#METABOLOMICS WORKBENCH Lu_Group_20210907_075703_mwtab.txt DATATRACK_ID:2828 STUDY_ID:ST001939 ANALYSIS_ID:AN003154 PROJECT_ID:000000 VERSION 1 CREATED_ON October 12, 2021, 6:32 pm #PROJECT PR:PROJECT_TITLE Metabolomics characterized concentration-dependent metabolic influence of PR:PROJECT_TITLE magnesium on biofilm formation in Escherichia coli PR:PROJECT_TYPE Targeted MS quantitative analysis PR:PROJECT_SUMMARY Metabolomics characterized concentration-dependent metabolic influence of PR:PROJECT_SUMMARY magnesium on biofilm formation in Escherichia coli PR:INSTITUTE Shanghai Center for Systems Biomedicine, Shanghai Jiaotong University PR:DEPARTMENT Shanghai Center for Systems Biomedicine PR:LABORATORY Lu Group PR:LAST_NAME Lu PR:FIRST_NAME Haitao PR:ADDRESS 800 Dongchuan RD. Minhang District, Shanghai, Shanghai, 200240, China PR:EMAIL haitao_lu@sjtu.edu.cn PR:PHONE 15221478139 #STUDY ST:STUDY_TITLE Metabolomics characterized concentration-dependent metabolic influence of ST:STUDY_TITLE magnesium on biofilm formation in Escherichia coli (Part 2) ST:STUDY_SUMMARY Biofilms are broadly formed by a diversity of microorganisms that enable them to ST:STUDY_SUMMARY adapt stressful environments. Biofilms often impose harmful influences in many ST:STUDY_SUMMARY niches, as they can cause food contamination, antibiotics resistance, and ST:STUDY_SUMMARY environmental issues. However, eradicating biofilms remains difficult since the ST:STUDY_SUMMARY formation mechanism of biofilms are still incompletely clarified. In this study, ST:STUDY_SUMMARY we aimed at exploring the regulatory role of magnesium (Mg2+) on biofilm ST:STUDY_SUMMARY formation in Escherichia coli (E. coli) using phenotype visualization combined ST:STUDY_SUMMARY with targeted metabolomics method. We found that Mg2+ could exert significant ST:STUDY_SUMMARY influence on biofilm formation in a concentration-dependent manner by regulating ST:STUDY_SUMMARY phenotypic morphology and triggering metabolic modifications of biofilm. ST:STUDY_SUMMARY Phenotypic imaging revealed that increasing concentration of Mg2+ gradually ST:STUDY_SUMMARY inhibited biofilm formation, Mg2+ was observed to restore the microstructure of ST:STUDY_SUMMARY E. coli strain in biofilms to that in the relevant planktonic cells. In ST:STUDY_SUMMARY addition, our metabolomics analysis characterized 20 differential metabolites ST:STUDY_SUMMARY and associated 2 metabolic pathways including nucleotide metabolism and amino ST:STUDY_SUMMARY acid metabolism that were notably modified during biofilm formation under the ST:STUDY_SUMMARY treatments of different concentrations of Mg2+. Altogether, our work provides a ST:STUDY_SUMMARY novel insight into the influence of Mg2+ on biofilm formation at a metabolic ST:STUDY_SUMMARY level, which are implicated in the novel solution to disturb biofilm formation ST:STUDY_SUMMARY through the regulation of Mg2+ and functional metabolite interaction, then ST:STUDY_SUMMARY biofilms associated harmful impacts in different niches could be well tangled ST:STUDY_SUMMARY accordingly. ST:INSTITUTE Shanghai Center for Systems Biomedicine, Shanghai Jiaotong University ST:DEPARTMENT Shanghai Center for Systems Biomedicine ST:LABORATORY Lu Group ST:LAST_NAME Lu ST:FIRST_NAME Haitao ST:ADDRESS 800 Dongchuan RD. Minhang District, Shanghai, Shanghai, 200240, China ST:EMAIL haitao_lu@sjtu.edu.cn ST:PHONE 