#METABOLOMICS WORKBENCH roser09_20230216_091653 DATATRACK_ID:3745 STUDY_ID:ST002497 ANALYSIS_ID:AN004101 PROJECT_ID:PR001612 VERSION 1 CREATED_ON March 1, 2023, 12:53 pm #PROJECT PR:PROJECT_TITLE Postnatal hyperglycemia alters amino acid profile in retinas (model of Phase I PR:PROJECT_TITLE ROP) PR:PROJECT_SUMMARY Nutritional deprivation occurring in most preterm infants postnatally, can PR:PROJECT_SUMMARY induce hyperglycemia, a significant and independent risk factor for suppressing PR:PROJECT_SUMMARY physiological retinal vascularization (Phase I retinopathy of prematurity PR:PROJECT_SUMMARY (ROP)), leading to compensatory but pathological neovascularization. Amino acid PR:PROJECT_SUMMARY supplementation reduces retinal neovascularization in mice. Little is known PR:PROJECT_SUMMARY about amino acid contribution to Phase I ROP. Significant changes in retinal PR:PROJECT_SUMMARY amino acids (including most decreased L-leucine, L-isoleucine and L-valine) were PR:PROJECT_SUMMARY found in mice modeling hyperglycemia-associated Phase I ROP, and parenteral PR:PROJECT_SUMMARY (i.p.) L-isoleucine suppressed physiological retinal vascularization. In PR:PROJECT_SUMMARY premature infants, severe ROP was associated with a higher mean intake of PR:PROJECT_SUMMARY parenteral versus enteral amino acids in the first two weeks of life after PR:PROJECT_SUMMARY adjustment for treatment group, gestational age at birth, birth weight and sex. PR:PROJECT_SUMMARY The number of days with parenteral amino acids support independently predicted PR:PROJECT_SUMMARY severe ROP. Further understanding and modulating amino acids may help improve PR:PROJECT_SUMMARY nutritional intervention and prevent Phase I ROP PR:INSTITUTE Boston Childrens Hospital PR:LAST_NAME Fu PR:FIRST_NAME Zhongjie PR:ADDRESS 1 Blackfan Circle, Boston, MA 02114 PR:EMAIL Zhongjie.Fu@childrens.harvard.edu PR:PHONE 617-919-2534 #STUDY ST:STUDY_TITLE Postnatal hyperglycemia alters amino acid profile in retinas ST:STUDY_SUMMARY Nutritional deprivation occurring in most preterm infants postnatally, can ST:STUDY_SUMMARY induce hyperglycemia, a significant and independent risk factor for suppressing ST:STUDY_SUMMARY physiological retinal vascularization (Phase I retinopathy of prematurity ST:STUDY_SUMMARY (ROP)), leading to compensatory but pathological neovascularization. Amino acid ST:STUDY_SUMMARY supplementation reduces retinal neovascularization in mice. Little is known ST:STUDY_SUMMARY about amino acid contribution to Phase I ROP. Significant changes in retinal ST:STUDY_SUMMARY amino acids (including most decreased L-leucine, L-isoleucine and L-valine) were ST:STUDY_SUMMARY found in mice modeling hyperglycemia-associated Phase I ROP, and parenteral ST:STUDY_SUMMARY (i.p.) L-isoleucine suppressed physiological retinal vascularization. In ST:STUDY_SUMMARY premature infants, severe ROP was associated with a higher mean intake of ST:STUDY_SUMMARY parenteral versus enteral amino acids in the first two weeks of life after ST:STUDY_SUMMARY adjustment for treatment group, gestational age at birth, birth weight and sex. ST:STUDY_SUMMARY The number of days with parenteral amino acids support independently predicted ST:STUDY_SUMMARY severe ROP. Further understanding and modulating amino acids may help improve ST:STUDY_SUMMARY nutritional intervention and prevent Phase I ROP ST:INSTITUTE Boston Childrens Hospital ST:LAST_NAME Fu ST:FIRST_NAME Zhongjie ST:ADDRESS 1 Blackfan Circle, Boston, MA 02114 ST:EMAIL Zhongjie.Fu@childrens.harvard.edu