#METABOLOMICS WORKBENCH amuirchicago_20250225_104023 DATATRACK_ID:5676 STUDY_ID:ST003759 ANALYSIS_ID:AN006172 PROJECT_ID:PR002342 VERSION 1 CREATED_ON February 28, 2025, 12:51 pm #PROJECT PR:PROJECT_TITLE Analysis of metabolite levels in the circulation of pancreatic tumor bearing PR:PROJECT_TITLE mice with or without depletion of myeloid cells PR:PROJECT_SUMMARY In this study, we assessed the impact of myeloid cells on the levels of PR:PROJECT_SUMMARY circulating metabolites in the plasma of mice bearing tumors. To do so, murine PR:PROJECT_SUMMARY pancreatic ductal adenocarcinoma (PDAC) cells were implanted into C57Bl6J PR:PROJECT_SUMMARY CD11b-DTR host animals. Myeloid cells were then ablated with 25 ng/g diptheria PR:PROJECT_SUMMARY toxin (DT). Serum from DT treated mice was collected for analysis. Additionally, PR:PROJECT_SUMMARY serum was collected from PDAC bearing mice that were vehicle treated. This data PR:PROJECT_SUMMARY allows for identification of how myeloid cells in tumor bearing mice contribute PR:PROJECT_SUMMARY to levels of circulating metabolites. PR:INSTITUTE University of Chicago PR:DEPARTMENT Ben May Department for Cancer Research PR:LAST_NAME Muir PR:FIRST_NAME Alexander PR:ADDRESS 929 E 57th St PR:EMAIL amuir@uchicago.edu PR:PHONE 7738346506 #STUDY ST:STUDY_TITLE Analysis of metabolite levels in the circulation of pancreatic tumor bearing ST:STUDY_TITLE mice with or without depletion of myeloid cells ST:STUDY_SUMMARY In this study, we assessed the impact of myeloid cells on the levels of ST:STUDY_SUMMARY circulating metabolites in the plasma of mice bearing tumors. To do so, murine ST:STUDY_SUMMARY pancreatic ductal adenocarcinoma (PDAC) cells were implanted into C57Bl6J ST:STUDY_SUMMARY CD11b-DTR host animals. Myeloid cells were then ablated with 25 ng/kg diptheria ST:STUDY_SUMMARY toxin (DT). Serum from DT treated mice was collected for analysis. Additionally, ST:STUDY_SUMMARY serum was collected from PDAC bearing mice that were vehicle treated. This data ST:STUDY_SUMMARY allows for identification of how myeloid cells in tumor bearing mice contribute ST:STUDY_SUMMARY to levels of circulating metabolites. ST:INSTITUTE University of Chicago ST:LAST_NAME Muir ST:FIRST_NAME Alexander ST:ADDRESS 929 E 57th St ST:EMAIL amuir@uchicago.edu ST:PHONE 7738346506 #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 - serum_DT_1 Sample source:Blood serum | Treatment:Diptheria_toxin RAW_FILE_NAME(Raw_data_file_name)=190723_QE2_AlexanderMuir_Job1623_35.raw SUBJECT_SAMPLE_FACTORS - serum_DT_2 Sample source:Blood serum | Treatment:Diptheria_toxin RAW_FILE_NAME(Raw_data_file_name)=190723_QE2_AlexanderMuir_Job1623_36.raw SUBJECT_SAMPLE_FACTORS - serum_DT_3 Sample source:Blood serum | Treatment:Diptheria_toxin RAW_FILE_NAME(Raw_data_file_name)=190723_QE2_AlexanderMuir_Job1623_37.raw SUBJECT_SAMPLE_FACTORS - serum_DT_4 Sample source:Blood serum | Treatment:Diptheria_toxin RAW_FILE_NAME(Raw_data_file_name)=190723_QE2_AlexanderMuir_Job1623_38.raw SUBJECT_SAMPLE_FACTORS - serum_DT_5 Sample source:Blood serum | Treatment:Diptheria_toxin RAW_FILE_NAME(Raw_data_file_name)=190723_QE2_AlexanderMuir_Job1623_39.raw SUBJECT_SAMPLE_FACTORS - serum_DT_6 Sample source:Blood serum | Treatment:Diptheria_toxin RAW_FILE_NAME(Raw_data_file_name)=190723_QE2_AlexanderMuir_Job1623_40.raw SUBJECT_SAMPLE_FACTORS - serum_noDT_1 Sample source:Blood serum | Treatment:vehicle RAW_FILE_NAME(Raw_data_file_name)=190723_QE2_AlexanderMuir_Job1623_47.raw SUBJECT_SAMPLE_FACTORS - serum_noDT_2 Sample source:Blood serum | Treatment:vehicle RAW_FILE_NAME(Raw_data_file_name)=190723_QE2_AlexanderMuir_Job1623_48.raw SUBJECT_SAMPLE_FACTORS - serum_noDT_3 Sample source:Blood serum | Treatment:vehicle RAW_FILE_NAME(Raw_data_file_name)=190723_QE2_AlexanderMuir_Job1623_49.raw SUBJECT_SAMPLE_FACTORS - serum_noDT_4 Sample source:Blood serum | Treatment:vehicle RAW_FILE_NAME(Raw_data_file_name)=190723_QE2_AlexanderMuir_Job1623_50.raw SUBJECT_SAMPLE_FACTORS - serum_noDT_5 Sample source:Blood serum | Treatment:vehicle RAW_FILE_NAME(Raw_data_file_name)=190723_QE2_AlexanderMuir_Job1623_51.raw SUBJECT_SAMPLE_FACTORS - serum_noDT_6 Sample source:Blood serum | Treatment:vehicle RAW_FILE_NAME(Raw_data_file_name)=190723_QE2_AlexanderMuir_Job1623_52.raw #COLLECTION CO:COLLECTION_SUMMARY Mice were euthanised 24 hours after treatment by CO2 asphyxiation and blood CO:COLLECTION_SUMMARY collected through cardiac puncture. Serum was isolated using BD microtainer CO:COLLECTION_SUMMARY serum separator tubes as per manufactures’ instructions and stored at -80 CO:COLLECTION_SUMMARY until analysis. CO:SAMPLE_TYPE Blood (serum) #TREATMENT TR:TREATMENT_SUMMARY CD11b-DTR mice were produced as an F1 cross between TR:TREATMENT_SUMMARY B6.FVB-Tg(ITGAM-DTR/EGFP)34Lan/J mice and C57BL/6J mice (Jackson Laboratory). TR:TREATMENT_SUMMARY Male mice between 8-12 weeks of age were used experimentally. Orthotopic PDAC TR:TREATMENT_SUMMARY tumours were established from 5x104 KPC-7940B PDAC cells injected into the TR:TREATMENT_SUMMARY pancreas were established as tumors for 14 days. Mice were then treated with TR:TREATMENT_SUMMARY diphtheria toxin (Enzo Life Science) at a concentration of 25 ng/kg or vehicle TR:TREATMENT_SUMMARY by intraperitoneal injection. Mice were euthanised 24 hours after treatment by TR:TREATMENT_SUMMARY CO2 asphyxiation and blood collected through cardiac puncture. Serum was TR:TREATMENT_SUMMARY isolated using BD microtainer serum separator tubes as per manufactures’ TR:TREATMENT_SUMMARY instructions and stored at -80 until analysis. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Metabolites were quantified as described in Sullivan et al eLife 2019 (PMID: SP:SAMPLEPREP_SUMMARY 30990168). Briefly, in order to quantitate metabolites in serum samples, we SP:SAMPLEPREP_SUMMARY first constructed a library of 149 chemical standards of plasma polar SP:SAMPLEPREP_SUMMARY metabolites. These compounds were selected to encompass a number of metabolic SP:SAMPLEPREP_SUMMARY processes and have