15221478139 #SUBJECT SU:SUBJECT_TYPE Bacteria SU:SUBJECT_SPECIES Escherichia coli SU:TAXONOMY_ID 562 #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 - 20210227-3-C-1 Concentration of MgSO4(w/v):0.01% RAW_FILE_NAME=20210227-3-C-1 SUBJECT_SAMPLE_FACTORS - 20210227-3-C-2 Concentration of MgSO4(w/v):0.01% RAW_FILE_NAME=20210227-3-C-2 SUBJECT_SAMPLE_FACTORS - 20210227-3-C-3 Concentration of MgSO4(w/v):0.01% RAW_FILE_NAME=20210227-3-C-3 SUBJECT_SAMPLE_FACTORS - 20210227-3-C-4 Concentration of MgSO4(w/v):0.01% RAW_FILE_NAME=20210227-3-C-4 SUBJECT_SAMPLE_FACTORS - 20210227-3-C-5 Concentration of MgSO4(w/v):0.01% RAW_FILE_NAME=20210227-3-C-5 SUBJECT_SAMPLE_FACTORS - 20210227-3-C-6 Concentration of MgSO4(w/v):0.01% RAW_FILE_NAME=20210227-3-C-6 SUBJECT_SAMPLE_FACTORS - 20210227-3-30-1 Concentration of MgSO4(w/v):0.15% RAW_FILE_NAME=20210227-3-30-1 SUBJECT_SAMPLE_FACTORS - 20210227-3-30-2 Concentration of MgSO4(w/v):0.15% RAW_FILE_NAME=20210227-3-30-2 SUBJECT_SAMPLE_FACTORS - 20210227-3-30-3 Concentration of MgSO4(w/v):0.15% RAW_FILE_NAME=20210227-3-30-3 SUBJECT_SAMPLE_FACTORS - 20210227-3-30-4 Concentration of MgSO4(w/v):0.15% RAW_FILE_NAME=20210227-3-30-4 SUBJECT_SAMPLE_FACTORS - 20210227-3-30-5 Concentration of MgSO4(w/v):0.15% RAW_FILE_NAME=20210227-3-30-5 SUBJECT_SAMPLE_FACTORS - 20210227-3-30-6 Concentration of MgSO4(w/v):0.15% RAW_FILE_NAME=20210227-3-30-6 SUBJECT_SAMPLE_FACTORS - 20210227-3-50-1 Concentration of MgSO4(w/v):0.25% RAW_FILE_NAME=20210227-3-50-1 SUBJECT_SAMPLE_FACTORS - 20210227-3-50-2 Concentration of MgSO4(w/v):0.25% RAW_FILE_NAME=20210227-3-50-2 SUBJECT_SAMPLE_FACTORS - 20210227-3-50-3 Concentration of MgSO4(w/v):0.25% RAW_FILE_NAME=20210227-3-50-3 SUBJECT_SAMPLE_FACTORS - 20210227-3-50-4 Concentration of MgSO4(w/v):0.25% RAW_FILE_NAME=20210227-3-50-4 SUBJECT_SAMPLE_FACTORS - 20210227-3-50-5 Concentration of MgSO4(w/v):0.25% RAW_FILE_NAME=20210227-3-50-5 SUBJECT_SAMPLE_FACTORS - 20210227-3-50-6 Concentration of MgSO4(w/v):0.25% RAW_FILE_NAME=20210227-3-50-6 SUBJECT_SAMPLE_FACTORS - 20210227-3-80-1 Concentration of MgSO4(w/v):0.40% RAW_FILE_NAME=20210227-3-80-1 SUBJECT_SAMPLE_FACTORS - 20210227-3-80-2 Concentration of MgSO4(w/v):0.40% RAW_FILE_NAME=20210227-3-80-2 SUBJECT_SAMPLE_FACTORS - 20210227-3-80-3 Concentration of MgSO4(w/v):0.40% RAW_FILE_NAME=20210227-3-80-3 SUBJECT_SAMPLE_FACTORS - 20210227-3-80-4 Concentration of MgSO4(w/v):0.40% RAW_FILE_NAME=20210227-3-80-4 SUBJECT_SAMPLE_FACTORS - 20210227-3-80-5 Concentration of MgSO4(w/v):0.40% RAW_FILE_NAME=20210227-3-80-5 SUBJECT_SAMPLE_FACTORS - 20210227-3-80-6 Concentration of MgSO4(w/v):0.40% RAW_FILE_NAME=20210227-3-80-6 SUBJECT_SAMPLE_FACTORS - 20210227-3-100-1 Concentration of MgSO4(w/v):0.50% RAW_FILE_NAME=20210227-3-100-1 SUBJECT_SAMPLE_FACTORS - 20210227-3-100-2 Concentration of MgSO4(w/v):0.50% RAW_FILE_NAME=20210227-3-100-2 SUBJECT_SAMPLE_FACTORS - 20210227-3-100-3 Concentration of MgSO4(w/v):0.50% RAW_FILE_NAME=20210227-3-100-3 SUBJECT_SAMPLE_FACTORS - 20210227-3-100-4 Concentration of MgSO4(w/v):0.50% RAW_FILE_NAME=20210227-3-100-4 SUBJECT_SAMPLE_FACTORS - 20210227-3-100-5 Concentration of MgSO4(w/v):0.50% RAW_FILE_NAME=20210227-3-100-5 SUBJECT_SAMPLE_FACTORS - 20210227-3-100-6 Concentration of MgSO4(w/v):0.50% RAW_FILE_NAME=20210227-3-100-6 #COLLECTION CO:COLLECTION_SUMMARY After 72 hours of culture, the planktonic cells were isolated, and the biofilms