#METABOLOMICS WORKBENCH hormel101_20160923_063958_mwtab.txt DATATRACK_ID:746 STUDY_ID:ST000483 ANALYSIS_ID:AN000749 PROJECT_ID:PR000367 VERSION 1 CREATED_ON October 3, 2016, 5:28 pm #PROJECT PR:PROJECT_TITLE Mechanism by Which Caloric Restriction Improves Insulin Sensitivity in Sedentary PR:PROJECT_TITLE Obese Adults PR:PROJECT_TYPE skeletal muscle ceramide, diacylglycerol, or amino acid metabolite levels PR:PROJECT_SUMMARY effect of caloric restriction on insulin sensitivity through skeletal muscle PR:PROJECT_SUMMARY ceramide, diacylglycerol, or amino acid metabolite levels PR:INSTITUTE Mayo Clinic PR:DEPARTMENT Endocrinology PR:LABORATORY Mayo Clinic Metabolomics Resource Core PR:LAST_NAME Nair PR:FIRST_NAME Sreekumaran PR:ADDRESS 200 First Street SW, Rochester, MN 55905 PR:EMAIL Nair.K@mayo.edu PR:PHONE 507-285-2415 #STUDY ST:STUDY_TITLE Amino Acid Quantifcation of obese patients on a 16 week caloric restriction from ST:STUDY_TITLE Plasma ST:STUDY_TYPE timecourse, quantitative measurements of amino acid ST:STUDY_SUMMARY Caloric restriction (CR) improves insulin sensitivity and reduces the incidence ST:STUDY_SUMMARY of diabetes in obese individuals. The underlying mechanisms whereby CR improves ST:STUDY_SUMMARY insulin sensitivity are not clear. We evaluated the effect of 16 weeks of CR on ST:STUDY_SUMMARY whole-body insulin sensitivity by pancreatic clamp before and after CR in 11 ST:STUDY_SUMMARY obese participants (BMI = 35 kg/m2) compared with 9 matched control subjects ST:STUDY_SUMMARY (BMI = 34 kg/m2). Compared with the control subjects, CR increased the glucose ST:STUDY_SUMMARY infusion rate needed to maintain euglycemia during hyperinsulinemia, indicating ST:STUDY_SUMMARY enhancement of peripheral insulin sensitivity. This improvement in insulin ST:STUDY_SUMMARY sensitivity was not accompanied by changes in skeletal muscle mitochondrial ST:STUDY_SUMMARY oxidative capacity or oxidant emissions, nor were there changes in skeletal ST:STUDY_SUMMARY muscle ceramide, diacylglycerol, or amino acid metabolite levels. However, CR ST:STUDY_SUMMARY lowered insulin-stimulated thioredoxin-interacting protein (TXNIP) levels and ST:STUDY_SUMMARY enhanced nonoxidative glucose disposal. These results support a role for TXNIP ST:STUDY_SUMMARY in mediating the improvement in peripheral insulin sensitivity after CR. ST:INSTITUTE Mayo Clinic ST:DEPARTMENT Endocrinology ST:LABORATORY Mayo Clinic Metabolomics Resource Core ST:LAST_NAME Nair ST:FIRST_NAME Sreekumaran ST:ADDRESS 200 First Street SW, Rochester, MN 55905 ST:EMAIL Nair.K@mayo.edu ST:PHONE 507-285-2415 #SUBJECT SU:SUBJECT_TYPE Animal SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS s1 ms5361-1 time:1 | Category:CR sample type=plasma; Sex=F SUBJECT_SAMPLE_FACTORS s1 ms5361-2 time:2 | Category:CR sample type=plasma; Sex=F SUBJECT_SAMPLE_FACTORS s2 ms5361-3 time:1 | Category:CR sample type=plasma; Sex=F SUBJECT_SAMPLE_FACTORS s2 ms5361-4 time:2 | Category:CR sample type=plasma; Sex=F SUBJECT_SAMPLE_FACTORS s3 ms5361-5 time:1 | Category:CR sample type=plasma; Sex=M SUBJECT_SAMPLE_FACTORS s3 ms5361-6 time:2 | Category:CR sample type=plasma; Sex=M SUBJECT_SAMPLE_FACTORS s4 ms5361-7 time:1 | Category:CR sample type=plasma; Sex=F SUBJECT_SAMPLE_FACTORS s4 ms5361-8 time:2 | Category:CR sample type=plasma; Sex=F SUBJECT_SAMPLE_FACTORS s5 ms5361-9 time:1 | Category:CR sample type=plasma; Sex=F SUBJECT_SAMPLE_FACTORS s5 ms5361-10 time:2 | Category:CR sample type=plasma; Sex=F SUBJECT_SAMPLE_FACTORS s6 ms5361-11 time:1 | Category:CR sample type=plasma; Sex=M SUBJECT_SAMPLE_FACTORS s6 ms5361-12 time:2 | Category:CR sample type=plasma; Sex=M SUBJECT_SAMPLE_FACTORS s7 ms5361-13 time:1 | Category:CR sample type=plasma; Sex=M SUBJECT_SAMPLE_FACTORS s7 ms5361-14 time:2 | Category:CR sample type=plasma; Sex=M SUBJECT_SAMPLE_FACTORS s8 ms5361-15 time:1 | Category:Con sample type=plasma; Sex=M SUBJECT_SAMPLE_FACTORS s8 ms5361-16 time:2 | Category:Con sample type=plasma; Sex=M SUBJECT_SAMPLE_FACTORS s9 ms5361-17 time:1 | Category:Con sample type=plasma; Sex=F SUBJECT_SAMPLE_FACTORS s9 ms5361-18 time:2 | Category:Con sample type=plasma; Sex=F SUBJECT_SAMPLE_FACTORS s10 ms5361-19 time:1 | Category:CR sample type=plasma; Sex=F