#METABOLOMICS WORKBENCH Tom_Metz_20140630_9826741_mwtab.txt DATATRACK_ID:133 STUDY_ID:ST000083 ANALYSIS_ID:AN000135 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 A Multi-Omic View of Host-Pathogen-Commensal Interplay in Salmonella-Mediated ST:STUDY_TITLE Infection ST:STUDY_TYPE Timecourse of Infection ST:STUDY_SUMMARY The potential for commensal microorganisms indigenous to a host (the ST:STUDY_SUMMARY or microbiota) to alter infection outcome by influencing host-pathogen ST:STUDY_SUMMARY is largely unknown. We used a multi-omics systems approach, ST:STUDY_SUMMARY proteomics, metabolomics, glycomics, and metagenomics, to explore the molecular ST:STUDY_SUMMARY between the murine host, the pathogen Salmonella enterica serovar Typhimurium ST:STUDY_SUMMARY Typhimurium), and commensal gut microorganisms during intestinal infection with ST:STUDY_SUMMARY Typhimurium. We find proteomic evidence that S. Typhimurium thrives within the ST:STUDY_SUMMARY 129/SvJ mouse gut without antibiotic pre-treatment, inducing inflammation and ST:STUDY_SUMMARY the intestinal microbiome (e.g., suppressing Bacteroidetes and Firmicutes while ST:STUDY_SUMMARY growth of Salmonella and Enterococcus). Alteration of the host microbiome ST:STUDY_SUMMARY structure was highly correlated with gut environmental changes, including the ST:STUDY_SUMMARY of metabolites normally consumed by commensal microbiota. Finally, the less ST:STUDY_SUMMARY phase of S. Typhimuriums lifecycle was investigated, and both proteomic and ST:STUDY_SUMMARY evidence suggests S. Typhimurium may take advantage of increased fucose ST:STUDY_SUMMARY to metabolize fucose while growing in the gut. The application of multiple ST:STUDY_SUMMARY measurements to Salmonella-induced intestinal inflammation provides insights ST:STUDY_SUMMARY complex molecular strategies employed during pathogenesis between host, ST:STUDY_SUMMARY and the microbiome. 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 4 ST:TOTAL_SUBJECTS 30 #SUBJECT SU:SUBJECT_TYPE Animal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:GENOTYPE_STRAIN 129/SvJ SU:AGE_OR_AGE_RANGE 6- to 8-week-old SU:GENDER Female SU:ANIMAL_ANIMAL_SUPPLIER Jackson Laboratories,Bar Harbor, ME SU:ANIMAL_HOUSING specific pathogen-free conditions in filter-top cages SU:ANIMAL_FEED food provided ad libitum SU:ANIMAL_WATER sterile water 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_Microbiome.002 Infection:Control | Experimental Group:Control Group 1 | Harvest Day:1 SUBJECT_SAMPLE_FACTORS - SBEP_Microbiome.001 Infection:Control | Experimental Group:Control Group 1 | Harvest Day:-1 SUBJECT_SAMPLE_FACTORS - SBEP_Microbiome.005 Infection:Control | Experimental Group:Control Group 1 | Harvest Day:10 SUBJECT_SAMPLE_FACTORS - SBEP_Microbiome.006 Infection:Control | Experimental Group:Control Group 1 | Harvest Day:14 SUBJECT_SAMPLE_FACTORS - SBEP_Microbiome.007 Infection:Control | Experimental Group:Control Group 1 | Harvest Day:21 SUBJECT_SAMPLE_FACTORS - SBEP_Microbiome.008 Infection:Control | Experimental Group:Control Group 1 | Harvest Day:28 SUBJECT_SAMPLE_FACTORS - SBEP_Microbiome.003 Infection:Control | Experimental Group:Control Group 1 | Harvest Day:3 SUBJECT_SAMPLE_FACTORS - SBEP_Microbiome.004 Infection:Control | Experimental Group:Control Group 1 | Harvest Day:6 SUBJECT_SAMPLE_FACTORS - SBEP_Microbiome.010 Infection:Control | Experimental Group:Control Group 2 | Harvest Day:1 SUBJECT_SAMPLE_FACTORS - SBEP_Microbiome.009 Infection:Control | Experimental Group:Control Group 2 | Harvest Day:-1 SUBJECT_SAMPLE_FACTORS - SBEP_Microbiome.013 Infection:Control | Experimental Group:Control Group 2 | Harvest Day:10 SUBJECT_SAMPLE_FACTORS - SBEP_Microbiome.014 Infection:Control | Experimental Group:Control Group 2 | Harvest Day:14 SUBJECT_SAMPLE_FACTORS - SBEP_Microbiome.015 Infection:Control | Experimental Group:Control Group 2 | Harvest Day:21 SUBJECT_SAMPLE_FACTORS - SBEP_Microbiome.016 Infection:Control | Experimental Group:Control Group 2 | Harvest Day:28 SUBJECT_SAMPLE_FACTORS - SBEP_Microbiome.011 Infection:Control | Experimental Group:Control Group 2 | Harvest Day:3 SUBJECT_SAMPLE_FACTORS - SBEP_Microbiome.012 Infection:Control | Experimental Group:Control Group 2 | Harvest Day:6 SUBJECT_SAMPLE_FACTORS - SBEP_Microbiome.018 Infection:Infected | Experimental Group:Experimental 1 | Harvest Day:1 SUBJECT_SAMPLE_FACTORS - SBEP_Microbiome.017 Infection:Infected | Experimental Group:Experimental 1 | Harvest Day:-1 SUBJECT_SAMPLE_FACTORS - SBEP_Microbiome.021 Infection:Infected | Experimental Group:Experimental 1 | Harvest Day:10 SUBJECT_SAMPLE_FACTORS - SBEP_Microbiome.022 Infection:Infected | Experimental Group:Experimental 1 | Harvest Day:14 SUBJECT_SAMPLE_FACTORS - SBEP_Microbiome.023 Infection:Infected | Experimental Group:Experimental 1 | Harvest Day:21 SUBJECT_SAMPLE_FACTORS - SBEP_Microbiome.024 Infection:Infected | Experimental Group:Experimental 1 | Harvest Day:28 SUBJECT_SAMPLE_FACTORS - SBEP_Microbiome.019 Infection:Infected | Experimental Group:Experimental 1 | Harvest Day:3 SUBJECT_SAMPLE_FACTORS - SBEP_Microbiome.020 Infection:Infected | Experimental Group:Experimental 1 | Harvest Day:6 SUBJECT_SAMPLE_FACTORS - SBEP_Microbiome.026 Infection:Infected | Experimental Group:Experimental 2 | Harvest Day:1 SUBJECT_SAMPLE_FACTORS - SBEP_Microbiome.025 Infection:Infected | Experimental Group:Experimental 2 | Harvest Day:-1 SUBJECT_SAMPLE_FACTORS - SBEP_Microbiome.029 Infection:Infected | Experimental Group:Experimental 2 | Harvest Day:10 SUBJECT_SAMPLE_FACTORS - SBEP_Microbiome.030 Infection:Infected | Experimental Group:Experimental 2 | Harvest Day:14 SUBJECT_SAMPLE_FACTORS - SBEP_Microbiome.027 Infection:Infected | Experimental Group:Experimental 2 | Harvest Day:3 SUBJECT_SAMPLE_FACTORS - SBEP_Microbiome.028 Infection:Infected | Experimental Group:Experimental 2 | Harvest Day:6 #COLLECTION CO:COLLECTION_SUMMARY Feces collected and frozen at seven time points post infection and one time CO:COLLECTION_SUMMARY pre-infection CO:COLLECTION_PROTOCOL_COMMENTS On the day prior to infection (day -1), and at seven time points CO:COLLECTION_PROTOCOL_COMMENTS mice from one cage were transferred to a clean cage and fecal samples produced CO:COLLECTION_PROTOCOL_COMMENTS that time were collected, pooled, and immediately frozen at -80° C. Samples CO:COLLECTION_PROTOCOL_COMMENTS collected on days -1, 1, 3, 6, 10, 14, 21, and 28. CO:SAMPLE_TYPE Feces CO:COLLECTION_METHOD Fecal matter collection CO:COLLECTION_LOCATION mice from one cage were transferred to a clean cage and fecal samples produced CO:COLLECTION_FREQUENCY days -1, 1, 3, 6, 10, 14, 21, and 28 relative to day of infection (day 0) CO:COLLECTION_TIME days -1, 1, 3, 6, 10, 14, 21, and 28 relative to day of infection (day 0) CO:VOLUMEORAMOUNT_COLLECTED four to 25 fecal pellets per pooled sample CO:STORAGE_CONDITIONS frozen at -80° C #TREATMENT TR:TREATMENT_SUMMARY Experimental: Infected with 1.6 x 10^8 CFU S. Typhimurium | mouse | Control: TR:TREATMENT_SUMMARY with equal volume saline solution TR:TREATMENT_PROTOCOL_COMMENTS A final inoculum of 1.6 x 10^8 CFU S. Typhimurium/mouse was delivered by oral TR:TREATMENT_PROTOCOL_COMMENTS to 10 mice (two cages of five mice each = Salmonella-infected). An equal number TR:TREATMENT_PROTOCOL_COMMENTS mock-infected animals (two cages of five mice each = control) were