ST:PHONE 617-919-2534 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 #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 - P10N ctrl_1 Group:control group mouse retinas RAW_FILE_NAME=S16192 SUBJECT_SAMPLE_FACTORS - P10N ctrl_2 Group:control group mouse retinas RAW_FILE_NAME=S16193 SUBJECT_SAMPLE_FACTORS - P10N ctrl_3 Group:control group mouse retinas RAW_FILE_NAME=S16194 SUBJECT_SAMPLE_FACTORS - P10N ctrl_4 Group:control group mouse retinas RAW_FILE_NAME=S16195 SUBJECT_SAMPLE_FACTORS - P10N ctrl_5 Group:control group mouse retinas RAW_FILE_NAME=S16196 SUBJECT_SAMPLE_FACTORS - P10N ctrl_6 Group:control group mouse retinas RAW_FILE_NAME=S16197 SUBJECT_SAMPLE_FACTORS - P10HAR_1 Group:HAR mouse retinas RAW_FILE_NAME=S16198 SUBJECT_SAMPLE_FACTORS - P10HAR_2 Group:HAR mouse retinas RAW_FILE_NAME=S16199 SUBJECT_SAMPLE_FACTORS - P10HAR_3 Group:HAR mouse retinas RAW_FILE_NAME=S16200 SUBJECT_SAMPLE_FACTORS - P10HAR_4 Group:HAR mouse retinas RAW_FILE_NAME=S16201 SUBJECT_SAMPLE_FACTORS - P10HAR_5 Group:HAR mouse retinas RAW_FILE_NAME=S16202 SUBJECT_SAMPLE_FACTORS - P10HAR_6 Group:HAR mouse retinas RAW_FILE_NAME=S16203 #COLLECTION CO:COLLECTION_SUMMARY Retinas were collected at P10 following a single incision across the sclera and CO:COLLECTION_SUMMARY immediately snap frozen in liquid nitrogen and stored at -80 ºC until sample CO:COLLECTION_SUMMARY processing 2. Samples were processed and analyzed by LC-MS/MS by the NYU CO:COLLECTION_SUMMARY Metabolomics Core Resource Laboratory, New York, NY, USA, as described CO:COLLECTION_SUMMARY previously 3,4. Briefly, samples were homogenized using a bead blaster for 10 CO:COLLECTION_SUMMARY cycles with 30 seconds on and 30 seconds off. Metabolites were extracted using CO:COLLECTION_SUMMARY 80% methanol and dried down using a speedvac. Next, samples were reconstituted CO:COLLECTION_SUMMARY in 50 µL MS-grade water and sonicated for two minutes. Samples were then spun CO:COLLECTION_SUMMARY down in a centrifuge at 10 G for 4 min and finally transferred to MS vials for CO:COLLECTION_SUMMARY analysis. Internal standards were used for correction of retention time and CO:COLLECTION_SUMMARY identification of metabolites. Six retinas were pooled as one replicate to CO:COLLECTION_SUMMARY reduce biological variability for metabolomics analysis in each group, n=6 per CO:COLLECTION_SUMMARY group (HAR vs. control). CO:SAMPLE_TYPE Retina #TREATMENT TR:TREATMENT_SUMMARY To study the metabolic alterations occurring in hyperglycemia-associated Phase I TR:TREATMENT_SUMMARY ROP, we applied quantitative metabolomics and proteomics on mouse retinas from TR:TREATMENT_SUMMARY HAR and normal control mice. C57BL/6J (Jackson Laboratory, Bar Harbor, ME) mice, TR:TREATMENT_SUMMARY of each sex, aged 10-12 weeks, were purchased, housed and bred in the TR:TREATMENT_SUMMARY institutional vivarium and maintained on a 12hour/12hour light/dark cycle with TR:TREATMENT_SUMMARY mouse chow provided ad libitum. Neonatal mice were randomly assigned to TR:TREATMENT_SUMMARY experimental groups. Induction of hyperglycemia was accomplished as previously TR:TREATMENT_SUMMARY described 1. Neonatal mice were intraperitoneally injected with 50mg/kg/day STZ TR:TREATMENT_SUMMARY consecutively from P1 to P9 using a 34-G needle (Hamilton syringe) (Fig. 1B). TR:TREATMENT_SUMMARY Vehicle control animals received equal volumes of vehicle phosphate-buffered TR:TREATMENT_SUMMARY saline (PBS, Gibco, Waltham, MA). Hyperglycemia is induced around P8 and delayed TR:TREATMENT_SUMMARY