#METABOLOMICS WORKBENCH Tom_Metz_20140707_9470461_mwtab.txt DATATRACK_ID:135 STUDY_ID:ST000084 ANALYSIS_ID:AN000136 PROJECT_ID:PR000075 VERSION 1 CREATED_ON 2016-09-17 #PROJECT PR:PROJECT_TITLE Systems Biology for EnteroPathogens PR:PROJECT_TYPE MS analysis PR:PROJECT_SUMMARY sysbep.org PR:INSTITUTE Pacific Northwest National Laboratory PR:DEPARTMENT Biological Separation and Mass Spectrometry PR:LAST_NAME Joshua PR:FIRST_NAME Adkins PR:ADDRESS - PR:EMAIL Joshua.Adkins@pnnl.gov PR:PHONE - #STUDY ST:STUDY_TITLE Model-driven multi-omic data analysis elucidates metabolic immunomodulators of ST:STUDY_TITLE activation ST:STUDY_TYPE growth condition, timecourse ST:STUDY_SUMMARY Macrophages are central players in immune response, manifesting divergent ST:STUDY_SUMMARY to control inflammation and innate immunity through release of cytokines and ST:STUDY_SUMMARY signaling factors. Recently, the focus on metabolism has been reemphasized as ST:STUDY_SUMMARY signaling and regulatory pathways of human pathophysiology, ranging from cancer ST:STUDY_SUMMARY aging, often converge on metabolic responses. Here, we used genome-scale ST:STUDY_SUMMARY and multi-omics (transcriptomics, proteomics, and metabolomics) analysis to ST:STUDY_SUMMARY metabolic features that are critical for macrophage activation. A genome-scale ST:STUDY_SUMMARY network for the RAW 264.7 cell line was constructed to determine metabolic ST:STUDY_SUMMARY of activation. Metabolites well-known to be associated with immunoactivation ST:STUDY_SUMMARY and arginine) and immunosuppression (tryptophan and vitamin D3) were among the ST:STUDY_SUMMARY critical effectors. Intracellular metabolic mechanisms were assessed, ST:STUDY_SUMMARY a suppressive role for de-novo nucleotide synthesis. Finally, underlying ST:STUDY_SUMMARY mechanisms of macrophage activation were identified by analyzing multi-omic ST:STUDY_SUMMARY obtained from LPS-stimulated RAW cells in the context of our flux-based ST:STUDY_SUMMARY This study demonstrates that the role of metabolism in regulating activation ST:STUDY_SUMMARY be greater than previously anticipated and elucidates underlying connections ST:STUDY_SUMMARY activation and metabolic effectors. This submission corresponds to the ST:STUDY_SUMMARY data from this study. ST:INSTITUTE Pacific Northwest National Laboratory ST:DEPARTMENT Biological Separation and Mass Spectrometry ST:LAST_NAME Metz ST:FIRST_NAME Thomas ST:ADDRESS - ST:EMAIL thomas.metz@pnnl.gov ST:PHONE - ST:NUM_GROUPS 2 ST:TOTAL_SUBJECTS 12 #SUBJECT SU:SUBJECT_TYPE Cells SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:GENOTYPE_STRAIN RAW 264.7 SU:CELL_STRAIN_DETAILS RAW 264.7 SU:CELL_PRIMARY_IMMORTALIZED immortalized SU:SPECIES_GROUP Mammal #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - SBEP_Metab_LPSactivated1 Lipopolysaccharide Treatment:100 ng/ml SUBJECT_SAMPLE_FACTORS - SBEP_Metab_LPSactivated2 Lipopolysaccharide Treatment:100 ng/ml SUBJECT_SAMPLE_FACTORS - SBEP_Metab_LPSactivated3 Lipopolysaccharide Treatment:100 ng/ml SUBJECT_SAMPLE_FACTORS - SBEP_Metab_Control1 Lipopolysaccharide Treatment:None SUBJECT_SAMPLE_FACTORS - SBEP_Metab_Control2 Lipopolysaccharide Treatment:None SUBJECT_SAMPLE_FACTORS - SBEP_Metab_Control3 Lipopolysaccharide Treatment:None #COLLECTION CO:COLLECTION_SUMMARY After stimulation, cells were washed twice with Dulbecco?s PBS, scraped out and CO:COLLECTION_SUMMARY into 15-mL centrifuge tubes. CO:SAMPLE_TYPE Cell CO:COLLECTION_METHOD After stimulation, cells were washed twice with Dulbecco?s PBS, scraped out and CO:COLLECTION_METHOD into 15-mL centrifuge tubes. CO:COLLECTION_TIME 24 hours CO:COLLECTION_VIALS 15-mL centrifuge tubes CO:TISSUE_CELL_QUANTITY_TAKEN All #TREATMENT TR:TREATMENT_SUMMARY Macrophages (RAW 264.7 cells) grown for two day with suplimentation of 100ng | TR:TREATMENT_SUMMARY lipopolysaccharide | Macrophages (RAW 264.7 cells) grown for two days without TR:TREATMENT_PROTOCOL_COMMENTS Macrophages (RAW 264.7 cells) were seeded at a density of 3.0E6 cells in 150 mm TR:TREATMENT_PROTOCOL_COMMENTS using Dulbecco's Modified Eagle's Medium supplemented with 10% fetal calf TR:TREATMENT_PROTOCOL_COMMENTS then grown for 2 days and stimulated for 24 hours with 100 ng/mL of TR:TREATMENT_PROTOCOL_COMMENTS diluted in fresh medium. A control culture was run in parallel by incubating TR:TREATMENT_PROTOCOL_COMMENTS the same period of time with fresh medium only. Three biological replicates TR:TREATMENT_PROTOCOL_COMMENTS performed per condition, and two dishes were used for each replicate. / TR:TREATMENT_PROTOCOL_COMMENTS (RAW 264.7 cells) were seeded at a density of 3.0E6 cells in 150 mm dishes TR:TREATMENT_PROTOCOL_COMMENTS Dulbecco's Modified Eagle's Medium supplemented with 10% fetal calf serum, then TR:TREATMENT_PROTOCOL_COMMENTS for 2 days and stimulated for 24 hours with 100 ng/mL of lipopolysaccharide TR:TREATMENT_PROTOCOL_COMMENTS in fresh medium. A control culture was run in parallel by incubating for the TR:TREATMENT_PROTOCOL_COMMENTS period of time with fresh medium only. Three biological replicates were TR:TREATMENT_PROTOCOL_COMMENTS per condition, and two dishes were used for each replicate. TR:TREATMENT Abiotic TR:TREATMENT_COMPOUND lipopolysaccharide / fresh medium TR:TREATMENT_DOSE 100 ng/mL /-- TR:TREATMENT_VEHICLE fresh medium TR:CELL_GROWTH_CONTAINER 150 mm dishes TR:CELL_INOC_PROC Macrophages (RAW 264.7 cells) were seeded at a density of 3.0E6 cells in 150 mm TR:CELL_MEDIA Dulbecco's Modified Eagle's Medium supplemented with 10% fetal calf serum TR:CELL_HARVESTING After stimulation, cells were washed twice with Dulbecco?s PBS, scraped out and TR:CELL_HARVESTING into 15-mL centrifuge tubes. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Suspentions softly centrifuged, buffer removed, amonium bicarbonate added, SP:SAMPLEPREP_SUMMARY extraxted with chloroform/methanol (2:1, v/v), vortexed, centrifuged, aqueous SP:SAMPLEPREP_SUMMARY dried in vacuum concentrator, derivatization with methoxyamine in pyridine, SP:SAMPLEPREP_SUMMARY (MSTFA), and 1% trimethylchlorosilane (TMCS) SP:SAMPLEPREP_PROTOCOL_COMMENTS Cell suspensions were softly centrifuged (230 × g for 5 min) and as much SP:SAMPLEPREP_PROTOCOL_COMMENTS as possible was removed. Then, 170 µL of 150 mM ammonium bicarbonate was added SP:SAMPLEPREP_PROTOCOL_COMMENTS the cell pellet and the cell suspensions were transferred to 2 mL SP:SAMPLEPREP_PROTOCOL_COMMENTS tubes for extraction. Subsequently, the water soluble metabolites were SP:SAMPLEPREP_PROTOCOL_COMMENTS with four volumes of chilled (-20°C) chloroform/methanol (2:1, v/v). After SP:SAMPLEPREP_PROTOCOL_COMMENTS the samples were centrifuged (12,000 × g for 5 min) and the upper (aqueous) SP:SAMPLEPREP_PROTOCOL_COMMENTS containing water-soluble metabolites were transferred into glass vials, SP:SAMPLEPREP_PROTOCOL_COMMENTS by drying in a vacuum concentrator. For the derivatization, 20 µL of SP:SAMPLEPREP_PROTOCOL_COMMENTS in pyridine (30 mg/mL) were added to each sample, followed by incubation at SP:SAMPLEPREP_PROTOCOL_COMMENTS with shaking for 90 min to protect carbonyl groups. Next, 80 µL of SP:SAMPLEPREP_PROTOCOL_COMMENTS (MSTFA) with 1% trimethylchlorosilane (TMCS) were added to each vial, followed SP:SAMPLEPREP_PROTOCOL_COMMENTS incubation at 37°C with shaking for 30 min to derivatize hydroxyl and amine SP:SAMPLEPREP_PROTOCOL_COMMENTS The samples were then allowed to cool to room temperature. SP:PROCESSING_METHOD Homogenization SP:EXTRACTION_METHOD The water soluble metabolites were extracted with four volumes of chilled SP:EXTRACTION_METHOD chloroform/methanol (2:1, v/v). After vortexing, the samples were centrifuged SP:EXTRACTION_METHOD × g for 5 min) and the upper (aqueous) layers containing water-soluble SP:EXTRACTION_METHOD were transferred into glass vials, followed by drying in a vacuum concentrator. SP:EXTRACT_ENRICHMENT Vacuum Concentrator SP:SAMPLE_RESUSPENSION 20 µL of methoxyamine in pyridine (30 mg/mL) SP:SAMPLE_DERIVATIZATION 20 µL of methoxyamine in pyridine (30 mg/mL), 80 µL of SP:SAMPLE_DERIVATIZATION (MSTFA) with 1% trimethylchlorosilane (TMCS), SP:CELL_TYPE Macrophage #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Agilent 7890A gas chromatograph with a HP-5MS gas chromatography column using CH:CHROMATOGRAPHY_TYPE GC CH:INSTRUMENT_NAME Agilent 7890A CH:COLUMN_NAME Agilent HP5-MS (30m × 0.25mm, 0.25 um) CH:CHROMATOGRAPHY_COMMENTS Chromatography was carried out on an Agilent 7890A gas chromatograph using the CH:CHROMATOGRAPHY_COMMENTS software (Chemstation) and a HP-5MS gas chromatography column (Agilent CH:CHROMATOGRAPHY_COMMENTS Santa Clara, CA; 30 m x 0.25 mm x 0.25 m film thickness). The sample injection CH:CHROMATOGRAPHY_COMMENTS was splitless, and 1 L of each sample was injected. The injection port CH:CHROMATOGRAPHY_COMMENTS was held at 250 C throughout