#METABOLOMICS WORKBENCH ryant_20201211_124513 DATATRACK_ID:2354 STUDY_ID:ST001628 ANALYSIS_ID:AN002663 PROJECT_ID:000000 VERSION 1 CREATED_ON December 15, 2020, 11:27 am #PROJECT PR:PROJECT_TITLE Tissue-specific 1H-NMR metabolomic profiling in mice with adenine-induced PR:PROJECT_TITLE chronic kidney disease PR:PROJECT_TYPE Metabolomic profiling of the heart, liver, kidney, and skeletal muscles obtained PR:PROJECT_TYPE from mice with and without CKD via 1H NMR PR:PROJECT_SUMMARY This project is focused on a metabolomic analyses of the heart, liver, kidney, PR:PROJECT_SUMMARY and skeletal muscles obtained from mice with and without CKD. To accomplish this PR:PROJECT_SUMMARY objective, we extracted tissues from mice with CKD induced by long-term (24 PR:PROJECT_SUMMARY week) adenine-supplemented diet as well as their control-diet fed counterparts PR:PROJECT_SUMMARY with normal kidney function. Metabolites were extracted from tissues and 1H PR:PROJECT_SUMMARY nuclear magnetic resonance (NMR) was performed and coupled with multivariate PR:PROJECT_SUMMARY statistical analysis. PR:INSTITUTE University of Florida PR:DEPARTMENT Applied Physiology and Kinesiology PR:LABORATORY Rm 42 and Rm 43 PR:LAST_NAME Ryan PR:FIRST_NAME Terence PR:ADDRESS 1864 Stadium Rd, FLG 114, Gainesville, FL, 32611, USA PR:EMAIL ryant@ufl.edu PR:PHONE 352-294-1700 PR:FUNDING_SOURCE This research was funded by grants from the National Institutes of Health and PR:FUNDING_SOURCE the National Heart, Lung, and Blood, Institute numbers R01-HL149704 (to T.E.R.) PR:FUNDING_SOURCE and the American Heart Association grant number 18CDA34110044 (to T.E.R.). PR:PROJECT_COMMENTS CKD metabolomic study via NMR using mice model PR:PUBLICATIONS MDPI PR:CONTRIBUTORS Ram B. Khattri, Trace Thome, and Terence E. Ryan #STUDY ST:STUDY_TITLE Tissue-specific 1H-NMR metabolomic profiling in mice with adenine-induced ST:STUDY_TITLE chronic kidney disease - aqueous phase Liver (part-V) ST:STUDY_TYPE Metabolomic profiling of the heart, liver, kidney, and skeletal muscles obtained ST:STUDY_TYPE from mice with and without CKD via 1H NMR ST:STUDY_SUMMARY This project is focused on a metabolomic analyses of the heart, liver, kidney, ST:STUDY_SUMMARY and skeletal muscles obtained from mice with and without CKD. To accomplish this ST:STUDY_SUMMARY objective, we extracted tissues from mice with CKD induced by long-term (24 ST:STUDY_SUMMARY week) adenine-supplemented diet as well as their control-diet fed counterparts ST:STUDY_SUMMARY with normal kidney function. Metabolites were extracted from tissues and 1H ST:STUDY_SUMMARY nuclear magnetic resonance (NMR) was performed and coupled with multivariate ST:STUDY_SUMMARY statistical analysis. ST:INSTITUTE University of Florida ST:DEPARTMENT Applied Physiology and Kinesiology ST:LABORATORY Rm 42 and Rm 43 ST:LAST_NAME Ryan ST:FIRST_NAME Terence ST:ADDRESS 1864 Stadium Rd, FLG 114, Gainesville, FL, 32611, USA ST:EMAIL ryant@ufl.edu ST:PHONE 352-294-1700 ST:NUM_GROUPS 2 ST:TOTAL_SUBJECTS 17 ST:NUM_MALES All ST:STUDY_COMMENTS CKD metabolomic study via NMR using mice model ST:PUBLICATIONS MDPI #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:GENOTYPE_STRAIN C57BL/6J SU:AGE_OR_AGE_RANGE 13 months SU:WEIGHT_OR_WEIGHT_RANGE (32.86±1.21 g (control mice) vs 23.57±1.27 g (CKD mice), P < 0.0001, SU:WEIGHT_OR_WEIGHT_RANGE N=7/group). SU:GENDER Male SU:ANIMAL_ANIMAL_SUPPLIER Jackson Labs (Stock # 000664) SU:ANIMAL_HOUSING Housed in a temperature of 22 oC SU:ANIMAL_LIGHT_CYCLE 12-hour light/12-hour dark SU:ANIMAL_FEED Ad libitum (Casein control diet vs. adenine-supplemented diet to induce CKD) SU:ANIMAL_WATER free access to food and water (3-5 animals per cage). SU:ANIMAL_INCLUSION_CRITERIA (3-5 animals per cage). #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 - CKD1Liver_Aq Group:CKD RAW_FILE_NAME=CKD1Liver_Aq SUBJECT_SAMPLE_FACTORS - CKD2Liver_Aq Group:CKD RAW_FILE_NAME=CKD2Liver_Aq SUBJECT_SAMPLE_FACTORS - CKD3Liver_Aq Group:CKD RAW_FILE_NAME=CKD3Liver_Aq SUBJECT_SAMPLE_FACTORS - CKD4Liver_Aq Group:CKD RAW_FILE_NAME=CKD4Liver_Aq SUBJECT_SAMPLE_FACTORS - CKD5Liver_Aq Group:CKD RAW_FILE_NAME=CKD5Liver_Aq SUBJECT_SAMPLE_FACTORS - CKD6Liver_Aq Group:CKD RAW_FILE_NAME=CKD6Liver_Aq SUBJECT_SAMPLE_FACTORS - CKD7Liver_Aq Group:CKD RAW_FILE_NAME=CKD7Liver_Aq SUBJECT_SAMPLE_FACTORS - Con1Liver_Aq Group:Control RAW_FILE_NAME=Con1Liver_Aq SUBJECT_SAMPLE_FACTORS - Con2Liver_Aq Group:Control RAW_FILE_NAME=Con2Liver_Aq SUBJECT_SAMPLE_FACTORS - Con3Liver_Aq Group:Control RAW_FILE_NAME=Con3Liver_Aq SUBJECT_SAMPLE_FACTORS - Con4Liver_Aq Group:Control RAW_FILE_NAME=Con4Liver_Aq SUBJECT_SAMPLE_FACTORS - Con5Liver_Aq Group:Control RAW_FILE_NAME=Con5Liver_Aq SUBJECT_SAMPLE_FACTORS - Con6Liver_Aq Group:Control RAW_FILE_NAME=Con6Liver_Aq SUBJECT_SAMPLE_FACTORS - Con7Liver_Aq Group:Control RAW_FILE_NAME=Con7Liver_Aq #COLLECTION CO:COLLECTION_SUMMARY While under isoflurane anesthesia, tissues were rapidly dissected and snap CO:COLLECTION_SUMMARY frozen in liquid nitrogen and stored at -80°C until metabolite extraction. The CO:COLLECTION_SUMMARY following tissues were used in this study: kidney, liver, heart (left CO:COLLECTION_SUMMARY ventricle), skeletal muscle (quadriceps). Euthanasia was carried out by CO:COLLECTION_SUMMARY thoracotomy followed by cervical dislocation. CO:SAMPLE_TYPE Liver CO:COLLECTION_METHOD While under isoflurane anesthesia, tissues were rapidly dissected and snap CO:COLLECTION_METHOD frozen in liquid nitrogen and stored at -80°C until metabolite extraction CO:COLLECTION_LOCATION University of Florida, Applied Physiology and Kinesiology, 1864 stadium RD, CO:COLLECTION_LOCATION Gainesville, FL 32611 CO:STORAGE_CONDITIONS -80℃ CO:STORAGE_VIALS cryovials #TREATMENT TR:TREATMENT_SUMMARY To induce CKD, we utilized an established adenine-diet model. Briefly, mice were TR:TREATMENT_SUMMARY assigned to a casein-base chow for 7-days, followed by induction of renal TR:TREATMENT_SUMMARY tubular injury by supplementing the diet with 0.2% adenine for 7-days, and TR:TREATMENT_SUMMARY subsequently maintained on a 0.15% adenine diet for 5 months and two weeks. CKD TR:TREATMENT_SUMMARY mice were then placed back on casein control diet for 2-weeks prior to TR:TREATMENT_SUMMARY euthanasia and terminal experiments, allowing for