CO:COLLECTION_SUMMARY were washed with PBS solvent for three times. Then, planktonic cells and biofilm CO:COLLECTION_SUMMARY solutions were centrifuged at 2000 rpm and 4°C for 15 minutes, to collect the CO:COLLECTION_SUMMARY strain pellets. CO:SAMPLE_TYPE Biofilm #TREATMENT TR:TREATMENT_SUMMARY Colony-forming antigen (CFA) medium (1% casamino acids, 0.15% yeast extract, TR:TREATMENT_SUMMARY 0.005% magnesium sulfate, and 0.0005% manganese chloride) was used to form TR:TREATMENT_SUMMARY biofilms. The E. coli strain was incubated in LB-agar plate for 12 hours, one TR:TREATMENT_SUMMARY colony was isolated to LB broth for further 4 hours incubation, then diluted the TR:TREATMENT_SUMMARY solution into CFA medium at a ratio of 1:1000 and the cultures were incubated TR:TREATMENT_SUMMARY for another 72 h at 30°C to form mature biofilms. To test the influence of TR:TREATMENT_SUMMARY MgSO4 on biofilm formation, the purified compound of MgSO4 was added to the CFA TR:TREATMENT_SUMMARY medium at different concentrations for the preparation of treated culture TR:TREATMENT_SUMMARY medium; The left cultural procedures were kept as the same as the protocol TR:TREATMENT_SUMMARY listed above. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Each pellet sample was mixed with 2 mL of 80% ice-cold methanol, and the mixed SP:SAMPLEPREP_SUMMARY solution was homogenized on ice for 2 minutes and repeated the same procedure SP:SAMPLEPREP_SUMMARY for three times. Next, the collected samples were centrifuged at 12000 rpm and SP:SAMPLEPREP_SUMMARY 4°C for 15 minutes, to collect the supernatants. The supernatants were further SP:SAMPLEPREP_SUMMARY mixed with 800 μL of ice-cold acetonitrile for 20 minutes before further SP:SAMPLEPREP_SUMMARY centrifuge at 12000 rpm and 4°C for 15 minutes. At last, the isolated SP:SAMPLEPREP_SUMMARY supernatants were gone through a 0.22 μm filter membrane (nylon) before the SP:SAMPLEPREP_SUMMARY lyophilization. The lyophilized samples were stored at -80°C for metabolomics SP:SAMPLEPREP_SUMMARY analysis. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY The chromatographic separation of biological samples was implemented on a Waters CH:CHROMATOGRAPHY_SUMMARY ACQUITY UPLC HSS T3 column (2.1×100 mm, 1.8 μm) with an optimized CH:CHROMATOGRAPHY_SUMMARY gradient-elution program (mobile phase A and B were 0.1% formic acid in water CH:CHROMATOGRAPHY_SUMMARY and acetonitrile (v/v)): 0-2 minutes, 98% A; 2-10 minutes, 98%-65% A; 10-12 CH:CHROMATOGRAPHY_SUMMARY minutes, 65%-20% A; 12-14 minutes, 20%-2% A; 14-30 minutes, 2% A. The column CH:CHROMATOGRAPHY_SUMMARY temperature was at 40°C and the flow rate was set at 0.3 mL/ minutes. CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Agilent 1290 Infinity CH:COLUMN_NAME Waters Acquity BEH HSS T3 (100 x 2.1mm, 1.8um) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Agilent 6495 QQQ MS:INSTRUMENT_TYPE Triple quadrupole MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS Agilent MassHunter Workstation Data Acquisition Agilent MassHunter MS:MS_COMMENTS QualitativeAnalysis B.07.00 Agilent MassHunter Quantitative Analysis (for QQQ) #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS - MS_METABOLITE_DATA_START Samples 20210227-3-C-1 20210227-3-C-2 20210227-3-C-3 20210227-3-C-4 20210227-3-C-5 20210227-3-C-6 20210227-3-30-1 20210227-3-30-2 20210227-3-30-3 20210227-3-30-4 20210227-3-30-5 