SUBJECT_SAMPLE_FACTORS s10 ms5361-20 time:2 | Category:CR sample type=plasma; Sex=F SUBJECT_SAMPLE_FACTORS s11 ms5361-21 time:1 | Category:Con sample type=plasma; Sex=F SUBJECT_SAMPLE_FACTORS s11 ms5361-22 time:2 | Category:Con sample type=plasma; Sex=F SUBJECT_SAMPLE_FACTORS s12 ms5361-23 time:1 | Category:CR sample type=plasma; Sex=M SUBJECT_SAMPLE_FACTORS s12 ms5361-24 time:2 | Category:CR sample type=plasma; Sex=M SUBJECT_SAMPLE_FACTORS s13 ms5361-25 time:1 | Category:Con sample type=plasma; Sex=M SUBJECT_SAMPLE_FACTORS s13 ms5361-26 time:2 | Category:Con sample type=plasma; Sex=M SUBJECT_SAMPLE_FACTORS s14 ms5361-27 time:1 | Category:Con sample type=plasma; Sex=M SUBJECT_SAMPLE_FACTORS s14 ms5361-28 time:2 | Category:Con sample type=plasma; Sex=M SUBJECT_SAMPLE_FACTORS s15 ms5361-29 time:1 | Category:Con sample type=plasma; Sex=M SUBJECT_SAMPLE_FACTORS s15 ms5361-30 time:2 | Category:Con sample type=plasma; Sex=M SUBJECT_SAMPLE_FACTORS s16 ms5361-31 time:1 | Category:CR sample type=plasma; Sex=M SUBJECT_SAMPLE_FACTORS s16 ms5361-32 time:2 | Category:CR sample type=plasma; Sex=M SUBJECT_SAMPLE_FACTORS s17 ms5361-33 time:1 | Category:CR sample type=plasma; Sex=F SUBJECT_SAMPLE_FACTORS s17 ms5361-34 time:2 | Category:CR sample type=plasma; Sex=F SUBJECT_SAMPLE_FACTORS s18 ms5361-35 time:1 | Category:Con sample type=plasma; Sex=M SUBJECT_SAMPLE_FACTORS s18 ms5361-36 time:2 | Category:Con sample type=plasma; Sex=M SUBJECT_SAMPLE_FACTORS s19 ms5361-37 time:1 | Category:Con sample type=plasma; Sex=F SUBJECT_SAMPLE_FACTORS s19 ms5361-38 time:2 | Category:Con sample type=plasma; Sex=F SUBJECT_SAMPLE_FACTORS s20 ms5361-39 time:1 | Category:Con sample type=plasma; Sex=F SUBJECT_SAMPLE_FACTORS s20 ms5361-40 time:2 | Category:Con sample type=plasma; Sex=F #COLLECTION CO:COLLECTION_SUMMARY Blood samples were collected in a heated hotbox (131°F) through a retrograde CO:COLLECTION_SUMMARY intravenous catheter at baseline for glucose and hormone levels, and every 10 CO:COLLECTION_SUMMARY min during the clamp to maintain euglycemia. In addition, blood samples were CO:COLLECTION_SUMMARY collected every 20 min from 0600 to 0700, 0900 to 1000, and 1200 to 1300 to CO:COLLECTION_SUMMARY measure plasma [6,62H2]glucose. At 1330 h, a percutaneous needle muscle biopsy CO:COLLECTION_SUMMARY specimen (350–400 mg) was obtained from the vastus lateralis muscle under CO:COLLECTION_SUMMARY local anesthesia, immediately frozen in liquid nitrogen, and stored at −80°F CO:COLLECTION_SUMMARY for future analysis (27). This biopsy sample was used for analysis of TXNIP mRNA CO:COLLECTION_SUMMARY and protein content. The participant remained in the CRU through the remainder CO:COLLECTION_SUMMARY of the day and was given a weight-maintenance diet until 2200 h. At 0700 h the CO:COLLECTION_SUMMARY following morning, a second muscle biopsy specimen was obtained under local CO:COLLECTION_SUMMARY anesthesia, and ∼100 mg was used immediately for mitochondrial function CO:COLLECTION_SUMMARY measurements of isolated mitochondria and mtH2O2 emissions (28). The remainder CO:COLLECTION_SUMMARY was immediately frozen in liquid nitrogen and stored at −80°F for future CO:COLLECTION_SUMMARY analysis, including DAG, ceramide, and amino acid measurements (Fig. 1). CO:SAMPLE_TYPE Blood. Plasma was isolated for MS analysis. #TREATMENT TR:TREATMENT_SUMMARY Before and after 16 weeks of CR or CON, two outpatient visits and one inpatient TR:TREATMENT_SUMMARY visit were scheduled. Before the outpatient visits, participants were instructed TR:TREATMENT_SUMMARY to fast overnight from 10:00 p.m. the evening before and to avoid strenuous TR:TREATMENT_SUMMARY exercise for 24 h preceding the visits. One outpatient visit consisted of an MRI TR:TREATMENT_SUMMARY to measure subcutaneous and visceral fat distribution and magnetic resonance TR:TREATMENT_SUMMARY spectroscopy to measure skeletal muscle oxidative capacity (25). The second TR:TREATMENT_SUMMARY outpatient visit was for measurements of resting energy expenditure (REE) for TR:TREATMENT_SUMMARY the calculation of a weight-maintenance diet (Parvo Medics TrueOne 2400 Canopy TR:TREATMENT_SUMMARY system), DEXA scan (Lunar DPX-L; Lunar Radiation, Madison, WI), and