a washout period of adenine TR:TREATMENT_SUMMARY based chow. Control mice received casein diet for the duration of the study. TR:TREATMENT_SUMMARY Total duration of CKD encompassed 6-months. TR:ANIMAL_VET_TREATMENTS none TR:ANIMAL_ANESTHESIA isoflurane TR:ANIMAL_FASTING non-fasted TR:ANIMAL_ENDP_EUTHANASIA Euthanasia was carried out by thoracotomy followed by cervical dislocation. TR:ANIMAL_ENDP_TISSUE_COLL_LIST kidney, liver, heart (left ventricle), skeletal muscle (quadriceps) #SAMPLEPREP SP:SAMPLEPREP_SUMMARY A modified form of FOLCH extraction protocol was used to extract metabolites SP:SAMPLEPREP_SUMMARY from the tissues. Wet weights of all tissue samples were recorded prior to SP:SAMPLEPREP_SUMMARY extraction. Tissue samples were immediately homogenized to prevent any possible SP:SAMPLEPREP_SUMMARY enzymatic action using 1 mL of ice-cold methanol in a PowerLyzer 24 Homogenizer SP:SAMPLEPREP_SUMMARY (QIAGEN Group, Hilden, Germany). The mixture was centrifuged using 13,200 rpm at SP:SAMPLEPREP_SUMMARY 4oC for 30 minutes and the resulting supernatant was transferred to a new glass SP:SAMPLEPREP_SUMMARY vial consisting 3 mL of ice cold chloroform:methanol (2:1, v/v) mixture. The SP:SAMPLEPREP_SUMMARY homogenate was vortexed and left in an ice bath for 15 minutes to allow for SP:SAMPLEPREP_SUMMARY phase separation. Next, 1 mL of 0.9% of saline was added, vortexed it for couple SP:SAMPLEPREP_SUMMARY of minutes followed by a second incubation in an ice bath for 30-45 min for SP:SAMPLEPREP_SUMMARY complete phase separation. The upper aqueous layer was transferred to a new SP:SAMPLEPREP_SUMMARY falcon tube. To the remaining organic phase sample, 1 mL of 0.9% of saline was SP:SAMPLEPREP_SUMMARY added again followed by vigorous mixing and letting it stand in ice bath (15 SP:SAMPLEPREP_SUMMARY minutes) for a second phase separation. This second aqueous phase was combined SP:SAMPLEPREP_SUMMARY with the first. The resulting aqueous and organic layers were dried separately. SP:SAMPLEPREP_SUMMARY The aqueous layer was dried overnight with a Labconco freezer dryer (Labconco SP:SAMPLEPREP_SUMMARY Corporation, MO, USA) and the organic layer was dried via inert nitrogen gas. SP:SAMPLEPREP_SUMMARY These two dried powders (aqueous and organic phases) were stored at -80oC until SP:SAMPLEPREP_SUMMARY performing NMR experiments. SP:PROCESSING_METHOD Lyophilization and Homogenization SP:PROCESSING_STORAGE_CONDITIONS -80℃ SP:EXTRACTION_METHOD Modified FOLCH extraction SP:EXTRACT_STORAGE -80℃ SP:SAMPLE_RESUSPENSION In 50 microliter of 50 mM phosphate buffer (pH 7.2) with 2 mM EDTA, 0.5 mM DSS SP:SAMPLE_RESUSPENSION and 0.2% sodium azide for aqueous phase samples. SP:SAMPLE_SPIKING 0.5 mM of DSS for aqueous phase samples #ANALYSIS AN:DATA_FORMAT fid, 1r #NMR NM:INSTRUMENT_NAME Bruker Avance Neo 600 MHz/54mm console NM:INSTRUMENT_TYPE FT-NMR NM:NMR_EXPERIMENT_TYPE 1D-1H NM:FIELD_FREQUENCY_LOCK Deuterium NM:STANDARD_CONCENTRATION 0.5 mM DSS NM:SPECTROMETER_FREQUENCY 600.2328273 MHz NM:NMR_PROBE 1.7 mm TXI CryoProbe NM:NMR_SOLVENT Phosphate buffer (pH 7.2) + 2 mM EDTA + 0.5 mM DSS + 0.2% of sodium azide in NM:NMR_SOLVENT