#METABOLOMICS WORKBENCH amuir_20190103_160816 DATATRACK_ID:1594 STUDY_ID:ST001119 ANALYSIS_ID:AN001831 PROJECT_ID:PR000750 VERSION 1 CREATED_ON January 14, 2019, 11:14 am #PROJECT PR:PROJECT_TITLE Quantification of microenvironmental metabolites in murine cancers reveals PR:PROJECT_TITLE determinants of tumor nutrient availability PR:PROJECT_SUMMARY Cancer cell metabolism is heavily influenced by microenvironmental factors, PR:PROJECT_SUMMARY including nutrient availability. Therefore, knowledge of microenvironmental PR:PROJECT_SUMMARY nutrient levels is essential to understand tumor metabolism. To measure the PR:PROJECT_SUMMARY extracellular nutrient levels available to tumors, we developed a quantitative PR:PROJECT_SUMMARY metabolomics method to measure the absolute concentrations of >118 metabolites PR:PROJECT_SUMMARY in plasma and tumor interstitial fluid, the extracellular fluid that perfuses PR:PROJECT_SUMMARY tumors. Comparison of nutrient levels in tumor interstitial fluid and plasma PR:PROJECT_SUMMARY revealed that the nutrients available to tumors differ from those present in PR:PROJECT_SUMMARY circulation. Further, by comparing interstitial fluid nutrient levels between PR:PROJECT_SUMMARY autochthonous and transplant models of murine pancreatic and lung PR:PROJECT_SUMMARY adenocarcinoma, we found that tumor type, anatomical location and animal diet PR:PROJECT_SUMMARY affect local nutrient availability. These data provide a comprehensive PR:PROJECT_SUMMARY characterization of the nutrients present in the tumor microenvironment of PR:PROJECT_SUMMARY widely used models of lung and pancreatic cancer and identify factors that PR:PROJECT_SUMMARY influence metabolite levels in tumors. PR:INSTITUTE University of Chicago PR:LAST_NAME Muir PR:FIRST_NAME Alexander PR:ADDRESS 929 E 57th St. GCIS W 306, Chicago, Illinois, 60637, USA PR:EMAIL muir.alexander@gmail.com PR:PHONE 5104950975 #STUDY ST:STUDY_TITLE Quantification of microenvironmental metabolites in murine cancers reveals ST:STUDY_TITLE determinants of tumor nutrient availability ST:STUDY_SUMMARY Cancer cell metabolism is heavily influenced by microenvironmental factors, ST:STUDY_SUMMARY including nutrient availability. Therefore, knowledge of microenvironmental ST:STUDY_SUMMARY nutrient levels is essential to understand tumor metabolism. To measure the ST:STUDY_SUMMARY extracellular nutrient levels available to tumors, we developed a quantitative ST:STUDY_SUMMARY metabolomics method to measure the absolute concentrations of >118 metabolites ST:STUDY_SUMMARY in plasma and tumor interstitial fluid, the extracellular fluid that perfuses ST:STUDY_SUMMARY tumors. Comparison of nutrient levels in tumor interstitial fluid and plasma ST:STUDY_SUMMARY revealed that the nutrients available to tumors differ from those present in ST:STUDY_SUMMARY circulation. Further, by comparing interstitial fluid nutrient levels between ST:STUDY_SUMMARY autochthonous and transplant models of murine pancreatic and lung ST:STUDY_SUMMARY adenocarcinoma, we found that tumor type, anatomical location and animal diet ST:STUDY_SUMMARY affect local nutrient availability. These data provide a comprehensive ST:STUDY_SUMMARY characterization of the nutrients present in the tumor microenvironment of ST:STUDY_SUMMARY widely used models of lung and pancreatic cancer and identify factors that ST:STUDY_SUMMARY influence metabolite levels in tumors. ST:INSTITUTE University of Chicago ST:LAST_NAME Muir ST:FIRST_NAME Alexander ST:ADDRESS 929 E 57th St. W GCIS 306, Chicago, Illinois, 60637, USA ST:EMAIL muir.alexander@gmail.com ST:PHONE 5104950975 #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]Additional sample data SUBJECT_SAMPLE_FACTORS - 1234 KP 1 TIF Sample Type:lung_subcutaneous Run number=2 SUBJECT_SAMPLE_FACTORS - 1234 KP 2 TIF Sample Type:lung_subcutaneous Run number=2 SUBJECT_SAMPLE_FACTORS - 1234 KP 3 TIF Sample Type:lung_subcutaneous Run number=2 SUBJECT_SAMPLE_FACTORS - 1234 KP 4 TIF Sample Type:lung_subcutaneous Run number=2 SUBJECT_SAMPLE_FACTORS - 1234 KP 5 TIF Sample Type:lung_subcutaneous Run number=2 SUBJECT_SAMPLE_FACTORS - 1234 KP 6 TIF Sample Type:lung_subcutaneous Run number=2 SUBJECT_SAMPLE_FACTORS - 1234 KPK 1 TIF Sample Type:Keap1_mutant_lung_subcutaneous Run number=2 SUBJECT_SAMPLE_FACTORS - 1234 KPK 3 TIF Sample Type:Keap1_mutant_lung_subcutaneous Run number=2 SUBJECT_SAMPLE_FACTORS - 1234 KPK 4 TIF Sample Type:Keap1_mutant_lung_subcutaneous Run number=2 SUBJECT_SAMPLE_FACTORS - 1234 KPK 5 TIF Sample Type:Keap1_mutant_lung_subcutaneous Run number=2 SUBJECT_SAMPLE_FACTORS - 1234 KPK 6 TIF Sample Type:Keap1_mutant_lung_subcutaneous Run number=2 SUBJECT_SAMPLE_FACTORS - 1233 KP 3 TIF Sample Type:lung_subcutaneous Run number=2 SUBJECT_SAMPLE_FACTORS - 1233 KP 4 TIF Sample Type:lung_subcutaneous Run number=2 SUBJECT_SAMPLE_FACTORS - 1233 KP 5 TIF Sample Type:lung_subcutaneous Run number=2 SUBJECT_SAMPLE_FACTORS - 1233 KP 6 TIF Sample Type:lung_subcutaneous Run number=2 SUBJECT_SAMPLE_FACTORS - 1233 KPK 1 TIF Sample Type:Keap1_mutant_lung_subcutaneous Run number=2 SUBJECT_SAMPLE_FACTORS - 1233 KPK 5 TIF Sample Type:Keap1_mutant_lung_subcutaneous Run number=2 SUBJECT_SAMPLE_FACTORS - 1233 KPK 6 TIF Sample Type:Keap1_mutant_lung_subcutaneous Run number=2 SUBJECT_SAMPLE_FACTORS - 1339 TIF Sample Type:pancreatic ductal adenocarcinoma Run number=2 SUBJECT_SAMPLE_FACTORS - 5197 TIF Sample Type:pancreatic ductal adenocarcinoma Run number=2 SUBJECT_SAMPLE_FACTORS - AL1 TIF Sample Type:pancreatic ductal adenocarcinoma Run number=2 SUBJECT_SAMPLE_FACTORS - 268 TIF Sample Type:pancreatic ductal adenocarcinoma Run number=2 SUBJECT_SAMPLE_FACTORS - 142 TIF Sample Type:pancreatic ductal adenocarcinoma Run number=2 SUBJECT_SAMPLE_FACTORS - 172 TIF Sample Type:pancreatic ductal adenocarcinoma Run number=2 SUBJECT_SAMPLE_FACTORS - TBt0 plasma 705 Sample Type:pancreatic ductal adenocarcinoma plasma Run number=2 SUBJECT_SAMPLE_FACTORS - TBt0 plasma 708 Sample Type:pancreatic ductal adenocarcinoma plasma Run number=2 SUBJECT_SAMPLE_FACTORS - TBt0 plasma 707 Sample Type:pancreatic ductal adenocarcinoma plasma Run number=2 SUBJECT_SAMPLE_FACTORS - TBt0 plasma 712 Sample Type:pancreatic ductal adenocarcinoma plasma Run number=2 SUBJECT_SAMPLE_FACTORS - TBt0 plasma 711 Sample