#METABOLOMICS WORKBENCH Tomoya_Tsukimi_20170309_001205 DATATRACK_ID:861 STUDY_ID:ST000570 ANALYSIS_ID:AN000878 PROJECT_ID:PR000418 VERSION 1 CREATED_ON March 13, 2017, 11:57 am #PROJECT PR:PROJECT_TITLE Metabolome analysis of the cecal contents of GF mice and GF mice colonized with PR:PROJECT_TITLE dominant gut microbes present in the ceca of neonatal and adult mice PR:PROJECT_SUMMARY The high susceptibility of neonates to infections has been assumed to be due to PR:PROJECT_SUMMARY immaturity of the immune system, but the mechanism remains unclear. By PR:PROJECT_SUMMARY colonizing adult germ-free mice with the cecal contents of neonatal and adult PR:PROJECT_SUMMARY mice, we show that the neonatal microbiota is impaired in mediating colonization PR:PROJECT_SUMMARY resistance against two major pathogens causing mortality in neonates. The lack PR:PROJECT_SUMMARY of colonization resistance was caused by the absence of Clostridiales in the PR:PROJECT_SUMMARY neonatal microbiota. Administration of Clostridiales, but not Bacteroidales, PR:PROJECT_SUMMARY restored colonization resistance and abrogated intestinal pathology upon PR:PROJECT_SUMMARY pathogen challenge. Conversely, depletion of Clostridiales abolished PR:PROJECT_SUMMARY colonization resistance in adult mice. Furthermore, intragastric administration PR:PROJECT_SUMMARY of Clostridiales protected neonatal mice from pathogen infection. The neonatal PR:PROJECT_SUMMARY bacteria enhanced the ability of these protective Clostridiales to colonize the PR:PROJECT_SUMMARY gut. These results identify the gut microbiota as a critical determinant of PR:PROJECT_SUMMARY increased susceptibility to enteric infection during the neonatal period. PR:INSTITUTE Keio University PR:DEPARTMENT Institute for Advanced Biosciences PR:LAST_NAME Fukuda PR:FIRST_NAME Shinji PR:ADDRESS Tsuruoka, Yamagata 997-0052, Japan PR:EMAIL sfukuda@sfc.keio.ac.jp PR:PHONE +81-235-29-0528 #STUDY ST:STUDY_TITLE Metabolome analysis of the cecal contents of GF mice and GF mice colonized with ST:STUDY_TITLE dominant gut microbes present in the ceca of neonatal and adult mice ST:STUDY_SUMMARY Metabolome profiles of GF or GF mice reconstituted with Esherichia coli (EC), ST:STUDY_SUMMARY Bacteroides acidifaciens (Bac), or Clostridia consortium (CL) were compared. ST:INSTITUTE Keio University ST:DEPARTMENT Institute for Advanced Biosciences ST:LAST_NAME Fukuda ST:FIRST_NAME Shinji ST:ADDRESS Tsuruoka, Yamagata 997-0052, Japan ST:EMAIL sfukuda@sfc.keio.ac.jp ST:PHONE +81-235-29-0528 ST:NUM_GROUPS 4 ST:TOTAL_SUBJECTS 17 #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 - 1_B1 colonized bacteria:Bacteroides acidifaciens SUBJECT_SAMPLE_FACTORS - 1_B2 colonized bacteria:Bacteroides acidifaciens SUBJECT_SAMPLE_FACTORS - 1_B3 colonized bacteria:Bacteroides acidifaciens SUBJECT_SAMPLE_FACTORS - 1_B4 colonized bacteria:Bacteroides acidifaciens SUBJECT_SAMPLE_FACTORS - 1_B5 colonized bacteria:Bacteroides acidifaciens SUBJECT_SAMPLE_FACTORS - 1_CL1 colonized bacteria:Clostridia SUBJECT_SAMPLE_FACTORS - 1_CL2 colonized bacteria:Clostridia