deuterated environment NM:NMR_TUBE_SIZE 1.7 mm O.D. NM:SHIMMING_METHOD Topshim NM:PULSE_SEQUENCE noesypr1d NM:WATER_SUPPRESSION presat NM:PULSE_WIDTH 90-degree NM:RECEIVER_GAIN 101 NM:OFFSET_FREQUENCY 2827.31 Hz NM:CHEMICAL_SHIFT_REF_CPD DSS (4,4-dimethyl-4-silapentane-1-sulfonic acid) NM:TEMPERATURE 25 o C NM:NUMBER_OF_SCANS 128 scans NM:DUMMY_SCANS 8 NM:ACQUISITION_TIME 4 s NM:RELAXATION_DELAY 1 s NM:SPECTRAL_WIDTH 7142.9 Hz NM:NUM_DATA_POINTS_ACQUIRED 28571 NM:REAL_DATA_POINTS 65536 NM:LINE_BROADENING 0.22 Hz NM:ZERO_FILLING 65,536 points NM:APODIZATION Exponential NM:BASELINE_CORRECTION_METHOD Spline NM:CHEMICAL_SHIFT_REF_STD 0 ppm for DSS #NMR_METABOLITE_DATA NMR_METABOLITE_DATA:UNITS A.U. NMR_METABOLITE_DATA_START Samples CKD1Liver_Aq CKD2Liver_Aq CKD3Liver_Aq CKD4Liver_Aq CKD5Liver_Aq CKD6Liver_Aq CKD7Liver_Aq Con1Liver_Aq Con2Liver_Aq Con3Liver_Aq Con4Liver_Aq Con5Liver_Aq Con6Liver_Aq Con7Liver_Aq Factors Group:CKD Group:CKD Group:CKD Group:CKD Group:CKD Group:CKD Group:CKD Group:Control Group:Control Group:Control Group:Control Group:Control Group:Control Group:Control Pantothenate? 4.249942567 2.530783109 2.48543602 2.769719847 4.873292366 2.595089694 1.832047479 3.084348491 2.27264492 4.458216985 2.292986714 2.767755415 1.905335243 3.877968432 Leucine 24.47028594 19.61823473 16.07667826 21.27953672 32.66695527 23.54718209 17.32243992 33.15210176 24.99024268 35.1598984 22.44020192 25.97721711 22.77709679 41.90298009 Valine1 10.9668263 6.846121194 6.396798417 7.985564309 13.56920701 8.854075943 7.654724169 14.03601598 11.6136629 16.61047053 8.940186418 10.62576066 8.06093437 15.38306181 Isoleucine 7.291543215 4.009439872 4.232342732 4.64445641 8.094475167 5.144406509 4.62157877 8.090774532 6.086445465 9.247209585 5.389824591 6.312519425 4.283887885 7.795632302 Valine2 10.92103641 6.794048795 6.532431328 8.319252856 13.61708315 9.139710665 7.778132403 14.36068625 11.02316182 16.7603789 9.375217344 10.55047389 7.806143562 14.89410583 Isobutyrate 5.560953732 2.488276924 3.185506668 3.548867287 6.939335056 4.134445976 3.046377358 4.919082373 3.369963163 6.101972443 4.031745514 4.931420309 3.199993999 4.683549033 Lactate1 282.5735317 215.8404584 156.1543643 239.0973171 337.1864615 279.6144294 246.4716442 369.9775116 355.8430072 380.9119254 320.1687157 373.8069672 335.9467347 395.9222774 Alanine 149.5545692 184.2926838 56.93330363 138.4804502 185.9545296 116.9885724 125.0962582 175.2233201 190.910721 88.56977759 136.8767971 193.0063284 141.9130729 242.6533615 Lysine 23.95625539 19.41020508 21.06953955 29.57263692 34.79472325 26.49194041 20.99283905 34.8503806 29.41668763 31.69909976 27.43382285 39.95136773 27.23644877 43.79579386 Acetate 6.111848731 6.977260243 6.985665877 10.1861933 13.48299078 12.53370051 8.042046417 9.776083879 11.35844696 8.170725309 9.904921948 12.29368527 8.013053882 9.810045182 Glutamate1 58.10469835 41.32299666 45.15770337 62.89578991 73.30669257 59.2315852 41.3173514 75.18703468 70.57363453 99.81097545 58.76856953 77.18191882 51.92831669 90.90779834 Methionine+O-Acetylcholine 117.429744 102.0302716 92.51771963 133.1539747 153.1587544 161.7484639 100.7874238 