325 124.1074 139.329 41.88681 45.99792 33.07016 193.7777 47.92835 30.63993 126.4498 140.5042 67.94419 89.45331 70.7674 68.94515 63.25307 68.67239 60.23431 77.98824 41.61708 47.17649 108.7066 73.76331 34.01742 97.11002 88.06571 138.8435 46.17961 10.49_98.9622m/z 753.3563 846.6685 1213.476 971.2864 841.1677 910.1693 1040.764 1111.492 904.4282 1091.629 902.5231 1166.512 1102.567 734.2083 1247.441 1123.22 838.0184 786.8043 998.1356 784.6606 801.4441 1288.944 950.8062 1013.256 903.8104 955.8846 950.8381 902.0306 970.5915 691.678 926.8286 814.8737 727.0844 671.8332 869.1007 876.2181 885.5143 1072.527 1078.709 1086.631 860.3046 1188.535 958.4125 935.3231 923.899 1027.404 1224.183 697.3955 956.8607 774.3928 809.517 813.1769 1105.223 979.3344 958.2047 427.1 1202.099 651.9607 647.4767 663.2919 909.9479 724.7495 949.4138 776.9609 776.2301 696.4287 948.5298 523.7933 671.2862 844.5187 589.4322 693.404 1004.975 815.4396 1054.015 1004.643 730.1707 913.5815 775.9878 1053.048 810.9684 852.5632 5.63_212.1654m/z 2978.17 4533.918 1618.063 2891.631 2560.192 3794.331 5404.583 2231.867 2490.889 3108.974 2749.791 3251.508 3347.982 2491.026 3159.335 2912.232 2887.709 2929.14 3021.636 2331.808 2762.418 5241.099 5277.938 4758.101 1270.964 1376.586 1514.149 1378.389 1318.974 1088.164 2019.143 1901.428 1724.035 757.051 852.6504 896.049 621.9843 844.2258 773.2092 1252.398 1268.665 994.558 525.6504 405.3517 476.571 465.7532 683.2729 585.6474 835.0481 798.2776 659.6682 780.9544 888.8552 658.8737 869.1577 551.779 571.543 649.5825 502.4265 588.9415 642.454 623.7062 715.0625 714.7568 382.9709 366.0038 533.9189 495.0587 689.361 379.5134 375.407 423.3633 554.1995 485.139 596.0257 535.1469 358.0123 556.9972 459.3804 681.9794 438.2105 401.6528 5.63_252.1583m/z 2092.545 3024.609 1455.148 2328.304 1658.751 2466.958 3892.197 1878.19 1878.48 2036.327 2044.023 2588.289 2649.679 722.6801 2610.698 2386.381 2322.001 2061.335 2347.789 1696.041 2057.771 3699.649 3618.373 3085.653 1041.486 1155.838 1187.39 965.5122 923.5105 864.5845 1461.009 1482.603 1413.401 515.7018 754.5693 702.5885 602.1301 809.6707 563.9121 1049.899 1082.074 825.7002 399.8538 386.6225 442.3802 378.4761 608.5303 388.9007 638.6584 707.5833 525.4163 630.3152 922.6602 646.6688 708.8618 151.3558 608.3929 396.4459 460.7109 403.539 634.7703 422.8165 676.6868 382.6219 293.7052 283.5761 364.1894 363.1625 487.5809 323.7453 344.2755 323.7661 494.8951 347.4622 416.4727 366.0448 262.8639 473.7565 366.7212 477.5677 374.5953 351.6099 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name retention index quantified m/z PubChem ID KEGG ID 1.02_152.0579m/z 1.02 152.0579 1.02_118.0865m/z 1.02 118.0865 1.02_208.9870m/z 1.02 208.987 1.02_113.9647m/z 1.02 113.9647 1.02_72.9373m/z 1.02 72.9373 1.03_136.0628m/z 1.03 136.0628 1.09_224.1289m/z 1.09 224.1289 1.09_160.1343m/z 1.09 160.1343 1.10_121.0404m/z 1.1 121.0404 1.12_137.0468m/z 1.12 137.0468 1.12_124.0402m/z 1.12 124.0402 1.15_123.0812m/z 1.15 123.0812 1.20_102.1278m/z 1.2 102.1278 1.20_133.0327m/z 1.2 133.0327 1.23_153.0418m/z 1.23 153.0418 