#METABOLOMICS WORKBENCH ijochmans_20230717_025226 DATATRACK_ID:4167 STUDY_ID:ST002786 ANALYSIS_ID:AN004533 PROJECT_ID:PR001736 VERSION 1 CREATED_ON July 19, 2023, 9:03 am #PROJECT PR:PROJECT_TITLE Metabolic changes during normothermic isolated kidney perfusion PR:PROJECT_TYPE Experimental PR:PROJECT_SUMMARY Normothermic isolated kidney perfusion is being developed as a method to PR:PROJECT_SUMMARY preserve donor kidneys and to assess their future function before they are PR:PROJECT_SUMMARY transplanted. Donor kidneys can be exposed to different types and degrees of PR:PROJECT_SUMMARY ischemic injury before they are preserved. This project studies how the PR:PROJECT_SUMMARY metabolome changes in relation to different types and degrees of ischemia. PR:INSTITUTE KU Leuven PR:DEPARTMENT Microbiology, Immunology and Transplantation PR:LABORATORY Lab of Abdominal Transplantation PR:LAST_NAME Jochmans PR:FIRST_NAME Ina PR:ADDRESS Herestraat 49, Leuven, 3000, Belgium PR:EMAIL ina.jochmans@kuleuven.be PR:PHONE None PR:FUNDING_SOURCE FTBO, KOOR, FWO PR:CONTRIBUTORS Julie De Beule, Bart Ghesquière, Ina Jochmans #STUDY ST:STUDY_TITLE Metabolic changes of the pig kidney during isolated normothermic perfusion with ST:STUDY_TITLE autologous whole blood - urine ST:STUDY_SUMMARY This study investigates how glucose, lactate and 20 amino acids in the urine of ST:STUDY_SUMMARY normothermically perfused pig kidneys changes over time. It also studies how ST:STUDY_SUMMARY type and severity of ischemia before perfusion (cold or warm ischemia) change ST:STUDY_SUMMARY this profile. ST:INSTITUTE KU Leuven ST:DEPARTMENT Microbiology, Immunology and Transplantation ST:LABORATORY Lab of Abdominal Transplantation ST:LAST_NAME Jochmans ST:FIRST_NAME Ina ST:ADDRESS Herestraat 49, Leuven, 3000, Belgium ST:EMAIL ina.jochmans@kuleuven.be ST:NUM_GROUPS 2 ST:TOTAL_SUBJECTS 12 ST:NUM_MALES 6 male pigs ST:STUDY_TYPE Experimental ST:PHONE None #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Sus scrofa SU:TAXONOMY_ID 9823 SU:AGE_OR_AGE_RANGE ~4 months SU:WEIGHT_OR_WEIGHT_RANGE 35-45 kg SU:GENDER Male SU:ANIMAL_ANIMAL_SUPPLIER TOPIGS TN70, Tojapigs, the Netherlands SU:ANIMAL_LIGHT_CYCLE 12 hours SU:ANIMAL_WATER ad libitum #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 19 19_T1 Timepoint:60 | Group:Control | Urine_volume:46 RAW_FILE_NAME=19_T1.mzML SUBJECT_SAMPLE_FACTORS 19 19_T2 Timepoint:120 | Group:Control | Urine_volume:90 RAW_FILE_NAME=19_T2.mzML SUBJECT_SAMPLE_FACTORS 19 19_T3 Timepoint:180 | Group:Control | Urine_volume:100 RAW_FILE_NAME=19_T3.mzML SUBJECT_SAMPLE_FACTORS 19 19_T4 Timepoint:240 | Group:Control | Urine_volume:50 RAW_FILE_NAME=19_T4.mzML SUBJECT_SAMPLE_FACTORS 31 31_T1 Timepoint:60 | Group:Control | Urine_volume:57 RAW_FILE_NAME=31_T1.mzML SUBJECT_SAMPLE_FACTORS 31 31_T2 Timepoint:120 | Group:Control | Urine_volume:60 RAW_FILE_NAME=31_T2.mzML SUBJECT_SAMPLE_FACTORS 31 31_T3 Timepoint:180 | Group:Control | Urine_volume:42 RAW_FILE_NAME=31_T3.mzML SUBJECT_SAMPLE_FACTORS 31 31_T4 Timepoint:240 | Group:Control | Urine_volume:17 RAW_FILE_NAME=31_T4.mzML SUBJECT_SAMPLE_FACTORS 34 34_T1 Timepoint:60 | Group:Control | Urine_volume:67 RAW_FILE_NAME=34_T1.mzML SUBJECT_SAMPLE_FACTORS 34 34_T2 Timepoint:120 | Group:Control | Urine_volume:29 RAW_FILE_NAME=34_T2.mzML SUBJECT_SAMPLE_FACTORS 34 34_T3 Timepoint:180 | Group:Control | Urine_volume:60 RAW_FILE_NAME=34_T3.mzML SUBJECT_SAMPLE_FACTORS 34 34_T4 Timepoint:240 | Group:Control | Urine_volume:28 RAW_FILE_NAME=34_T4.mzML SUBJECT_SAMPLE_FACTORS 67 67_T1 Timepoint:60 | Group:Control | Urine_volume:70 RAW_FILE_NAME=67_T1.mzML SUBJECT_SAMPLE_FACTORS 67 67_T2 Timepoint:120 | Group:Control | Urine_volume:8 RAW_FILE_NAME=67_T2.mzML SUBJECT_SAMPLE_FACTORS 67 67_T3 Timepoint:180 | Group:Control | Urine_volume:3 RAW_FILE_NAME=67_T3.mzML SUBJECT_SAMPLE_FACTORS 67 67_T4 Timepoint:240 | Group:Control | Urine_volume:12 RAW_FILE_NAME=67_T4.mzML SUBJECT_SAMPLE_FACTORS 70 70_T1 Timepoint:60 | Group:Control | Urine_volume:80 RAW_FILE_NAME=70_T1.mzML SUBJECT_SAMPLE_FACTORS 70 70_T2 Timepoint:120 | Group:Control | Urine_volume:47 RAW_FILE_NAME=70_T2.mzML SUBJECT_SAMPLE_FACTORS 70 70_T3 Timepoint:180 | Group:Control | Urine_volume:90 RAW_FILE_NAME=70_T3.mzML SUBJECT_SAMPLE_FACTORS 70 70_T4 Timepoint:240 | Group:Control | Urine_volume:42 RAW_FILE_NAME=70_T4.mzML SUBJECT_SAMPLE_FACTORS 76 76_T1 Timepoint:60 | Group:Control | Urine_volume:105 RAW_FILE_NAME=76_T1.mzML SUBJECT_SAMPLE_FACTORS 76 76_T2 Timepoint:120 | Group:Control | Urine_volume:80 RAW_FILE_NAME=76_T2.mzML SUBJECT_SAMPLE_FACTORS 76 76_T3 Timepoint:180 | Group:Control | Urine_volume:110 RAW_FILE_NAME=76_T3.mzML SUBJECT_SAMPLE_FACTORS 76 76_T4 Timepoint:240 | Group:Control | Urine_volume:105 RAW_FILE_NAME=76_T4.mzML SUBJECT_SAMPLE_FACTORS 29 29_T1 Timepoint:60 | Group:CI | Urine_volume:67 RAW_FILE_NAME=29_T1.mzML SUBJECT_SAMPLE_FACTORS 29 29_T2 Timepoint:120 | Group:CI | Urine_volume:19 RAW_FILE_NAME=29_T2.mzML SUBJECT_SAMPLE_FACTORS 29 29_T3 Timepoint:180 | Group:CI | Urine_volume:33 RAW_FILE_NAME=29_T3.mzML SUBJECT_SAMPLE_FACTORS 29 29_T4 Timepoint:240 | Group:CI | Urine_volume:32 RAW_FILE_NAME=29_T4.mzML SUBJECT_SAMPLE_FACTORS 30 30_T1 Timepoint:60 | Group:CI | Urine_volume:84 RAW_FILE_NAME=30_T1.mzML SUBJECT_SAMPLE_FACTORS 30 30_T2 Timepoint:120 | Group:CI | Urine_volume:33 RAW_FILE_NAME=30_T2.mzML SUBJECT_SAMPLE_FACTORS 30 30_T3 Timepoint:180 | Group:CI | Urine_volume:20 RAW_FILE_NAME=30_T3.mzML SUBJECT_SAMPLE_FACTORS 30 30_T4 Timepoint:240 | Group:CI | Urine_volume:15 RAW_FILE_NAME=30_T4.mzML SUBJECT_SAMPLE_FACTORS 33 33_T1 Timepoint:60 | Group:CI | Urine_volume:21 RAW_FILE_NAME=33_T1.mzML SUBJECT_SAMPLE_FACTORS 33 33_T2 Timepoint:120 | Group:CI | Urine_volume:45 RAW_FILE_NAME=33_T2.mzML SUBJECT_SAMPLE_FACTORS 33 33_T3 Timepoint:180 | Group:CI | Urine_volume:80 RAW_FILE_NAME=33_T3.mzML SUBJECT_SAMPLE_FACTORS 33 33_T4 Timepoint:240 | Group:CI | Urine_volume:26 RAW_FILE_NAME=33_T4.mzML SUBJECT_SAMPLE_FACTORS 71 71_T1 Timepoint:60 | Group:CI | Urine_volume:30 RAW_FILE_NAME=71_T1.mzML SUBJECT_SAMPLE_FACTORS 71 71_T2 Timepoint:120 | Group:CI | Urine_volume:50 RAW_FILE_NAME=71_T2.mzML SUBJECT_SAMPLE_FACTORS 71 71_T3 Timepoint:180 | Group:CI | Urine_volume:24 RAW_FILE_NAME=71_T3.mzML SUBJECT_SAMPLE_FACTORS 71 71_T4 Timepoint:240 | Group:CI | Urine_volume:10 RAW_FILE_NAME=71_T4.mzML SUBJECT_SAMPLE_FACTORS 72 72_T1 Timepoint:60 | Group:CI | Urine_volume:35 RAW_FILE_NAME=72_T1.mzML SUBJECT_SAMPLE_FACTORS 72 72_T2 Timepoint:120 | Group:CI | Urine_volume:85 RAW_FILE_NAME=72_T2.mzML SUBJECT_SAMPLE_FACTORS 72 72_T3 Timepoint:180 | Group:CI | Urine_volume:60 RAW_FILE_NAME=72_T3.mzML SUBJECT_SAMPLE_FACTORS 72 72_T4 Timepoint:240 | Group:CI | Urine_volume:50 RAW_FILE_NAME=72_T4.mzML SUBJECT_SAMPLE_FACTORS 75 75_T1 Timepoint:60 | Group:CI | Urine_volume:120 RAW_FILE_NAME=75_T1.mzML SUBJECT_SAMPLE_FACTORS 75 75_T2 Timepoint:120 | Group:CI | Urine_volume:27 RAW_FILE_NAME=75_T2.mzML SUBJECT_SAMPLE_FACTORS 75 75_T3 Timepoint:180 | Group:CI | Urine_volume:3 RAW_FILE_NAME=75_T3.mzML SUBJECT_SAMPLE_FACTORS 75 75_T4 Timepoint:240 | Group:CI | Urine_volume:0 RAW_FILE_NAME=75_T4.mzML #COLLECTION CO:COLLECTION_SUMMARY Urine production over 1 hour of perfusion was diverted to a collection bag. At CO:COLLECTION_SUMMARY the end of the hour, urine samples were taken in dry tubes from this collection CO:COLLECTION_SUMMARY bag. One milliliter perfusate aliquots (centrifuged 1000g, 10 minutes, 4°C) CO:COLLECTION_SUMMARY were snap frozen in liquid nitrogen and stored at -80°C until analysis. CO:SAMPLE_TYPE Urine CO:COLLECTION_FREQUENCY Every hour during perfusion, so T1 a sample from urine produced during the first CO:COLLECTION_FREQUENCY hour of perfusion, T2 for the second hour, T3 for the third hour, and T4 for the CO:COLLECTION_FREQUENCY fourth hour CO:STORAGE_CONDITIONS -80℃ CO:ADDITIVES None #TREATMENT TR:TREATMENT_SUMMARY Three experimental conditions were investigated (n=6/group): (a) Controls; (b) TR:TREATMENT_SUMMARY warm ischemia (WI) simulating hypoxic acute kidney injury; (c) cold ischemia TR:TREATMENT_SUMMARY (CI) replicating clinical cold storage. Control kidneys were retrieved, flushed TR:TREATMENT_SUMMARY with cold IGL-1, and immediately reperfused. CI group, kidneys were exposed to TR:TREATMENT_SUMMARY 22h of CI by submerging them in IGL- 1 (a clinical preservation solution for TR:TREATMENT_SUMMARY cold storage) and storing them on ice. In WI, the renal artery and vein were TR:TREATMENT_SUMMARY clamped for 60 min before retrieval. In the All kidneys were flushed with 200 ml TR:TREATMENT_SUMMARY of Ringer’s solution before mounting them on the ex situ circuit to wash out TR:TREATMENT_SUMMARY IGL-1. Because little urine was produced by WI kidneys, too few samples were TR:TREATMENT_SUMMARY available for a meaningful analysis. TR:ANIMAL_ANESTHESIA Pigs were sedated by an intramuscular injection of Tiletamine/Zolazepam (8 TR:ANIMAL_ANESTHESIA mg/kg, Zoletil®, Virbac, Belgium) and Xylazine (2 mg/kg, Xylazine®, VMD TR:ANIMAL_ANESTHESIA pharma, Belgium) to allow orotracheal intubation. Anesthesia was maintained by TR:ANIMAL_ANESTHESIA inhalation of isoflurane (1% Isovet®, Piramal Critical Care B.V., Belgium) and TR:ANIMAL_ANESTHESIA continuous infusion of fentanyl (8 µg/kg, Fentanyl®, Janssen Pharmaceutica, TR:ANIMAL_ANESTHESIA Belgium). TR:ANIMAL_ACCLIMATION_DURATION Minimum of 2 days TR:ANIMAL_FASTING Overnight TR:ANIMAL_ENDP_EUTHANASIA Non-recovery study #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Samples were extracted in an 80% methanol (80:20 methanol:water) (Methanol SP:SAMPLEPREP_SUMMARY ≥99.9%, HiPerSolv CHROMANORM®, ULTRA for LC-MS, suitable for UPLC/UHPLC-MS SP:SAMPLEPREP_SUMMARY instruments, VWR, Belgium) extraction buffer containing 1 µM of deuterated D27 SP:SAMPLEPREP_SUMMARY myristic acid, 5 µM D12 glucose, 3 µM 13C5-D5-15N Glutamic acid and 3 µM SP:SAMPLEPREP_SUMMARY D7-15N4-Arginine as internal standards. 