Type:pancreatic ductal adenocarcinoma plasma Run number=2 SUBJECT_SAMPLE_FACTORS - TBt0 plasma 710 Sample Type:pancreatic ductal adenocarcinoma plasma Run number=2 SUBJECT_SAMPLE_FACTORS - 922 TIF Sample Type:pancreatic ductal adenocarcinoma Run number=1 SUBJECT_SAMPLE_FACTORS - 58 TIF Sample Type:pancreatic ductal adenocarcinoma Run number=1 SUBJECT_SAMPLE_FACTORS - 5197 TIF repeat Sample Type:pancreatic ductal adenocarcinoma Run number=1 SUBJECT_SAMPLE_FACTORS - 172 TIF repeat Sample Type:pancreatic ductal adenocarcinoma Run number=1 SUBJECT_SAMPLE_FACTORS - 142 TIF repeat Sample Type:pancreatic ductal adenocarcinoma Run number=1 SUBJECT_SAMPLE_FACTORS - 1339 TIF repeat Sample Type:pancreatic ductal adenocarcinoma Run number=1 SUBJECT_SAMPLE_FACTORS - 268 TIF repeat Sample Type:pancreatic ductal adenocarcinoma Run number=1 SUBJECT_SAMPLE_FACTORS - AL1 TIF repeat Sample Type:pancreatic ductal adenocarcinoma Run number=1 SUBJECT_SAMPLE_FACTORS - X1 TIF Sample Type:pancreatic ductal adenocarcinoma subcutaneous Run number=1 SUBJECT_SAMPLE_FACTORS - X2 TIF Sample Type:pancreatic ductal adenocarcinoma subcutaneous Run number=1 SUBJECT_SAMPLE_FACTORS - X3 TIF Sample Type:pancreatic ductal adenocarcinoma subcutaneous Run number=1 SUBJECT_SAMPLE_FACTORS - X4 TIF Sample Type:pancreatic ductal adenocarcinoma subcutaneous Run number=1 SUBJECT_SAMPLE_FACTORS - X5 TIF Sample Type:pancreatic ductal adenocarcinoma subcutaneous Run number=1 SUBJECT_SAMPLE_FACTORS - 922_Plasma Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Run number=1 SUBJECT_SAMPLE_FACTORS - 58_Plasma Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Run number=1 SUBJECT_SAMPLE_FACTORS - 5197_Plasma Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Run number=1 SUBJECT_SAMPLE_FACTORS - 172_Plasma Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Run number=1 SUBJECT_SAMPLE_FACTORS - 142_Plasma Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Run number=1 SUBJECT_SAMPLE_FACTORS - 268_Plasma Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Run number=1 SUBJECT_SAMPLE_FACTORS - AL1_Plasma Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Run number=1 SUBJECT_SAMPLE_FACTORS - X2_Plasma Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Run number=1 SUBJECT_SAMPLE_FACTORS - X3_Plasma Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Run number=1 SUBJECT_SAMPLE_FACTORS - X4_Plasma Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Run number=1 SUBJECT_SAMPLE_FACTORS - 3731_Plasma Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Run number=1 SUBJECT_SAMPLE_FACTORS - 4198_Plasma Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Run number=1 SUBJECT_SAMPLE_FACTORS - 4300_Plasma Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Run number=1 SUBJECT_SAMPLE_FACTORS - A0_Plasma Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Run number=1 SUBJECT_SAMPLE_FACTORS - A1_Plasma Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Run number=1 SUBJECT_SAMPLE_FACTORS - B0_Plasma Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Run number=1 SUBJECT_SAMPLE_FACTORS - B1_Plasma Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Run number=1 SUBJECT_SAMPLE_FACTORS - B2_Plasma Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Run number=1 #COLLECTION CO:COLLECTION_SUMMARY Isolation of tumor interstitial fluid (TIF) TIF was isolated from tumors using a CO:COLLECTION_SUMMARY previously described centrifugal method (Eil et al., 2016; Haslene-Hox et al., CO:COLLECTION_SUMMARY 2011; Ho et al., 2015; Wiig et al., 2003). Briefly, tumor bearing animals were CO:COLLECTION_SUMMARY euthanized by cervical dislocation and tumors were rapidly dissected from the CO:COLLECTION_SUMMARY animals. Dissections took <1 min. to complete. Blood was collected from the same CO:COLLECTION_SUMMARY animal via cardiac puncture, and was immediately placed in EDTA-tubes (Sarstedt, CO:COLLECTION_SUMMARY North Rhine-Westphalia, Germany) and centrifuged at 845 x g for 10 minutes at CO:COLLECTION_SUMMARY 4°C to separate plasma. Plasma was frozen in liquid nitrogen and stored at CO:COLLECTION_SUMMARY -80°C until further analysis. Tumors were then weighed and briefly rinsed in CO:COLLECTION_SUMMARY room temperature saline (150mM NaCl) and blotted on filter paper (VWR, Radnor, CO:COLLECTION_SUMMARY PA, 28298-020). The entire process of preparing the tumor prior to isolation of CO:COLLECTION_SUMMARY TIF took ~2 min. The tumors were then put onto 20µm nylon filters (Spectrum CO:COLLECTION_SUMMARY Labs, Waltham, MA, 148134) affixed atop 50mL conical tubes, and centrifuged for CO:COLLECTION_SUMMARY 10 min. at 4°C at 106 x g. TIF was then collected from the conical tube, frozen CO:COLLECTION_SUMMARY in liquid nitrogen and stored at -80°C until further analysis. CO:SAMPLE_TYPE Interstitial Fluid #TREATMENT TR:TREATMENT_SUMMARY N/A #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Quantification of metabolite levels in TIF and plasma In order to quantitate SP:SAMPLEPREP_SUMMARY metabolites in TIF and plasma samples, we first constructed a library of 149 SP:SAMPLEPREP_SUMMARY chemical standards of plasma polar metabolites (see Supplementary File 1 for SP:SAMPLEPREP_SUMMARY suppliers for each chemical standard). These compounds were selected to SP:SAMPLEPREP_SUMMARY encompass a number of metabolic processes and have previously been included in SP:SAMPLEPREP_SUMMARY efforts to profile plasma polar metabolites by LC/MS (Cantor et al., 2017; Evans SP:SAMPLEPREP_SUMMARY et al., 2009; Lawton et al., 2008; Mazzone et al., 2016). We pooled these SP:SAMPLEPREP_SUMMARY metabolites into 7 separate chemical standard pools (Supplementary File 1). To SP:SAMPLEPREP_SUMMARY do this, each metabolite in a given pool was weighed and then mixed (6 cycles of SP:SAMPLEPREP_SUMMARY 1 min. mixing at 25 Hz followed by 3 min. resting) using a Mixer Mill MM301 SP:SAMPLEPREP_SUMMARY (Retsch, Düsseldorf, Germany), and mixed metabolite powder stocks were stored SP:SAMPLEPREP_SUMMARY at -20°C prior to resuspension and analysis. Stock solutions of the mixed SP:SAMPLEPREP_SUMMARY standards pools containing ~5mM, ~1mM, ~300µM, ~100µM, ~30µM, ~10µM, ~3µM SP:SAMPLEPREP_SUMMARY and ~1µM of each metabolite were made in HPLC grade water and were stored at SP:SAMPLEPREP_SUMMARY -80°C (see Supplementary File 1 for the concentration of each metabolite in the SP:SAMPLEPREP_SUMMARY external standard pools). 