1.23_136.0149m/z 1.23 136.0149 1.23_110.0357m/z 1.23 110.0357 1.26_229.1554m/z 1.26 229.1554 1.29_160.1340m/z 1.29 160.134 1.34_204.1356m/z 1.34 204.1356 1.34_150.0789m/z 1.34 150.0789 1.38_230.1865m/z 1.38 230.1865 1.44_119.0500m/z 1.44 119.05 1.44_136.0766m/z 1.44 136.0766 1.44_123.0450m/z 1.44 123.045 1.44_91.0542m/z 1.44 91.0542 1.44_164.0483n 1.44 165.0557 164.0483 1.44_118.0866m/z 1.44 118.0866 1.44_136.2041m/z 1.44 136.2041 1.44_119.1698m/z 1.44 119.1698 1.44_123.1670m/z 1.44 123.167 1.44_94.0415n 1.44 95.0492 94.0415 1.44_91.1588m/z 1.44 91.1588 1.50_204.1349m/z 1.5 204.1349 1.52_491.2482m/z 1.52 491.2482 1.56_94.0650m/z 1.56 94.065 1.68_132.1025m/z 1.68 132.1025 1.69_267.1348m/z 1.69 267.1348 1.70_252.9765m/z 1.7 252.9765 1.71_118.0860m/z 1.71 118.086 1.77_86.0597m/z 1.77 86.0597 1.77_141.9961m/z 1.77 141.9961 1.77_185.9861m/z 1.77 185.9861 1.83_132.1026m/z 1.83 132.1026 1.85_127.0507m/z 1.85 127.0507 1.94_215.1398m/z 1.94 215.1398 1.94_70.0645m/z 1.94 70.0645 1.98_145.0507m/z 1.98 145.0507 1.98_85.0281m/z 1.98 85.0281 1.98_149.0461m/z 1.98 149.0461 1.99_253.0835m/z 1.99 253.0835 2.01_190.0624m/z 2.01 190.0624 2.09_180.0903m/z 2.09 180.0903 2.10_72.0798m/z 2.1 72.0798 2.11_200.0537m/z 2.11 200.0537 2.12_224.0798m/z 2.12 224.0798 2.13_434.1908m/z 2.13 434.1908 2.13_173.0799m/z 2.13 173.0799 2.13_434.4167m/z 2.13 434.4167 2.14_195.0779m/z 2.14 195.0779 2.16_70.0647m/z 2.16 70.0647 2.17_362.9668m/z 2.17 362.9668 2.17_375.1171n 2.17 358.1138 375.1171 2.18_174.1497m/z 2.18 174.1497 2.18_179.0346m/z 2.18 179.0346 2.20_122.0972m/z 2.2 122.0972 2.20_310.1765m/z 2.2 310.1765 2.23_224.0775m/z 2.23 224.0775 2.25_72.0800m/z 2.25 72.08 2.25_217.1553m/z 2.25 217.1553 2.25_118.1236m/z 2.25 118.1236 2.28_167.0225n 2.28 168.0303 167.0225 2.28_161.0488n 2.28 162.0559 161.0488 2.28_133.0540n 2.28 116.0507 133.054 2.28_162.1938m/z 2.28 162.1938 2.28_164.0376m/z 2.28 164.0376 2.28_122.0247m/z 2.28 122.0247 2.28_246.0385m/z 2.28 246.0385 2.32_127.0505m/z 2.32 127.0505 2.34_186.1131m/z 2.34 186.1131 2.35_114.0659m/z 2.35 114.0659 2.36_146.0608m/z 2.36 146.0608 2.36_72.0796m/z 2.36 72.0796 2.38_120.0824m/z 2.38 120.0824 2.38_103.0546m/z 2.38 103.0546 2.38_166.0872m/z 2.38 166.0872 2.38_77.0382m/z 2.38 77.0382 2.38_120.1995m/z 2.38 120.1995 2.38_93.0696m/z 2.38 93.0696 2.38_91.0541m/z 2.38 91.0541 2.38_148.0538n 2.38 131.0505 148.0538 2.38_103.1647m/z 2.38 103.1647 2.38_198.0769m/z 2.38 198.0769 2.38_79.0535m/z 2.38 79.0535 2.38_118.0674m/z 2.38 118.0674 2.38_166.2278m/z 2.38 166.2278 2.38_77.1345m/z 2.38 77.1345 2.38_121.1367m/z 2.38 121.1367 2.38_93.1763m/z 2.38 93.1763 2.38_107.0497m/z 2.38 107.0497 2.38_138.0560m/z 2.38 138.056 2.39_93.0182m/z 2.39 93.0182 2.39_269.1149m/z 2.39 269.1149 2.42_310.1293m/z 2.42 310.1293 2.42_175.0875m/z 2.42 175.0875 2.44_268.0795m/z 2.44 268.0795 2.44_147.0600m/z 2.44 147.06 2.45_297.1453m/z 2.45 297.1453 2.45_252.0740m/z 2.45 252.074 2.48_200.1291m/z 2.48 200.1291 2.48_281.1501m/z 2.48 281.1501 2.50_233.1501m/z 2.5 233.1501 2.50_298.0926m/z 2.5 298.0926 2.50_142.0872m/z 2.5 