199.1313905 187.8684571 173.2895786 151.8852668 186.6189092 140.9749281 179.0881375 Glutamine1+Glutamate 8.028236004 7.493687526 5.558752776 9.442973012 10.08981121 11.17523576 6.372660226 9.636738536 10.11208737 6.651805227 6.399464851 8.705024459 6.356149832 9.074318429 2-Aminoadipate 17.28298757 10.15449933 14.64439777 17.2000859 24.92263939 18.1806351 12.84775438 24.24953081 21.82647891 60.86007263 20.72824352 26.68639417 20.42012805 25.76735062 Glutamate 26.47747279 23.12104133 21.30486341 29.1033411 38.65495518 36.07744286 21.59194883 41.93053047 52.03209014 42.8703396 29.15308545 37.84650447 25.31757391 45.17291106 Pyruvate+AM20:CM20 3.349865135 4.020968638 4.589982896 6.168842056 10.21375608 6.764162812 4.399345226 5.912793995 6.330641543 4.842097946 5.92240167 8.214690254 4.6042043 5.26775359 Succinate 25.20009733 20.14934319 15.54891144 19.87223252 31.86074605 28.17176944 26.97333641 35.41098105 28.09152638 23.77855891 25.95613675 37.07309992 26.5246467 36.0283028 Glutamine2 42.5842519 35.30106168 35.96366153 42.51091025 53.81837992 56.58003755 34.77815415 63.5572855 56.63069072 34.74062482 43.71278427 64.26254556 41.87064342 55.92411753 Isocitrate+Glutathione 88.69335078 71.13706492 80.51496786 110.4936572 123.2456897 119.9242416 81.44913629 165.3376624 136.9452393 141.0655909 142.4154275 160.8595167 120.9772537 142.1159463 Sarcosine? 4.815779315 3.300902212 3.954251611 4.857162361 6.636041371 5.727603228 3.753685928 5.699658298 5.97761087 3.836689124 5.37643282 7.078611367 5.165656955 7.07363084 Aspartate1 7.317306676 4.589806876 6.168290153 6.73427273 9.258193881 5.939257971 5.903352236 10.5516147 6.596765623 8.800761591 8.277570644 10.44301336 6.661179659 10.42302442 Aspartate2 6.619891258 3.75383731 5.828495551 6.172744441 8.243430899 5.418076277 4.975133521 9.431586541 6.008132333 7.782911599 7.590347308 9.54409415 6.118117941 8.851352343 Glutathione2 47.61251991 32.29410282 40.68962795 51.76086223 63.21076686 57.5220808 38.01912399 83.37339757 67.38252423 68.46845302 69.57910707 78.30534399 60.8149269 69.19968817 Creatine1 6.562500501 6.07348838 5.069256136 8.384934771 8.137205248 5.034081948 4.391377409 6.934537546 6.318458755 7.927879936 5.589937267 8.125128515 6.491887403 8.776958052 Creatinine 2.550528711 1.561082173 2.274737284 2.665513307 3.075926889 2.366636126 1.676329212 3.273581381 2.430081676 2.165531528 2.682472586 3.402942154 2.467617947 3.060109937 Glucose1 165.5801616 110.2707858 108.5788334 152.5952606 181.0800272 132.9309102 103.475091 156.2284007 143.2766111 112.5634122 157.3046969 162.1289666 119.0165035 132.1798075 Glycine 19.09761156 14.89644733 15.21481544 22.63327518 23.55705258 18.35861197 18.9212929 20.55582554 15.93417904 16.36982191 16.42627203 19.91554766 15.17931306 19.70837032 Glucose2 273.890835 189.5556291 171.293342 257.5384456 303.0938856 227.6956921 172.7146732 277.6313158 248.7764112 197.2544394 273.3599271 289.9953924 214.6212781 239.1017909 Creatine2 8.955968981 7.276730361 5.987970585 9.807664391 11.0345632 8.099648135 4.913630503 9.941309738 10.1980301 10.88151183 9.868759335 13.31391066 9.849237098 12.00659723 Lactate2 61.73857042 42.72606434 