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 TIF, plasma 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) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE NEGATIVE 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 (see Supplementary File 1 MS:MS_COMMENTS for the m/z, retention time and limit of detection for each metabolite MS:MS_COMMENTS analyzed). Metabolite quantification was performed by two separate methods. MS:MS_COMMENTS Where internal standards were available, first, comparison of the peak areas of MS:MS_COMMENTS the stable isotope labeled internal standards with the external standard pools MS:MS_COMMENTS allowed for quantification of the concentration of labeled internal standards in MS:MS_COMMENTS the extraction buffer. Subsequently, we compared the peak area of a given MS:MS_COMMENTS metabolite in the TIF and plasma samples with the peak area of the internal MS:MS_COMMENTS standard to quantitate the concentration of that metabolite in the TIF or plasma MS:MS_COMMENTS sample. 70 metabolites were quantitated using this internal standard method (see MS:MS_COMMENTS Supplementary File 1 for the metabolites quantitated with internal standards). MS:MS_COMMENTS For metabolites without internal standards, the peak area of each analyte was MS:MS_COMMENTS normalized to the peak area of a labeled amino acid internal standard that MS:MS_COMMENTS eluted at roughly the same retention time to account for differences in recovery MS:MS_COMMENTS between samples (see Supplementary File 1 for the labeled amino acid paired to MS:MS_COMMENTS each metabolite analyzed without an internal standard). From the normalized peak MS:MS_COMMENTS areas of metabolites in the external standard pools, we generated a standard MS:MS_COMMENTS curve describing the relationship between metabolite concentration and MS:MS_COMMENTS normalized peak area. The standard curves were linear with fits typically at or MS:MS_COMMENTS above r2=0.95. Metabolites which did not meet these criteria were excluded from MS:MS_COMMENTS further analysis. These equations were then used to convert normalized peak MS:MS_COMMENTS areas of analytes in the TIF or plasma samples into analyte concentration in the MS:MS_COMMENTS samples. 74 metabolites were quantitated using this method. The relationship MS:MS_COMMENTS between metabolite concentration and normalized peak area is matrix dependent, MS:MS_COMMENTS and the external standards are prepared in water, which is a different matrix MS:MS_COMMENTS than either TIF or plasma. Therefore, we consider metabolite measurements using MS:MS_COMMENTS this external standard method semi-quantitative. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS micromoles/L MS_METABOLITE_DATA_START Samples 1234 KP 1 TIF 1234 KP 2 TIF 1234 KP 3 TIF 1234 KP 4 TIF 1234 KP 5 TIF 1234 KP 6 TIF 1234 KPK 1 TIF 1234 KPK 3 TIF 1234 KPK 4 TIF 1234 KPK 5 TIF 1234 KPK 6 TIF 1233 KP 3 TIF 1233 KP 4 TIF 1233 KP 5 TIF 1233 KP 6 TIF 1233 KPK 1 TIF 1233 KPK 5 TIF 1233 KPK 6 TIF 1339 TIF 5197 TIF AL1 TIF 268 TIF 142 TIF 172 TIF TBt0 plasma 705 TBt0 plasma 708 TBt0 plasma 707 TBt0 plasma 712 TBt0 plasma 711 TBt0 plasma 710 922 TIF 58 TIF 5197 TIF repeat 172 TIF repeat 142 TIF repeat 1339 TIF repeat 268 TIF repeat AL1 TIF repeat X1 TIF X2 TIF X3 TIF X4 TIF X5 TIF 922_Plasma 58_Plasma 5197_Plasma 172_Plasma 142_Plasma 268_Plasma AL1_Plasma X2_Plasma X3_Plasma X4_Plasma 3731_Plasma 4198_Plasma 4300_Plasma A0_Plasma A1_Plasma B0_Plasma B1_Plasma B2_Plasma Factors Sample Type:lung_subcutaneous Sample Type:lung_subcutaneous Sample Type:lung_subcutaneous Sample Type:lung_subcutaneous Sample Type:lung_subcutaneous Sample Type:lung_subcutaneous Sample Type:Keap1_mutant_lung_subcutaneous Sample Type:Keap1_mutant_lung_subcutaneous Sample Type:Keap1_mutant_lung_subcutaneous Sample Type:Keap1_mutant_lung_subcutaneous Sample Type:Keap1_mutant_lung_subcutaneous Sample Type:lung_subcutaneous Sample Type:lung_subcutaneous Sample Type:lung_subcutaneous Sample Type:lung_subcutaneous Sample Type:Keap1_mutant_lung_subcutaneous Sample Type:Keap1_mutant_lung_subcutaneous Sample Type:Keap1_mutant_lung_subcutaneous Sample Type:pancreatic ductal adenocarcinoma Sample Type:pancreatic ductal adenocarcinoma Sample Type:pancreatic ductal adenocarcinoma Sample Type:pancreatic ductal adenocarcinoma Sample Type:pancreatic ductal adenocarcinoma Sample Type:pancreatic ductal adenocarcinoma Sample Type:pancreatic ductal adenocarcinoma plasma Sample Type:pancreatic ductal adenocarcinoma plasma Sample Type:pancreatic ductal adenocarcinoma plasma Sample Type:pancreatic ductal adenocarcinoma plasma Sample Type:pancreatic ductal adenocarcinoma plasma Sample Type:pancreatic ductal adenocarcinoma plasma Sample Type:pancreatic ductal adenocarcinoma Sample Type:pancreatic ductal adenocarcinoma Sample Type:pancreatic ductal adenocarcinoma Sample Type:pancreatic ductal adenocarcinoma Sample Type:pancreatic ductal adenocarcinoma Sample Type:pancreatic ductal adenocarcinoma Sample Type:pancreatic ductal adenocarcinoma Sample Type:pancreatic ductal adenocarcinoma Sample Type:pancreatic ductal adenocarcinoma subcutaneous Sample Type:pancreatic ductal adenocarcinoma subcutaneous Sample Type:pancreatic ductal adenocarcinoma subcutaneous Sample Type:pancreatic ductal adenocarcinoma subcutaneous Sample Type:pancreatic ductal adenocarcinoma subcutaneous Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture Sample Type:pancreatic ductal adenocarcinoma plasma cardiac puncture 2-hydroxybutyric acid 13.93842777 18.64102785 28.21010286 25.03352557 13.49074631 21.55703017 28.00516324 18.85515839 18.00313588 23.75490631 24.92813617 20.65547237 18.35727434 26.11065024 16.05841447 11.78269045 17.93706992 22.55495163 7.914836285 10.14143993 5.707765534 27.72041921 22.69937963 22.08359968 23.80501425 31.60251436 22.26951212 25.64194841 19.92308528 21.63316723 81.37011885 66.82147952 70.89991417 79.87501234 81.77136921 69.44699214 83.11273369 68.36303762 67.34010233 69.37001401 69.45613324 69.57912068 69.97001567 18.89084602 12.87585285 10.69146873 11.98306052 22.1186564 38.11458117 8.603517525 4.218133569 6.154834417 13.80910251 19.65271973 45.73522947 15.08584909 13.40133468 