SUBJECT_SAMPLE_FACTORS - 1_CL3 colonized bacteria:Clostridia SUBJECT_SAMPLE_FACTORS - 1_CL4 colonized bacteria:Clostridia SUBJECT_SAMPLE_FACTORS - 1_CL5 colonized bacteria:Clostridia SUBJECT_SAMPLE_FACTORS - 2_EB1 colonized bacteria:Esherichia coli SUBJECT_SAMPLE_FACTORS - 2_EB2 colonized bacteria:Esherichia coli SUBJECT_SAMPLE_FACTORS - 2_EB3 colonized bacteria:Esherichia coli SUBJECT_SAMPLE_FACTORS - 5_C1 colonized bacteria:none (GF) SUBJECT_SAMPLE_FACTORS - 5_C2 colonized bacteria:none (GF) SUBJECT_SAMPLE_FACTORS - 5_C3 colonized bacteria:none (GF) SUBJECT_SAMPLE_FACTORS - 5_C4 colonized bacteria:none (GF) #COLLECTION CO:COLLECTION_SUMMARY Feces of GF or GF mice reconstituted with Esherichia coli (EC), Bacteroides CO:COLLECTION_SUMMARY acidifaciens (Bac), or Clostridia consortium (CL) CO:SAMPLE_TYPE Feces CO:STORAGE_CONDITIONS -80℃ #TREATMENT TR:TREATMENT_SUMMARY Animals Specific-pathogen-free (SPF) C57BL/6 mice were originally purchased from TR:TREATMENT_SUMMARY Jackson Laboratories. Wild-type mice on GF background were bred and maintained TR:TREATMENT_SUMMARY at the Germ-free Animal Core Facility of the University of Michigan. GF mice TR:TREATMENT_SUMMARY were maintained in flexible film isolators and were checked weekly for germ-free TR:TREATMENT_SUMMARY status by aerobic and anaerobic culture. The absence of microbiota was verified TR:TREATMENT_SUMMARY by microscopic analysis of stained cecal contents to detect unculturable TR:TREATMENT_SUMMARY contamination. All animal studies were performed according to approved protocols TR:TREATMENT_SUMMARY by the University of Michigan Committee on the Use and Care of Animals. TR:TREATMENT_SUMMARY Reconstitution of GF mice with cecal contents and defined bacteria Cecal TR:TREATMENT_SUMMARY contents from single neonatal and 7-week old SPF mice were diluted into 50 ml TR:TREATMENT_SUMMARY (per one adult mouse) or 5 ml (per one 4, 12, 16-day old mouse) of PBS, and TR:TREATMENT_SUMMARY passed through a 70 μm cell strainer to eliminate clumps and debris under TR:TREATMENT_SUMMARY sterile conditions) under an atmosphere of 5% H2, 5% CO2 and 90% N2 in an TR:TREATMENT_SUMMARY anaerobic growth chamber (Coy Manufacturing, Grass Lake, MI). Then 200 μl of TR:TREATMENT_SUMMARY the suspension was processed by intragastric gavage into each of the recipient TR:TREATMENT_SUMMARY GF mice. Bacteroides species were anaerobically grown at 37°C overnight in TR:TREATMENT_SUMMARY pre-reduced chopped-meat broth (Anaerobe systems, Morgan Hill, CA). E. coli were TR:TREATMENT_SUMMARY grown aerobically overnight in LB broth with shaking. TR:TREATMENT_PROTOCOL_FILENAME Treatment_Protocol.pdf TR:TREATMENT Biotic TR:TREATMENT_COMPOUND Suspension of diluted cecal contents TR:TREATMENT_ROUTE Intragastric gavage TR:TREATMENT_DOSE 200 μl #SAMPLEPREP SP:SAMPLEPREP_SUMMARY CE-TOFMS-based metabolome analysis(sample preparation) Fecal samples were freeze SP:SAMPLEPREP_SUMMARY dried, and disrupted by vigorous shaking at 1,500 rpm for 10 min with four 3 mm SP:SAMPLEPREP_SUMMARY zirconia beads by Shake Master NEO (Bio Medical Science Inc.). 