previously been included in efforts to profile plasma polar SP:SAMPLEPREP_SUMMARY metabolites by LC/MS (Cantor et al., 2017; Evans et al., 2009; Lawton et al., SP:SAMPLEPREP_SUMMARY 2008; Mazzone et al., 2016). We pooled these metabolites into 7 separate SP:SAMPLEPREP_SUMMARY chemical standard pools. To do this, each metabolite in a given pool was weighed SP:SAMPLEPREP_SUMMARY and then mixed (6 cycles of 1 min. mixing at 25 Hz followed by 3 min. resting) SP:SAMPLEPREP_SUMMARY using a Mixer Mill MM301 (Retsch, Düsseldorf, Germany), and mixed metabolite SP:SAMPLEPREP_SUMMARY powder stocks were stored at -20°C prior to resuspension and analysis. Stock SP:SAMPLEPREP_SUMMARY solutions of the mixed standards pools containing ~5mM, ~1mM, ~300µM, ~100µM, SP:SAMPLEPREP_SUMMARY ~30µM, ~10µM, ~3µM and ~1µM of each metabolite were made in HPLC grade water SP:SAMPLEPREP_SUMMARY and were stored at -80°C. We refer to these stock solutions as “external SP:SAMPLEPREP_SUMMARY standard pools” throughout. External standard pools were used to confirm the SP:SAMPLEPREP_SUMMARY retention time and m/z for each analyte and provide standards to quantitate SP:SAMPLEPREP_SUMMARY concentrations of stable isotope labeled internal standards used in downstream SP:SAMPLEPREP_SUMMARY analysis, as well as to quantitate metabolite concentrations in TIF and plasma SP:SAMPLEPREP_SUMMARY samples directly where internal standards were not available (see below for SP:SAMPLEPREP_SUMMARY details). To extract polar metabolites from plasma, TIF or the external standard SP:SAMPLEPREP_SUMMARY pools, 5µL of serum or external sample pools was mixed with 45uL of SP:SAMPLEPREP_SUMMARY acetonitrile:methanol:formic acid (75:25:0.1) extraction buffer including the SP:SAMPLEPREP_SUMMARY following isotopically labeled internal standards: 13C labeled yeast extract SP:SAMPLEPREP_SUMMARY (Cambridge Isotope Laboratory, Andover, MA, ISO1), 13C3 lactate (Sigma Aldrich, SP:SAMPLEPREP_SUMMARY Darmstadt, Germany, 485926), 13C3 glycerol (Cambridge Isotope Laboratory, SP:SAMPLEPREP_SUMMARY Andover, MA, CLM-1510), 13C6 15N2 cystine (Cambridge Isotope Laboratory, SP:SAMPLEPREP_SUMMARY Andover, MA, CNLM-4244), 2H9 choline (Cambridge Isotope Laboratory, Andover, MA, SP:SAMPLEPREP_SUMMARY DLM-549), 13C4 3-hydroxybutyrate (Cambridge Isotope Laboratory, Andover, MA, SP:SAMPLEPREP_SUMMARY CLM-3853), 13C6 glucose (Cambridge Isotope Laboratory, Andover, MA, CLM-1396), SP:SAMPLEPREP_SUMMARY 13C2 15N taurine (Cambridge Isotope Laboratory, Andover, MA, CNLM-10253), 2H3 SP:SAMPLEPREP_SUMMARY creatinine (Cambridge Isotope Laboratory, Andover, MA, DLM-3653), 8-13C adenine SP:SAMPLEPREP_SUMMARY (Cambridge Isotope Laboratory, Andover, MA, CLM-1654), 13C5 hypoxanthine SP:SAMPLEPREP_SUMMARY (Cambridge Isotope Laboratory, Andover, MA, CLM-8042), 8-13C guanine (Cambridge SP:SAMPLEPREP_SUMMARY Isotope Laboratory, Andover, MA, CLM-1019), 13C3 serine (Cambridge Isotope SP:SAMPLEPREP_SUMMARY Laboratory, Andover, MA, CLM-1574) and 13C2 glycine (Cambridge Isotope SP:SAMPLEPREP_SUMMARY Laboratory, Andover, MA, CLM-1017). All solvents used in the extraction buffer SP:SAMPLEPREP_SUMMARY were HPLC grade. Samples were then vortexed for 10 min. at 4°C and insoluble SP:SAMPLEPREP_SUMMARY material was sedimented by centrifugation at 15kg for 10 min. at 4°C. 20µL of SP:SAMPLEPREP_SUMMARY the soluble polar metabolite extract was taken for LC/MS analysis. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Thermo Dionex Ultimate 3000 CH:COLUMN_NAME SeQuant ZIC-pHILIC (150 x 2.1mm,5um) CH:SOLVENT_A 100% water; 20 mM ammonium carbonate, 0.1% ammonium hydroxide CH:SOLVENT_B 100% acetonitrile CH:FLOW_GRADIENT linear gradient from 80% to 20% B; 20-20.5 min: linear gradient from 20% to 80% CH:FLOW_GRADIENT B; 20.5-28min: hold at 80% B. CH:FLOW_RATE 0.150 mL/min CH:COLUMN_TEMPERATURE 25 #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS LC/MS analysis was performed using a QExactive orbitrap mass spectrometer using MS:MS_COMMENTS an Ion Max source and heated electrospray ionization (HESI) probe coupled to a MS:MS_COMMENTS Dionex Ultimate 3000 UPLC system (Thermo Fisher Scientific, Waltham, MA). MS:MS_COMMENTS External mass calibration was performed every 7 days. 2μL of each sample was MS:MS_COMMENTS injected onto a ZIC-pHILIC 2.1 × 150 mm analytical column equipped with a 2.1 MS:MS_COMMENTS × 20 mm guard column (both 5 μm particle size, EMD Millipore). The autosampler MS:MS_COMMENTS and column oven were held at 4°C and 25°C, respectively. Buffer A was 20 mM MS:MS_COMMENTS ammonium carbonate, 0.1% ammonium hydroxide; buffer B was acetonitrile. The MS:MS_COMMENTS chromatographic gradient was run at a flow rate of 0.150 mL/min as follows: 0-20 MS:MS_COMMENTS min: linear gradient from 80% to 20% B; 20-20.5 min: linear gradient from 20% to MS:MS_COMMENTS 80% B; 20.5-28min: hold at 80% B. The mass spectrometer was operated in full MS:MS_COMMENTS scan, polarity-switching mode with the spray voltage set to 3.0 kV, the heated MS:MS_COMMENTS capillary held at 275°C, and the HESI probe held at 350°C. The sheath gas flow MS:MS_COMMENTS rate was set to 40 units, the auxiliary gas flow was set to 15 units, and the MS:MS_COMMENTS sweep gas flow was set to 1 unit. The MS data acquisition was performed in a MS:MS_COMMENTS range of 70-1000 m/z, with the resolution set to 70,000, the AGC target at 1e6, MS:MS_COMMENTS and the maximum injection time at 20 msec. Metabolite identification and MS:MS_COMMENTS quantification was performed with XCalibur 2.2 software (Thermo Fisher MS:MS_COMMENTS Scientific, Waltham, MA) using a 5ppm mass accuracy and a 0.5 min. retention MS:MS_COMMENTS time window. For metabolite identification, external standard pools were used MS:MS_COMMENTS for assignment of metabolites to peaks at given m/z and retention time, and to MS:MS_COMMENTS determine the limit of detection for each metabolite. Metabolite quantification MS:MS_COMMENTS was performed by two separate methods. Where internal standards were available, MS:MS_COMMENTS