retinal vascularization is found at P10 1. Mice with weight range 4 to 5 grams TR:TREATMENT_SUMMARY were used for further metabolomics and proteomics analysis. Mouse litters were TR:TREATMENT_SUMMARY randomly assigned to HAR or control groups, both sexes were used. The cages were TR:TREATMENT_SUMMARY located at close spots to minimize the potential housing influences. All TR:TREATMENT_SUMMARY procedures were approved by our Institutional Animal Care and Use Committee and TR:TREATMENT_SUMMARY adhered to ARRIVE guidelines and the NIH Guide for the Care and Use of TR:TREATMENT_SUMMARY Laboratory Animals. With conditions tested with β=0.8 and α=0.05, at least n=6 TR:TREATMENT_SUMMARY per group will be needed for the analysis. Control was re-named as group 1 and TR:TREATMENT_SUMMARY HAR was re-named as group 2 for analysis. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Both retinas from each mouse were collected, pooled, and prepared for targeted SP:SAMPLEPREP_SUMMARY MS-based proteomics as described previously 1. After sample preparation, SP:SAMPLEPREP_SUMMARY LC-tandem MS analysis was performed using selected reaction monitoring (SRM) 5 SP:SAMPLEPREP_SUMMARY on a Thermo Scientific TSQ Vantage mass spectrometer equipped with an Eksigent SP:SAMPLEPREP_SUMMARY splitless nanoflow HPLC system. 7 µL aliquots of each sample were injected onto SP:SAMPLEPREP_SUMMARY a 10 cm x 75 µm i.d. capillary column packed with Phenomenex Jupiter C18 SP:SAMPLEPREP_SUMMARY reversed phase beads. The column was eluted at 150 nL/min with a 60 min linear SP:SAMPLEPREP_SUMMARY gradient of acetonitrile in 0.1% formic acid. The SRM assays were developed and SP:SAMPLEPREP_SUMMARY validated to monitor two peptides per protein. Each peptide was monitored in a SP:SAMPLEPREP_SUMMARY 6-min window centered on the known elution time of the peptide 6-8. SP:SAMPLEPREP_SUMMARY Approximately 30 protein assays are grouped into a panel of proteins that are SP:SAMPLEPREP_SUMMARY measured in a single LC-tandem MS run. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Thermo Ultimate 3000 CH:COLUMN_NAME SeQuant ZIC-pHILIC (150 x 2.1mm, 5um) CH:SOLVENT_A 10 mM ammonium carbonate in water, pH 9.0 CH:SOLVENT_B acetonitrile CH:FLOW_GRADIENT 80-20%B (0-30 min), 20-80%B (30-31 min), 80-80%B (31-42 min) CH:FLOW_RATE 0.1mL/min CH:COLUMN_TEMPERATURE 25 #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive HF hybrid Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE UNSPECIFIED MS:MS_COMMENTS Initial data analysis (metabolite identification & quantification) was performed MS:MS_COMMENTS by the NYU metabolomics facility 3,9-11. Subsequent downstream bioinformatic MS:MS_COMMENTS analysis was performed using MetaboAnalyst R-based statistical and pathway MS:MS_COMMENTS analysis (v5.0) as described by Petrova et al. 12 using updates as described by MS:MS_COMMENTS Pang et al. 13. Differences in metabolite levels between groups were assessed MS:MS_COMMENTS using unpaired t test and considered statistically significant if P<0.05. For MS:MS_COMMENTS pathway analysis SMPDB database was reviewed, applying Fisher’s Exact Test for MS:MS_COMMENTS mapping. Only metabolites meeting P-value criteria were loaded for the analysis. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Absolute Intensity MS_METABOLITE_DATA_START Samples P10N ctrl_1 P10N ctrl_2 P10N ctrl_3 P10N ctrl_4 P10N ctrl_5 P10N ctrl_6 P10HAR_1 P10HAR_2 P10HAR_3 P10HAR_4 P10HAR_5 P10HAR_6 Factors Group:control group mouse retinas Group:control group mouse retinas Group:control group mouse retinas Group:control group mouse retinas Group:control group mouse retinas Group:control group mouse retinas Group:HAR mouse retinas Group:HAR mouse retinas Group:HAR mouse retinas Group:HAR mouse retinas Group:HAR mouse retinas Group:HAR mouse retinas 1-Methylnicotinamide 74424 55542 39353 55080 45061 24939 28818 29671 28901 - - - 2,3-Diphosphoglyceric acid - - - - - - - - - - - - 3-Hydroxybutyric acid 5829096 4217463 3564105 1959991 4372168 2630588 2777449 2212004 3247281 1479458 1238824 1143429 3-Phosphoglyceric acid 1350567 225077 443820 80495 202968 238768 393623 236586 629741 257605 187584 134339 4,5-Dihydroorotic acid - - 36633 - - 59429 - - - - - - 4-Hydroxyproline 8320392 3417044 7980858 1133076 2824433 11159321 2440154 3271149 2936808 2625923 1132481 1617831 5'-Methylthioadenosine 5084993 7863038 6421994 6453999 8928390 4177312 6693165 8303954 7511064 6448350 1148801 5556908 5-Phosphoribosylamine - - 15945 - - - - 16145 - 19401 16189 16097 6-Phosphogluconic acid 32324 15585 34606 12791 39362 - 20826 19029 37220 23930 14526 - Acetoacetic acid 869963 1076040 984089 939645 980591 541802 883981 948128 1036553 843808 745092 631270 Acetyl-CoA 26548 27531 37472 22354 13604 20176 43414 42381 33558 43979 41174 53316 Acetylcysteine 179724 108313 67336 151908 119877 72154 327703 334086 151894 269617 267402 550952 Adenine 5758434 30049850 26261822 8396303 35233712 14655877 39718484 55746204 50029552 39250700 34390176 19425504 Adenosine 6852134 - 313868256 148176800 324890752 143369008 270621920 787186 58729196 395674144 237038208 217139376 Adenosine diphosphate ribose - - 124743 - - 118486 10677 - - 50976 - 78343 Adenosine monophosphate 4723206 6239572 56838716 - 6149556 35366016 6316092 13306863 18100448 25446486 6280457 34587224 Adenosine triphosphate 103868 109078 691084 95800 148917 301989 236921 235236 451475 433979 186176 227057 Adenylsuccinic acid - - 483348 - - 228950 80499 147338 220177 253986 84290 347020 ADP - - 4951094 - 831142 2018722 898520 921235 1505300 3322492 968118 1660468 Allantoin 3709603 2145311 1793117 3361808 2843926 1198740 3521870 2826786 3595673 4160772 3987260 2522536 Aminoadipic acid 915357 866410 865594 634063 809033 486994 1029545 1171518 1196172 750878 659072 718612 Argininosuccinic acid 222406 181694 144217 168861 212233 95678 203820 172763 164461 82587 133534 107016 Beta-Alanine 9982906 13675458 8253806 9602075 10630598 3100616 23875546 22077874 19989592 18130792 15261805 13839588 Betaine 285468576 285325472 281922976 221320416 293656576 191050336 501515040 530948384 596173056 432128672 415874432 314912832 Butyrylcarnitine 10413293 17398456 28210674 17060726 21969332 12040268 16428207 14892954 24715472 21787560 14852670 13159949 Carbamoyl phosphate - - - - - - - - - - - - Carnosine 46323856 12923925 8217734 10182651 10488677 5213395 7707806 7796622 8407556 6943406 8872372 6472540 CDP 89641 123799 222990 93163 112766 141003 163687 206239 268060 170482 155555 154276 Choline 462099584 387719936 280240704 423939136 365537792 124584776 452118816 358908160 400445280 313440800 362788896 311253472 cis-Aconitic acid 3755808 3100678 1835657 2150224 2640225 1327903 3424520 2066428 3331442 2328886 2979821 1967510 Citric acid 23833854 21999042 17293716 14629866 20978898 10101768 24968428 20732702 27388292 18217672 18636246 16038016 Citrulline 46586852 25515704 29583142 24614394 31983458 13923658 42965116 42248592 45750828 33266248 31716896 33598664 Creatine 2299613952 2280146432 2286970624 2263857920 