VO2peak test TR:TREATMENT_SUMMARY on a bicycle ergometer (Fig. 1). Participants were admitted to the Clinical TR:TREATMENT_SUMMARY Research Unit (CRU) on the evening of the fifth day of the weight-maintaining TR:TREATMENT_SUMMARY diet provided by the CRU metabolic kitchen (Supplementary Fig. 1). The TR:TREATMENT_SUMMARY weight-maintenance meals (diet composition: 20% protein, 30% fat, 50% TR:TREATMENT_SUMMARY carbohydrate) were monitored daily to ensure that the correct calorie level was TR:TREATMENT_SUMMARY achieved. Upon admission to the CRU, no calories were consumed after 2100 h to TR:TREATMENT_SUMMARY achieve a 10-h fast before the two-stage insulin euglycemic pancreatic clamp the TR:TREATMENT_SUMMARY following morning, as previously published (26), with modifications as follows: TR:TREATMENT_SUMMARY the following morning at 0400 h, a primed [6,62H2]glucose bolus (6 mg ⋅ kg TR:TREATMENT_SUMMARY fat-free mass[FFM]−1) was administered, followed by a 9-h continuous infusion TR:TREATMENT_SUMMARY of [6,62H2]glucose (started at 4 mg ⋅ kgFFM−1 ⋅ h−1 then titrated TR:TREATMENT_SUMMARY downward over the infusion time period to match anticipated changes in TR:TREATMENT_SUMMARY endogenous glucose production [EGP]). At 0600 h, gas exchange was measured by TR:TREATMENT_SUMMARY indirect calorimetry for 30 min for REE determination. Then at 0700 h, glucagon TR:TREATMENT_SUMMARY (0.001 μg ⋅ kgFFM−1 ⋅ min−1), somatostatin (0.093 μg ⋅ kgFFM−1 ⋅ TR:TREATMENT_SUMMARY min−1), and growth hormone (0.0047 μg ⋅ kgFFM−1 ⋅ min−1) were infused TR:TREATMENT_SUMMARY for 6 h. Insulin was infused from 0700 to 1000 h at 0.62 mU ⋅ kgFFM−1 ⋅ TR:TREATMENT_SUMMARY min−1 and then from 1000 to 1300 h at 2.3 mU ⋅ kgFFM−1 ⋅ min−1. A 40% TR:TREATMENT_SUMMARY dextrose with 2% enrichment of [6,62H2]glucose was infused as needed to maintain TR:TREATMENT_SUMMARY blood glucose above 4.7 mmol/L from 0700 to 1000 h and then between 4.7 and 5.3 TR:TREATMENT_SUMMARY mmol/L from 1000 to 1300 h. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Plasma samples and amino acid calibration standards were prepared with MassTrak SP:SAMPLEPREP_SUMMARY Amino Acid Analysis Solution (AAA) kit from Waters according to instructions SP:SAMPLEPREP_SUMMARY with slight modifications for detection on a mass spectrometer. A 10 point SP:SAMPLEPREP_SUMMARY standard concentration curve was made from the calibration standard solution to SP:SAMPLEPREP_SUMMARY calculate amino acid concentrations in plasma samples. A solution containing SP:SAMPLEPREP_SUMMARY U-13C4-L-aspartic acid, U-13C3-L-alanine, U-13C4-L-threonine, U-13C5-L-proline, SP:SAMPLEPREP_SUMMARY U-13C5-L-valine, U-13C6-leucine, U-13C6-phenylalanine all from Cambridge Isotope SP:SAMPLEPREP_SUMMARY Laboratories, 13C6-tyrosine from Isotec, L-arginine (15N2, 2H2) from MassTrace, SP:SAMPLEPREP_SUMMARY norvaline from Sigma dissolved in 0.01N HCl was used as the internal standard SP:SAMPLEPREP_SUMMARY solution. Frozen plasma samples were thawed, spiked with internal standard then SP:SAMPLEPREP_SUMMARY deproteinized with cold MeOH followed by centrifugation at 10,000 g for 5 SP:SAMPLEPREP_SUMMARY minutes prior to derivatization according to MassTrak instructions. The amino SP:SAMPLEPREP_SUMMARY acid derivatizing reagent used was 6-aminoquinolyl-N-hydroxysuccinimidyl SP:SAMPLEPREP_SUMMARY carbamate. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY High resolution separation was done using an Acquity UPLC system, injecting 1 CH:CHROMATOGRAPHY_SUMMARY µl of derviatized solution, with a UPLC BEH C18 1.7 micron 2.1×150 mm column CH:CHROMATOGRAPHY_SUMMARY from Waters. Column flow was set to 400 µl/min with a gradient from 99.9%A to CH:CHROMATOGRAPHY_SUMMARY 98%B where buffer A is 1% acetonitrile in 0.1% formic acid and buffer B is 100% CH:CHROMATOGRAPHY_SUMMARY acetonitrile. A column temp of 43 degrees Celsius and a sample tray temp of 6% CH:CHROMATOGRAPHY_SUMMARY Celsius. Mass detection was completed on a TSQ Ultra Quantum from Thermo CH:CHROMATOGRAPHY_SUMMARY Finnigan running in ESI positive mode. A scan width of 0.002, scan time of 0.04 CH:CHROMATOGRAPHY_SUMMARY seconds per transition mass, collision energy of 25, collision gas pressure of CH:CHROMATOGRAPHY_SUMMARY 