10 mg (±0.5 mg) SP:SAMPLEPREP_SUMMARY fecal samples were homogenized with 500 μl MeOH containing internal standards SP:SAMPLEPREP_SUMMARY (20 μM each of methionine sulfone, and D-camphor-10-sulfonic acid (CSA)) and SP:SAMPLEPREP_SUMMARY 100 mg of 0.1 mm and four of 3 mm zirconia/silica beads (BioSpec Products). SP:SAMPLEPREP_SUMMARY After vigorous shaking (1,500 rpm for 5 min) by Shake Master NEO (Bio Medical SP:SAMPLEPREP_SUMMARY Science Inc.), 200 μl of Milli-Q water and 500 μl of chloroform was added and SP:SAMPLEPREP_SUMMARY then shaking in a same manner as before. After centrifugation at 4,600 × g for SP:SAMPLEPREP_SUMMARY 15 min at 4°C, the supernatant was transferred to a 5kDa cutoff centrifugal SP:SAMPLEPREP_SUMMARY filter tube. The filtrate was centrifugally concentrated at 40°C and SP:SAMPLEPREP_SUMMARY reconstituted with 40 μl of Milli-Q water. SP:SAMPLEPREP_PROTOCOL_FILENAME Sample_Prep_Protocol.pdf SP:PROCESSING_METHOD Homogenized with 500 μl MeOH containing internal standards and 100 mg of 0.1 mm SP:PROCESSING_METHOD and four of 3 mm zirconia/silica beads SP:PROCESSING_STORAGE_CONDITIONS On ice SP:EXTRACTION_METHOD Chloroform SP:EXTRACT_CLEANUP 5kDa cutoff centrifugal filter tube SP:SAMPLE_RESUSPENSION 40 μl of Milli-Q water #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE CE CH:INSTRUMENT_NAME Agilent CE CH:COLUMN_NAME COSMO(+), i.d.50 µm(anion), Fused-silica, i.d. 50 µm(cation) CH:INTERNAL_STANDARD 20 μM each of methionine sulfone, and D-camphor-10-sulfonic acid (CSA) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:MS_COMMENTS - MS:INSTRUMENT_NAME Agilent CE-TOFMS MS:INSTRUMENT_TYPE TOF MS:MS_TYPE ESI MS:ION_MODE NEGATIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS nmol/g MS_METABOLITE_DATA_START Samples 1_B1 1_B2 1_B3 1_B4 1_B5 1_CL1 1_CL2 1_CL3 1_CL4 1_CL5 2_EB1 2_EB2 2_EB3 5_C1 5_C2 5_C3 5_C4 Factors colonized bacteria:Bacteroides acidifaciens colonized bacteria:Bacteroides acidifaciens colonized bacteria:Bacteroides acidifaciens colonized bacteria:Bacteroides acidifaciens colonized bacteria:Bacteroides acidifaciens colonized bacteria:Clostridia colonized bacteria:Clostridia colonized bacteria:Clostridia colonized bacteria:Clostridia colonized bacteria:Clostridia colonized bacteria:Esherichia coli colonized bacteria:Esherichia coli colonized bacteria:Esherichia coli colonized bacteria:none (GF) colonized bacteria:none (GF) colonized bacteria:none (GF) colonized bacteria:none (GF) Acetylcholine 2 1 1 1 0 3 0 0 1 0 1 1 1 1 1 0 0 Adenine 18 41 109 86 31 20 28 15 18 11 2 2 2 1 2 2 2 Adenosine 10 10 15 15 16 23 18 10 15 7 3 5 3 3 4 2 3 ADMA 50 83 57 117 57 76 92 69 85 54 41 50 40 151 196 94 84 Agmatine 4 7 6 7 5 21 20 5 16 7 27 16 18 1 2 2 1 Ala 582 597 826 788 711 3057 2863 2129 2673 2780 2058 1537 4591 1162 1004 1127 4027 Ala-Ala 17 19 28 32 22 304 419 110 259 256 39 51 25 6 19 10 12 Allantoin 2968 4558 2010 3607 2558 592 993 3427 1391 721 595 393 770 4811 3546 2279 1373 alpha-Aminoadipate 12 13 13 14 11 26 36 15 30 22 6 5 3 17 27 11 21 Alpha-Methylserine 6 14 10 16 14 21 21 14 20 11 9 9 6 7 6 5 Anserine 