administered TR:TREATMENT_PROTOCOL_COMMENTS equal volume of sterile saline. Our infecting dose (10^8 CFU/mouse) aimed to TR:TREATMENT_PROTOCOL_COMMENTS a persistent infection that would ensure observation of S. Typhimurium proteins TR:TREATMENT_PROTOCOL_COMMENTS downstream analyses. TR:TREATMENT Biotic / Abiotic TR:TREATMENT_COMPOUND S. Typhimurium / Saline TR:TREATMENT_DOSE 1.6 x 10^8 CFU S. Typhimurium/mouse / equal volume saline solution TR:TREATMENT_VEHICLE Saline TR:ANIMAL_FASTING 14 h before orogastric inoculation TR:ANIMAL_ENDP_EUTHANASIA Carbon Dioxide Asphixiation followed by Cervical Dislocaton TR:ANIMAL_ENDP_TISSUE_COLL_LIST Feces #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Feces thawed, buffer added, vortexed, filtered and centrifuged after which SP:SAMPLEPREP_SUMMARY subjected to further centrifugation and chemical derivatization SP:SAMPLEPREP_PROTOCOL_COMMENTS After thawing, 150 mM ammonium bicarbonate buffer was added to the sample SP:SAMPLEPREP_PROTOCOL_COMMENTS 1-2.5 ml based upon starting weight; volumes were recorded and used for SP:SAMPLEPREP_PROTOCOL_COMMENTS normalization), which was subsequently vortexed to disrupt fecal pellets. The SP:SAMPLEPREP_PROTOCOL_COMMENTS slurry was filtered through a 70 mm sieve to separate and remove large debris SP:SAMPLEPREP_PROTOCOL_COMMENTS undigested food particles). Filtrate was centrifuged (900 x g for 10 min), and SP:SAMPLEPREP_PROTOCOL_COMMENTS protein-rich pellet thought to contain cellular material was retained as P1. SP:SAMPLEPREP_PROTOCOL_COMMENTS supernatant was centrifuged to further clarify the sample (15,000 x g for 10 SP:SAMPLEPREP_PROTOCOL_COMMENTS The pellet was retained as P2 and the supernatant retained as SN2. All SP:SAMPLEPREP_PROTOCOL_COMMENTS and reagents used in metabolomics analyses were purchased from Sigma-Aldrich SP:SAMPLEPREP_PROTOCOL_COMMENTS Louis, MO), except for ammonium bicarbonate (Merck, Darmstadt, Germany), SP:SAMPLEPREP_PROTOCOL_COMMENTS of fatty acid methyl esters (FAMEs) and deuterated myristic acid (Agilent SP:SAMPLEPREP_PROTOCOL_COMMENTS Santa Clara, CA). Deionized and purified water was used to prepare buffer and SP:SAMPLEPREP_PROTOCOL_COMMENTS solutions (Nanopure Infinity ultrapure water system, Barnstead, Newton, WA). SP:SAMPLEPREP_PROTOCOL_COMMENTS samples (see Fecal sample preparation) were transferred to 0.6 ml SP:SAMPLEPREP_PROTOCOL_COMMENTS tubes, and water soluble metabolites were extracted with four volumes of SP:SAMPLEPREP_PROTOCOL_COMMENTS (-20° C) chloroform: methanol mixture (2:1). After separating the two phases SP:SAMPLEPREP_PROTOCOL_COMMENTS centrifugation (12,000 x g, 5 min), the upper aqueous layers were transferred SP:SAMPLEPREP_PROTOCOL_COMMENTS glass vials and dried under a vacuum concentrator (SpeedVac; Thermo Scientific, SP:SAMPLEPREP_PROTOCOL_COMMENTS MA). All extracted metabolites were subjected to chemical derivatization to SP:SAMPLEPREP_PROTOCOL_COMMENTS their stability and volatility during GC-MS analysis. Methoxyamine in pyridine SP:SAMPLEPREP_PROTOCOL_COMMENTS mg/ml) was added to each dried sample, and incubated at 37° C with shaking for SP:SAMPLEPREP_PROTOCOL_COMMENTS min to protect carbonyl groups and reduce the number of tautomeric peaks. SP:SAMPLEPREP_PROTOCOL_COMMENTS (trimethylsilyl) trifluoroacetamide (MSTFA) with 1% trimethylchlorosilane SP:SAMPLEPREP_PROTOCOL_COMMENTS was then added, followed by incubation at 37° C with shaking for 30 min to SP:SAMPLEPREP_PROTOCOL_COMMENTS hydroxyl and amine groups to trimethylsilyated (TMS) forms. The samples were SP:SAMPLEPREP_PROTOCOL_COMMENTS allowed to cool to room temperature and were analyzed using gas chromatography SP:PROCESSING_METHOD