1.5 mTorr, tube lens value set to 90, monitoring a signature ion of the CH:CHROMATOGRAPHY_SUMMARY derivitized amines at m/z 171.04 by selected reaction monitoring. CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Waters Acquity CH:COLUMN_NAME C18 #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:MS_COMMENTS - MS:INSTRUMENT_NAME Thermo Quantum Ultra MS:INSTRUMENT_TYPE Triple quadrupole MS:MS_TYPE ESI MS:ION_MODE POSITIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS micromolar MS_METABOLITE_DATA_START Samples ms5361-1 ms5361-2 ms5361-3 ms5361-4 ms5361-5 ms5361-6 ms5361-7 ms5361-8 ms5361-9 ms5361-10 ms5361-11 ms5361-12 ms5361-13 ms5361-14 ms5361-15 ms5361-16 ms5361-17 ms5361-18 ms5361-19 ms5361-20 ms5361-21 ms5361-22 ms5361-23 ms5361-24 ms5361-25 ms5361-26 ms5361-27 ms5361-28 ms5361-29 ms5361-30 ms5361-31 ms5361-32 ms5361-33 ms5361-34 ms5361-35 ms5361-36 ms5361-37 ms5361-38 ms5361-39 ms5361-40 Factors time:1 | Category:CR time:2 | Category:CR time:1 | Category:CR time:2 | Category:CR time:1 | Category:CR time:2 | Category:CR time:1 | Category:CR time:2 | Category:CR time:1 | Category:CR time:2 | Category:CR time:1 | Category:CR time:2 | Category:CR time:1 | Category:CR time:2 | Category:CR time:1 | Category:Con time:2 | Category:Con time:1 | Category:Con time:2 | Category:Con time:1 | Category:CR time:2 | Category:CR time:1 | Category:Con time:2 | Category:Con time:1 | Category:CR time:2 | Category:CR time:1 | Category:Con time:2 | Category:Con time:1 | Category:Con time:2 | Category:Con time:1 | Category:Con time:2 | Category:Con time:1 | Category:CR time:2 | Category:CR time:1 | Category:CR time:2 | Category:CR time:1 | Category:Con time:2 | Category:Con time:1 | Category:Con time:2 | Category:Con time:1 | Category:Con time:2 | Category:Con Histidine 79.73221787 79.98740894 82.87217308 74.67935017 76.45689505 75.25402917 66.14653339 73.88879442 83.31408594 74.11787242 76.78948308 57.42705439 55.33787291 63.15736423 64.13947428 71.10874676 49.67504046 55.09664355 68.41682592 61.58077139 76.4188013 78.09771914 66.686533 60.18589589 66.38475041 49.28291046 67.79968078 50.45981839 63.01652835 53.62181589 71.77178837 55.48121674 67.40996644 64.24259079 55.71008731 62.71578451 56.60224659 60.1986178 62.0603712 51.50277394 Hydroxyproline 7.081764986 8.908969173 6.826616893 7.553177376 8.554088361 9.486287348 5.273203127 6.457924233 9.043135589 11.69326851 9.451227717 7.683165367 8.559965301 8.33587927 11.01336054 9.413349474 10.78461633 9.224586705 6.782332396 7.517495935 8.860754617 8.25022671 11.11680114 13.13010425 9.356472125 7.510148727 13.7600327 10.34923152 8.274952169 9.132692224 9.319659517 7.715252631 8.144461286 12.40230554 8.873473658 11.07594496 6.722739414 6.668187471 7.248919145 7.381637584 1-Methylhistidine 20.52775068 19.96052677 14.17241118 0 0 0 17.20459793 25.0883942 26.67520255 5.855412765 5.079467049 12.96228818 6.035044228 22.01613448 8.521564915 17.92789398 17.97331407 19.33898765 7.650906525 5.356256113 40.73060818 26.5074773 11.14427106 10.60824609 20.15996349 8.000651477 14.04340689 14.76660911 15.21570174 12.63946572 12.24999467 18.368861 18.80761064 6.610085291 15.19743589 6.817551077 15.24481852 15.96149908 12.64547196 14.10183029 3-Methylhistidine 4.870458091 2.884068267 4.120473898 3.090970282 2.186147452 2.430040679 2.153081334 5.28069182 7.935247706 14.47516883 4.2933041 3.864388269 4.109588928 3.909906248 3.28707989 5.902174118 6.180728831 5.877655749 2.358916238 2.139208104 3.334675728 3.469173505 2.615558264 2.917335887 3.138994737 2.705395954 4.356211278 3.819252583 4.435265751 3.422989749 7.0830281 4.457352264 5.503106941 3.203208904 1.978898182 3.859826308 3.607204419 3.766323059 2.915073915 3.075453144 Phosphoethanolamine 1.070211627 1.139464473 0.568438222 0.757735316 1.064049453 1.404440987 0.841093306 0.512088242 1.001527881 1.607176039 1.518247615 1.29025045 1.121525226 0.972077027 1.39286799 0.93992408 1.150058237 1.262165549 0.747495775 1.110856899 1.18993572 0.957477973 0.763908956 0.866652163 0.909439385 0.680895126 0.679584134 0.679331028 0.939554067 0.983952644 1.32596114 0.881111984 0.767397431 0.888296076 0.868746658 0.926925779 0.958925902 1.245590603 1.074772591 0.600123736 Carnosine 