10 µl of sample was added to 990 µl of SP:SAMPLEPREP_SUMMARY the extraction buffer and stored overnight at -80 °C. Insolubilities and SP:SAMPLEPREP_SUMMARY precipitated proteins were removed by centrifugation at 20.000 g, for 15 min at SP:SAMPLEPREP_SUMMARY 4 °C. 200 µL of the supernatant was transferred to an appropriate SP:SAMPLEPREP_SUMMARY mass-spectrometry vial. 50 µl of samples was added to 950 µl of the extraction SP:SAMPLEPREP_SUMMARY buffer and stored overnight at -80°C. SP:EXTRACT_STORAGE -80℃ #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY 10 µl of each sample was loaded into a Dionex UltiMate 3000 LC System (Thermo CH:CHROMATOGRAPHY_SUMMARY Scientific Bremen, Germany) equipped with a Waters ACQUITY UPLC BEH C18 (150 x CH:CHROMATOGRAPHY_SUMMARY 2.1mm,1.7um) column coupled to a Q Exactive Orbitrap mass spectrometer (Thermo CH:CHROMATOGRAPHY_SUMMARY Scientific) operating in negative ion mode. A step gradient was carried out CH:CHROMATOGRAPHY_SUMMARY using solvent A (10 mM TBA and 15 mM acetic acid) and solvent B (100% methanol). CH:CHROMATOGRAPHY_SUMMARY The gradient started with 5% of solvent B and 95% solvent A and remained at 5% B CH:CHROMATOGRAPHY_SUMMARY until 2 min post injection. A linear gradient to 37% B was carried out until 7 CH:CHROMATOGRAPHY_SUMMARY min and increased to 41% until 14 min. Between 14 and 26 minutes the gradient CH:CHROMATOGRAPHY_SUMMARY increased to 95% of B and remained at 95% B for 4 minutes. At 30 min the CH:CHROMATOGRAPHY_SUMMARY gradient returned to 5% B. The chromatography was stopped at 40 min. The flow CH:CHROMATOGRAPHY_SUMMARY was kept constant at 0.25 mL/min and the column was placed at 40°C throughout CH:CHROMATOGRAPHY_SUMMARY the analysis. The MS operated in full scan mode (m/z range: [70.0000-1050.0000]) CH:CHROMATOGRAPHY_SUMMARY using a spray voltage of 4.80 kV, capillary temperature of 300°C, sheath gas at CH:CHROMATOGRAPHY_SUMMARY 40.0, auxiliary gas at 10.0. The AGC target was set at 3.0E+006 using a CH:CHROMATOGRAPHY_SUMMARY resolution of 140000, with a maximum IT fill time of 512 ms. Data collection was CH:CHROMATOGRAPHY_SUMMARY performed using the Xcalibur software (Thermo Scientific). The data analyses CH:CHROMATOGRAPHY_SUMMARY were performed by integrating the peak areas (El-Maven - Polly - Elucidata). CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Thermo Dionex Ultimate 3000 CH:COLUMN_NAME Waters ACQUITY UPLC BEH C18 (150 x 2.1mm,1.7um) CH:SOLVENT_A 100% water; 10 mM TBA; 15 mM acetic acid CH:SOLVENT_B 100% methanol CH:FLOW_GRADIENT The gradient started with 5% of solvent B and 95% solvent A and remained at 5% B CH:FLOW_GRADIENT until 2 min post injection. A linear gradient to 37% B was carried out until 7 CH:FLOW_GRADIENT min and increased to 41% until 14 min. Between 14 and 26 minutes the gradient CH:FLOW_GRADIENT increased to 95% of B and remained at 95% B for 4 minutes. At 30 min the CH:FLOW_GRADIENT gradient returned to 5% B. The chromatography was stopped at 40 min. CH:FLOW_RATE 0.25 ml/min CH:COLUMN_TEMPERATURE 40°C #ANALYSIS AN:ANALYSIS_TYPE MS AN:LABORATORY_NAME Laboratory of Applied Mass Spectrometry, KU Leuven #MS MS:INSTRUMENT_NAME Thermo Q Exactive Focus MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS Raw MS data were converted into mzML using the MSConvert tool of Proteowizard MS:MS_COMMENTS (version 3.0.20247). Peak picking was performed with El_Maven (El_Maven, MS:MS_COMMENTS Elucidata, Massachusetts, USA, version 0.12.0). Metabolites were identified MS:MS_COMMENTS using an in-house library containing exact mass and retention time. The mass MS:MS_COMMENTS accuracy during data processing in El Maven was set at 10 ppm. Calculation of MS:MS_COMMENTS abundances was done in the LC-MS Workflow of El_Maven. Raw abundances (peak area MS:MS_COMMENTS values) for each metabolite were corrected for internal standard (Myristic acid MS:MS_COMMENTS d27). These were corrected for urine volume. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Peak area MS_METABOLITE_DATA_START Samples 19_T1 19_T2 19_T3 19_T4 31_T1 31_T2 31_T3 31_T4 34_T1 34_T2 34_T3 34_T4 67_T1 67_T2 67_T3 67_T4 70_T1 70_T2 70_T3 70_T4 76_T1 76_T2 76_T3 76_T4 29_T1 29_T2 29_T3 29_T4 30_T1 30_T2 30_T3 30_T4 33_T1 33_T2 33_T3 33_T4 71_T1 71_T2 71_T3 71_T4 72_T1 72_T2 72_T3 72_T4 75_T1 75_T2 75_T3 75_T4 Factors Timepoint:60 | Group:Control | Urine_volume:46 Timepoint:120 | Group:Control | Urine_volume:90 Timepoint:180 | Group:Control | Urine_volume:100 Timepoint:240 | Group:Control | Urine_volume:50 Timepoint:60 | Group:Control | Urine_volume:57 Timepoint:120 | Group:Control | Urine_volume:60 Timepoint:180 | Group:Control | Urine_volume:42 Timepoint:240 | Group:Control | Urine_volume:17 Timepoint:60 | Group:Control | Urine_volume:67 Timepoint:120 | Group:Control | Urine_volume:29 Timepoint:180 | Group:Control | Urine_volume:60 Timepoint:240 | Group:Control | Urine_volume:28 Timepoint:60 | Group:Control | Urine_volume:70 Timepoint:120 | Group:Control | Urine_volume:8 Timepoint:180 | Group:Control | Urine_volume:3 Timepoint:240 | Group:Control | Urine_volume:12 Timepoint:60 | Group:Control | Urine_volume:80 Timepoint:120 | Group:Control | Urine_volume:47 Timepoint:180 | Group:Control | Urine_volume:90 Timepoint:240 | Group:Control | Urine_volume:42 Timepoint:60 | Group:Control | Urine_volume:105 Timepoint:120 | Group:Control | Urine_volume:80 Timepoint:180 | Group:Control | Urine_volume:110 Timepoint:240 | Group:Control | Urine_volume:105 Timepoint:60 | Group:CI | Urine_volume:67 Timepoint:120 | Group:CI | Urine_volume:19 Timepoint:180 | Group:CI | Urine_volume:33 Timepoint:240 | Group:CI | Urine_volume:32 Timepoint:60 | Group:CI | Urine_volume:84 Timepoint:120 | Group:CI | Urine_volume:33 Timepoint:180 | Group:CI | Urine_volume:20 Timepoint:240 | Group:CI | Urine_volume:15 Timepoint:60 | Group:CI | Urine_volume:21 Timepoint:120 | Group:CI | Urine_volume:45 Timepoint:180 | Group:CI | Urine_volume:80 Timepoint:240 | Group:CI | Urine_volume:26 Timepoint:60 | Group:CI | Urine_volume:30 Timepoint:120 | Group:CI | Urine_volume:50 Timepoint:180 | Group:CI | Urine_volume:24 Timepoint:240 | Group:CI | Urine_volume:10 Timepoint:60 | Group:CI | Urine_volume:35 Timepoint:120 | Group:CI | Urine_volume:85 Timepoint:180 | Group:CI | Urine_volume:60 Timepoint:240 | Group:CI | Urine_volume:50 Timepoint:60 | Group:CI | Urine_volume:120 Timepoint:120 | Group:CI | Urine_volume:27 Timepoint:180 | Group:CI | Urine_volume:3 Timepoint:240 | Group:CI | Urine_volume:0 Glucose 0.949752687 0.307711313 0.072097865 0.047302157 0.295942339 0.186947969 0.07347158 0.213602659 0.586316782 0.326465557 0.209735144 0.140667034 0.377920646 0.532937871 0.710223549 0.54227837 0.414942192 0.252096492 0.176520272 0.079840557 0.309594657 0.098865694 0.073575579 0.060705494 0.664826297 0.858580533 