the analysis. The GC oven was held at 60 C for 1 CH:CHROMATOGRAPHY_COMMENTS after injection, and the temperature was then increased to 325 C by 10 C/min, CH:CHROMATOGRAPHY_COMMENTS by a 5 min hold at 325 C. The helium gas flow rates for each Experiment were CH:CHROMATOGRAPHY_COMMENTS by the Agilent Retention Time Locking function based on analysis of deuterated CH:CHROMATOGRAPHY_COMMENTS acid and were in the range of 0.450.5 mL/min. CH:FLOW_RATE 0.45-0.5mL/min CH:INJECTION_TEMPERATURE 250 C CH:SAMPLE_INJECTION 1 L, Splitless CH:ANALYTICAL_TIME 37.5 min CH:OVEN_TEMPERATURE 60 C for 1 min, then increased to 325 C by 10 C/min, followed by a 5 min hold CH:OVEN_TEMPERATURE 325 C CH:SAMPLE_SYRINGE_SIZE 10 L #ANALYSIS AN:ANALYSIS_TYPE MS AN:LABORATORY_NAME Biological Separations & Mass Spectrometry group, Pacific Northwest National AN:LABORATORY_NAME (www.omics.pnl.gov) AN:ACQUISITION_DATE 2012 AN:SOFTWARE_VERSION Metabolite Detector vs. 2.0.6 beta AN:DATA_FORMAT Raw .D.Zip; Processed .CDF #MS MS:INSTRUMENT_NAME Agilent 5975C MS:INSTRUMENT_TYPE Single quadrupole MS:MS_TYPE EI MS:ION_MODE POSITIVE MS:MS_COMMENTS An Agilent GC 7890A coupled with a single quadrupole MSD 5975C (Agilent MS:MS_COMMENTS Inc.; Santa Clara, CA, USA) was used, and the samples were blocked and analyzed MS:MS_COMMENTS random order for each experiment. Data were collected over the mass range MS:MS_COMMENTS m/z. A mixture of FAMEs (C8-C28) was analyzed once per day together with the MS:MS_COMMENTS for retention index alignment purposes during subsequent data analysis. MS:SCAN_RANGE_MOVERZ 50-550 m/z MS:MS_COMMENTS After converting raw data to netCDF format, the data were processed by the MS:MS_COMMENTS software for peak deconvolution and chromatographic alignment. Retention MS:MS_COMMENTS (RI) were calculated based on the analysis of a mixture of fatty acid methyl MS:MS_COMMENTS (C8 - C30) (Agilent Technologies) as external retention time standards, then MS:MS_COMMENTS retention index information was subsequently applied to all experimental MS:MS_COMMENTS for retention time alignment. MetaboliteDetector parameters for peak detection MS:MS_COMMENTS deconvolution are as follows: Peak threshold, 7; minimum peak height, 7; MS:MS_COMMENTS width, 8. Deconvoluted features were identified by matching to the Agilent MS:MS_COMMENTS Metabolomics Retention Time Locked Library, which contains mass spectral and MS:MS_COMMENTS indix information for approximately 700 metabolites. Each initial match to the MS:MS_COMMENTS was manually inspected to confirm a confident identification. The data were MS:MS_COMMENTS matched against the NIST 08 library to identify additional peaks not included MS:MS_COMMENTS the Fiehn library. MetaboliteDetector software was used for database matching MS:MS_COMMENTS batch identification/quantification parameters are as follows: required score, MS:MS_COMMENTS ?RI, 25; minimum S/N, 20; maximum peak discrepancy index, 100. Ions 