first, comparison of the peak areas of the stable isotope labeled internal MS:MS_COMMENTS standards with the external standard pools allowed for quantification of the MS:MS_COMMENTS concentration of labeled internal standards in the extraction buffer. MS:MS_COMMENTS Subsequently, we compared the peak area of a given metabolite in the TIF and MS:MS_COMMENTS plasma samples with the peak area of the internal standard to quantitate the MS:MS_COMMENTS concentration of that metabolite in the serum sample. For metabolites without MS:MS_COMMENTS internal standards, the peak area of each analyte was normalized to the peak MS:MS_COMMENTS area of a labeled amino acid internal standard that eluted at roughly the same MS:MS_COMMENTS retention time to account for differences in recovery between samples. From the MS:MS_COMMENTS normalized peak areas of metabolites in the external standard pools, we MS:MS_COMMENTS generated a standard curve describing the relationship between metabolite MS:MS_COMMENTS concentration and normalized peak area. The standard curves were linear with MS:MS_COMMENTS fits typically at or above r2=0.95. Metabolites which did not meet these MS:MS_COMMENTS criteria were excluded from further analysis. These equations were then used to MS:MS_COMMENTS convert normalized peak areas of analytes in the serum samples into analyte MS:MS_COMMENTS concentration in the samples. The relationship between metabolite concentration MS:MS_COMMENTS and normalized peak area is matrix dependent, and the external standards are MS:MS_COMMENTS prepared in water, which is a different matrix than serum. Therefore, we MS:MS_COMMENTS consider metabolite measurements using this external standard method MS:MS_COMMENTS semi-quantitative. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS micromole/L MS_METABOLITE_DATA_START Samples serum_DT_1 serum_DT_2 serum_DT_3 serum_DT_4 serum_DT_5 serum_DT_6 serum_noDT_1 serum_noDT_2 serum_noDT_3 serum_noDT_4 serum_noDT_5 serum_noDT_6 Factors Sample source:Blood serum | Treatment:Diptheria_toxin Sample source:Blood serum | Treatment:Diptheria_toxin Sample source:Blood serum | Treatment:Diptheria_toxin Sample source:Blood serum | Treatment:Diptheria_toxin Sample source:Blood serum | Treatment:Diptheria_toxin Sample source:Blood serum | Treatment:Diptheria_toxin Sample source:Blood serum | Treatment:vehicle Sample source:Blood serum | Treatment:vehicle Sample source:Blood serum | Treatment:vehicle Sample source:Blood serum | Treatment:vehicle Sample source:Blood serum | Treatment:vehicle Sample source:Blood serum | Treatment:vehicle 2-aminobutyrate 16.58206841 10.40826316 8.797613955 11.18974295 7.000373744 10.46246429 22.80401203 21.81586947 33.89294099 24.19108908 28.005384 9.26986781 N6-acetyllysine 0.553791277 0.594841806 0.576745977 1.105778364 0.689065803 0.643398049 0.640196393 0.358792331 0.824000462 0.965811647 0.853440137 0.536461239 gamma-glutamyl-alanine 0.921868417 1.831219108 0.889718035 1.228854241 1.078860516 1.596024465 0.169631525 1.025482231 