26.88483435 12.09300383 14.5713545 12.9932472 2-hydroxyglutaric acid 27.38874293 17.99558517 29.67730809 38.41025571 17.57179878 50.52621678 15.61092539 18.14061604 22.94220159 17.7765516 16.05440308 18.88900796 19.51254946 23.19372008 29.76784389 20.0716448 16.94696079 26.20880636 7.133464859 11.20813188 7.261696353 7.826124415 9.37400269 10.06355586 12.36303717 15.95383669 9.866476258 6.464092745 6.915003148 7.876329674 13.20372178 1.887099677 8.486314046 8.967073805 7.542683917 5.902585357 6.901099976 6.47534253 13.05209997 25.64265064 21.58088266 23.73939723 19.51633804 10.28074502 9.82112015 1.736452125 3.804115664 6.204662756 6.475507663 4.196681576 8.340767472 7.566616919 12.99314904 3.291054803 2.227931717 3.436673785 3.675934025 16.2163891 4.472081746 17.98138772 10.13624995 beta-hydroxybutyric acid 51.15500469 30.51568195 50.68385122 47.7769565 25.54667849 26.39853282 45.73007444 30.13382672 34.79335643 27.65044282 33.93167133 53.7351163 35.88591059 37.04981631 30.17664237 25.74682758 41.48782508 28.2328469 11.10479828 61.50914146 24.65601478 168.4256472 94.10920245 57.40052129 106.3105755 123.4748275 105.6551738 168.3692065 174.700444 142.6510751 43.80468209 21.9649185 72.96396313 70.37579504 119.6288594 12.01758286 218.5137297 27.64439355 11.34697487 21.06286779 29.4520748 33.02376121 27.38287896 114.5785273 207.1978726 180.1370789 69.88269643 175.3390619 475.9606134 89.18580064 55.27270081 92.43028434 145.7593027 107.8380624 151.7782643 100.3498669 101.9829462 133.0552504 103.8845813 106.0416676 41.91223024 3-hydroxyisobutyric acid 10.14800682 17.9413946 25.37784742 32.30029983 15.44136815 16.58957437 20.03133955 12.55449904 15.5747188 20.40384058 18.25844573 12.84166264 15.7502072 14.30789823 13.45323177 9.416047848 14.12152146 16.1364908 6.895876388 18.71983308 11.99142138 18.16250548 7.874611159 27.07582615 30.27834848 29.849395 22.20481669 29.60315476 23.15302845 26.25676363 19.53300613 12.40469181 21.64662971 29.77624562 10.88178239 9.649814801 25.11818521 17.63190566 8.733465175 20.25136278 14.40206408 18.84669933 10.56730799 18.22267062 31.52971161 25.86962843 32.24790439 15.89115789 51.20281448 35.15548523 8.434135215 12.1660766 21.07129969 78.28132276 41.49616218 25.88596288 16.87791468 27.39124611 16.49550042 18.96375166 20.67535388 Ketoisovaleric acid 11.75142916 15.32297179 18.40415007 20.95370243 12.05402743 22.589705 14.27129807 17.39802279 11.99028504 17.0849155 28.62327889 11.95974303 17.8572455 20.7635237 16.69867374 15.36388455 12.62474166 10.98294003 2.789890973 5.130581218 2.576457395 3.261842296 2.630997908 5.211619841 7.443144872 9.80833999 7.307166928 6.96529696 7.436158879 7.241497181 18.93392356 7.607656135 12.78776629 22.78409962 8.029735113 12.96112027 8.045743668 8.056939764 11.8411599 12.90208868 8.628517206 7.656226063 9.881655185 13.28276847 16.94057841 10.44193542 10.76357315 11.11903427 13.48019002 13.07600216 6.061084447 3.38814698 8.263310488 9.603004215 6.65211761 11.78637897 12.99448302 8.337344654 14.07598501 6.469578493 7.799951157 3-Methyl-2-oxovaleric acid 28.44626727 42.97103725 45.73535113 55.24032426 22.98272224 64.64469978 32.75141654 42.7608132 27.24671766 48.11384722 75.41718955 32.42923818 33.66934045 48.03354782 31.30871059 33.57826581 35.71601106 24.74335172 4.1444189 11.86034877 2.928174757 4.854915376 3.425144379 6.771483293 20.35484078 18.85459923 19.56276841 19.88051508 15.60532816 16.60450942 15.46869941 3.4568735 12.29456109 6.614143966 4.003184386 4.553039069 4.474437677 3.353173627 7.42393709 14.27572674 17.56615017 14.27374153 20.57307346 17.60436509 24.14004031 12.03290958 9.555497871 8.714882278 26.65448572 9.642077998 3.969033309 5.907981583 15.12158202 10.99267586 11.45503184 14.22433071 10.51037985 16.76144353 9.034751531 10.26135349 13.57847277 3-Phosphoglyceric acid 44.83920192 67.8419378 74.75194348 93.53630603 56.97933351 49.17991024 43.7132241 57.32851857 21.24025028 51.80985904 41.54989438 16.34472 42.11691519 30.15919867 10.91501851 38.13463442 37.55380456 32.23353434 31.39619424 48.00967618 22.48446203 29.27247189 90.03349615 106.5756026 0.023000591 1.916540668 3.364741474 1.912195576 2.868465565 1.011835694 31.13004846 1.805162707 18.4717333 54.85374704 51.77387347 22.53598061 18.95037352 18.32883468 33.04852065 42.24201252 16.33566346 13.70086072 17.5500394 0.276775646 0.02458168 0.255276024 -0.10682577 1.152741245 1.439891784 6.436889993 1.865075028 1.118478848 1.746128415 -0.10682577 0.285593498 1.374286496 0.909133807 1.844866954 1.309958799 12.1998703 2.313595121 3-methyl pyruvic acid 1.365123608 2.849746088 3.103457128 3.5815633 -0.475783204 3.042266869 2.430564291 1.73233685 1.510969659 1.113495652 3.358805104 3.640272927 1.690178584 2.036134915 1.910509708 2.133819594 2.349026657 2.592498205 0.353751356 0.487431147 0.779489211 2.224240141 2.187023174 2.185653312 3.064403929 3.19915658 3.063476772 1.371794588 3.05262066 3.643217937 0.897550948 0.492350873 1.186591912 2.060812833 3.572749725 4.471817098 1.753389502 0.317156611 1.140274983 0.33711239 3.239635829 -1.214963297 2.791421297 1.687493127 1.653709144 1.583023312 0.673697679 1.872987132 8.784757451 5.343111489 -0.451325618 6.656508218 3.506183479 3.111998353 -1.102069357 0.210046022 6.305877266 11.91270299 6.581330161 12.82708816 6.833741655 Oxoglutaric acid 22.47133791 22.95404864 23.04829768 10.39062582 8.479504241 82.17912307 19.06343219 10.65955804 27.70280613 5.164577893 26.45962537 25.33457232 28.98087854 30.46423974 39.77153103 12.95665111 15.70623383 8.94090758 2.498105034 6.638875514 4.628588526 7.823783078 0.927724644 3.40387663 54.12850667 48.55619386 50.90969428 31.99357042 34.0345022 32.91427594 11.25292716 5.385041327 13.13053303 11.21809945 2.50646414 3.927586538 8.397043423 5.45302171 11.80263873 5.337210504 22.83454763 11.61772854 18.93578203 12.66414957 13.22053147 8.436090514 16.64692032 25.30541028 11.69011152 16.25465886 8.435991067 27.28578178 18.81932397 14.03755384 9.826076228 12.25570438 11.83923609 10.99794364 11.86904566 4.849055321 19.02948344 Allantoin 191.8553424 0 342.4561992 0 134.1070701 1097.149248 0 0 0 0 149.4419311 138.5071228 0 123.6669988 0 140.2609877 124.9357794 0 0 0 0 189.0533788 193.3605005 142.9183299 117.6241502 0 0 109.9310202 79.63982252 0 153.2892439 62.78925813 102.0691615 100.7275409 196.1829297 