73 and 143 MS:MS_COMMENTS excluded from use as metabolite quantification ions, since these are due to MS:MS_COMMENTS of the trimethylsilyl groups. Otherwise, three unique fragment ions were MS:MS_COMMENTS to each metabolite for quantification and used for each individual GC-MS MS:MS_COMMENTS when processing the data in batch mode. The summed areas of the three MS:MS_COMMENTS ions were exported from MetaboliteDetector and used in further statistical MS:MS_COMMENTS All identifications were manually validated by inspection of retention index MS:MS_COMMENTS spectrum matches. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Peak area MS_METABOLITE_DATA_START Samples SBEP_Metab_LPSactivated1 SBEP_Metab_LPSactivated2 SBEP_Metab_LPSactivated3 SBEP_Metab_Control1 SBEP_Metab_Control2 SBEP_Metab_Control3 Factors Lipopolysaccharide Treatment:100 ng/ml Lipopolysaccharide Treatment:100 ng/ml Lipopolysaccharide Treatment:100 ng/ml Lipopolysaccharide Treatment:None Lipopolysaccharide Treatment:None Lipopolysaccharide Treatment:None 1-methyl nicotinamide 1128844.9000 990557.0975 1142586.6960 1117011.1816 1315764.9690 1609434.2165 2-aminoethanethiol 941833.5000 868973.5000 953831.6860 1600093.6015 1964054.9380 2213228.8040 3-hydroxybenzaldehyde 501242.7073 431011.4386 477214.9231 3-phosphoglyceric acid 1303610.3515 1504818.0300 1740486.5540 877204.8435 1211525.0710 1272797.0990 4-guanidinobutyric acid 1859373.2670 1685591.6650 1858273.8870 2645271.9460 2600277.8205 3854828.0225 4-hydroxyquinoline-2-carboxylic acid 177836.4865 168974.9107 193569.5687 2110118.1290 2221603.0990 2535250.0470 5^-deoxy-5^-(methylthio)adenosine 2882511.8800 2957775.4975 2724568.3550 464864.2507 589460.9126 780203.1709 adenosine 1537097.8750 1214250.8015 1316169.5090 4053242.8525 4148655.7665 4784051.0780 adenosine-5-monophosphate 9454142.7335 8121689.6410 8463618.3950 27543792.1050 34940220.5400 42146640.0400 alpha ketoglutaric acid 2092664.4335 2059916.6520 2437121.8085 arabitol 390153.2435 387118.4070 454665.9208 beta- alanine 12036820.1650 10851261.8450 13599141.0650 23432641.8050 24002849.8650 30832474.5100 citraconic acid 6589380.9000 6127515.1420 7514006.2660 citric acid 682651.2431 498260.2384 538841.7364 1070709.0001 948590.2074 1063573.1426 creatinine 10452669.1560 10339577.5660 11304334.0600 14319437.0950 12841146.2800 16519702.7700 Cysteinylglycine 747331.7561 634460.2773 848180.1665 D-galactose 523849.8309 545109.2323 811281.7084 D-glucosaminic acid 26639733.4550 28680203.6300 29834683.9500 1869625.0015 2704562.6160 3391397.5005 D-glucose-6-phosphate 1721774.7810 1643900.4280 1763549.7790 DL-3-aminoisobutyric acid 172434.6981 92082.6045 141553.3294 1511728.4585 1461268.2315 2514955.0150 DL-threo-beta-Hydroxyaspartic acid 95146.8605 103992.0491 120882.9009 170726.0954 335916.9590 425773.9757 D-malic acid 1147617.8690 1135687.1810 1415029.3205 4107528.9195 3486786.6035 4118208.6520 D-mannitol 2946232.9380 2897797.4370 