0 0.761911066 pantothenate 2.433377291 3.590881119 2.764160155 2.825014459 2.23656454 3.475354039 4.714815782 4.446637484 3.211570513 4.441922265 5.604054452 1.978618784 glycerophosphocholine 27.36178057 8.930696976 27.10518938 34.44467288 12.37919919 16.55067531 654.5172655 1024.280043 542.002414 745.8400036 1569.936761 154.6471897 aminoadipate 9.541266419 5.057720982 6.594377594 11.3957707 12.57823715 12.80687666 21.92524036 20.34622188 19.74723015 28.3593168 20.30044331 10.28526151 cystathionine 1.778018851 1.3277972 2.134931637 2.840104911 2.079586818 2.118562662 6.276123121 7.321769109 10.95049747 8.117833929 10.93936533 4.791443744 niacinamide 6.576988383 4.553413699 6.026329097 9.481170161 5.621145859 7.16778238 66.17522048 100.0437354 68.40451919 73.04007657 119.5749752 19.02425624 methylthioadenosine 0.010952009 0.013680545 0.013986713 0.02184472 0.010110219 0.018764162 0.214906831 0.534265703 0.371770746 0.421547422 0.446626501 0.080773581 SAH 0.063912433 0.151959743 0 0.16882824 0 0.707254059 0.070260335 0.35551796 0.032184404 0.14156361 0.099431255 0 methionine sulfoxide 2.006015087 2.645910612 2.678561032 4.333297843 3.577822417 1.810846295 4.632365534 2.898114402 5.417978786 5.462493236 3.467551248 1.476162568 acetylglutamate 0.363548652 0.352242223 0.157599773 0.468821242 0.178783924 0.555043221 0.638828606 0.54196071 0.727423224 0.86164877 0.971261012 0.248441695 asymmetric dimethylarginine 0.909507393 1.931551419 1.108846067 0.888622968 0.78274502 2.544688054 0.84228 0.578350491 1.634995289 0.905484833 0.875561497 1.4806692 trimethyllysine 0.816340745 0.802510252 1.20613032 1.310868489 1.415009616 1.264636302 1.704423408 0.906038238 1.410257952 2.593566706 2.0386802 1.299055851 creatinine 6.904493612 8.192232234 9.653837157 11.88348617 12.59013677 9.994774245 26.55051999 13.34401373 16.38811656 21.12444243 30.54032865 7.86556262 acetylalanine 0.327405877 1.516412352 0 2.574038865 0 0 7.216375906 0 6.674269723 0 1.976552781 4.786887817 3-hydroxybutyrate 155.3051279 161.409914 159.9282278 183.1314695 197.3921599 447.4164091 21.63571328 22.07958817 27.41639114 71.67442351 79.55329344 35.21790871 hypoxanthine 0.02386804 0.016306091 0.042243435 0.101605121 6.698904751 0.114315073 45.92013553 197.2033046 17.57073038 10.27122802 205.2447652 7.066315851 acetylglutamine 1.228685994 0.635126045 0.566382491 1.878659603 0.630897139 2.152175671 0.567654495 0.857547076 0.904260798 0.759544783 0.986038472 0.497761683 NAD 0 0 0 0 0 0 0 0 0 0 0 0 gaba 0 0.170729934 0 0 0 0.114684096 0.83689396 0 0 1.242254494 6.096206803 0.140836672 phosphocholine 3.24795529 2.99545579 2.789018026 3.530169947 2.758232729 6.030024923 381.4130896 884.9933323 397.2791976 480.9045921 791.8220878 69.21551607 gssg 13.61146912 14.72704248 5.245865523 42.78405993 7.317904748 17.53140884 171.6677236 384.7492953 129.7301866 156.7802848 334.7067768 41.55930891 choline 69.95580677 64.41025134 71.03574 88.89487317 43.41053457 140.101752 252.1209757 289.0483058 214.5479322 238.3381602 426.1847314 