Homogenization, Filtration, Centrifugation SP:EXTRACTION_METHOD SN2 samples (see Fecal sample preparation) were transferred to 0.6 ml SP:EXTRACTION_METHOD tubes, and water soluble metabolites were extracted with four volumes of SP:EXTRACTION_METHOD (-20° C) chloroform: methanol mixture (2:1). After separating the two phases SP:EXTRACTION_METHOD centrifugation (12,000 x g, 5 min), the upper aqueous layers were transferred SP:EXTRACTION_METHOD glass vials and dried under a vacuum concentrator (SpeedVac; Thermo Scientific, SP:EXTRACTION_METHOD MA). SP:EXTRACT_ENRICHMENT dried under a vacuum concentrator SP:EXTRACT_STORAGE dried under a vacuum concentrator SP:SAMPLE_RESUSPENSION Methoxyamine in pyridine (30 mg/ml) SP:SAMPLE_DERIVATIZATION Methoxyamine in pyridine (30 mg/ml), N-methyl-N- (trimethylsilyl) SP:SAMPLE_DERIVATIZATION (MSTFA) with 1% trimethylchlorosilane (TMCS) #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 0.450.5 mL/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 6/24/11 to 6/25/11 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 GC-MS raw data files from each Experiment were processed using the Metabolite MS:MS_COMMENTS software, version 2.0.6 beta. Briefly, Agilent.D files were converted to netCDF MS:MS_COMMENTS using Agilent Chemstation, followed by conversion to binary files using MS:MS_COMMENTS Detector. Retention indices of detected metabolites were calculated based on MS:MS_COMMENTS analysis of the FAMEs mixture, followed by their chromatographic alignment MS:MS_COMMENTS all analyses after deconvolution. Metabolites were initially identified by MS:MS_COMMENTS experimental spectra to an augmented version of FiehnLib (i.e., the Agilent MS:MS_COMMENTS Metabolomics Retention Time Locked (RTL) Library, containing spectra and MS:MS_COMMENTS retention indices for over 700 metabolites), using a Metabolite Detector match MS:MS_COMMENTS threshold of 0.6 (combined retention index and spectral probability). All MS:MS_COMMENTS identifications were manually validated to reduce deconvolution errors during MS:MS_COMMENTS data-processing and to eliminate false identifications. The NIST 08 GC-MS MS:MS_COMMENTS was also used to cross validate the spectral matching scores obtained using the MS:MS_COMMENTS library and to provide identifications of unmatched metabolites. The three most MS:MS_COMMENTS fragment ions in the spectra of each identified metabolite were automatically MS:MS_COMMENTS by Metabolite Detector, and their summed abundances were integrated across the MS:MS_COMMENTS elution profile; fragment ions due to trimethylsilylation (i.e. m/z 73 and 147) MS:MS_COMMENTS excluded from the determination of metabolite abundance. Features resulting MS:MS_COMMENTS GC column bleeding were removed from the data matrices prior to further data MS:MS_COMMENTS and analysis. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Peak area MS_METABOLITE_DATA_START Samples SBEP_Microbiome.002 SBEP_Microbiome.001 SBEP_Microbiome.005 SBEP_Microbiome.006 SBEP_Microbiome.007 SBEP_Microbiome.008 SBEP_Microbiome.003 SBEP_Microbiome.004 SBEP_Microbiome.010 SBEP_Microbiome.009 SBEP_Microbiome.013 SBEP_Microbiome.014 SBEP_Microbiome.015 SBEP_Microbiome.016 SBEP_Microbiome.011 SBEP_Microbiome.012 SBEP_Microbiome.018 SBEP_Microbiome.017 SBEP_Microbiome.021 SBEP_Microbiome.022 SBEP_Microbiome.023 SBEP_Microbiome.024 SBEP_Microbiome.019 SBEP_Microbiome.020 SBEP_Microbiome.026 SBEP_Microbiome.025 SBEP_Microbiome.029 SBEP_Microbiome.030 SBEP_Microbiome.027 SBEP_Microbiome.028 Factors Infection:Control | Experimental Group:Control Group 1 | Harvest Day:1 Infection:Control | Experimental Group:Control Group 1 | Harvest Day:-1 