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Anserine 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0.178235413 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0.108475313 0 0 0 0 0 Ethanolamine 4.938094821 2.817826762 3.93813896 4.017968316 4.843148139 4.517940679 4.541671017 3.549844197 4.074631308 3.848754311 5.615620883 5.384265052 3.993750157 4.988556725 5.815821941 5.311006346 4.847481794 4.17932158 5.436070072 4.699247163 4.405725281 5.121047053 6.303023679 4.735601891 6.04267745 4.95380833 7.221565409 6.798330145 4.089690085 4.399244234 4.386909001 3.240015707 5.127935237 3.901441413 5.708268601 5.250897607 4.016600089 4.217785691 4.611161096 3.845758582 Aspartic Acid 2.288611039 3.001865724 1.116896508 1.417144135 2.225574123 1.660808072 1.19890684 2.054552488 1.510406052 2.224515675 1.25670334 1.562994669 2.139547585 2.200032731 3.251394659 1.518529513 1.949641731 2.157185899 2.484551697 1.193618044 2.265365429 2.296964455 2.201779116 1.369224659 1.89369755 1.930685107 4.53832506 2.211987038 2.479950777 2.399138777 2.136478957 1.956495825 1.384956614 2.384419015 2.184527397 2.075512084 1.278559009 2.268690863 2.254570161 1.847283274 Sarcosine 0.922031807 0.733445942 1.031249477 0.846148892 1.294369556 1.049857207 0.880696675 0.587135655 0.909406948 1.041558615 1.622289357 1.561204609 1.892155648 1.637275714 1.172197887 1.168244777 1.128358882 1.106593676 0.869032544 0.688796262 1.357381384 1.438852585 2.193155769 2.234609555 1.147907687 1.035888439 2.247319305 2.005520975 1.115426715 1.078292371 1.23220461 0.774730648 1.285090526 1.27924367 1.43666826 1.636454422 1.063113006 1.176576518 1.287431497 1.222850059 beta-Alanine 2.776569772 3.073019491 2.893312709 2.483226151 1.919534311 1.265808435 2.603198577 3.253568893 1.774418207 1.903898552 3.456765651 3.379889884 2.408368193 3.931138312 3.396387574 2.894259753 1.984395756 2.33525946 2.138800334 1.516904427 3.185912749 3.900356459 3.555063447 3.861296845 2.676560849 1.285519918 3.245485726 3.274818498 2.06379355 2.419410221 3.938837499 2.676393691 1.865769948 1.441400099 4.256012718 3.420687812 2.229939427 2.129284301 1.940482524 2.250678547 gamma-Amino-N-butyric-acid 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 alpha-Aminoadipic-acid 1.47100932 1.308486465 0.635408677 0.556389318 1.778367124 0.951704644 1.731731061 1.402668847 1.040467633 2.020905393 1.334044508 1.550587326 1.528408533 1.66612679 1.727671005 1.601253761 0.741772762 0.615741998 0.591757363 0.727058678 1.364891968 1.178652776 1.979330548 1.456489291 1.332806071 1.431496094 1.77801642 1.998660273 2.315499788 2.24399757 1.727492289 1.111473152 0.875314676 0.972007538 1.710827214 1.316539378 0.881439232 1.150059726 1.007629477 0.915139822 beta-Aminoisobutyric-acid 0.852028043 1.682594253 1.682155272 1.638683889 0.752305745 1.423055741 0.893771641 1.960397121 1.086231826 2.358538497 0.848852146 1.665344219 1.21142386 1.266372644 1.529427874 0.718318178 1.503144371 1.367595488 0.810869501 1.221246349 1.675243244 1.3893428 0.983243424 0.884977061 0.702676847 0.664564887 1.029022949 1.253563666 2.121463752 2.441997092 1.065877186 1.986214526 1.019013876 1.372475071 0.574027558 0.609250284 2.116168556 2.296260111 1.561772093 1.574103892 Hydroxylysine 1 0.011263353 0.017766371 0.035866936 0.046790397 0.036917747 0.002206914 0.019018027 0.02139762 0.040041753 0.064054995 0.018040783 0 0.013967788 0.014683775 0.009180288 0.025620133 0.015362185 0.00096482 0.066443706 0.05532699 0.00478919 0.00432154 0.013674681 0.009604978 0.006211529 0 0.0049705 0.02199275 0.01738541 0.011365766 0 0 0.020745534 0.004371305 0.040622235 0.041216747 0.004846478 0.013979982 0.001553705 0 Hydroxylysine 2 0.671476749 0.546522237 0.595688864 0.672730629 0.419460727 0.270534756 0.462084427 0.531116236 0.372829866 0.498523062 0.230555635 0.285796753 0.514350934 0.365050817 0.425606108 0.386427931 0.464462452 0.343038617 0.586269873 0.63532953 0.486424817 0.605748859 0.426160332 0.558451964 0.280742222 0.251956627 0.435586952 0.44881462 0.487596365 0.464488267 0.279735112 0.285913567 0.759664087 0.705210386 0.355314128 0.524757372 0.305593308 0.306938921 0.816513983 0.519317718 alpha-Amino-N-butyric-acid 20.36239677 48.50635937 19.51573274 