0.551121731 0.218682948 0.878998657 0.756497721 0.60004047 0.303012241 0.595616659 0.311470862 0.11936302 0.156621223 0.50789365 0.647701599 0.49589895 0.54580424 0.550789055 0.727878449 0.555185302 0.349070174 0.620338612 0.546282469 0.702515668 n.a. Lactate 6.04177336 1.689937093 1.201038663 2.18740033 3.703943811 2.195559883 3.050557069 6.649451029 8.805944791 6.670920491 8.183487197 8.508561 3.742233151 2.349633094 3.395725736 3.489689969 3.959425019 0.932111148 0.991810128 1.158693698 1.687682446 0.441398363 0.737889915 0.47516663 6.461498896 6.18275215 7.202565263 6.782522712 7.338586448 6.059743393 7.27589037 8.138732448 4.568789232 4.551396835 4.987790835 6.603765476 4.716748396 2.02106002 2.221023709 4.362181614 4.543642993 2.77440423 4.187767976 4.847093565 4.225105178 1.941771896 3.321099476 n.a. Gly 0.065195438 0.033847819 0.043768925 0.056468933 0.060095542 0.030659644 0.013770293 0.035611385 0.097192788 0.052635567 0.0596145 0.0762999 0.054965559 0.060431482 0.085092628 0.07643032 0.045115188 0.025158376 0.026889511 0.015466792 0.042105395 0.022224931 0.019663274 0.031872349 0.067295744 0.082586628 0.075276444 0.058748871 0.096277807 0.082503653 0.083376567 0.081416384 0.092230102 0.047598497 0.032698741 0.053523341 0.071945633 0.104118564 0.088065563 0.069649565 0.085118927 0.138651021 0.095255492 0.060354036 0.094685911 0.086275911 0.146680878 n.a. Ala 0.270454042 0.080084021 0.050802694 0.113521607 0.055453747 0.022832406 0.016698085 0.062788768 0.204633677 0.149360585 0.187238626 0.276114274 0.063043917 0.043750869 0.042379848 0.05438016 0.053366389 0.015432684 0.023852091 0.014791321 0.033658074 0.006334372 0.007756278 0.007879355 0.130603857 0.22047631 0.151202257 0.121565048 0.187740776 0.214449942 0.176266062 0.148905248 0.115551657 0.048120392 0.082402444 0.158638778 0.112056036 0.05444024 0.047715851 0.093253659 0.130958276 0.076278961 0.090998368 0.119326008 0.118422238 0.058748715 0.103222127 n.a. Arg 0.000697344 0.000180568 0.00017857 0.00017723 0.000553421 0.000215966 0.000176964 0.000189495 0.003906645 0.000436107 0.000302871 0.000248876 0.000617188 0.0018846 0.004189849 0.002760123 0.000425463 0.000185071 0.000194903 0.000182241 0.000183611 0.000183283 0.000171233 0.000187417 0.001273988 0.002684322 0.000778007 0.000299636 0.001549839 0.00263595 0.001430094 0.000786956 0.002073756 0.000933133 0.000186782 0.000321726 0.000727729 0.00042999 0.000245801 0.000537794 0.00167341 0.00025286 0.000403796 0.000467389 0.001660667 0.000341721 0.002712927 n.a. Asn 0.038649936 0.033316103 0.035113002 0.055474311 0.012789418 0.015298644 0.006987117 0.021281046 0.039689772 0.030446678 0.032058505 0.036839767 0.017773151 0.042149664 0.076750675 0.071239302 0.015887584 0.014704551 0.019321217 0.011936911 0.01095757 0.011264085 0.011347954 0.013240607 0.031222806 0.077914292 0.052030204 0.03160273 0.042757353 0.099827631 0.065274252 0.042912488 0.024060451 0.028787939 0.019654085 0.03950366 0.035750812 0.06239002 0.035414535 0.033676231 0.048563163 0.077292846 0.04166835 0.031268119 0.046335141 0.064414798 0.095015815 n.a. Glu 1.04156642 0.247800189 0.939888881 1.144192611 0.847298549 0.184916447 0.067254942 0.163814497 1.768098683 0.65307321 0.615298893 0.712761657 0.777393655 0.383234663 0.394074685 0.381101207 0.652577739 0.205383377 0.171731225 0.082136598 0.452798731 0.06402126 