12 9 9 11 6 Anthranilate 1 1 1 0 Arg 1114 1045 1018 1512 821 4287 4741 3113 3442 2811 4780 5484 5640 2888 7669 3011 4201 Argininosuccinate 41 53 31 63 40 9 6 14 9 2 26 28 28 70 94 50 45 Asn 69 198 195 174 255 1281 1475 669 909 923 119 201 321 3045 8219 4156 4958 Asp 1051 948 1360 1306 1490 3821 4322 1713 2849 2334 1026 1422 1148 1677 2256 1282 3812 beta-Ala 14 25 16 23 17 329 271 163 290 173 18 11 27 38 37 31 51 beta-Imidazolelactate 5 10 8 11 9 3 7 13 4 3 17 20 11 8 7 6 6 Betaine 89 92 72 136 88 930 66 178 643 78 99 130 126 138 305 149 171 Betaine aldehyde 24 44 35 22 27 118 97 70 162 77 30 38 35 44 37 22 23 Betonicine 7 13 6 19 7 4 2 7 8 2 11 19 16 10 14 10 14 Cadaverine 3 6 45 8 11 5 5 26 8 10 2929 3690 2723 6 17 22 22 Carnitine 29 49 35 60 38 14 7 17 24 11 7 15 8 78 144 86 83 Carnosine 1 1 1 1 1 2 3 5 3 2 2 2 3 3 6 4 4 Choline 1085 1111 1213 1167 994 607 172 243 358 221 2148 3436 3119 712 1073 877 684 Citrulline 68 71 64 99 70 35 49 61 36 50 367 480 324 143 491 188 244 Creatine 304 1343 206 521 431 812 406 804 891 568 433 476 454 1873 569 1801 1800 Creatinine 631 1141 588 1208 725 1133 1712 1815 1202 1177 491 728 719 2865 3258 1727 1506 Cyclohexylamine 5 11 6 8 5 46 17 50 173 31 6 6 5 24 36 54 12 Cys 3 2 Cystathionine 16 16 13 24 15 14 4 7 8 5 6 10 9 8 Cysteine-glutathione disulphide -Divalent 7 5 5 Cystine 3 5 2 5 3 3 3 5 4 2 42 42 26 415 663 466 396 Cytidine 14 9 9 11 16 64 61 32 88 93 4 7 7 111 110 167 190 Cytosine 8 23 12 17 13 6 4 8 12 6 2 5 13 15 21 14 12 Diethanolamine 47 66 84 125 61 71 42 95 70 88 41 55 125 40 65 43 105 DOPA 2 2 4 4 4 4 5 2 4 6 15 21 14 2 8 4 2 Dopamine 2 2 1 2 2 2 2 2 3 4 2 2 1 Ectoine 2 9 1 6 2 6 6 9 10 5 2 3 3 49 59 51 39 GABA 17 29 23 41 28 265 182 319 222 196 38 56 46 52 67 65 60 gamma-Butyrobetaine 5 5 6 6 5 99 57 46 107 54 60 80 37 12 17 8 11 gamma-Guanidinobutyrate 25 153 34 114 72 48 44 149 83 45 76 67 152 168 171 133 184 Gln 362 490 529 416 553 1414 1119 864 1137 1438 5086 7290 7052 3899 13749 4496 6763 Glu 15569 15127 15968 22102 15733 20777 18200 5842 16067 9430 21934 26820 22379 9236 20994 8726 13117 Glucosaminate 8 8 9 10 8 1 2 2 2 2 5 8 6 7 10 7 7 Glucosamine 40 77 63 67 62 136 138 79 118 128 71 75 73 404 810 473 499 Glu-Glu 6 5 7 8 7 6 5 5 6 4 16 21 50 3 15 4 5 Glutathione(ox) 1 5 5 3 1 2 1 0 Gly 405 478 567 557 626 1670 2203 1210 1534 1286 1377 1113 3513 1653 1252 1641 5793 Glycerophosphorylcholine 43 39 41 60 37 72 19 36 41 16 16 22 16 54 60 21 20 Gly-Gly 14 14 19 18 14 7 8 13 7 11 17 15 36 11 14 13 20 Gly-Leu 25 49 32 49 40 95 93 107 103 83 41 35 61 101 142 87 89 Guanidinoacetate 31 204 5 276 89 18 47 215 126 29 41 20 89 1561 1228 583 490 Guanidinosuccinate 8 17 5 12 9 3 5 10 3 4 5 6 4 39 33 17 12 Guanine 47 40 53 55 55 70 57 70 78 46 41 33 38 89 103 112 124 Guanosine 17 14 18 22 24 61 41 21 48 38 9 9 11 239 320 243 277 His 298 260 322 327 270 552 625 223 351 375 3001 3561 2917 547 2145 795 1460 Histamine 1 3 1 1 1 7 9 5 10 6 6 6 5 11 32 4 8 Histidinol 1 1 1 1 1 1 1 0 1 1 0 0 0 0 Homocystine 2 6 6 4 Homoserine 28 25 43 37 27 107 113 38 96 61 7 4 7 7 11 12 10 Hydroxyproline 218 194 202 