3412819.1040 1900178.9300 1597070.3660 1223019.2760 D-Ribose 5-phosphate 2229996.4555 2427192.3445 2573543.4410 3592947.2585 5229012.1725 8143540.4925 D-sorbitol 589958.5707 435463.2498 518026.4727 249105.8004 207333.6954 190275.6371 D-threitol 1081927.9775 1084599.8660 1233440.0695 1162887.3080 1628252.2225 1772900.8970 fructose 2037004.2305 2114549.7710 2326119.2610 481045.2352 709047.3964 789425.1007 fumaric acid 3276814.9275 3400546.9545 4137408.8530 glucoheptonic acid 2634651.2925 2829935.9925 2869015.6730 1288067.9260 1342649.3530 1229431.2395 gluconic acid 494827.0838 699589.9422 617827.3551 glycerol 1-phosphate 10264178.5300 10798000.4200 9612604.6905 glycine 37465350.1100 33155757.6900 40856466.0700 53534675.3200 55540492.2750 69796232.4650 guanosine 975057.3367 1245361.9060 1440699.5980 guanosine-5^-monophosphate 32649491.4100 29851123.1350 33411172.9700 66815175.9950 72595108.8700 81348058.1400 hypotaurine 3802828.5805 3079498.0710 4089572.1595 5823187.2135 6589528.6090 9111528.9630 hypoxanthine 46279.6424 46244.4621 47779.0448 54423.4678 77548.7990 113037.4879 inosine 3807795.3400 3671602.6305 4020506.4480 5453101.7045 6577889.8230 7621978.0705 L-alanine 2966422.2705 2527379.4385 2874097.5210 9763688.5645 10716948.6250 14565213.9750 L-aspartic acid 1611386.4610 854706.8746 1149295.5377 17758208.6450 11004554.3600 13441915.2450 L-cysteine 6594124.7115 6239553.3880 6916586.0315 4363129.8585 4647791.9710 5261679.3860 L-glutamic acid 19048241.4500 17688103.9950 22072682.5300 32429901.4500 31172798.7500 37958720.7050 L-leucine 921691.1742 927836.8475 1013165.8040 1733967.8160 1628935.4350 2075679.0600 L-lysine 3397988.6500 3137480.4855 4116308.9395 4663940.2620 5255705.5005 6985776.7045 L-methionine 778737.0805 823831.7756 869545.9695 449913.1699 L-ornithine 895489.0345 1786521.5760 338658.3193 2462425.0610 2398847.8915 3367301.2360 L-serine 1567014.7625 1635960.1275 1790927.5215 2503343.5225 2284952.7000 3016039.1185 L-sorbose 1445182.5500 926870.9442 1067504.9490 2227343.3225 2465901.1845 3205120.3090 L-threonine 4206167.4105 3813468.4815 4105387.5070 6327888.6420 7315329.1770 10419036.5800 L-tyrosine 2494203.6000 3139766.7630 4027543.5960 L-valine 2032012.8800 1984586.5770 2182315.5285 1921864.9465 1993920.9160 2503465.9570 maltitol 1017471.4548 923455.5432 982604.0935 1232980.4005 1316691.3385 1617773.4420 mucic acid 218055.6989 207827.6139 195215.3010 oxalic acid 22672.0000 27759.5000 26053.0000 31231.8201 37874.1906 35845.2230 palmitic acid 321087.4780 251727.9914 290664.8260 1144873.4435 1430828.4635 1885948.7900 pantothenic acid 1494386.4495 1474212.0750 1509237.7320 2246904.2570 2593580.9985 2915574.7765 porphine 487431.0139 489637.8323 510780.0168 425471.3115 889348.9603 1129465.8540 putrescine 24086631.4450 23407531.2800 26341519.8150 6759718.3870 7405081.2425 9908081.7660 pyruvic acid 728753.8819 862734.1556 926361.1350 1394193.6825 1288652.5820 1390265.1600 ribonic acid gamma-lactone 