0.078614204 0.450773325 0.989470559 0.36223941 0.246695377 0.182623522 1.294318721 0.400511945 0.330150716 0.27165667 0.617766052 0.16663002 0.196923053 0.362451202 1.506166156 1.588604436 0.959261751 0.927914418 1.971153722 1.92095652 1.424083845 1.046150452 1.562940747 1.413268758 1.871824894 n.a. Gln 1.293820058 0.377514657 0.161714048 0.140546637 0.301288087 0.033794216 0.006115644 0.02095357 0.666215108 0.126009971 0.090419291 0.123876256 0.529517546 0.415857072 0.188571742 0.104707309 0.483257573 0.13732509 0.030665912 0.021391842 0.309614118 0.031010696 0.012359812 0.015738984 0.39803013 0.219447039 0.087873131 0.04300982 0.685903456 0.309720451 0.11216694 0.053478085 0.812122093 0.056181895 0.012883917 0.027979553 0.265625817 0.097096612 0.043295205 0.065839813 0.370179496 0.125963479 0.05360409 0.03694925 0.875972205 0.735425101 0.748769485 n.a. (Iso)Leu 0.158459226 0.07400103 0.053597212 0.111748132 0.077331378 0.052288395 0.030007738 0.164828456 0.353678619 0.19322892 0.249605269 0.396139231 0.096328951 0.149674926 0.380145884 0.329190931 0.098342568 0.055659343 0.080779998 0.044721743 0.045073767 0.026099123 0.033700863 0.044118285 0.158012916 0.306972855 0.294613227 0.205985777 0.264344381 0.296328456 0.30553564 0.268361535 0.149346011 0.080428148 0.089608661 0.259938196 0.191299058 0.237722753 0.203220771 0.383811702 0.207828818 0.3457931 0.297581247 0.264789873 0.183785748 0.205329894 0.354060927 n.a. Lys 0.003516426 0.005601905 0.003254739 0.00495453 0.003194671 0.003964287 0.002724275 0.009486716 0.013420333 0.012645687 0.011128669 0.015434736 0.003707108 0.010818496 0.024748897 0.022771059 0.002063081 0.003792425 0.006540094 0.003707738 0.001855345 0.001820718 0.001908566 0.00174144 0.006381778 0.01776862 0.016282478 0.011725313 0.006764062 0.015157639 0.016142106 0.016461721 0.004309839 0.006130978 0.005949688 0.014694708 0.008074846 0.014970108 0.009560201 0.013681874 0.009744026 0.014933027 0.010538932 0.009875446 0.007670993 0.012487933 0.019529543 n.a. Met 0.009329151 0.005758576 0.005330173 0.012110785 0.003645616 0.003975783 0.002241078 0.01375336 0.016712747 0.012480583 0.01852823 0.029027597 0.003948949 0.010050036 0.031105419 0.028466988 0.003419435 0.004325917 0.008471699 0.004752525 0.003182373 0.002517478 0.003909205 0.004879405 0.010391967 0.024543033 0.028161215 0.023297729 0.017215114 0.028472171 0.027043539 0.030058534 0.006978918 0.006154853 0.007938786 0.024254367 0.011923912 0.019501734 0.017067387 0.0353524 0.01468137 0.028908585 0.030004612 0.025967627 0.012934597 0.016601277 0.031551198 n.a. Phe 0.022153878 0.018746045 0.015354373 0.032072782 0.011833079 0.013755001 0.008925747 0.060671276 0.054141536 0.043093298 0.060257984 0.099167995 0.014082268 0.029329008 0.112075155 0.106769898 0.013077758 0.018010237 0.036022248 0.024049642 0.008595023 0.01155319 0.014404148 0.018319064 0.031927103 0.073478985 0.092558378 0.072462799 0.043087953 0.078726647 0.092641547 0.087540246 0.014797411 0.020583701 0.027208294 0.097120636 0.037530881 0.061805005 0.053988323 0.105616997 0.04424707 0.077882035 0.075694813 0.078189918 0.038453251 0.047378055 0.103333743 n.a. Pro 0.108346871 0.062801744 0.063397609 0.107287047 0.043327284 0.036597693 0.04326144 0.13596642 0.1262218 0.098669112 0.148310464 0.265768357 0.035250573 0.047280284 0.11410502 0.131016142 0.035047678 