2310536960 1836752640 2279457792 2342175232 2313855488 2256781056 2286203904 2227392256 Creatinine 47942808 53079648 34877416 31570988 41328584 16361488 54508456 60658640 61693852 35601328 39372204 34667420 Cyclic AMP 54525 69626 62639 51730 64531 34471 105953 105619 124329 66630 76153 63527 Cytidine 30771816 25519862 8368654 29157468 30171404 3990729 29815842 24473682 18951510 12191864 20873106 9321221 Cytidine monophosphate 434176 832034 3630395 391311 614672 1383258 773546 1461873 1294368 2033827 921276 2119914 Cytidine triphosphate - - 21513 - 16802 - 31592 21950 36723 - 15216 - Cytosine - - - - - - - - - - - - D-2-Hydroxyglutaric acid 6802162 7685722 6876000 7471324 7857198 5304651 8031076 7906252 9798706 7444526 6783862 5617596 D-Glucose 2669474 2251900 1943508 1377953 2132646 1240309 2639045 2463142 2424578 1456383 1757500 1504549 D-Glyceraldehyde 3-phosphate 196302 278728 489138 366785 452660 340061 398486 353631 510929 461311 338145 318472 Dihydrofolic acid - - - - - - - - - - - - Dihydroxyacetone phosphate 470693 47057 146844 87719 296281 130770 350703 275471 236903 133941 186523 247583 D-Ribose 5-phosphate 579077 305955 709355 439400 337056 443316 439034 720354 585036 870794 725165 694543 D-Sedoheptulose 7-phosphate 690508 144006 552604 156479 335881 548612 270967 540549 491543 558825 242195 502920 dTDP - - 13724 - - - 13684 - - - - - FAD 95446 68777 77424 63529 65040 20424 85925 85936 85502 66644 68904 55689 Folic acid - - - - - - - - - - - - Fructose 1,6-bisphosphate 339959 - 198527 - 119420 60291 171948 119522 355678 99039 82177 169625 Fructose 6-phosphate 1314122 218158 382848 193454 1373871 393010 552257 1096189 554852 457133 804068 1015715 Fumaric acid 4894758 5771458 3611338 4981901 5610847 1905685 6154914 5349179 5588434 3867779 5051024 3292064 Glucose 6-phosphate 2516088 267667 1181165 217776 615870 1224182 827463 1073987 1398293 565426 402286 1258896 Glutathione 4651765 15857486 24692964 10850323 18901012 15432597 15612209 17058298 13649790 19541726 23043616 9570486 Glycerol 3-phosphate 2153820 3234974 6613538 3619756 3120231 3796392 4347180 6624924 7151427 3779073 2989274 4330920 Glycine 24508592 26597042 21010144 25393602 24198062 11745676 29779084 30384524 30028178 25225042 25574664 22542046 Guanidoacetic acid 15558102 10664781 5768602 7863641 8133622 2408196 23126628 29286558 14781516 19140158 15913050 18100952 Guanine 90413776 112324840 27577416 128235144 112753736 12659457 95862632 79388792 76168496 46695084 86912608 35750500 Guanosine 30022178 47261112 11048927 53359048 52501852 5752413 43062872 38283968 37891876 24185884 51181352 19366528 Guanosine diphosphate - 257695 883132 - 248334 372854 314418 435420 757488 527118 415714 447307 Guanosine monophosphate 2061893 2898971 14447220 - 2454648 6792932 3226018 5650928 7239320 11281357 3818333 10845138 Guanosine triphosphate - 32201 76444 28580 33338 33717 60373 62817 130582 44424 45987 34318 Homocysteine 25643 42823 44700 45180 32435 49155 40941 47162 44880 33351 32421 28295 Hypotaurine 15420142 15258946 10716103 8527672 13881359 4949526 58398568 53788332 35192972 55712356 53795312 67218664 Hypoxanthine 533042144 503050528 174532912 456518752 412852000 92520912 451924864 398087840 342721216 270488672 429108352 237202224 Inosine 113806240 148969952 44580104 142046544 157850320 29956122 137604704 159184416 136407360 79398968 132058688 74704032 Inosinic acid 1375556 183985 1310027 - 122317 484905 348445 296656 694898 760427 150496 610674 L-Acetylcarnitine 547971264 660282368 385052896 