64.70083612 118.8702335 93.32575077 62.39698618 90.92130747 94.19227924 75.70809517 46.12229958 227.161193 203.4975352 151.0591868 340.0182743 331.563161 347.9874207 167.0339776 43.97285923 157.2807033 178.8502753 369.7984455 205.6938197 172.2374148 109.0814495 193.5578761 89.25043771 73.84596422 50.40349092 AMP 0.298603133 0.410962277 0 0.518258922 0.353749696 0 10.83886029 3.382618491 1.168158511 0.62976784 43.05850127 0.313377957 1.02084626 0.480890239 129.4680877 197.6593952 130.0087206 152.9664792 4.675648218 41.26487139 9.514092122 1.922389189 53.35835644 36.36307447 3.811908817 34.46392131 3.112664122 3.041384382 7.495908875 2.194112459 27.58784036 6.483492414 42.89763779 58.53255431 71.64015521 8.02348669 6.009457432 23.4171304 2.042556238 10.72084899 4.970734226 2.619758496 3.219839012 0.703597178 2.424144198 5.025780526 22.31319098 8.863653028 27.0134669 28.71085292 22.99495802 31.69845804 43.89240597 5.029309911 23.32472136 12.68702119 26.68978354 10.68704692 51.53105375 19.67376055 Argininosuccinic acid 0.923497585 0.682290444 0.98705335 0.655186368 0.782217358 0.827512132 0.901094644 0.581229614 0.899785866 0.511767554 0.568635153 0.332718431 0.47175574 0.599667994 0.779825804 0.486599655 0.510135047 0.783263953 0.575509608 0.977400582 0.6863793 0.381061161 0.504344416 0.851152501 0 0.171500975 0.328049282 0 0.257490706 0 0.198598383 0 1.304183652 1.242117033 0.669110619 0.623810388 0.430928232 0.772339804 1.414518006 2.081754705 0.945213187 0.826413867 0.210470103 0.299413938 0.13063853 0.236039898 0 0.389919541 0 0.228396791 0 0 0.341828835 0 0.073199334 0.252711774 0 0 0.120035108 0 0 Vitamin C -0.053288431 1.22700843 47.22979805 -0.053288431 0.210020001 -0.053288431 19.00190703 -0.053288431 -0.053288431 -0.042948526 4.954340007 0.00344017 -0.053288431 -0.053288431 -0.053288431 0.040142167 108.1673916 -0.053288431 0.031474389 0.885576784 1.777111304 109.9253903 151.3200588 135.7940052 0.022302955 -0.053288431 -0.053288431 -0.053288431 -0.038243914 -0.053288431 133.5099204 25.12039236 167.5017181 136.7565624 152.5048389 132.3823634 118.3444235 183.0740493 126.2548455 171.5316514 137.8526305 107.003677 4.142087657 54.70291513 4.965653088 45.62751322 0 27.22906903 0 8.95606872 0 0 0 54.29349954 7.849106207 0 9.073778701 16.41167666 0 0 0 Aspartic acid 464.0070188 292.0312321 403.9686134 130.5245695 260.8255222 341.1614313 170.5197237 109.2659702 291.9565556 106.3744023 149.8890519 250.8969362 161.5905128 258.8126909 247.9904109 237.6652215 224.1338709 188.0406631 362.8217552 395.114879 312.7778826 278.2725227 370.440002 204.581986 14.77076297 10.60949546 10.38074866 9.557575391 13.13224424 9.625486412 454.5265022 60.42264977 463.5546061 246.549426 403.678867 519.1047635 317.1154238 385.529005 264.2556345 351.0880038 237.0557694 163.477879 265.6484165 20.98891813 24.87715549 28.11837466 27.80945652 23.56995192 66.31928771 68.59733739 55.6930688 48.329595 13.6096256 34.29471422 45.58681146 64.66518734 21.72731632 121.9164172 40.27260519 Cis-Aconitic acid 4.944889257 4.240839806 4.495479158 8.399482592 3.587486104 9.850493251 3.790812491 3.265524113 3.312552193 3.425940417 3.023481991 3.047830892 3.74336844 4.054931724 3.110118047 4.317826267 3.547675006 3.99069666 1.336119549 3.171804181 1.446701605 2.633562954 4.610014963 4.158798558 6.173776741 6.749532223 5.655483236 3.780505654 4.110734484 4.767965862 2.058183482 0.702909278 3.227387468 4.253535076 5.458778565 1.380062453 2.808641557 1.578186754 8.263406118 9.631129573 4.728234876 4.257578887 2.32663493 3.833808734 5.287087337 5.205207537 4.423428078 12.74001119 8.835356862 5.051814396 3.7525691 6.279216024 6.43390373 9.561486074 5.497651542 7.680517706 4.563992535 6.346302557 3.556475575 4.857249134 3.54587402 Citric acid 383.5590238 337.165113 427.7007367 678.6777188 255.2846183 935.8151066 365.3174986 312.4466296 297.5139113 270.0870114 350.8869586 288.7480912 354.6152818 364.2222077 295.5086415 411.9224228 380.3314208 344.9591419 155.1513418 404.8609486 175.0106114 263.740296 548.9603979 382.4152347 459.7424959 494.4986788 436.4624412 280.1706431 286.4577218 342.902721 278.1655586 85.3320305 476.6579168 414.9613255 585.297198 164.7550217 262.9935328 192.705684 593.8516675 800.7126131 631.1913866 415.6367613 265.9437065 242.3350867 368.0493467 320.5692736 356.8653308 986.7841255 557.4573708 321.4927804 307.9053578 377.0911561 414.458349 734.6093027 413.2096021 557.1382917 286.2748211 389.2432363 237.13803 249.7853765 237.2574302 CTP 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0.085647074 0 0 0 0 0 0.011129665 0 0 0 0 0 0 0 0 0 9.925069649 9.492157883 9.492157883 9.548742503 9.492157883 9.492157883 9.492157883 9.492157883 9.492157883 9.492157883 9.492157883 9.492157883 9.492157883 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Cysteic acid 0.157980914 0.158282271 0.118623513 0.095917353 0 0 0.164644264 0.279457452 0.338076813 0.415596186 0 0.111024026 0.157363599 0 0.175819091 0.153697997 0.093801202 0.401212766 0 0 0 0 0 0 0 0 0 0 0 0 2.454471956 2.454471956 2.454471956 2.454471956 2.487978529 2.454471956 2.482560276 2.454471956 2.650017057 2.809966561 2.534648723 2.454471956 2.491724788 -0.022781375 -0.022781375 -0.022781375 -0.022781375 -0.022781375 -0.022781375 -0.022781375 -0.022781375 -0.022781375 -0.022781375 -0.022781375 -0.022781375 -0.022781375 -0.022781375 -0.022781375 -0.022781375 -0.022781375 -0.022781375 Dihydroxyacetone phosphate 357.7201798 337.7876291 563.6413944 277.1933964 373.0690837 560.0791669 399.8691022 130.2691255 661.1350933 189.0744219 132.6750147 289.6204157 190.1633027 306.7087231 609.5990577 189.9307179 130.0099099 123.7867132 179.8877868 223.9572181 227.254766 141.8948745 228.305867 219.0130701 10.32441651 4.731936291 17.68955029 11.68838921 13.49566142 10.86038454 955.4098722 81.22239248 638.1566014 833.9944746 838.8558919 594.9946828 657.7063004 701.2278709 1007.637431 1108.936305 590.0479593 635.7304208 528.3661818 10.81216031 10.34007935 3.343739317 111.0339064 11.04570919 93.6675249 103.9032194 113.2833736 314.702846 88.14692256 4.144996531 20.84381509 71.89366825 43.15752221 254.0391108 30.5193348 356.8382607 96.28472092 dTMP 0 0 0 0.049616455 0 0 0.029023745 0 0.006489663 0 0.156822116 0 0 0 0.409207528 0.304156539 0.260084408 0.62517592 0 0.016836826 0 0 0.039557848 