368 348 62 193 85 75 68 167 157 230 235 143 217 629 Hypotaurine 13 77 31 40 10 19 29 21 6 10 5 21 46 33 22 27 Hypoxanthine 19 15 23 20 26 144 108 53 212 235 4 6 5 74 74 67 65 Ile 133 182 172 162 185 1024 2736 2096 660 1857 3140 2995 3631 768 1485 846 2281 Imidazole-4-acetate 1 13 2 17 5 5 6 8 57 35 30 29 Indole-3-acetaldehyde 1869 2240 1846 1795 1810 1003 1244 1676 771 985 997 569 1014 1420 1319 866 567 Indole-3-acetamide 53 53 58 29 52 112 141 107 90 106 35 33 34 30 40 38 26 Indole-3-acetate 38 36 42 34 37 43 43 36 38 37 28 20 26 34 32 23 23 Inosine 103 40 99 108 141 306 393 131 537 739 39 59 29 275 343 202 287 Isoamylamine 1 1 2 4 2 2 Isopropanolamine 3 6 7 8 5 7 13 6 11 9 15 23 18 6 7 4 4 Kynurenine 2 4 2 3 3 3 4 Leu 241 365 271 329 316 2378 3577 3253 1669 2933 4072 3708 4829 1183 2204 1271 3065 Lys 1874 1893 1968 2742 1688 4558 5150 2629 3735 3007 2609 1524 2478 5389 12734 5483 7249 Mannosamine 98 123 146 140 106 30 46 93 46 36 133 197 146 142 287 155 187 Melamine 2 3 7 Met 168 174 226 267 161 863 923 573 616 563 1003 918 1059 248 546 313 568 Methionine sulfoxide 57 128 84 120 72 1044 736 566 810 472 95 123 153 173 320 221 412 Methylguanidine 8 38 17 8 16 14 22 32 9 4 11 39 56 27 21 N,N-Dimethylglycine 7 23 38 6 17 28 104 91 18 8 5 27 13 184 35 42 N1,N12-Diacetylspermine 16 18 10 20 12 N1,N8-Diacetylspermidine 1 1 1 1 1 103 99 82 84 64 8 6 6 1 2 1 1 N1-Acetylspermidine 13 26 17 21 17 64 72 25 71 45 46 40 43 44 63 24 19 N1-Acetylspermine 2 2 2 N6,N6,N6-Trimethyllysine 52 64 50 106 54 16 16 31 26 18 75 93 63 126 196 101 95 N8-Acetylspermidine 2 4 4 3 3 4 6 3 5 4 19 18 18 5 8 3 3 N-Acetylglucosamine 42 46 61 72 70 155 150 251 137 151 50 36 37 251 314 176 186 N-Acetylglucosylamine 64 72 27 10 10 N-Acetylhistidine 30 52 33 48 31 49 70 61 46 47 35 45 35 48 111 41 52 N-Acetylornithine 5 8 8 8 8 239 243 94 192 165 7 6 12 6 16 9 9 N-Acetylputrescine 16 26 27 30 25 570 722 452 501 663 223 173 165 36 41 29 30 N-Acetylvaline 16 69 10 61 34 24 53 151 64 41 17 16 22 183 225 117 101 N-alpha,N-alpha-Dimethylhistidine 2 2 3 2 2 2 3 2 2 4 3 4 N-epsilon-Acetyllysine 20 19 25 29 20 84 82 53 73 47 73 91 67 24 71 29 45 Nicotinamide 47 46 45 64 54 78 100 52 83 66 9 10 13 50 58 38 41 N-Methylalanine 2 3 2 Noradrenaline 5 5 6 10 5 9 9 10 8 8 6 7 5 10 16 8 10 o-Acetylcarnitine 9 7 7 7 7 2 1 2 1 1 2 2 2 9 16 9 10 O-Acetylserine 20 11 20 17 26 51 57 4 50 30 3 5 Ophthalmate 41 42 45 54 47 11 13 18 10 11 93 119 76 154 230 162 180 Ornithine 65 86 96 145 135 123 108 69 129 123 57 78 65 108 430 110 177 o-Succinylhomoserine 16 17 25 22 20 13 14 15 46 13 13 24 8 13 26 15 17 p-Aminobenzoate 2 1 3 3 3 3 3 1 Phe 352 416 434 459 329 1136 1582 1403 809 1310 2751 2662 3089 566 1590 596 1612 Phenethylamine 2 2 Phosphorylcholine 5 3 162 67 35 113 48 5 8 Pipecolate 24 101 26 88 44 118 82 153 150 70 22 22 30 176 174 89 73 Pro 850 970 1198 1491 1376 1432 1449 743 1287 1199 1338 1061 4398 990 502 1364 4582 Proline betaine 45 268 5 242 96 84 70 246 235 46 33 19 82 271 341 243 234 Pseudopelletierine 1 0 Pterin 5 5 2 4 Putrescine(1,4-Butanediamine) 8 19 87 21 26 33 57 84 59 39 361 415 