69.09686671 gsh 0.194121023 0.156030337 0.130188133 0.176090905 0.073176267 0.156456204 1.056979569 12.93405434 0.585151367 0.351093216 4.210889917 2.641063139 UMP 0 0 0 0 0 0 44.52067157 77.95674414 68.18578789 74.61440851 88.86688341 13.82647387 CMP 0 0 0 0 0 0 14.53422116 19.94607564 19.03830469 22.23093118 23.43119493 4.922055739 phenylalanine 117.7058749 160.4937958 127.8572526 173.83861 101.6037433 155.2187652 141.9252023 110.9786649 162.1474704 180.8273427 175.1320259 109.0978284 leucine 290.3664773 305.4109804 318.2549728 449.7186174 200.4109316 325.6982804 380.4528715 297.5540052 406.9534499 462.8105462 383.8202716 296.7924931 methionine 71.67479401 89.27718133 107.5872104 104.4983945 90.72091545 71.4190615 136.1323636 104.5356058 152.0016261 158.2260982 133.5228458 75.07103523 tryptophan 110.798161 102.3252978 107.1217146 129.2691453 81.23862266 106.0918156 110.0373878 51.27751757 75.97122499 117.0475326 105.3382284 53.91219867 valine 304.4737664 339.1615472 326.7038854 430.3817347 237.3426702 339.7924487 370.7764096 324.3272781 497.1842233 501.8346726 363.9440374 300.8244377 proline 86.84299935 138.9722274 115.0173409 122.5672023 102.9732806 88.48479683 286.6094858 304.3340273 347.9441478 323.8690954 392.0234986 157.5586823 tyrosine 93.22945255 134.0614888 85.42853819 123.7170421 82.17040103 101.1883383 164.9191054 110.6924331 177.1011622 171.8363383 237.0968764 84.56116029 threonine 171.9693614 182.8800807 216.2774844 185.6777109 182.8086426 151.570958 396.3117155 334.8560494 435.6112429 408.6750466 371.6286821 206.9322863 histidine 85.24093507 124.1763113 118.869703 140.6127544 94.81402899 96.49291106 147.5013642 148.235936 167.6848855 169.3015812 184.1496002 156.9406138 alanine 447.8084778 755.1773225 608.6514628 546.6275583 458.1140048 591.2069414 1410.933163 1495.63889 1261.653327 1407.546212 2369.731479 645.8904399 isoleucine 168.9898402 169.3224022 169.7572562 252.2496474 118.5231478 208.9515648 203.1564022 159.3161333 229.7469553 247.4684718 207.8138844 144.2249672 glutamine 1187.94565 1376.471018 1767.25918 1717.187257 1180.784692 1411.888659 1068.386103 944.036859 1001.389767 1057.246616 1202.90923 859.3459131 asparagine 43.06883637 106.6047347 56.33323331 50.92651089 42.96179117 48.02743125 148.4491069 134.8620582 134.2679817 140.7812958 259.6156289 81.78077521 glycine 304.1010581 434.8457489 412.1254199 367.4467913 330.3174614 365.0538523 1187.021805 1494.231468 1818.944297 1468.910138 1921.037769 633.388181 citrulline 68.26234406 82.75077816 82.02820536 101.4801354 80.39815966 57.55068203 103.3090189 84.52580949 92.15788694 90.77249796 94.72732235 63.12795945 cystine 2.681833799 2.197004783 3.327847825 4.610249062 9.275564219 6.157629541 16.14776252 10.53865802 28.35859955 23.08870064 12.44330138 33.7864684 ornithine 106.9807828 192.778119 161.2691093 181.3117584 145.0074077 111.4061745 414.9906027 242.2405921 372.7841733 352.763636 314.7542017 224.1817948 lysine 393.611208 455.9314728 594.5171758 609.5928814 403.1729341 406.8224185 569.9721103 