0.068566568 0 0 0 0 0 0 2.992225476 2.886365818 3.005907948 3.299717741 3.048360224 2.886365818 2.886365818 2.886365818 2.987916795 3.33247665 2.993592948 2.954622405 3.013687715 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Fumaric acid 0.738897394 0 0.383380206 2.005018102 0 5.672718031 0.661788056 0.719074427 1.323934927 0.624049236 0.448622042 0.287230127 1.080759876 0.726317783 1.522981957 0.66823944 0.792315146 1.607843977 0.366563505 2.007817032 1.579656452 0.198604807 0.430789786 0.634706182 0 0 0 0 0 0 12.26460427 8.355163928 15.95693316 10.45463203 9.342852713 9.082110713 8.355163928 10.75107142 8.355163928 9.367798991 9.537290078 9.44194723 10.22876323 0 0 0 0 0 0 0 0 0 0 0.805797798 0 0 0 0 0 0 0 Glucose 5071.641502 5257.489926 2980.402864 3956.933829 1175.780288 4483.951893 6698.994786 5187.112675 4404.436004 3351.144139 4003.014922 2342.148821 3170.949566 2734.315229 3737.220277 7414.229366 8229.27843 4262.40815 4092.747029 8792.028071 2496.404467 914.5677893 3555.763015 3480.872121 15085.05666 16018.16278 16237.42337 15407.55528 14302.45908 15766.98184 3818.196238 912.0422802 10096.15621 4129.382096 3961.808317 4600.147418 808.175768 2851.431134 3592.156698 8876.385506 7850.854011 5718.575758 3936.818168 11088.48631 8367.216535 16734.63795 4450.094009 8787.822492 3449.912959 8749.439029 4002.853021 5578.993884 11606.52422 5860.013315 9328.640526 5338.077993 10180.03101 11014.42155 10688.9855 5663.797756 7065.157769 glucose-1-phosphate/fructose-6 -hosphate 93.91737293 43.07869339 129.2499316 75.46404379 65.55227762 72.0513854 48.52016429 29.12085147 89.52422507 60.49318507 31.70347332 74.6731059 39.53025873 98.07370449 43.53688879 23.76982801 26.55865778 15.75898679 105.5301892 54.43524591 129.836898 129.9296399 124.8000784 153.5486877 0 1.912607316 1.715785853 1.27022187 2.019335255 0.353758652 113.1420971 5.335239016 31.9347521 87.95241957 71.93996414 71.7962044 91.98468459 91.03184057 92.27793657 89.85220312 50.5227464 59.88295733 80.46151694 3.986011473 3.628711402 2.03977202 0 2.063875861 3.153899692 7.170940627 2.592554632 7.879545106 10.62055608 1.83417904 2.794654064 5.559668251 3.240286637 6.285614028 3.663753264 7.064737248 10.15552488 Glucose-6-phosphate 111.7450099 43.57136238 134.9216286 86.85807873 70.87374728 92.72269632 56.06584083 35.51786041 98.38539193 72.99010376 40.30080892 88.76596007 45.25037768 121.2302361 60.51820709 42.20526787 26.29396518 19.93577216 122.8525734 58.20817441 150.9239627 146.6481429 152.5802913 160.598536 0 1.633631503 0.961849232 0.539518756 1.217352746 0 153.443476 7.1601646 39.62852116 102.588507 94.24296663 99.21184386 136.728283 126.2779477 123.5139928 116.2054846 65.76545661 88.46570547 118.6530715 5.726950742 4.411497348 2.277420665 2.596222156 4.019541031 10.23269984 4.050364674 8.006083429 14.91260838 2.848516243 3.651225019 8.246470443 3.822553835 7.081841778 4.30966538 6.813882514 11.93850548 Glutamic acid 1632.694866 1327.14649 1731.284182 1373.664693 1266.300067 1490.789518 777.575862 722.7595273 1013.566484 737.7033433 682.2735146 958.0053483 1011.933406 1182.271034 1370.699459 1054.50476 1026.119083 1103.616153 985.0834599 1181.690986 852.0245172 736.3607739 886.6930983 938.8217559 50.16202667 30.22328551 45.79504126 38.13242791 56.97026763 40.26351275 1300.490358 146.5755016 1259.001113 1029.348379 949.4809683 1095.404181 789.1325345 957.7043704 964.6373356 1272.074515 942.866659 606.7068856 887.5166505 48.37915027 48.3521646 57.38594854 55.29310954 51.77690614 72.57254868 138.6683713 118.1827359 125.5401694 123.287269 53.33220676 52.82133286 65.96693326 55.65938091 74.74937249 50.75605189 169.5561988 88.66978419 Glyceric acid 4.607668148 4.50532622 13.99497646 9.421926938 2.997671755 3.962902557 2.631628596 2.261906082 1.782748136 2.69976528 4.621399867 2.867391691 2.609447545 2.890751894 2.767290058 1.93307191 1.774989352 4.935659868 12.78294923 3.675791538 6.368338273 4.314076779 4.348603335 10.0465665 1.599445924 1.526640845 1.585077589 1.182638586 1.435865583 1.367556038 3.674178484 1.330711748 4.788845595 3.388188486 2.500935592 10.14292255 2.992017994 3.082806895 5.072384378 3.489559772 4.449962414 2.221742967 2.532169846 1.674224737 2.647886598 1.871534302 0.453421755 0.829800235 1.195402186 2.426302023 1.882919249 1.832832466 1.741262102 1.661362499 1.388704494 1.151444679 1.60360491 2.612049107 1.32401982 2.332365416 2.018575721 GMP 0 0.028065022 0 0 0.1296476 0.036068695 1.636278466 1.03532912 0.580837521 0.060622108 3.862950025 0 0.100272699 0 10.19314842 11.77535194 7.874810533 8.568974777 1.888922631 5.130110418 3.495491913 2.940214139 10.36563846 9.711478044 0 1.751429016 0.086847581 0.061428983 0.278150502 0 3.912577286 1.894163539 4.222218204 12.45260821 13.07267991 2.342866378 3.765128042 5.566165076 -0.314270616 0.563959883 -0.088740487 -0.072889727 0.170341199 -0.441785581 -0.346787729 -0.24413066 -0.189428255 -0.063583155 1.00789363 1.696050621 0.489229897 0.996753641 0.834633303 -0.429863252 -0.197404164 0.138956445 0.118216154 1.976398132 -0.200652806 3.301675831 0.459057945 Itaconic acid 140.484952 82.29631463 131.0520249 131.5762462 61.98670155 151.3308354 55.16239002 65.49900032 107.2936728 83.31140619 46.21590419 47.30885739 68.4726898 63.99364705 87.52548212 89.43600413 57.98510341 99.69859693 41.87185192 45.48382306 44.86962831 46.12485103 33.96639131 30.20602964 142.2018946 168.8445309 -1.808163481 99.0994752 76.09034089 84.94538474 32.51402102 5.890256207 20.8887609 13.46952169 15.82442649 18.27078704 22.56946588 15.37110153 34.50049405 70.33481032 47.75124593 18.97123702 53.59127197 9.400026131 10.20512543 14.29773106 145.1280259 10.17247152 38.95804212 21.07074151 25.44889722 62.54727936 12.33753943 3.858735447 18.85050009 23.29623668 44.47133454 61.08418958 9.574039507 54.82855324 16.86981699 Lactic acid 8486.821565 9739.061817 14960.91556 9879.193202 8108.914167 10254.6386 8764.486014 9123.791279 9868.625083 8896.390498 12003.34599 10860.28419 9432.205273 12293.28156 9436.474679 5588.538832 6205.585268 8080.210603 4170.448786 7416.446128 3148.190107 2995.542526 4979.440242 5259.199908 4067.348393 4126.725706 4201.881925 4477.138405 4728.727888 4042.852831 6754.072921 928.8789205 5052.266373 3601.482632 3514.817709 2968.358016 2008.985642 2660.939809 4733.214319 5701.715483 7506.295605 