273 39 54 26 17 Pyridoxal 8 11 5 12 11 23 21 21 20 30 6 7 6 9 12 9 13 Pyridoxamine 2 2 3 3 2 3 5 1 2 2 2 3 2 1 1 1 1 Pyridoxamine 5'-phosphate 5 4 15 19 9 8 2 Pyridoxine 1 0 1 1 1 3 2 2 2 1 1 1 1 1 1 1 1 Riboflavin 21 35 18 34 28 21 26 21 24 21 11 12 18 30 31 24 22 Saccharopine 8 9 9 16 7 10 15 10 8 7 20 28 23 21 SAH 2 2 4 3 2 3 4 1 2 2 1 1 1 2 1 1 SAM+ 12 21 23 23 16 119 199 17 113 82 7 8 6 22 18 8 7 Sarcosine 9 10 12 33 12 SDMA 8 16 9 22 13 9 10 14 13 7 5 5 5 35 35 17 15 Ser 527 488 696 674 727 751 692 824 494 553 100 138 271 2213 6400 3141 5056 Serotonin 7 6 6 5 4 7 5 6 5 4 8 6 5 5 4 3 3 Spermidine 15 39 36 39 26 40 62 7 50 21 10 8 7 9 27 10 9 Spermine 1 1 1 1 1 1 0 2 1 Taurine 895 3245 218 1234 2166 827 972 1451 985 690 122 76 184 1434 534 652 448 Thiamine 6 3 6 4 7 18 19 14 23 8 28 23 19 58 103 88 76 Thiamine monophosphate 8 4 8 9 9 5 11 5 5 4 3 4 Thr 241 287 306 212 495 1581 2044 982 1231 1459 213 336 490 7674 20437 8454 11562 Thymidine 217 185 244 233 201 262 266 176 208 206 166 179 147 332 482 322 362 Thymine 193 157 231 199 183 260 109 105 160 103 437 470 383 533 620 626 540 TRH 6 6 3 4 4 Trigonelline 10 116 1 86 23 59 29 105 117 20 10 6 28 263 365 253 189 Trimethylamine N-oxide 2 28 9 5 6 88 1 5 1 1 3 28 43 23 21 Tropinone 1 1 1 Trp 153 143 190 192 114 111 137 85 59 92 12 11 24 177 457 196 483 Tryptamine 1 2 1 1 Tyr 424 462 524 517 387 1311 1663 888 911 1438 4232 4558 4541 769 2713 868 2009 Tyramine 2 2 1 2 2 6 7 5 6 6 2 2 2 4 5 3 3 Uracil 48 123 94 60 902 1411 181 805 604 104 64 Uridine 279 300 294 299 299 297 363 226 339 345 204 191 194 652 673 443 407 Urocanate 166 191 161 263 138 19 48 57 29 57 27 33 39 39 51 43 75 Val 441 503 528 520 480 1905 4436 3372 1365 3310 5741 6466 6332 1401 3593 1622 3727 Xanthine 192 223 334 322 200 188 235 54 175 156 217 324 200 232 Xanthosine 6 5 4 6 7 12 13 11 11 8 3 2 4 23 29 16 21 1-Amino-1-cyclopentanecarboxylate 1 1 1-Aminocyclopropane-1-carboxylate 1 5 3 2 2 13 6 3 5 1 2 7 7 6 5 1-Methyladenosine 14 20 12 21 16 3 14 4 3 4 3 6 61 49 20 19 1-Methylhistamine 3 13 8 4 2 6 15 8 2 2 2 5 50 104 18 25 1-Methylnicotinamide 62 141 33 117 66 15 33 69 33 25 27 29 36 258 204 110 96 1,3-Diaminopropane 2 3 2 3 4 2 3 1 1 1 1 1 2,3-Diaminopropionate 2 2 2 2,4-Diaminobutyrate 33 53 7 33 25 2AB 18 20 41 37 26 96 69 58 93 56 251 245 200 7 14 10 17 2-Aminophenol 2 1 2 2 1 2 2 2 1 3 1 1 1 2'-Deoxycytidine 7 7 7 8 7 12 8 10 7 6 2 2 117 161 111 151 2'-Deoxyguanosine 8 8 9 12 8 34 21 16 12 11 18 14 7 205 302 180 208 2'-Deoxyinosine 313 366 280 231 3-Aminoisobutyrate 3 11 8 5 5 4 14 8 5 2 1 4 23 20 10 10 3-Aminopropane-1,2-diol 10 8 10 11 10 6 3-Methoxytyramine 1 3-Methylguanine 11 13 18 17 12 4 9 3 4 5 15 20 15 2 8 5 3-Methylhistidine 5 8 4 8 4 13 13 24 22 9 9 10 10 15 30 13 16 4-(beta-Acetylaminoethyl)imidazole 2 3 2 3 2 9 12 7 8 9 2 2 1 3 5 3 3 4-Hydroxymethylimidazole 2 2 2 4 6 5 5 4 1 5-Aminolevulinate 3 11 3 7 6 12 30 16 14 10 3 4 6 16 24 10 9 5-Aminovalerate 65 47 48 44 28441 32934 33696 24719 26729 108 59 162 38 38 38 74 5-Hydroxyindoleacetate 384 491 332 569 493 326 1077 1133 355 1120 69 57 70 166 129 97 