20210227-3-30-6 20210227-3-50-1 20210227-3-50-2 Factors Concentration of MgSO4(w/v):0.01% Concentration of MgSO4(w/v):0.01% Concentration of MgSO4(w/v):0.01% Concentration of MgSO4(w/v):0.01% Concentration of MgSO4(w/v):0.01% Concentration of MgSO4(w/v):0.01% Concentration of MgSO4(w/v):0.15% Concentration of MgSO4(w/v):0.15% Concentration of MgSO4(w/v):0.15% Concentration of MgSO4(w/v):0.15% Concentration of MgSO4(w/v):0.15% Concentration of MgSO4(w/v):0.15% Concentration of MgSO4(w/v):0.25% Concentration of MgSO4(w/v):0.25% 2,4-Diaminobutyric acid 135.2053976 140.7351377 85.1929598 168.019519 70.283462 110.4866633 49.65029948 54.62838613 59.80065456 49.30062208 60.92534865 99.17902136 29.91396049 43.16970693 Isoleucine 40054.51465 42595.15645 19038.69369 43595.81686 18240.90012 27998.62116 21183.07985 19558.04977 14398.06035 13839.79292 18720.41664 24829.28095 20300.31775 17398.17759 L-Homoserine 14.80389798 12.04998072 7.222661995 14.87922947 6.497929105 8.317436738 6.409355527 7.871985101 5.118214314 4.520306141 9.195353861 7.370498034 5.943012135 5.461500949 Acetylcholine 7.798872785 8.176974294 3.395951075 9.821156868 3.94508085 5.053630357 3.453954653 2.380623699 1.644342087 2.048776758 2.810452204 3.307595813 4.230498347 2.649259941 L-Norleucine 39027.5309 40610.56375 18103.74136 44856.15654 17762.42764 28167.29457 20245.49299 20619.82718 15060.78877 14709.11475 18791.5553 22975.69884 19137.06965 16750.52705 L-Citrulline 36.24943691 39.92858669 11.76625615 49.57704141 16.22184156 22.31529927 8.568297311 23.39289116 18.24307645 15.8497066 15.62506267 20.02260381 13.20374335 12.38328116 Normetanephrine 354.4387003 450.8554624 201.9915632 438.7058445 166.6875123 250.0801212 221.4986509 76.53549691 203.3951825 154.5963906 58.8640952 264.8185397 35.03657576 86.70155992 Nicotinic acid 70.00912529 51.47932023 26.78886003 69.35053467 27.16363753 42.11303549 70.67061761 88.60831459 63.66738104 49.6499685 83.27500508 86.03342416 43.43789229 41.64422776 N-Acetyl-L-methionine 8.649428676 10.54618205 4.904388495 9.613191257 4.058782742 6.091529347 2.522028181 3.109939761 3.980925706 3.46386509 3.079590375 3.900131658 3.61140651 3.594802492 Guanidineacetic acid 6307.750655 6643.370917 2912.510063 8103.245068 3311.29809 4662.747443 2718.85746 3677.050515 1908.554228 1626.217697 3066.152966 2578.382735 3767.406271 2680.779141 Pyridoxal 11.53616908 11.28477421 7.125588607 15.91421366 6.587190585 10.36890917 4.054442117 5.610808844 3.684701576 3.256900097 4.554104591 4.72054407 4.839437497 3.949074656 Gamma-Aminobutyric acid 97.29953073 108.7471807 86.24433687 85.90359963 35.50633463 56.32158845 17.14011926 34.71748199 17.58481297 13.22253232 37.04744224 39.77297793 14.2934727 64.67566565 L-Proline 598.5445245 596.6009274 396.0460559 743.5315106 321.8755827 473.4111505 133.5890447 87.58294662 204.118945 160.3841228 76.22504203 398.162233 60.82568976 84.76360537 L-Histidine 339.3712573 368.2599399 200.953805 440.7462249 217.8908849 341.9606549 541.9085871 270.5135966 243.2650584 213.8362361 429.9976448 395.506589 208.5018623 251.6123188 L-Methionine 2886.287828 3082.371636 1310.735264 3530.053899 1425.429533 2127.311235 749.0925419 1047.977704 1181.693428 1037.249579 897.4741708 814.6220332 773.4907171 730.158672 L-Leucine 72524.98597 75524.77549 34122.51172 83718.74994 34181.69227 50692.36724 41580.99882 38278.33433 27913.1481 27694.86757 