4608.135529 5425.354634 7165.318872 4690.23253 3309.035576 2877.40917 1255.332347 1834.894167 2718.747045 5350.165332 7843.960882 8648.067325 3769.901815 3776.211367 5472.510523 7206.872189 7565.996164 8181.967436 8098.546704 5362.066902 Malic acid 216.6273749 175.8719633 391.6717471 265.1462583 222.6494774 237.839133 131.405865 164.3450998 156.1981046 143.7773138 121.7656545 110.5166297 118.0774095 168.4589825 141.9397987 115.4906171 123.1255576 153.5374008 92.11271976 127.2288705 108.895346 105.0164167 183.5142951 190.3054945 99.38966028 73.95776924 69.97391608 60.24219883 63.35082073 67.14734198 204.0305344 17.60531413 133.8914599 208.8218196 197.9379586 105.1186429 114.1080478 125.3323978 134.0742082 178.9935888 149.3131887 84.4820247 117.576808 18.267879 20.02292974 13.10514553 45.63948678 32.81334162 25.5653732 48.85381732 50.88310008 87.47393105 56.2068761 20.92017167 21.64286248 29.61788597 23.01760246 40.21134113 31.80252551 98.28142854 47.14959288 5'-Methylthioadenosine 0.171603552 0.188562248 0.271846543 0.189156354 0.193492928 0.148055563 0.061416933 0.07181766 0.050218301 0.08319556 0.03312457 0.05922733 0.076949508 0.101800038 0.136463533 0.08203304 0.075838755 0.074091627 0.050316764 0.054980246 0.048036587 0.068016546 0.066803567 0.099813733 0 0.009467091 0.00472874 0 0 0 0.077952927 0.008260108 0.04287359 0.083054525 0.078494297 0.051395615 0.046163443 0.048720155 0.022024403 0.025789237 0.03112836 0.015670143 0.012120379 0.008833344 0.007110373 0.02937732 0.010969617 0.013579288 0.005577949 0.009092294 0.014433758 0.017756079 0.013595446 0.014752968 0.010945966 0.012212917 0.016079488 0.027172368 0.050482038 Phosphoethanolamine 1385.542848 1033.212036 1773.815418 1215.418553 1195.463786 1430.376207 562.9948594 547.3014697 783.5656669 505.1000309 746.580658 931.0975348 980.6592037 1064.99003 1206.352242 1373.353358 782.6129132 1090.305043 725.2028224 629.7389016 752.665347 694.5472803 1159.181404 941.394483 39.44255902 34.265719 27.32762966 27.50575152 34.97502837 26.53015563 553.4335926 64.87349558 442.2857353 618.8433182 584.0409044 600.2672059 437.3082184 512.4631456 251.3788615 333.6998482 234.991209 188.9899635 396.7845949 17.5795275 17.17371617 14.87581533 23.38929993 10.69213836 86.19223295 66.26885136 24.26536862 73.8420274 38.88580482 15.45400084 30.75229188 31.14778571 48.76218319 121.1227672 23.67502498 85.71646041 39.09121738 Orotic acid 0.991203969 1.555981032 1.341917254 3.808650224 0.437265663 4.587353148 2.075458154 2.474472921 1.736658815 2.705348632 2.999059625 2.139665624 2.013724404 2.032221279 2.799482684 1.027161586 1.143188754 2.110676212 0.281292083 0.748486961 0.497487907 0.568289172 0.811157969 0.879261429 0.764675779 0.488426655 0.630803062 0.410747554 0.437832248 0.504296432 0.801142022 0.456829071 0.783542419 0.961464816 0.697353248 0.611048475 0.698017313 1.079446621 0.383467574 0.483766817 1.159170213 2.551321441 0.756135051 1.503520566 1.427408084 0.286249513 0.560651281 0.750957224 0.907323168 1.12054828 0.672421421 0.440947924 0.582184656 0.773633191 P-Aminobenzoic acid 0.037404467 -0.091083263 0.129753661 0.009349474 0.310305185 -0.091083263 -0.091083263 0.094586147 -0.091083263 -0.091083263 0.025238567 0.133306861 0.031223389 0.037651558 -0.091083263 -0.003673596 0.126055517 -0.091083263 -0.091083263 -0.091083263 0.012097935 -0.091083263 0.048250757 0.117309128 -0.091083263 0.013137758 -0.091083263 -0.007812987 -0.007012559 -0.091083263 3.74602874 2.240342255 3.885550438 2.630466735 8.662069893 2.760936421 2.819855272 1.917381053 2.7388056 4.430197118 3.410798221 3.025366097 2.092253864 3.458759442 7.07978342 3.751968246 2.738678754 12.54214936 4.677414637 3.440040739 5.770878332 4.358727063 5.649284111 6.08013322 6.956201726 5.29191974 -0.013893842 -0.013893842 0.038651403 0.044253085 0.071506846 Pyruvic acid 32.55540157 41.29540102 43.06398167 17.26030543 18.34444864 148.735765 39.91192771 7.131290151 90.89240375 5.800903471 46.4221336 207.0014004 143.7518935 153.9245119 269.5684181 81.61446581 68.38733918 86.77559364 33.00583661 28.65765337 59.64264355 37.68250666 8.354831929 14.59232584 105.8167908 119.0003495 147.5099522 97.26921553 128.2285352 111.2076215 100.5853926 130.9718405 54.57460011 10.54110314 10.63694656 30.00339495 29.48566602 55.67015802 23.38667666 13.86041081 92.54305143 47.66816241 122.6833361 330.6890585 152.5354813 155.8377197 77.04357408 213.2463832 55.72231506 167.7539906 64.29373988 50.049783 206.4759435 140.5083487 44.50106231 40.34047124 220.5923636 195.4486481 257.3205307 106.5476943 307.8609451 Ribose-5-phosphate 329.0725624 255.9142565 205.0832324 177.8768083 188.7017986 350.6712364 97.35482295 59.98416586 159.6121685 69.20141515 63.59827446 188.9980975 241.3861653 204.8147825 219.1789374 139.9516614 105.6474541 75.22480674 115.8304085 114.8938031 83.86831428 49.10392578 174.3511761 182.4587331 3.729774154 9.764851628 8.050694135 6.435266799 5.657051716 3.258017809 103.494295 15.27907318 99.80848797 181.4541696 143.3734717 118.7686042 62.88895197 72.22940322 67.29324109 96.87536159 66.25431972 68.50456798 36.83131133 2.992857739 3.004010354 2.189813923 0.435768532 1.649122191 19.07215842 18.04399525 9.57504513 21.07712146 11.22327109 1.117798352 6.330760936 9.580892404 12.34142282 48.76918571 6.239452666 46.12835329 10.85800305 Sorbitol 6.861985395 6.580886261 11.28599044 9.802387056 5.470719825 14.75781419 7.018598327 4.235628798 6.012238854 4.969896339 4.325719725 6.416961763 6.533506515 7.120472788 9.760412148 16.14547346 4.900499267 9.914125984 4.653499648 6.687920486 6.043604981 3.360730297 8.987071433 9.486589418 6.228362253 4.177892258 4.27447097 4.74594515 4.722270171 3.24535666 2.142191504 1.096163902 1.551428016 2.355379022 2.615451348 1.649677623 1.255362942 2.122763388 1.504042356 1.730802945 1.551735999 1.093703492 2.186543644 0.535145167 0.763119956 0.275391833 0.434854213 0.169491696 0.256616232 0.464869756 5.450824252 1.330985424 0.551687419 0.258863211 0.434925976 0.318642573 0.08602651 0.574457637 0.220804895 1.265651914 0.505848755 Succinic acid 153.2361557 147.6970753 214.5536066 185.6227219 143.6851801 183.1857154 91.08881159 116.127678 95.71693068 94.0311951 120.1867853 127.4031386 132.2725309 148.0895028 191.5927582 81.71803337 89.22073119 147.8900439 60.93663628 113.1241389 69.50074846 