114 5-Hydroxylysine 35 28 40 58 31 5 6 7 6 6 18 25 17 16 38 18 20 5-Methyl-2'-deoxycytidine 8 13 12 15 10 18 20 15 19 23 7 11 8 13 19 15 17 5-Methylcytosine 1 3 2 1 7 3 4 4 4 1 2 2 2 2 2 1 5-Methylthioadenosine 3 4 5 5 4 26 37 6 21 21 2 2 2 4 3 3 3 6-Hydroxynicotinate 6 7 22 27 16 16 13 7-Methylguanine 25 37 23 33 27 21 29 35 21 23 16 16 15 59 57 28 24 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name retention index quantified m/z PubChem ID KEGG ID RefMet name Acetylcholine C01996 Acetylcholine Adenine C00147 Adenine Adenosine C00212 Adenosine ADMA C03626 Dimethylarginine Agmatine C00179 Agmatine Ala C00041 Alanine Ala-Ala C00993 Ala-Ala Allantoin C01551 Allantoin alpha-Aminoadipate C00956 Aminoadipic acid Alpha-Methylserine C02115 Anserine C01262 Anserine Anthranilate C00108 2-Aminobenzoic acid Arg C00062 Arginine Argininosuccinate C03406 Argininosuccinic acid Asn C00152 Asparagine Asp C00049 Aspartic acid beta-Ala C00099 beta-Imidazolelactate C05568 Betaine C00719 Betaine Betaine aldehyde C00576 Betaine aldehyde Betonicine C08269 Betonicine Cadaverine C01672 Cadaverine Carnitine C00318 Carnitine Carnosine C00386 Carnosine Choline C00114 Choline Citrulline C00327 Citrulline Creatine C00300 Creatine Creatinine C00791 Creatinine Cyclohexylamine C00571 Cyclohexylamine Cys C00097 Cysteine Cystathionine C02291 Cystathionine Cysteine-glutathione disulphide -Divalent Cystine C00491 Cystine Cytidine C00475 Cytidine Cytosine C00380 Cytosine Diethanolamine C06772 Diethanolamine DOPA C00355 DOPA Dopamine C03758 Dopamine Ectoine C06231 Ectoine GABA C00334 Gamma-Aminobutyric acid gamma-Butyrobetaine C01181 3-Dehydroxycarnitine gamma-Guanidinobutyrate C01035 Gln C00064 Glutamine Glu C00025 Glutamic acid Glucosaminate C03752 Glucosaminic acid Glucosamine C00329 Glucosamine Glu-Glu C01425 Glu-Glu Glutathione(ox) C00127 Gly C00037 Glycine Glycerophosphorylcholine C00670 sn-glycero-3-phosphocholine Gly-Gly C02037 Gly-Gly Gly-Leu C02155 Gly-Leu Guanidinoacetate C00581 Guanidoacetic acid Guanidinosuccinate C03139 Guanidinosuccinic acid Guanine C00242 Guanine Guanosine C00387 Guanosine His C00135 Histidine Histamine C00388 Histamine Histidinol C00860 L-Histidinol Homocystine C01817 DL-Homocystine Homoserine C00263 Homoserine Hydroxyproline C01015 cis-3-hydroxyproline Hypotaurine C00519 Hypotaurine Hypoxanthine C00262 Hypoxanthine Ile C00407 Isoleucine Imidazole-4-acetate C02835 Indole-3-acetaldehyde C00637 Indoleacetaldehyde Indole-3-acetamide C02693 1H-Indole-3-acetamide Indole-3-acetate C00954 Indoleacetic acid Inosine C00294 Inosine Isoamylamine C02640 Isopropanolamine C05771 Kynurenine C00328 Kynurenine Leu C00123 Leucine Lys C00047 Lysine Mannosamine C03570 Melamine C08737 Melamine Met C00073 Methionine Methionine sulfoxide C02989 Methionine sulfoxide Methylguanidine C02294 Methylguanidine N,N-Dimethylglycine C01026 Dimethylglycine N1,N12-Diacetylspermine C03413 N1,N12-Diacetylspermine N1,N8-Diacetylspermidine N1-Acetylspermidine C00612 N1-Acetylspermidine N1-Acetylspermine C02567 N1-Acetylspermine N6,N6,N6-Trimethyllysine C03793 N8-Acetylspermidine