448285248 506535360 216815488 430986944 390851232 566026496 358856416 419205984 237121520 L-Alanine 33525762 28003754 26177372 25101452 26395966 22816418 27423168 28217544 24173214 24349784 24400044 24500772 L-Arginine 56594440 34482300 47643760 41566132 53762060 32483532 50896872 50377912 47234556 40139868 45655348 42876940 L-Asparagine 7382712 6612106 5664902 5592848 6261762 3416669 6866344 7543106 5463115 5319884 6022606 5473298 L-Aspartic acid 50835972 58527404 44317420 56216036 53999408 27469540 55807236 56552564 54921608 51791984 57203672 49788904 L-Carnitine 704010496 872086848 794830016 784153216 798777856 437406432 727978688 778844800 844843648 724435328 660654272 544163392 L-Cystathionine 1698012 873014 1310766 717609 1041099 1082898 2597147 2183875 1935381 1671745 1455190 1706278 L-Cysteine - 66122 - 107733 - - 35717 29346 - - 31383 - L-Cystine 148773 214731 - 329387 - - 44465 - - - - - L-Glutamic acid 157486480 187612192 175537360 184143792 193127120 99246600 160147408 175363472 176467776 166005376 158000592 147468688 L-Glutamine 493523904 514676224 474576288 504982784 512917664 317449600 573097152 601376832 621097856 579472192 548956096 560833536 L-Histidine 26415450 23058146 26412290 19208996 23818522 25334646 26810722 26799984 23508982 25732410 25030868 22120740 L-Isoleucine 74465600 58104968 54401564 54623860 52429048 41701244 54767920 48564652 37417724 36983172 46171032 35612356 L-Kynurenine 174428 146565 119199 86271 158344 88613 297571 268122 286446 215620 229672 190226 L-Lactic acid 17923564 47282680 30147306 28573572 41566276 10385160 48666740 35546356 19315728 29900442 44991252 23132682 L-Leucine 68931584 49177436 43866628 46736068 48524984 35334480 44560396 43844968 31132190 35591224 38933992 31622194 L-Lysine 48423940 36428328 42647424 31629564 43754824 30959102 44583420 41569192 47996800 33191310 37069172 32297768 L-Malic acid 68376192 79111272 51177296 64987792 74014680 30823266 82052152 70802960 72280872 52120168 67780672 48230320 L-Methionine 104496352 113156592 127243224 103136304 128679616 92615424 109597384 136391936 106901232 82815488 97272392 98818256 L-Phenylalanine 75306488 76803088 107816256 73930736 102845320 71474920 156941328 220044912 144449920 108622584 119857712 143750240 L-Proline 477426848 476488480 384833312 395605376 460802016 229700368 609696384 678577792 622210560 525580064 505508064 523207552 L-Serine 27608502 23025942 18255436 19623856 20230024 9140521 24618652 28763696 24187968 20657720 25256396 21182448 L-Threonine 6285436 5677345 3153330 5143922 5726839 2153743 5415972 5834447 5065099 4370462 5470772 3737836 L-Tryptophan 65351448 49692612 58264772 43805028 56482400 32422018 101065264 82866952 77136712 47642504 45631492 50527764 L-Tyrosine 68592248 80378816 99177520 96088304 120578096 68451264 167765472 178706768 188362960 125286136 122308488 138503632 L-Valine 98125832 75107840 68649104 68618888 78468016 44023080 77522608 71341552 71025152 51085792 63613284 51108124 N6-Acetyl-L-lysine 169298 - - 144932 437079 - 433015 601529 669867 1363711 251028 1174914 N-Acetylglutamic acid 819003 844248 667554 847923 859763 490196 1163824 892774 973382 695316 559256 523611 N-Acetylglutamine 691967 818466 978123 755569 789075 421254 1151658 1231130 1589880 1075846 997372 1165555 N-Acetyl-L-alanine 2174312 1935380 1109319 1190778 1443328 586513 2553728 2284102 2450318 1237921 1371239 1046178 N-Acetyl-L-methionine 662941 658747 484487 610140 749811 308068 1324988 1151444 832987 668288 883437 353679 N-Acetyl-L-phenylalanine 