586224.1754 764691.0749 863848.2198 spermidine 7597352.8600 4974057.2150 8640064.2905 11585311.2750 14707324.2150 17969750.7550 spermine 1297630.8355 554069.7708 1612147.8085 5256107.3915 7576976.7175 11210265.5350 stearic acid 1102517.7490 1011344.8775 1270094.6065 1759707.1405 1897270.9630 2297010.1180 tagatose 1463995.6225 1649932.2850 1673612.2225 thymine 722755.5389 1000376.5401 2537650.6245 tyramine 5108178.3740 5342080.0920 4457680.0675 1121906.5485 758655.2526 766216.4064 uridine 5^-monophosphate 1193945.2940 1132687.3275 1274190.3945 3593946.2695 3647580.9585 4667846.3210 xanthine 1545967.6430 1433454.6040 1812530.9525 3681925.3400 5000439.1170 5864402.9620 xylitol 654501.6410 657450.6469 678320.9415 298665.5000 440508.5775 618706.6942 5-deoxy-5-(methylthio)adenosine 2731021 2843049 2631001 397555 576958 585647 guanosine-5-monophosphate 31644175 29383169 32762698 63475745 69081290 77607095 uridine 5-monophosphate 1228226 1179639 1279919 3472964 3654297 4544097 Unidentified_01 667852 503774 691478 847455 1012036 1294021 Unidentified_02 3871892 2507673 2860159 16526784 21532416 29651772 Unidentified_03 1898544 1469469 1691181 2300752 2357821 2473606 Unidentified_04 10059442 8389064 7407525 2864880 4723176 7005910 Unidentified_05 534036 611976 659420 6285391 6906213 8899782 Unidentified_06 9427201 9919212 8469552 30604478 37724103 44658661 Unidentified_07 4118621 2335308 2673643 5588379 7580617 9610814 Unidentified_08 569459 529109 548205 492286 426787 504992 Unidentified_09 1175370 1127782 1262448 1953747 1686602 1886616 Unidentified_10 872574 865073 871272 403826 435012 464220 Unidentified_11 438110 319398 400521 1346161 1653397 2208702 Unidentified_12 155837 536296 487104 1181248 1128942 1477804 Unidentified_13 3618923 3406644 3513440 3126937 2882708 2950750 Unidentified_14 10612748 10141421 11406521 47435112 49258838 50256978 Unidentified_15 642425 759665 962899 131519 166594 301300 Unidentified_16 3742451 3812143 4161443 1582753 1704383 2040883 Unidentified_17 680163 601501 667084 711982 673185 890280 Unidentified_18 3510703 3494722 4093260 3788163 4946855 6167557 Unidentified_19 409948 315735 630997 652759 1031449 1822350 Unidentified_20 640052 392679 759300 1034596 1556482 2608259 Unidentified_21 3149606 3614620 3387828 3510301 3895706 4760973 Unidentified_22 3907777 3521515 3758783 992634 907774 1286177 Unidentified_23 1535597 1590410 1536950 3315486 3990111 4478393 Unidentified_24 581187 515310 726577 649528 833006 887258 ribonic acid ?-lactone N N N 609096 744730 729124 pyrophosphate 10011647 5910492 8783192 N N N MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name moverz_quant ri ri_type pubchem_id inchi_key kegg_id other_id other_id_type 1-methyl nicotinamide 10129985 CHEBI:16797 PNNL_ID 2-aminoethanethiol 6058 CHEBI:17141 PNNL_ID 3-hydroxybenzaldehyde 101 CHEBI:16207 PNNL_ID 3-phosphoglyceric acid 724 CHEBI:17794 PNNL_ID 4-guanidinobutyric acid 500 CHEBI:15728 PNNL_ID 