425.4964091 613.7122878 649.999406 542.4498774 407.9282905 arginine 269.4821649 214.4408662 375.0749735 400.2240465 266.4652428 269.3347424 4.395211891 3.757559975 4.96165094 8.09232496 6.10607161 17.95285607 indolelactate 0 0 0 0 0 0 0 0 0 0 0 0 picolinate 0 0 0 0 0 0 0 0 0 0 0 0 homocysteine 0 0 0 0 0 0 0 0 0 0 0 0 cys-gly 0 0 0 0 0 0 0 0 0 0 0 0 sarcosine 0.44 0.61 1.21 0.02 0.24 0.32 1.41 1.81 3.2 1.47 1.5 0.28 homocitrulline 0 0 0 0 0 0 0 0 0 0 0 0 riboflavin 0 0 0 0 0 0 0 0 0 0 0 0 inosine 0 0 0 0 20.77 0 35.52 48.64 4.88 39.24 31.84 8.8 beta-alanine 0 0 0 0 0 0 26.45 34.48 22.6 35.02 47.68 0 kynurenine 0.71 2.16 1.58 2.56 2.08 1 6.82 6.95 17.28 10.11 17.28 7.64 guanidinoacetate 0 0 0 0 0 3.9 0 0 58.07 4239.87 0 0 glycerol-3-phosphate 0 0 0 0 0 0 0 0 0 0 0 0 NADP 0 0 0 0 0 0 0 0 0 0 0 0 fructose-1,6-bisphosphate 0 0 0 0 0 0 0 0 0 0 0 0 phenylacetylglutamine 0 0 0 0 0 0 0 0 0 0 0 0 pyridoxine 0 0 0 0 0 0 0 0 0 0 0 0 deoxycytidine 0.83 0.78 1.37 0.84 0.78 0.72 0.79 0.18 0.94 0.8 0.75 1 hypotaurine 0 0 0 0 0 0 0 0 0 0 0 0 phosphocreatine 0 0 0 0 0.53 0 151.33 56.48 111.32 156.59 86.06 38.97 dATP 0 0 0 0 0 0 0 0 0 0 0 0 folate 0 0 0 0 0 0 0 0 0 0 0 0 acetylcarnitine 29.8 33.01 40.31 39.53 18.82 34.64 39.51 28.15 34.21 45.17 55.4 18.46 CoA 0 0 0 0 0 0 0 0 0 0 0 0 acetylaspartate 0 0 0 0 0 0 0 0 0 0 0 0 ADP 0 0 0 0 0 0 0 0 0 0 0.61 0 CDP-choline 0 0 0 0 0 0 0 0 0 0 0 0 UDP 0 0 0 0 0 0 0 0 0 0 0 0 CDP 0 0 0 0 0 0 0 0 0 0 0 0 GDP 0 0 0 0 0 0 0 0 0 0 0 0 pyridoxal 0.11 0.17 0.06 0.2 0.04 0.28 0.1 0.04 0.28 0.22 0 0.14 biotin 0.01 0.03 0.02 0.01 0 0 0 0 0 0 0 0 betaine 107.82 81.6 129.44 75.38 41.95 81.45 124.13 70.62 133.12 116.18 146.06 69.03 cytidine 0 0 0 0 0 0 0 0 0 0 0 0 trans-urocanate 0 0.48 0.44 1.36 0.49 0.17 1.8 1.71 0.79 2.3 2.1 0.58 creatine 52.25 76.91 92.16 61.1 86.99 72.33 806.5 527.36 321.25 318.19 1158.15 114.79 carnosine 0 0 0 0 0 0 0 0 0 0 0 0 IMP 0 0 0 0 0 0 0 0 0 0 0 0 hydroxyproline 1.28 2.48 1.84 4.16 1.24 4.05 0 0 8.68 5.68 0 5.16 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name 2-aminobutyrate N6-acetyllysine gamma-glutamyl-alanine pantothenate glycerophosphocholine aminoadipate cystathionine niacinamide methylthioadenosine SAH methionine sulfoxide acetylglutamate asymmetric dimethylarginine trimethyllysine creatinine acetylalanine 3-hydroxybutyrate hypoxanthine acetylglutamine NAD gaba phosphocholine gssg choline gsh UMP CMP phenylalanine leucine methionine tryptophan valine proline tyrosine threonine histidine alanine isoleucine glutamine asparagine glycine citrulline cystine ornithine lysine arginine indolelactate picolinate homocysteine cys-gly sarcosine homocitrulline riboflavin inosine beta-alanine kynurenine guanidinoacetate glycerol-3-phosphate NADP fructose-1,6-bisphosphate phenylacetylglutamine pyridoxine deoxycytidine hypotaurine phosphocreatine dATP folate acetylcarnitine CoA acetylaspartate ADP CDP-choline UDP CDP GDP pyridoxal biotin betaine cytidine trans-urocanate creatine carnosine IMP hydroxyproline METABOLITES_END #END