C01029 N8-Acetylspermidine N-Acetylglucosamine C00140 N-acetyl-D-glucosamine N-Acetylglucosylamine C01239 N-Acetylhistidine C02997 N-acetylhistidine N-Acetylornithine C00437 N-Acetylornithine N-Acetylputrescine C02714 N-Acetylputrescine N-Acetylvaline N-Acetylvaline N-alpha,N-alpha-Dimethylhistidine C04259 N-epsilon-Acetyllysine C02727 Nicotinamide C00153 Niacinamide N-Methylalanine C02721 N-methylalanine Noradrenaline C00547 Xi-Norepinephrine o-Acetylcarnitine C02571 CAR(2:0) O-Acetylserine C00979 O-Acetylserine Ophthalmate Ophthalmic acid Ornithine C00077 Ornithine o-Succinylhomoserine C01118 O-succinylhomoserine p-Aminobenzoate C00568 P-Aminobenzoic acid Phe C00079 Phenylalanine Phenethylamine C05332 Phenylethylamine Phosphorylcholine C00588 Choline phosphate Pipecolate C00408 Pipecolic acid Pro C00148 Proline Proline betaine C10172 Proline betaine Pseudopelletierine C10865 Pterin C00715 Pterin Putrescine(1,4-Butanediamine) C00134 Pyridoxal C00250 Pyridoxal Pyridoxamine C00534 Pyridoxamine Pyridoxamine 5'-phosphate C00647 Pyridoxamine 5'-phosphate Pyridoxine C00314 Pyridoxine Riboflavin C00255 Riboflavin Saccharopine C00449 Saccharopine SAH C00021 SAM+ C00019 Sarcosine C00213 Sarcosine SDMA Symmetric dimethyl-L-arginine Ser C00065 Serine Serotonin C00780 Serotonin Spermidine C00315 Spermidine Spermine C00750 Spermine Taurine C00245 Taurine Thiamine C00378 Thiamine Thiamine monophosphate C01081 Thiamine monophosphate Thr C00188 Threonine Thymidine C00214 Thymidine Thymine C00178 Thymine TRH C03958 Trigonelline C01004 Trigonelline Trimethylamine N-oxide C01104 Trimethylamine N-oxide Tropinone C00783 Tropinone Trp C00078 Tryptophan Tryptamine C00398 Tryptamine Tyr C00082 Tyrosine Tyramine C00483 Tyramine Uracil C00106 Uracil Uridine C00299 Uridine Urocanate C00785 (E)-Urocanic acid Val C00183 Valine Xanthine C00385 Xanthine Xanthosine C01762 Xanthosine 1-Amino-1-cyclopentanecarboxylate C03969 1-Aminocyclopropane-1-carboxylate C01234 1-Aminocyclopropanecarboxylic acid 1-Methyladenosine C02494 1-Methyladenosine 1-Methylhistamine C05127 1-Methylhistamine 1-Methylnicotinamide C02918 1-methyl nicotinamide 1,3-Diaminopropane C00986 1,3-Diaminopropane 2,3-Diaminopropionate C06393 2,4-Diaminobutyrate C03283 2AB C02356 2-Aminophenol C01987 2-aminophenol 2'-Deoxycytidine C00881 Deoxycytidine 2'-Deoxyguanosine C00330 Deoxyguanosine 2'-Deoxyinosine C05512 3-Aminoisobutyrate C05145 3-amino-isobutanoic acid 3-Aminopropane-1,2-diol C06057 3-Methoxytyramine C05587 3-Methoxytyramine 3-Methylguanine C02230 3-Methylguanine 3-Methylhistidine C01152 3-Methylhistidine 4-(beta-Acetylaminoethyl)imidazole C05135 N-Acetylhistamine 4-Hydroxymethylimidazole C05562 5-Aminolevulinate C00430 5-amino-levulinic acid 5-Aminovalerate C00431 5-Aminopentanoic acid 5-Hydroxyindoleacetate C05635 5-Hydroxyindoleacetic acid 5-Hydroxylysine C16741 5-hydroxylysine 5-Methyl-2'-deoxycytidine C03592 5-Methylcytosine C02376 5-Methylcytosine 5-Methylthioadenosine C00170 6-Hydroxynicotinate C01020 6-Hydroxynicotinic acid 7-Methylguanine C02242 7-Methylguanine METABOLITES_END #END