67.28906566 120.4375781 100.2927657 37.80769837 18.38896007 25.78422291 21.27018128 23.91644969 23.00445134 164.9306536 18.22973396 115.8872874 118.8936616 132.3141267 72.83539267 69.84231382 69.72103682 113.3090763 130.6214493 111.7063512 59.65802112 63.28832831 94.76899438 37.72617558 55.91286014 61.75066272 24.70458742 82.20553496 59.55142208 416.4911036 714.2792281 236.717914 75.69404249 103.8587336 124.4933622 260.6705837 405.7594562 349.9547145 455.0057364 192.9414876 Thiamine 12.96505054 19.54789353 12.96015097 18.7881685 13.16658494 9.697477732 7.155083454 7.075247754 11.84355085 14.04449311 2.431265835 16.38916242 17.28925113 16.19503338 11.81458574 4.231350508 6.334810362 7.427581392 9.134034421 14.05799665 9.957305561 8.13583078 3.768707909 5.434763151 0.886491366 1.714633935 2.216158567 1.692226642 0.973195327 1.226913959 9.309002595 2.415762516 9.773717865 7.779669813 7.439375165 11.8487921 10.85584206 9.133101514 18.00306864 18.04487827 15.90644743 16.5004933 7.092154655 2.359532285 2.462296204 2.805064435 1.446366729 0.355589125 2.426618724 2.774304618 3.225702785 5.953339807 4.866306232 1.970980468 3.715958326 2.256018776 4.204191132 3.955504624 4.71235995 3.10919194 1.395718353 Thymidine 0.169604512 0.210720684 0.325274506 0.317537546 0.180781292 0.293897011 0.302357604 0.194572653 0.201155588 0.260490854 0.241262547 0.227463393 0.161841187 0.242906578 0.181223483 0.1912747 0.166936715 0.214503494 0.085434605 0.202357037 0.173258756 0.085010661 0.067988329 0.142752336 0.095576548 0.077675249 0.222796203 0.230274969 0.08600732 0.104836339 4.049277715 -0.066758365 2.857816458 2.605774687 1.436427168 1.111186981 0.979080867 2.054228022 13.51572267 16.40245071 9.049452841 8.661945448 5.108697646 6.387109586 1.709318865 3.376196634 1.590771032 2.173255473 1.199725438 4.647420749 2.959042428 3.490308298 7.579025592 11.38717139 6.753025734 3.919787765 7.253003055 14.01059881 3.23167682 2.368214831 2.073856797 N-6-trimethyllysine 2.770148358 1.700094133 2.739927881 3.886750692 1.704344154 6.650247047 1.441963937 1.758224645 1.847252773 2.152365901 2.239923783 1.86351392 1.872722567 1.877393971 2.354705369 2.269201951 1.49630182 2.342263141 0.748659673 1.394254512 0.791788984 0.958665972 1.061676713 1.296582954 1.910631401 1.799376294 2.086104137 1.4649511 1.371630601 1.309271322 1.717702246 0.336637247 1.290455724 1.496518146 1.15186942 0.889753885 0.945357891 0.907489967 1.932245845 2.288824754 1.744992649 1.534569206 1.166739329 1.843032243 0.914474617 1.141559874 4.616036816 1.248673367 1.591867823 1.753825525 0.377777807 0.688930875 0.908437019 1.4229805 1.083189894 0.779989878 0.654877974 0.757895991 0.503311559 0.507661925 0.426678874 UDP-N-acetylglucosamine 47.72976893 29.21290633 22.01621733 19.57438195 32.91034873 33.4009212 33.76442882 31.66444308 30.68844656 20.40309533 33.20599703 24.04347224 44.07958498 35.65611181 61.64483877 74.85782293 48.04954416 43.43639128 13.96409836 16.29644844 7.95535308 5.741074021 19.17598456 28.59051255 0.226310725 0.626646676 1.032507885 0.700138056 1.254413879 0.911423063 25.7568155 4.418765247 33.55694913 49.64172114 32.16729707 19.41011834 11.3579289 20.97862007 7.586903876 16.11645774 7.492029838 3.450884144 15.49743831 0.509926225 0.396112192 0.68620504 2.17634826 5.652407007 0.606489848 2.652570181 0.293707934 0.753869828 1.137577179 0.620358456 3.557863169 0.527624827 4.277184177 0.94538507 UDP-glucose 23.98148328 15.93030041 12.02068896 13.77936563 24.44867201 19.03279846 13.78171583 16.92616416 10.68793616 10.70311181 16.63924963 13.1078311 20.74199528 12.29349846 43.02964994 31.63831628 22.98092264 28.31516999 4.785850615 4.433591187 1.834201804 0.609418809 6.12209744 7.563774623 0 0.301111959 0.858212776 0.477794876 0.832638594 0.363688737 10.39317688 1.708434066 6.716809979 16.02049645 11.88929268 6.948276815 1.514314284 5.822485921 2.085392407 3.820672471 1.516761626 1.067932613 3.101899799 0.169739959 0.347129594 0.977945739 2.881807362 1.143104275 0.365516013 0.476296235 0.813794439 3.727791366 0.455264245 Uracil 95.48566199 62.42446898 145.8497167 79.41711153 91.95410917 78.880118 38.86449128 36.14806892 56.51024691 45.20287127 36.35586897 57.95050353 50.44849748 82.68982024 46.95332796 29.7267697 27.87394546 52.51281966 53.27521329 56.25894071 52.1820528 61.21965038 77.34425329 93.42216225 1.037356708 3.371155252 4.125171769 6.774905158 1.492833348 2.007863881 114.024014 4.636616098 67.47417622 130.3832901 111.2122208 67.00955294 89.13421849 67.9270078 76.03105038 74.02880058 55.70334431 44.04966791 53.80551915 5.327750804 3.545444061 2.858117008 3.194176757 0.884023662 4.088030911 4.163247955 8.862160637 14.7202732 5.945652287 7.429370787 11.19942798 7.837093021 7.076106471 18.88304341 5.180917831 11.32133535 3.442194984 Xanthine 66.34179766 48.05852659 115.023652 47.44433302 68.49618794 50.06021586 31.29790216 33.73012577 47.3445078 42.01408131 44.63075836 60.44232323 33.81900334 71.745812 33.87343202 45.1788031 34.64000009 42.46680977 36.7655377 44.73776721 48.22699409 64.57429409 60.28173345 71.38881817 -0.001132568 0.002942889 0.011043552 -0.005164005 0.057791151 0.014316382 26.75372183 5.80986772 21.71812528 22.23820513 22.41612635 21.77144642 27.87204145 27.67350335 31.45682029 39.6907204 22.70455047 25.75850308 -0.232983545 5.463082348 0.225091648 4.238328833 7.227121445 0.984597178 9.540348818 13.59997968 14.66286874 21.11110909 16.2721249 -0.073778491 12.93861189 10.36231609 20.0844056 25.28844313 17.77147229 19.74083246 10.25119055 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name 2-hydroxybutyric acid 2-hydroxyglutaric acid beta-hydroxybutyric acid 3-hydroxyisobutyric acid Ketoisovaleric acid 3-Methyl-2-oxovaleric acid 3-Phosphoglyceric acid 3-methyl pyruvic acid Oxoglutaric acid Allantoin AMP Argininosuccinic acid Vitamin C Aspartic acid Cis-Aconitic acid Citric acid CTP Cysteic acid Dihydroxyacetone phosphate dTMP Fumaric acid Glucose glucose-1-phosphate/fructose-6 -hosphate Glucose-6-phosphate Glutamic acid Glyceric acid GMP Itaconic acid Lactic acid Malic acid 5'-Methylthioadenosine Phosphoethanolamine Orotic acid P-Aminobenzoic acid Pyruvic acid Ribose-5-phosphate Sorbitol Succinic acid Thiamine Thymidine N-6-trimethyllysine UDP-N-acetylglucosamine UDP-glucose Uracil Xanthine METABOLITES_END #END