343832 293178 421823 257736 420330 225325 803354 1055247 837719 489077 484793 626548 N-Acetylornithine 911941 990431 1620214 640163 754405 807348 268333 250665 222930 104855 171265 130873 N-Acetylserine 4616142 4498558 4093713 3215373 3912806 2620328 5481708 5267473 5467670 3270348 3477976 3330961 NAD 234036 684548 1610771 436710 661098 1116672 287312 636867 694241 793814 460811 608795 NADH 19615 21190 88196 15370 42943 12533 22006 49691 74041 34287 - 37469 NADP 49458 128688 132459 94579 122879 67475 98167 128788 160484 111205 72426 77212 NADPH - - 20990 - - - - - - - - - Niacinamide 391533120 433248800 264832384 385152960 439334912 136379984 438508064 415959456 402544352 335559552 412049728 337765728 Nicotinic acid 33896 - - - - - - - - - - - O-Phosphoethanolamine 22231880 27457940 27208738 24753958 31505470 13675547 31108580 30089694 33103408 28380268 28394856 22140204 Ornithine 6529854 4499498 5057702 4283104 5095832 2956180 6098319 5569252 5309936 4496906 4520204 4683472 Orotic acid 141146 180415 227420 34714 241873 174003 216700 277735 162445 143865 189742 142524 Orotidylic acid - - - - - - - - - 77612 - - Oxidized glutathione 4785508 6029732 9930793 2733409 10440958 10193603 7072078 11310409 8947306 6999720 6939988 11587501 Oxoadipic acid 90749 36757 67825 22136 90303 32352 53805 49822 56472 41197 61053 50669 Oxoglutaric acid 482970 826189 786545 1105771 1426016 701894 791152 486524 719697 793102 1118046 471139 Palmitic acid - - - - - - - 5338527 - - - - Pantothenic acid 44127692 28983386 33945852 30330092 34129764 14234115 47913300 42708784 47838892 35016500 33857784 30347890 Phenol - - - - - - - - - - - - Phosphocreatine 1060263 1686043 1198882 529418 1133035 312175 1405686 2088002 2909610 981294 837628 1048081 Phosphoenolpyruvic acid 1593460 209356 689234 - 240018 303589 396591 275595 666669 345311 - 181070 Phosphoribosyl pyrophosphate - - - - - - - - - - - - Phosphorylcholine 589578752 706113472 716356032 644077824 765386112 524020608 730194176 802144320 753726464 623609280 680368512 618710144 Phosphoserine 35360 16581 21598 17734 19391 - 19524 31455 21986 23904 20929 25090 Pipecolic acid 19002530 29647782 24401212 17587852 19394282 10774268 28904150 32022386 19793568 13731009 18158274 15855254 Propionylcarnitine 14503268 24077098 13771973 14154539 12386124 5693138 17801582 18501888 22236308 11457389 9620194 8150996 Pyruvic acid 361200 327292 292032 474870 471772 254956 639988 549048 550735 296892 473044 147038 Ribose 1-phosphate 982210 1057607 1404300 2435992 3295298 522099 2092062 1318574 1760015 2848965 1845222 1181396 Saccharopine 24349 38253 40927 47567 53199 22848 68382 63926 95316 63813 68321 50333 S-Adenosylhomocysteine 287992 380480 357646 398812 554393 93919 375082 338267 385920 335879 251079 352563 S-Adenosylmethionine 4746423 7195950 4956370 8105812 6899174 5382145 5925698 6733835 5493724 7656568 8155568 6162154 Sarcosine 24314674 18812992 17276576 16705685 23028220 14465328 19316918 20994684 16439711 15265718 20267356 15961950 Succinic acid 3326920 2859638 10423596 1945737 5042460 6046704 2352028 6167818 7037566 6268853 1976090 3283751 Taurine 204718624 251329248 223235296 245123840 249499488 134817584 271643584 293133248 298757984 261473984 258785280 243660192 Thymidine 483075 511553 353745 387480 423578 189671 1389812 1034157 1258950 624847 789455 499544 Uracil 3730480 4629784 1052917 4520302 4641842 464307 4417652 3481070 2772704 2023764 3495430 1248325 Ureidopropionic acid 1926786 1978204 1476267 1566294 1910012 802718 2097368 