142.0872 2.51_132.1022m/z 2.51 132.1022 2.52_196.0973m/z 2.52 196.0973 2.52_279.1343m/z 2.52 279.1343 2.53_197.1288m/z 2.53 197.1288 2.54_217.1052m/z 2.54 217.1052 2.54_89.0595m/z 2.54 89.0595 2.54_195.1229m/z 2.54 195.1229 2.54_84.0440m/z 2.54 84.044 2.55_219.1116n 2.55 220.1191 219.1116 2.55_201.1010n 2.55 184.0977 201.101 2.55_116.0713m/z 2.55 116.0713 2.55_166.0871m/z 2.55 166.0871 2.55_144.0810m/z 2.55 144.081 2.55_257.1614m/z 2.55 257.1614 2.55_201.1239m/z 2.55 201.1239 2.55_226.1070m/z 2.55 226.107 2.55_181.0509m/z 2.55 181.0509 2.56_143.1190m/z 2.56 143.119 2.58_162.0559m/z 2.58 162.0559 2.58_212.0925m/z 2.58 212.0925 2.59_251.1009m/z 2.59 251.1009 2.60_267.0754m/z 2.6 267.0754 2.63_132.1025m/z 2.63 132.1025 2.63_133.0328m/z 2.63 133.0328 2.63_167.0465m/z 2.63 167.0465 2.63_185.1654m/z 2.63 185.1654 2.63_72.0795m/z 2.63 72.0795 2.63_143.1178m/z 2.63 143.1178 2.64_229.1556m/z 2.64 229.1556 2.64_181.0500m/z 2.64 181.05 2.66_188.0720m/z 2.66 188.072 2.66_146.0608m/z 2.66 146.0608 2.68_201.1240m/z 2.68 201.124 2.68_197.1289m/z 2.68 197.1289 2.68_243.1344m/z 2.68 243.1344 2.68_141.1030m/z 2.68 141.103 2.68_86.0960m/z 2.68 86.096 2.68_84.0443m/z 2.68 84.0443 2.69_270.1345m/z 2.69 270.1345 2.69_231.1714m/z 2.69 231.1714 2.69_72.0798m/z 2.69 72.0798 2.69_213.1598m/z 2.69 213.1598 2.69_185.1659m/z 2.69 185.1659 2.74_344.1246n 2.74 327.1213 344.1246 2.74_311.1607m/z 2.74 311.1607 2.74_207.0417m/z 2.74 207.0417 2.78_145.0536n 2.78 146.061 145.0536 2.77_267.1712m/z 2.77 267.1712 2.77_250.1448m/z 2.77 250.1448 2.77_277.1760m/z 2.77 277.176 2.79_405.1157m/z 2.79 405.1157 2.79_389.1423m/z 2.79 389.1423 2.79_326.0387m/z 2.79 326.0387 2.82_229.1557m/z 2.82 229.1557 2.82_206.0459m/z 2.82 206.0459 2.82_237.1611m/z 2.82 237.1611 2.83_259.1776m/z 2.83 259.1776 2.83_304.1757m/z 2.83 304.1757 2.84_238.1419n 2.84 261.1313 238.1419 2.84_133.0333m/z 2.84 133.0333 2.84_72.0800m/z 2.84 72.08 2.86_375.1166n 2.86 358.1133 375.1166 2.86_315.0965n 2.86 316.1033 315.0965 2.86_238.0716m/z 2.86 238.0716 2.86_181.0502m/z 2.86 181.0502 2.86_179.0357m/z 2.86 179.0357 2.87_145.9363m/z 2.87 145.9363 2.87_187.0646n 2.87 188.0732 187.0646 2.87_146.0618m/z 2.87 146.0618 2.87_118.0661m/z 2.87 118.0661 2.87_144.0816m/z 2.87 144.0816 2.87_159.0925m/z 2.87 159.0925 2.87_132.0816m/z 2.87 132.0816 2.87_188.2193m/z 2.87 188.2193 2.87_146.1917m/z 2.87 146.1917 2.87_205.0983m/z 2.87 205.0983 2.87_119.0740n 2.87 142.0659 119.074 2.87_144.0431m/z 2.87 144.0431 2.87_118.1853m/z 2.87 118.1853 2.87_144.2130m/z 2.87 144.213 2.87_145.9961m/z 2.87 145.9961 2.87_117.0641m/z 2.87 117.0641 2.87_91.0540m/z 2.87 91.054 2.87_116.0711m/z 2.87 116.0711 2.87_130.0658m/z 2.87 130.0658 2.87_115.0546m/z 2.87 115.0546 2.87_146.2600m/z 2.87 146.26 2.88_354.2027m/z 2.88 354.2027 2.89_240.1240m/z 2.89 240.124 2.90_171.1265n 2.9 154.1232 171.1265 2.90_281.1008m/z 2.9 281.1008 2.92_197.1294m/z 2.92 197.1294 2.93_281.1865m/z 2.93 281.1865 2.93_98.0603m/z 2.93 98.0603 2.93_156.1028m/z 2.93 156.1028 2.93_86.0595m/z 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