0.036376446 0.069378337 0.04625584 0.035411161 0.036977601 0.053420262 0.06300115 0.089909144 0.163299745 0.174518733 0.181223952 0.129954951 0.166005019 0.166057494 0.209322689 0.034131128 0.044465157 0.0871744 0.257226177 0.086654871 0.129013416 0.106467966 0.184371488 0.112220388 0.183858298 0.204200858 0.204221184 0.094854854 0.108254608 0.174323299 n.a. Ser 0.185499454 0.132434656 0.099450747 0.151584567 0.069837735 0.077523127 0.031247764 0.086955037 0.219791404 0.211843647 0.236914887 0.309684761 0.075401913 0.223735808 0.394355507 0.302457279 0.070797599 0.07077066 0.070385102 0.033745683 0.04665026 0.027938941 0.032282132 0.029039773 0.13996801 0.512344593 0.318839881 0.169423031 0.203045779 0.612357454 0.433652166 0.246523069 0.121869123 0.197719203 0.093144433 0.183441865 0.162510946 0.366074077 0.243347837 0.269080434 0.261858432 0.653568282 0.258650909 0.171058826 0.236546698 0.315258469 0.594215389 n.a. Thr 0.283729533 0.106463902 0.107157032 0.180100344 0.084872452 0.066603502 0.019208023 0.104271659 0.275359538 0.186744368 0.202435628 0.316024745 0.077295206 0.228937833 0.358009567 0.273964759 0.072232396 0.048266744 0.052177045 0.034895773 0.044491445 0.021587842 0.029726087 0.044670065 0.097585146 0.38849325 0.289594947 0.15310667 0.149744759 0.433861151 0.329215092 0.228278536 0.262832501 0.174616849 0.066811831 0.175165465 0.158850952 0.319235787 0.244941781 0.273352145 0.235738949 0.628369447 0.35750957 0.189287798 0.211780141 0.259463045 0.586793399 n.a. Trp 0.016362945 0.008336652 0.005519825 0.011092146 0.010926879 0.008006485 0.006222302 0.018722349 0.032961633 0.017741394 0.01611795 0.02355515 0.017448804 0.036758872 0.080898125 0.0737388 0.016267453 0.008733722 0.017585653 0.014461733 0.010910579 0.005533463 0.006668672 0.007576331 0.019837914 0.033263355 0.026763458 0.016590629 0.023489401 0.030129118 0.024731292 0.019938352 0.016300534 0.012697363 0.007650093 0.017993314 0.029346053 0.032809188 0.023887678 0.043348616 0.034478777 0.036427134 0.027042575 0.027944076 0.034299107 0.033263481 0.051599147 n.a. Tyr 0.023196686 0.014555287 0.022620336 0.049043673 0.011014199 0.009117919 0.005723836 0.036263778 0.053025285 0.035043765 0.049982507 0.082498361 0.013051499 0.027819484 0.079222784 0.078825741 0.009581648 0.010297865 0.021086917 0.013083321 0.006935121 0.005688708 0.010040137 0.01603972 0.024067462 0.05428259 0.064642027 0.047967644 0.038213554 0.059032464 0.066133654 0.063418287 0.020611206 0.014990392 0.019388826 0.062445693 0.029167971 0.052997657 0.04857716 0.087417921 0.038775836 0.089471139 0.080251871 0.078003043 0.033905166 0.045029605 0.080454976 n.a. Val 0.127783135 0.066943298 0.060819587 0.135505162 0.069208613 0.054130793 0.037040188 0.170519709 0.290541009 0.17228534 0.253293786 0.37981749 0.072618594 0.108341738 0.298491673 0.287912826 0.080790065 0.051953454 0.088791094 0.06461218 0.043100107 0.031129283 0.039306993 0.055770241 0.134528622 0.252210007 0.26094122 0.233610915 0.180855228 0.249849017 0.264036345 0.28992913 0.112936005 0.072803644 0.0969272 0.289252801 0.162731978 0.206311688 0.167930424 0.324244934 0.186164525 0.259129469 0.251985601 0.267816781 0.133533413 0.167869941 0.276966236 n.a. MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Glucose Lactate Gly Ala Arg Asn Glu Gln (Iso)Leu Lys Met Phe Pro Ser Thr Trp Tyr Val METABOLITES_END #END