2260192 1684877 1562930 1789342 1648697 Ureidosuccinic acid 18173 20167 66515 18741 34096 111452 17729 23508 19693 27298 17499 19505 Uric acid - - - - - - - - - - - - Uridine 10300753 11812453 4045802 10830614 10993867 2373719 11425749 10394955 8879564 6862205 10295603 4872922 Uridine 5'-diphosphate 272313 677849 690672 394778 587179 448800 567366 747455 644002 516768 549785 452963 Uridine 5'-monophosphate 797068 667352 8176756 566860 1225775 5431478 1078538 2501845 2466965 7034058 1151498 5664139 Uridine diphosphate glucose 2682746 7312104 6781222 5230522 6173646 4070464 7718596 7828482 6708224 5205920 6690853 4625156 Uridine diphosphate-N-acetylglucosamine 724178 1283049 1246120 964131 1223496 664333 1185380 1370507 1172057 1042580 978211 823614 Uridine triphosphate - - 21212 - - - - - 24383 - - - Valerylcarnitine 7634060 9542991 9216338 7818882 8002795 3774792 7621232 8229996 9202791 5615314 5862458 4795469 Xanthine 15201529 3588166 1458481 3128406 4838310 699575 4914390 3739425 4479017 3107543 3903325 2571544 Xanthosine 70745 - - - - - - - - - - - Xanthylic acid 23090 66436 170559 51915 55296 58394 90856 113144 139765 73399 48532 70477 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name 1-Methylnicotinamide 2,3-Diphosphoglyceric acid 3-Hydroxybutyric acid 3-Phosphoglyceric acid 4,5-Dihydroorotic acid 4-Hydroxyproline 5'-Methylthioadenosine 5-Phosphoribosylamine 6-Phosphogluconic acid Acetoacetic acid Acetyl-CoA Acetylcysteine Adenine Adenosine Adenosine diphosphate ribose Adenosine monophosphate Adenosine triphosphate Adenylsuccinic acid ADP Allantoin Aminoadipic acid Argininosuccinic acid Beta-Alanine Betaine Butyrylcarnitine Carbamoyl phosphate Carnosine CDP Choline cis-Aconitic acid Citric acid Citrulline Creatine Creatinine Cyclic AMP Cytidine Cytidine monophosphate Cytidine triphosphate Cytosine D-2-Hydroxyglutaric acid D-Glucose D-Glyceraldehyde 3-phosphate Dihydrofolic acid Dihydroxyacetone phosphate D-Ribose 5-phosphate D-Sedoheptulose 7-phosphate dTDP FAD Folic acid Fructose 1,6-bisphosphate Fructose 6-phosphate Fumaric acid Glucose 6-phosphate Glutathione Glycerol 3-phosphate Glycine Guanidoacetic acid Guanine Guanosine Guanosine diphosphate Guanosine monophosphate Guanosine triphosphate Homocysteine Hypotaurine Hypoxanthine Inosine Inosinic acid L-Acetylcarnitine L-Alanine L-Arginine L-Asparagine L-Aspartic acid L-Carnitine L-Cystathionine L-Cysteine L-Cystine L-Glutamic acid L-Glutamine L-Histidine L-Isoleucine L-Kynurenine L-Lactic acid L-Leucine L-Lysine L-Malic acid L-Methionine L-Phenylalanine L-Proline L-Serine L-Threonine L-Tryptophan L-Tyrosine L-Valine N6-Acetyl-L-lysine N-Acetylglutamic acid N-Acetylglutamine N-Acetyl-L-alanine N-Acetyl-L-methionine N-Acetyl-L-phenylalanine N-Acetylornithine N-Acetylserine NAD NADH NADP NADPH Niacinamide Nicotinic acid O-Phosphoethanolamine Ornithine Orotic acid Orotidylic acid Oxidized glutathione Oxoadipic acid Oxoglutaric acid Palmitic acid Pantothenic acid Phenol Phosphocreatine Phosphoenolpyruvic acid Phosphoribosyl pyrophosphate Phosphorylcholine Phosphoserine Pipecolic acid Propionylcarnitine Pyruvic acid Ribose 1-phosphate Saccharopine S-Adenosylhomocysteine S-Adenosylmethionine Sarcosine Succinic acid Taurine Thymidine Uracil Ureidopropionic acid Ureidosuccinic acid Uric acid Uridine Uridine 5'-diphosphate Uridine 5'-monophosphate Uridine diphosphate glucose Uridine diphosphate-N-acetylglucosamine Uridine triphosphate Valerylcarnitine Xanthine Xanthosine Xanthylic acid METABOLITES_END #END