39.59040546 56.86418009 93.20880653 64.90501431 46.27732322 78.24573907 76.81404016 68.57593049 72.08577235 79.02986933 72.57071799 85.74264394 ATP+AMP1 13.17392428 3.22620999 5.548550165 13.57130992 23.8917096 13.50906671 2.959498188 17.7266564 19.23725113 17.85782866 17.20302735 19.3911953 18.76296604 21.58067554 Glucose3 27.75138057 16.24723448 16.29884073 23.16952447 29.99486343 19.75901483 15.82938371 24.77922428 23.34814411 16.89364856 26.27203023 24.96193599 19.14899145 22.14823099 Mannose 0.229943439 1.050777482 0.36680616 1.519709704 0.753730721 0.74353124 0.714668009 0.125963194 0.585832112 0.729636609 0.062293475 0.994415202 0.460056823 0.25064071 Glucose4 90.34351963 59.77241314 61.00890838 79.9049382 102.0084223 69.32682704 55.11044033 81.97974416 78.66041752 59.26984089 85.23688846 83.70660825 63.28972222 70.21128032 Sucrose+Maltose 3.856423758 6.606194932 3.572194198 4.848099633 10.59935566 9.083764415 3.640320495 1.387272038 10.59628487 2.44496216 5.614880719 9.088746896 4.656520197 11.6760813 UDP-N-Acetylglucosamine1 1.108344959 2.178603678 0.196640697 1.987423809 0.774811089 1.259146143 0.48181274 0.227371972 0.691986391 0.077692174 0.029066244 0.918132801 0.747538924 0.776020169 Uridine triphosphate 1.124491857 1.375925145 1.124158049 3.780817014 2.064060055 2.330052099 0.666791026 1.636436076 1.306712886 2.141954102 1.606523904 3.762152258 1.147654982 1.506319988 Unknown5.95to6.0ppm 1.379142418 4.024088891 2.286845435 13.05445023 3.349297984 4.365323165 2.795901482 6.674046175 5.527284466 7.988424988 5.362254685 11.65563309 4.707433961 5.635979219 NAD+1 0.186729947 0.43929728 0.363118419 0.810350458 0.330072984 0.516415889 0.055137905 0.562772194 0.372427661 0.520368415 0.479857019 0.848894677 0.32827383 0.34111997 NAD+2 1.137121293 1.340835031 0.573794981 1.3346232 0.913590028 1.178614854 0.315454467 0.692137486 0.760229299 1.530005497 0.753756434 0.798607466 0.624570594 0.648866793 ATP+AMP3 3.136305956 4.379022327 3.413439805 5.797388849 5.687175822 5.690396887 3.769878523 4.659289216 5.469291553 5.644408317 4.305088858 6.521119274 4.526121789 4.520635432 Fumarate 0.475712053 0.902844631 0.649692314 1.170112573 1.229960315 0.681792124 1.133069315 0.558001631 0.423267311 0.306595513 0.433659283 0.581727626 0.262305119 0.621010035 Tyrosine1 1.377044412 1.303082028 0.776509848 1.211601676 1.684695806 1.845059408 1.37738321 2.029221169 2.208943008 2.0505455 1.605212923 2.116286906 1.934513306 3.843191406 Histidine 1.749446918 2.408792215 0.98570771 2.87314418 2.465526196 2.086438141 1.962121461 2.296047305 2.101339389 1.518466752 1.724648068 2.988883288 1.519103641 2.055232549 Tyrosine2 1.88884124 1.852199161 1.100526575 1.698647293 2.436392633 2.618530989 1.926633443 2.86017017 3.148284166 2.914719607 2.258168782 2.979308268 2.510392668 5.195335367 Tryptophan1 1.059700438 2.087856403 1.744083741 2.705177261 4.833024561 2.883685268 1.826634035 1.764664228 2.514683746 1.780211137 2.13055321 3.12849498 1.453564871 1.642613082 Phenylalanine 1.311594352 1.450798704 0.870546541 1.358909926 1.94416649 1.789627296 1.306431549 1.697814493 1.724541362 1.873681968 1.534011073 1.903338755 