19.00467004 26.99773069 35.9568443 23.60795497 49.23677868 15.62016723 25.91385441 17.38974348 17.10665671 22.77396509 20.79582714 25.0466936 29.55717225 25.09656606 23.57470473 28.38592025 26.3043912 34.65143677 29.45963659 29.51614918 25.76049847 28.58029187 28.05729992 32.00284807 36.15275921 21.76346452 23.42864605 21.53144493 25.71450356 26.70153797 32.14427505 17.5769041 16.48080309 20.70484337 27.32539005 32.93363227 35.49891506 Cystathionine 1 0.296021283 0.186267139 0.392782012 0.35363199 0.373752363 0.116687816 0.696375923 0.364795612 0.244324047 0.721574603 0.392572218 0.508538695 0.292142715 0.3235238 0.389384109 0.21817271 0.240671097 0.203951202 0.143907214 0.066764982 0.259263525 0.44889145 0.527333791 0.504964605 0.226589378 0.240686486 0.777375662 0.577346417 0.461403594 0.527144617 0.544502786 0.290142846 0.372304901 0.422008522 0.770693321 0.776111152 0.502751571 0.345049972 0.092837025 0.105706224 Cystathionine 2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Methionine 17.28642971 16.21979217 16.82687227 18.50349723 23.6624222 22.87535792 16.49323081 13.20568377 17.98159035 24.27743589 20.90319741 17.22667047 19.30333605 20.90368962 21.70100542 20.71005594 20.91649018 19.82089093 19.91144919 16.84536078 17.15979857 22.58222735 26.46713766 23.23407753 19.78245798 18.54101504 25.2160547 26.26801042 22.95577538 20.25934114 19.66170531 13.71983749 17.43642378 16.33350574 20.45169109 19.69046161 19.35886258 16.8716057 19.66381256 17.41707864 allo-Isoleucine 2.528767037 2.500050802 1.272987156 1.215325476 1.917047754 1.805312001 1.985951282 2.451526555 1.407913751 0 3.154319146 2.756022088 2.074676929 2.170410331 2.847787954 2.280883139 1.847660524 1.808970428 1.529067215 1.547592512 1.953706766 1.809236341 2.511103669 1.897060961 1.939599058 2.197236244 1.9319022 2.572704734 1.485560299 1.61918829 0 0.898156937 0 0 1.890754856 0 0 0 1.697838715 1.65972884 Homocystine 0.009674813 0.022095332 0.019507577 0 0 0 0.059938429 0.024016963 0.01411381 0.024203893 0.015385175 0.01637328 0 0.017036009 0.027889303 0.052992714 0.031464211 0.005333614 0 0.018847844 0.030310627 0 0 0.015035431 0.039323172 0.050483413 0.042892849 0 0.009915403 0.014546233 0.048941283 0.050978732 0.055639706 0.016515652 0.050840546 0.06022693 0.008686506 0.026967769 0.006773838 0 Asparagine 40.77356274 48.20682419 44.21704773 47.23084585 45.45720366 52.69925076 38.13961938 34.55929534 47.18658809 61.91510255 51.34207128 46.9510832 42.3407204 44.18239517 45.75556526 44.61661957 48.68098371 50.34064811 58.22643976 60.58151712 50.39302753 56.68857465 49.06034962 45.24789759 44.6973013 42.60759312 49.13604302 50.20176369 51.14860918 51.3739173 49.05581475 45.4860292 53.44614443 55.89539426 48.74063383 51.94088409 47.6459464 46.16820586 52.03982682 47.71464697 Arginine 63.41990277 79.87924234 68.81012947 80.82777039 66.13360875 63.96504654 63.37805362 49.2971512 58.74894554 53.92439405 70.2689545 63.88724162 61.54679567 55.90576473 73.02393647 66.01026601 67.98851671 81.22867838 70.62668866 63.04398277 53.48126839 68.71808095 72.48870982 71.19918526 56.45759517 62.40614017 77.68664256 79.85562447 68.61641464 77.06800616 69.18740878 57.3681949 67.695599 56.97531436 73.17943827 75.11906472 58.69488841 73.1291768 62.87654688 62.90396879 Taurine 31.22879437 31.15604315 28.30362599 23.83203579 44.78908661 35.12260245 26.73544728 22.27664203 36.10722803 51.19989115 33.71686873 36.45204467 25.39024214 30.4587164 33.08547614 25.39025506 30.33058406 34.87981788 34.58206335 38.59277664 30.65905276 34.37023865 21.97551765 19.68501674 23.75974026 27.5974468 24.13580382 21.50889545 26.13608405 25.44749085 39.6018455 22.52820714 34.58535167 36.08185908 26.19129365 24.31815474 27.6085523 29.58193896 36.74409979 30.44366725 Serine 71.06779956 112.6753639 88.97452677 108.9786388 89.11278222 101.2684362 70.54378459 103.3402306 94.26764783 119.6166327 69.49938833 70.91664432 84.08303494 90.77140492 76.91219532 78.16469275 80.73517901 74.28478022 117.1794919 112.5748547 100.1762202 96.69747263 84.9659436 97.3469608 78.91311661 82.74966323 75.62294301 77.42373076 79.43265561 78.92755618 80.35646154 101.6049101 68.59126298 93.35672853 90.61579396 99.52726462 85.9061911 104.3867675 128.0067248 