35439.54052 45074.76601 40015.91086 34458.07951 L-Phenylalanine 12682.52848 13156.02368 5399.4043 13898.9799 5722.354431 8557.835139 5933.862424 6413.171225 4736.373836 4316.21156 5973.233579 6820.232006 6762.293765 5974.739968 L-Glutamate 1906.490775 1851.308206 744.511905 3350.474734 1030.268517 1711.856608 887.94329 1610.652494 784.2947617 676.7736689 1072.639982 683.6108284 865.4620236 722.1072821 his 147.5272945 118.5100983 82.0257037 132.7663105 49.99319718 81.62123282 66.67250737 159.3372773 105.2608139 112.7434588 144.5527497 122.1875466 166.947117 143.8404507 L-Valine 13060.03672 12998.96141 5785.336578 14098.99997 6163.42721 8703.86236 5049.515145 7375.499145 3368.174857 3161.082276 6294.955938 5293.931936 6836.869125 4735.776399 Cholesterol 2600.508409 2103.434432 672.8500126 2321.334655 1071.304055 1293.606353 966.6636377 1369.869731 659.1230359 460.4431031 1174.365907 1157.696877 764.4162934 633.6377915 Beta-Sitosterol 88.25704042 84.33744792 46.34316537 78.02945035 33.42452748 37.07762001 23.19396924 37.97183759 28.19522727 11.98206205 28.20862177 30.02682167 24.3084042 29.37443202 Trans-4-Hydroxy-L-proline 72418.98716 75277.66921 34129.4268 81624.34671 34068.28748 50692.36724 41558.32103 38222.45487 27918.94549 27695.1958 35330.9665 45431.36001 40039.08206 34437.67851 Sarcosine 15.01222549 16.2312796 11.1319398 19.99721644 9.564991488 13.574977 7.641849642 7.912524522 6.813587172 7.361917479 9.589640442 13.1295797 4.58963278 6.518644491 Dopamine 14.80696659 12.75529967 8.986247396 16.90189104 6.483790206 6.793464378 2.337166661 4.225437254 3.924890688 3.287287141 3.493699344 4.971100588 3.693487732 2.319146986 Isonicotinic acid 73.8540502 51.47932023 26.7742452 72.58530264 27.89605312 43.91043605 70.67061761 88.60831459 63.66738104 49.6499685 83.27500508 86.03342416 43.43789229 41.64422776 N-Acetyl-L-cysteine 44.00486741 56.36373821 40.89898283 58.4256344 25.3950891 37.14749213 9.669900856 12.76823157 8.606175144 11.8601357 20.62766904 18.57870286 12.51111166 34.63052556 Reduced glutathione 4673.661784 4247.776358 2340.563904 5478.552687 2433.121346 3287.224726 1973.106027 3178.056389 1441.537189 1744.670312 2535.028515 1598.36538 363.9950498 1902.221915 2-Deoxyadenosine 10.20349552 7.58819585 3.679103329 12.60561845 5.018557623 7.600894539 29.20116837 14.87621679 13.86608657 5.139843832 4.433152505 18.25137399 50.64443815 3.150980382 CMP 257.5485551 287.7916552 117.8356386 411.7326973 167.450262 221.8182852 179.0439403 140.2219023 93.23655901 98.92457617 168.465765 222.6829639 81.94228317 81.36009658 Biotin 6.072796321 6.268168417 3.608167033 5.087566914 2.044391354 2.928072206 4.193366873 1.790774451 3.914971309 1.574999839 3.327529158 3.60768198 1.542480476 1.486518909 Adenosine 716.0050664 474.8854164 174.1952985 531.3980345 285.1620966 358.3166276 349.022864 197.0165674 251.7764835 155.9817215 70.3384933 297.9590874 864.3850062 127.9596049 L-Homocysteine 56.5672838 68.2322956 50.21371232 92.25771326 35.35341563 56.38451302 22.38176008 28.8206507 18.56087627 22.22611071 32.36429678 37.3448125 14.45625282 22.28042494 IMP 89.19027387 92.79033999 30.17666642 166.6199112 44.39158318 80.90794204 44.9615053 33.57938644 34.03499436 32.41708065 29.9594829 46.40037976 28.83865587 25.03629108 Cytidine 13.11340478 11.46341072 3.639214849 13.849837 5.754217742 10.87708303 4.127656326 4.188267551 3.173066296 3.054573241 5.557265248 