1.0170495 1.961199268 Benzoate 0.532685725 1.594437523 1.083773062 1.609846402 3.060375197 2.288041058 0.884846163 1.666460964 1.97405846 1.433166462 1.636786544 1.962195468 0.794541104 1.10507742 Nicotinurate1 2.537720076 2.713085191 1.722561527 2.928362068 3.142833031 3.04114888 2.030220145 2.431222855 2.605843904 2.598831876 2.109961719 2.890250701 1.729256942 2.036240697 Tryptophan2 0.781165924 1.570683226 1.397300309 2.245996998 3.960656126 2.156546891 1.332946251 1.190389301 1.916242271 1.453982597 1.640073543 2.26244859 1.021063743 1.019390851 UDP-N-Acetylglucosamine2 0.905461827 1.170837089 0.811490029 3.729922891 1.945607241 1.341366918 1.170823261 2.322735342 1.763215492 3.219387406 2.012642488 3.051527641 1.360595879 2.158561893 ATP+AMP4 0.821037486 1.462784833 0.866890675 3.82936946 1.50492594 1.335723553 1.127810251 1.765872545 2.188208402 1.816978576 1.814500215 3.277399243 1.756563447 1.639905257 Unknown7.96to7.94 1.200148479 1.942047789 1.685255894 2.696440762 2.650013745 2.26407347 1.538118815 1.558750469 1.842731159 1.873741283 1.467653222 2.296102383 1.403926891 1.609056996 Inosine_or_Adenosine 1.310009844 1.493949842 0.739417635 1.254002736 0.973643906 1.037190406 0.482340881 0.481876837 0.559739923 1.318241867 0.594821125 0.244922785 0.45610751 0.528381438 Formate 0.154515795 0.544798982 0.761613371 0.715393238 1.4284691 1.088148422 0.409508227 0.34111473 0.438230964 0.174822485 0.430795796 0.476491383 0.194100717 0.160361314 Nicotinurate 2 1.571899157 1.599288073 1.097969686 1.865840656 1.855559377 1.68471881 1.170553097 1.350376861 1.404338706 1.495807638 1.110036419 1.410074142 1.020052943 1.321392408 NAD+3 0.121914424 0.350018569 0.476728825 0.526091714 0.470541648 0.290738424 0.214228148 0.407889051 0.367919166 0.386986487 0.483351843 0.385467971 0.316890928 0.38287126 Nicotinurate3 1.09029707 1.164364328 0.874475419 1.426376213 1.435703978 1.205442411 0.844530735 0.847737484 0.824776236 0.943326895 0.696533881 1.032801427 0.599659474 0.886231904 NAD+4 0.143243986 0.368360438 0.586421439 0.46181357 0.456049924 0.438099162 0.348591242 0.477785846 0.429313509 0.47328135 0.427600913 0.548980494 0.394976949 0.455283035 NAD+5 0.21358695 0.352117794 0.497588604 0.548235711 0.556545523 0.414662728 0.338056864 0.466190752 0.466410325 0.441751574 0.438593153 0.537254626 0.409973986 0.439605852 NMR_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Pantothenate? Leucine Valine1 Isoleucine Valine2 Isobutyrate Lactate1 Alanine Lysine Acetate Glutamate1 Methionine+O-Acetylcholine Glutamine1+Glutamate 2-Aminoadipate Glutamate Pyruvate+AM20:CM20 Succinate Glutamine2 Isocitrate+Glutathione Sarcosine? Aspartate1 Aspartate2 Glutathione2 Creatine1 Creatinine Glucose1 Glycine Glucose2 Creatine2 Lactate2 ATP+AMP1 Glucose3 Mannose Glucose4 Sucrose+Maltose UDP-N-Acetylglucosamine1 Uridine triphosphate Unknown5.95to6.0ppm NAD+1 NAD+2 ATP+AMP3 Fumarate Tyrosine1 Histidine Tyrosine2 Tryptophan1 Phenylalanine Benzoate Nicotinurate1 Tryptophan2 UDP-N-Acetylglucosamine2 ATP+AMP4 Unknown7.96to7.94 Inosine_or_Adenosine Formate Nicotinurate 2 NAD+3 Nicotinurate3 NAD+4 NAD+5 METABOLITES_END #END