118.6372459 Glutamine 444.8841791 533.8893503 582.6181055 541.399931 575.9019593 610.9655342 476.5793872 503.3534121 532.5575039 603.0159993 492.5604607 512.0821722 478.8542359 514.8499743 542.7627828 474.3934938 444.7600891 449.3801254 539.8274473 523.2689677 486.662249 528.4645748 576.4987113 518.7690495 507.8899917 469.0169776 555.0568809 534.522019 484.0994724 505.735383 531.4836835 495.6768366 471.0060719 438.0360392 456.8780091 461.8221041 499.5299102 545.8686005 586.6539091 517.7719592 Glycine 183.012098 247.6580325 205.8264342 260.1804689 151.7613786 178.6282115 140.5731229 151.6546464 391.8197611 386.4208842 153.5467012 140.0432988 244.4454564 248.5365388 159.1675874 180.7784369 177.4672604 164.3188785 371.8969667 366.4470206 183.3691261 207.9324803 148.0993791 172.4498804 183.1630793 175.069837 188.7211281 175.0723624 160.2795864 175.153452 140.1858711 125.2436038 196.693576 225.0308127 172.9178544 191.3031342 165.0709462 177.6517148 317.8553563 327.7269434 Citrulline 32.14438038 36.55643355 33.29337631 26.28664026 38.18159439 28.60619336 18.98328053 19.44813035 36.58967537 32.36191886 31.02359137 27.04755887 28.70203953 27.06668216 33.14127622 24.29051282 32.24308789 27.7589378 26.72236878 25.13540191 23.32296416 26.16309168 33.10943179 29.4386618 25.13252693 23.49678088 35.89479201 34.03312439 28.91062804 29.90381443 37.72696767 27.00044999 28.01155061 29.58115586 32.85523398 39.14574478 39.97617936 41.84566509 23.30296828 24.40981821 Glutamic Acid 94.02269152 100.4575436 62.94616865 47.02051645 88.6641707 57.59876381 83.47021267 73.85638636 61.40618934 52.85450085 73.28601764 72.70505155 82.99077903 66.5057314 100.3440053 56.59115913 49.25638742 63.07488262 48.1084441 35.44427322 80.97744919 77.06461794 106.802563 84.16467605 59.48709723 83.16209886 91.04458528 94.79937643 117.5407118 132.1956877 68.42776818 55.79058444 74.91293595 77.32637766 95.13323224 93.54914458 46.4654887 55.31732107 62.78577456 58.34559701 Threonine 93.73254223 95.71168169 113.164849 158.6780118 131.5362853 124.2593796 80.9719838 96.59409835 97.31743785 131.0059495 99.15884461 88.96549383 103.3919857 99.07754126 132.7632529 118.3142365 119.9840456 139.0123848 112.1771536 109.0215953 112.8346295 141.1381006 139.9426824 164.1909799 112.2337197 106.0855491 119.1018229 121.6556142 98.37905626 109.4286197 136.1646979 87.80532155 82.72262695 92.63249291 139.0612988 128.4743349 140.9819632 146.6964326 119.2841078 119.8311005 Alanine 329.4485479 237.9595739 313.4960948 227.444168 269.3279413 217.5957302 391.4996985 174.2320496 363.7173192 205.987371 374.946258 238.1923892 376.1586236 357.8438337 385.7415486 393.3093614 224.5852241 272.9908167 258.6646271 200.3041215 350.4031252 473.4944811 404.6925494 356.0975652 382.1592278 300.9712448 453.199898 402.6940513 382.6845715 386.9640181 284.2223928 171.7885897 413.3714282 255.9675272 392.907269 408.509406 262.1199866 232.6147573 205.9481044 206.4313223 Proline 149.318225 155.2918538 140.9760726 121.9210039 157.886992 133.671115 170.7729477 141.6939719 142.0502438 188.6848331 182.6698197 162.449566 168.2048691 151.5446299 199.6281306 175.9754761 124.3410332 147.4841924 113.9449739 100.9078594 196.5469309 224.5645387 169.260937 174.7142541 168.1791978 165.5560783 227.9067484 203.5861599 164.0648556 168.3944355 206.1531158 163.0280554 188.9836311 272.025339 159.2346606 198.971405 200.659708 196.7185506 115.8477321 103.4168445 Ornithine 46.2953977 51.77699136 49.84946561 51.57832988 61.31737831 43.84031094 45.63572419 38.03526071 46.19946097 77.88572186 31.71157469 32.66546967 45.47365994 43.88302472 39.76207582 39.75743254 31.69951792 34.22742212 33.17804826 33.4500264 41.65085921 42.72997911 43.39797381 40.5971213 39.59950932 32.77929525 55.69342367 46.53027126 44.96716084 40.56606818 53.03611051 45.82139297 36.25032703 42.42848233 67.78631835 78.74626199 51.61715803 56.58776363 39.75376239 40.91150879 Lysine 195.2126673 210.3061416 212.2773828 204.7006388 197.0528171 225.0536251 260.8347772 212.7536626 183.3921163 244.7399214 195.0630574 164.0332361 198.8930144 213.9298694 174.0745077 175.9694373 212.1573587 217.5754996 154.4784543 154.8241725 207.4865639 246.2161589 215.2789752 202.9651351 186.387579 175.923257 235.588494 