6.926678501 6.950662999 4.471672526 N-Acetylglutamine 231.251642 277.020139 105.0557389 295.4910674 111.0476987 180.205265 87.94628676 106.4381354 74.18586661 93.47733565 90.91996983 114.2484846 72.332675 84.36521356 Betaine 2820.708039 2507.246189 1703.949858 2991.304721 1184.135174 1809.949734 428.9623044 1274.963559 457.7040917 414.2319408 654.2642575 507.8842603 788.3844569 663.0844273 PLP 214.8566614 165.0766451 70.97249174 219.5982278 91.9280064 121.2475145 107.3367035 168.7217912 137.6903395 97.52829038 100.2701558 108.3733997 159.6753655 78.15653101 Histamine 50.28875141 45.833356 19.8120768 59.53367353 24.77103305 36.27272202 27.52974357 21.22611603 15.56595596 16.42790237 21.00353153 24.71148219 15.7816808 13.6242841 Tryptamine 5.219660235 5.427012507 2.313672002 5.868473888 2.38081893 3.796208388 2.194415318 2.426974346 1.587325767 1.86218563 1.812905946 2.616849291 2.553811583 1.960746212 L-Lysine 246.5300827 234.495688 121.393156 213.0907236 106.0144511 142.8035047 116.5885972 179.0574436 104.2347117 87.92990592 204.1204224 135.3431247 123.1816891 141.1892967 Thymine 63.0848585 47.95269482 33.44190819 75.08202287 33.15451428 46.35562867 27.79370462 32.66781662 27.81118423 28.06908116 29.91760964 47.05572532 27.94959845 33.53844636 Putrescine 206.1888638 194.2000966 122.962947 213.2230141 91.81226037 124.3719737 82.66120582 73.26027471 82.40746837 65.96968479 83.0970069 126.7960916 88.08935948 59.97257647 L-Carnitine hydrochloride 7.656031273 7.376718107 3.081116968 6.727984613 2.76190048 4.238955655 2.66586967 5.042718554 2.651160155 2.320600712 2.552020677 2.307281893 2.482078905 2.020192288 5-Aminolevulinic acid 13.53318245 10.71205319 6.932986139 11.94530764 4.891215233 7.463091265 7.856026 6.260427814 5.305606004 4.431056005 8.527165484 8.526664586 6.10549225 4.332782827 5-aminosalicylic acid 30.43431924 10.24008849 10.30267256 22.03234023 9.602009461 10.50878505 9.664107996 10.84812037 10.6241382 8.652958625 8.587271752 12.02281222 5.896901228 6.570582361 Adenylosuccinic acid 165.220294 107.2024739 56.45497523 201.5611728 80.97730795 122.0265775 218.0467418 41.90321585 110.2798124 210.1882571 104.8560771 199.1049774 44.3040336 25.60894851 3-Guanidinopropionic acid? 79.13624379 77.72345706 45.5032445 73.47572104 30.75170993 45.23357628 56.84085143 38.36952286 33.66070074 33.8272389 57.5970101 67.71246411 54.47869583 34.13814491 AMP 2804.199206 2219.553351 995.237676 3038.817371 1057.475224 2051.538749 1905.390939 1471.086811 1211.88193 953.0368133 1073.647168 1496.131891 790.9469992 988.4441468 Oxidized glutathione 1892.931585 1757.695356 778.6144612 1907.243744 849.4034085 1077.468537 989.6098248 1126.239508 1279.229409 1023.730778 1286.083509 1490.977782 134.1087978 452.9281457 2'-Deoxyguanosine 5'-monophosphate 5.01927127 4.83397643 0.946426413 7.762843119 2.811326058 3.288378121 2.100944741 1.774882789 1.525805776 1.165485973 0.69996041 1.251257545 0.973740837 0.643881519 2'-Deoxycytidine 5'-monophosphate 12.47782778 13.89347805 4.65402396 17.84333303 8.436750346 10.29295327 12.84785463 8.718457124 5.85454127 5.436261791 4.940184223 11.10136265 4.990859357 3.356115097 Uridine 5'-monophosphate 38.37880995 44.49764906 8.852275492 45.61474899 22.11994841 32.45955636 17.69006383 30.66983365 10.76306354 7.539506495 12.57453646 9.996976289 8.485806617 6.073548761 S-(5'-Adenosyl)-L-homocysteine 16.01141028 14.7885878 