4-hydroxyquinoline-2-carboxylic acid 3845 CHEBI:18344 PNNL_ID 5'-deoxy-5'-(methylthio)adenosine 149 CHEBI:17509 PNNL_ID adenosine CHEBI:16335 PNNL_ID adenosine-5-monophosphate CHEBI:16027 PNNL_ID alpha ketoglutaric acid CHEBI:30915 PNNL_ID arabitol 94154 CHEBI:18333 PNNL_ID beta- alanine CHEBI:16958 PNNL_ID citraconic acid CHEBI:17626 PNNL_ID citric acid CHEBI:30769 PNNL_ID creatinine CHEBI:16737 PNNL_ID Cysteinylglycine 65270 CID65270 PNNL_ID D-galactose 439353 CHEBI:4139 PNNL_ID D-glucosaminic acid 235915 CHEBI:17784 PNNL_ID D-glucose-6-phosphate 439427 CID44134741 PNNL_ID DL-3-aminoisobutyric acid 64956 CHEBI:27389 PNNL_ID DL-threo-beta-Hydroxyaspartic acid 443239 CHEBI:10696 PNNL_ID D-malic acid 92824 CHEBI:30796 PNNL_ID D-mannitol CHEBI:16899 PNNL_ID D-Ribose 5-phosphate 77982 CHEBI:52742 PNNL_ID D-sorbitol 453 CHEBI:17924 PNNL_ID D-threitol 169019 CHEBI:48300 PNNL_ID fructose 5984 CHEBI:48095 PNNL_ID fumaric acid 444972 CHEBI:18012 PNNL_ID glucoheptonic acid CID25588 PNNL_ID gluconic acid 10690 CHEBI:33198 PNNL_ID glycerol 1-phosphate 754 CHEBI:14336 PNNL_ID glycine 750 CHEBI:15428 PNNL_ID guanosine 6802 CHEBI:16750 PNNL_ID guanosine-5'-monophosphate 6804 CHEBI:17345 PNNL_ID hypotaurine 107812 CHEBI:16668 PNNL_ID hypoxanthine CHEBI:17368 PNNL_ID inosine CHEBI:17596 PNNL_ID L-alanine 5950 CHEBI:16977 PNNL_ID L-aspartic acid 5960 CHEBI:17053 PNNL_ID L-cysteine 5862 CHEBI:17561 PNNL_ID L-glutamic acid 33032 CHEBI:16015 PNNL_ID L-leucine 6106 CHEBI:15603 PNNL_ID L-lysine 5962 CHEBI:18019 PNNL_ID L-methionine 6137 CHEBI:16643 PNNL_ID L-ornithine CHEBI:15729 PNNL_ID L-serine 5951 CHEBI:17115 PNNL_ID L-sorbose 6904 CHEBI:13172 PNNL_ID L-threonine 6288 CHEBI:16857 PNNL_ID L-tyrosine 6057 CHEBI:17895 PNNL_ID L-valine 6287 CHEBI:16414 PNNL_ID maltitol 493591 HMDB02928 PNNL_ID mucic acid 607 CHEBI:30852 PNNL_ID oxalic acid 18676629 CHEBI:16995 PNNL_ID palmitic acid 5326436 CHEBI:15756 PNNL_ID pantothenic acid 11777341 CHEBI:46905 PNNL_ID porphine 5481287 CHEBI:8337 PNNL_ID putrescine 1045 CHEBI:17148 PNNL_ID pyruvic acid CHEBI:32816 PNNL_ID ribonic acid gamma-lactone HMDB01900 PNNL_ID spermidine CHEBI:16610 PNNL_ID spermine 1103 CHEBI:15746 PNNL_ID stearic acid CHEBI:28842 PNNL_ID tagatose 92092 CHEBI:47693 PNNL_ID thymine 1135 CHEBI:17821 PNNL_ID tyramine 5610 CHEBI:15760 PNNL_ID uridine 5'-monophosphate 45109815 CHEBI:16695 PNNL_ID xanthine CHEBI:17712 PNNL_ID xylitol CHEBI:17151 PNNL_ID 5-deoxy-5-(methylthio)adenosine guanosine-5-monophosphate uridine 5-monophosphate Unidentified_01 Unidentified_02 Unidentified_03 Unidentified_04 Unidentified_05 Unidentified_06 Unidentified_07 Unidentified_08 Unidentified_09 Unidentified_10 Unidentified_11 Unidentified_12 Unidentified_13 Unidentified_14 Unidentified_15 Unidentified_16 Unidentified_17 Unidentified_18 Unidentified_19 Unidentified_20 Unidentified_21 Unidentified_22 Unidentified_23 Unidentified_24 ribonic acid ?-lactone pyrophosphate METABOLITES_END #END