232.2112593 238.8500371 204.630289 194.1964698 160.5216735 213.0075469 209.9130835 226.7024694 198.0804695 218.3873996 182.7918089 217.5042939 207.0187443 Cystine 76.49236842 78.24155348 81.00886036 64.78105934 91.03907679 76.97754346 67.50617614 61.96332527 78.60981921 84.97190854 72.66762628 67.93292822 78.87676129 75.51971511 69.69157575 75.84027992 83.22628693 80.88147548 51.59051228 75.76146456 68.79710869 58.33775892 75.90890754 96.53769376 70.59962637 68.38398846 90.4389688 86.5609955 91.84921301 96.94576169 78.60887723 77.04324342 93.65058026 98.88653764 75.11695967 85.71422593 77.28079919 83.93297933 73.9399356 73.74381094 Tyrosine 65.70390306 65.14477975 66.32056361 52.75914712 86.85470333 74.56556812 57.23563915 45.46982855 47.6930251 64.92804045 81.15890579 69.89381704 72.82944605 69.53843511 74.81952541 60.24955339 44.54918904 41.38265291 52.47993543 44.31804657 47.21611551 59.7358583 88.08631535 87.64753316 58.57401588 54.41124836 90.34080594 77.39223591 58.0557112 57.18819433 80.74771011 58.22471224 60.8037498 58.55746354 77.27203874 79.87919166 57.14574755 50.26187612 63.27750645 59.32990307 Valine 298.6733584 342.7737709 238.8336349 240.8627454 276.983004 284.7484948 267.9790305 330.1202257 274.7960671 471.6329966 328.5968129 394.7521331 270.0674364 291.0967788 280.7706266 292.3080769 243.3029571 266.4508244 213.4057283 215.8230883 293.3550494 284.5227827 327.5904829 308.9507389 291.0930236 265.0029947 285.1294009 288.9317379 268.2672427 259.4223645 259.9205318 266.0312296 262.6049699 310.8056791 294.0770155 309.7226193 297.2942988 298.8395292 254.0359972 234.2530288 Isoleucine 84.07692861 90.65799784 61.96445929 54.27235569 89.24198627 83.23998023 82.92302067 99.8971061 66.75813103 153.8362543 97.70299238 112.1072599 79.37501083 78.67870188 92.35346431 78.77468175 62.78889174 59.95355101 57.70139094 51.62664429 71.09166816 75.4328041 101.3908104 80.13683665 77.90486932 74.90904227 95.11762043 95.10920653 78.36816573 85.10970684 75.26058978 72.74049475 65.74158249 72.15134715 83.30991451 98.56974354 87.46586493 87.96342701 68.65584626 60.14815401 Leucine 145.3183562 161.8405098 114.596246 111.3061889 146.1739916 146.7483674 137.0548273 176.0981744 122.5056271 255.4098461 167.1864522 196.8429672 138.5548527 146.9467172 154.536031 144.5874706 110.4060512 114.4876525 109.1031147 104.2369026 129.1064934 132.2354549 172.50287 145.5166893 142.9497581 133.9441591 155.6672995 150.7687806 146.6260073 146.9297549 132.7076206 134.8437048 123.9667677 137.7337065 153.1080298 161.6979078 157.1237291 153.756322 127.2569154 114.2808161 Phenylalanine 57.89425726 55.39961735 49.76431001 49.48534395 64.72673182 63.73456715 66.10817731 80.37948048 52.32495941 63.97721815 70.63919556 71.23327292 56.53366568 63.63132006 65.82672305 59.09571354 48.53378689 48.32372059 54.25828247 50.28339831 53.82639321 57.7802439 72.40733696 67.92231624 58.1400312 53.99850521 70.37530681 66.05979238 71.94257756 68.84223816 65.10652555 56.26106229 63.38828193 62.26137347 74.16647624 77.27748639 59.44236237 59.07038818 59.1173141 54.64987139 Tryptophan 49.91466444 45.82037119 38.28485115 34.06676499 57.19452373 59.46238915 56.06156874 45.06492054 47.75988234 54.31905305 59.61105708 55.43741272 56.55504662 61.19588032 58.41554907 58.69045366 46.3738283 39.92992789 46.68920789 42.26599633 50.41368082 55.75636387 61.29362989 55.3015449 58.62706757 49.97935206 76.62713846 69.10476773 65.19780398 64.65717672 62.52403838 50.13328615 49.50089395 49.46495099 58.96129814 53.19259846 52.93028787 45.98528501 47.49846801 45.70714438 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name retention index quantified m/z PubChem ID KEGG ID Histidine Hydroxyproline 1-Methylhistidine 3-Methylhistidine Phosphoethanolamine Carnosine Anserine Ethanolamine Aspartic Acid Sarcosine beta-Alanine gamma-Amino-N-butyric-acid alpha-Aminoadipic-acid beta-Aminoisobutyric-acid Hydroxylysine 1 Hydroxylysine 2 alpha-Amino-N-butyric-acid Cystathionine 1 Cystathionine 2 Methionine allo-Isoleucine Homocystine Asparagine Arginine Taurine Serine Glutamine Glycine Citrulline Glutamic Acid Threonine Alanine Proline Ornithine Lysine Cystine Tyrosine Valine Isoleucine Leucine Phenylalanine Tryptophan METABOLITES_END #END