10.21643404 15.49884801 6.112941137 8.798312603 11.84242968 10.10890164 9.730739867 12.33525343 12.72363287 11.34538425 3.500883044 7.229374724 Dephospho-CoA 1040.279782 842.1137039 1039.394557 1056.897478 459.2044545 600.9600282 272.6284061 388.8503393 144.8805924 196.670316 386.0347702 215.9679803 476.4921581 644.2131224 Choline 3.247267501 2.565148634 1.359172605 3.64933348 1.611131176 2.80094398 0.866439903 1.082850054 1.375487652 1.1967452 1.045147448 1.059051543 1.101522272 0.702001991 Creatine 4.108463142 3.735934791 2.101547615 4.492471936 2.121902055 2.890809075 1.579251236 1.356862133 1.581953827 1.285218716 2.29946706 2.730780111 0.829991932 1.44807576 Hypoxanthine 146.9052255 242.4765948 73.67664952 355.9445023 159.1466304 170.62377 29.50540602 60.85671931 57.25398081 61.5030026 72.37043365 176.9774548 53.5053996 37.23195741 Spermidine 969.2334969 935.0524523 348.8999654 1016.689154 361.4129951 572.9051285 356.138696 463.7690941 375.9391626 404.6799218 368.9819123 542.9293704 422.8629652 373.4035501 Thiamine 5.867293714 3.935702533 0.894112685 4.625228063 1.545254354 4.42626638 1.511751322 4.780860425 2.764929363 2.613402495 2.640564614 2.701410708 2.823000905 1.613848546 Inosine 65.60998498 59.31052278 23.95970988 68.22017537 25.31926633 39.12391235 45.35427057 22.28235763 22.89046155 21.59455728 24.21879371 64.41839033 27.14208297 27.79665181 Agmatine 2248.842755 2038.608389 1383.622 2651.013888 999.5167869 1686.223763 2207.718093 1538.73116 1153.157399 940.8561953 1721.472311 1911.468976 1434.105246 975.3007797 Guanosine 5'-monophosphate 57.32337715 72.48926766 20.96693269 66.34605707 27.87575711 49.87438081 31.28285682 19.38099551 20.86301167 17.0343285 17.01942614 28.04596754 13.97891369 11.03919753 Alanine 36.986378 34.43928885 22.77125379 45.94477456 18.06788604 28.0317747 18.35375405 15.33465756 14.60083751 15.59965587 21.13837783 30.65319542 8.828321071 14.74709532 Uracil 46.96524466 56.6168356 17.10039324 71.33540041 25.83039143 39.82629197 17.27793371 14.57418276 24.8350894 24.29103478 20.64453995 36.28603469 12.24197711 15.56106144 Tryptophan 2112.852722 1852.108071 891.7110204 1880.937427 741.3542634 1278.187844 1029.229958 560.7603557 839.0343023 693.8500905 1039.849524 1410.572153 548.2790682 837.6259566 Adenine 314.3271148 288.8769157 101.2201658 443.4229253 154.6235143 245.2392188 139.194143 126.2901991 103.8773505 116.5498614 100.0257015 174.1862688 159.4908246 114.2684524 Indolepyruvic acid 2.524217433 3.421338677 2.140294778 5.210834703 2.002443933 3.029619445 0.58828358 0.405105661 0.288328118 0.641092317 0.759255423 0.797707103 0.673735443 1.75034984 Pyrogallol 16.39885438 14.21224514 9.692853438 17.71798364 8.790067815 13.76301773 11.29981711 8.159784378 7.697513085 8.940906286 9.104408218 15.48943788 9.704812963 9.92689851 5-Dodecenoic acid 11.02533181 11.1501676 4.033844839 14.89952847 5.203468468 8.579039983 3.291341353 5.371237908 3.948101301 5.133034925 2.798088496 5.162460861 3.301846263 3.634038254 Ethylmethylacetic acid 2.463486463 2.553041685 1.309430909 3.059218455 1.1845613 1.673597672 0.865183886 1.024254963 1.01739762 0.769913754 1.025914904 1.536169512 0.578878434 0.585803767 Pentadecanoic acid 7.994732715 5.195169215 3.804230326 4.85438599 1.929597902 2.880027963 1.604202305 5.33963369 2.377003305 4.377776367 2.85687726 2.478935131 3.639690924 4.148794371 Stearic acid 42.02036138 38.2832962 25.42312304 76.31468848 28.81278946 43.15176668 23.12962498 19.97930104 8.326051173 22.06274741 34.71234462 25.31931334 12.95093885 17.87087804 Aminoadipic acid 3958.460097 4179.840664 2273.497558 4872.054581 2268.890937 3121.639747 3298.061023 853.5916805 3127.066122 2982.478181 670.7209649 5166.232211 358.6379357 1077.654451 Norvaline 13060.03672 12998.96141 5785.336578 14098.99997 6163.42721 8703.86236 5051.226087 7115.535667 3368.174857 3158.218573 6123.079511 5293.931936 6837.652489 4735.776399 Pyroglutamic acid 9252.73864 9565.689944 3288.904562 11757.45285 4304.09061 7021.704663 3310.012974 4412.214838 3445.138256 3694.950992 4303.1766 4156.514604 3654.651573 3258.258468 Pipecolic acid 7135.093512 7685.972554 2943.128199 9652.363424 3621.608415 6080.909923 2604.581673 3768.231898 2842.184473 2674.302905 3345.955333 3459.830226 3114.406727 2540.420005 L-Glutamine 313.1606166 216.9617263 115.0537959 264.3951302 100.066335 160.1755267 122.8133733 122.4626532 101.851182 97.4896721 186.1739384 121.0042881 96.21958603 134.2845913 L-Ornithine 140.5928084 164.5509461 77.08132704 214.5333196 69.51736916 104.9991277 98.30220211 76.69038601 64.79334617 63.20526779 74.35434097 104.7565638 89.90261559 79.21530546 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name m/z RT 2,4-Diaminobutyric acid 119.11 1.82 Isoleucine 132.18 2.42 L-Homoserine 120.07 1.08 Acetylcholine 147.13 1.39 L-Norleucine 132.1 2.42 L-Citrulline 176.2 1.35 Normetanephrine 184.1 0.96 Nicotinic acid 124.12 1.32 N-Acetyl-L-methionine 192.26 5.86 Guanidineacetic acid 118.12 1.26 Pyridoxal 168.13 1.36 Gamma-Aminobutyric acid 104.13 0.91 L-Proline 116.14 0.95 L-Histidine 156.16 1.32 L-Methionine 150.22 1.43 L-Leucine 132.18 2.41 L-Phenylalanine 166.2 4.32 L-Glutamate 148.06 0.91 his 134.11 2.28 L-Valine 118.09 1.26 Cholesterol 387.3 12.61 Beta-Sitosterol 415.72 12.94 Trans-4-Hydroxy-L-proline 132.14 2.2 Sarcosine 90.06 0.95 Dopamine 154.15 1.55 Isonicotinic acid 124.12 1.32 N-Acetyl-L-cysteine 164.2 2.95 Reduced glutathione 308.09 1.41 2-Deoxyadenosine 252.1 2.61 CMP 324.21 1 Biotin 245.32 7.19 Adenosine 268.25 2.32 L-Homocysteine 136.05 1.02 IMP 349.06 1.26 Cytidine 244.23 1.24 N-Acetylglutamine 189.19 1.68 Betaine 118.16 0.9 PLP 248.15 1.44 Histamine 112.09 1.09 Tryptamine 161.23 6.23 L-Lysine 147.2 1.47 Thymine 127.12 2.46 Putrescine 89.08 0.7 L-Carnitine hydrochloride 162.1 0.92 5-Aminolevulinic acid 132.07 1.05 5-aminosalicylic acid 154.15 1.54 Adenylosuccinic acid 464.08 3.49 3-Guanidinopropionic acid? 132.14 1.06 AMP 348.07 1.25 Oxidized glutathione 613.2 1.59 2'-Deoxyguanosine 5'-monophosphate 348.07 1.48 2'-Deoxycytidine 5'-monophosphate 308.07 1.13 Uridine 5'-monophosphate 325.05 1.09 S-(5'-Adenosyl)-L-homocysteine 385.1 1.57 Dephospho-CoA 688.2 4.88 Choline 105.1 1.43 Creatine 132.08 2.03 Hypoxanthine 137 1.45 Spermidine 146.17 0.67 Thiamine 266.12 1.43 Inosine 269.09 2.86 Agmatine 131.1 1.06 Guanosine 5'-monophosphate 364.07 1.29 Alanine 90.1 0.95 Uracil 113.1 1.3 Tryptophan 205.1 5.75 Adenine 136.1 1.23 Indolepyruvic acid 204.07 8.56 Pyrogallol 127.04 2.47 5-Dodecenoic acid 199.17 13.58 Ethylmethylacetic acid 103.08 1.82 Pentadecanoic acid 243.23 14.49 Stearic acid 285.28 12.78 Aminoadipic acid 162.08 0.97 Norvaline 118.09 1.25 Pyroglutamic acid 130.05 1.64 Pipecolic acid 130.09 1.63 L-Glutamine 147.08 1.47 L-Ornithine 133.1 2.18 METABOLITES_END #END