Infection:Control | Experimental Group:Control Group 1 | Harvest Day:10 Infection:Control | Experimental Group:Control Group 1 | Harvest Day:14 Infection:Control | Experimental Group:Control Group 1 | Harvest Day:21 Infection:Control | Experimental Group:Control Group 1 | Harvest Day:28 Infection:Control | Experimental Group:Control Group 1 | Harvest Day:3 Infection:Control | Experimental Group:Control Group 1 | Harvest Day:6 Infection:Control | Experimental Group:Control Group 2 | Harvest Day:1 Infection:Control | Experimental Group:Control Group 2 | Harvest Day:-1 Infection:Control | Experimental Group:Control Group 2 | Harvest Day:10 Infection:Control | Experimental Group:Control Group 2 | Harvest Day:14 Infection:Control | Experimental Group:Control Group 2 | Harvest Day:21 Infection:Control | Experimental Group:Control Group 2 | Harvest Day:28 Infection:Control | Experimental Group:Control Group 2 | Harvest Day:3 Infection:Control | Experimental Group:Control Group 2 | Harvest Day:6 Infection:Infected | Experimental Group:Experimental 1 | Harvest Day:1 Infection:Infected | Experimental Group:Experimental 1 | Harvest Day:-1 Infection:Infected | Experimental Group:Experimental 1 | Harvest Day:10 Infection:Infected | Experimental Group:Experimental 1 | Harvest Day:14 Infection:Infected | Experimental Group:Experimental 1 | Harvest Day:21 Infection:Infected | Experimental Group:Experimental 1 | Harvest Day:28 Infection:Infected | Experimental Group:Experimental 1 | Harvest Day:3 Infection:Infected | Experimental Group:Experimental 1 | Harvest Day:6 Infection:Infected | Experimental Group:Experimental 2 | Harvest Day:1 Infection:Infected | Experimental Group:Experimental 2 | Harvest Day:-1 Infection:Infected | Experimental Group:Experimental 2 | Harvest Day:10 Infection:Infected | Experimental Group:Experimental 2 | Harvest Day:14 Infection:Infected | Experimental Group:Experimental 2 | Harvest Day:3 Infection:Infected | Experimental Group:Experimental 2 | Harvest Day:6 2-ketohexanoic acid 215391.9474 147693.0000 260416.9507 238578.9051 258215.9919 141766.0000 204192.2647 226763.5252 232633.7018 138502.0000 273115.2449 281097.1564 159066.0000 253941.8655 279212.0127 235987.2872 280557.2236 337277.4385 227069.3030 255327.2767 199692.2513 215791.4762 255169.8928 238490.9722 216317.5659 235695.5436 182947.6058 317155.8569 178254.7055 186970.0384 Galactinol 30667.0000 45616.0000 7624.0000 3084.0000 33044.5000 53577.5000 4758.5000 40715.0000 4413.5000 9398.5000 4874.0000 3103.5000 2294.5000 3581.0000 28288.5000 512994.0000 1849351.5000 269076.0000 1285834.5000 188388.0000 745325.5000 4592.5000 23543.0000 898379.5000 595965.0000 597117.0000 341873.0000 gluconic acid lactone 5653506.2990 3954397.4950 4593253.3080 152455.1188 323058.1516 7090810.9320 8772390.8390 4663944.4520 8767443.3850 4522412.1070 hypoxanthine 1106298.6810 1228340.0490 1146394.9600 950377.2619 939610.5010 973915.3610 358305.3504 16625.0000 1101904.7930 1067096.0420 962346.5277 1873103.7950 1995752.9040 1645528.8580 419270.2829 1170363.9010 1472080.6270 1330519.7250 642958.1822 956476.0000 304158.0000 173300.0000 317387.0809 227853.5737 305774.0000 254906.0000 124738.0000 511432.0000 292674.0000 308420.0000 inositol 64017.0070 221000.1798 90048.0000 84091.6119 109104.9448 48309.1262 625507.8506 21524.8943 87592.0000 72076.0000 75778.0000 64629.3682 60817.1177 66933.0000 74417.4396 75648.7001 70985.7348 67450.0000 6726258.8960 21797994.2500 24839341.7800 10804141.3100 55563.9908 60454.5841 22443627.9800 19461391.6800 14069969.5600 29916434.2100 33090137.6300 24045791.0500 lactose 5488.0000 49563.0000 7383.0000 3374.0000 6198.0000 14521.5000 2487.5000 50578.5000 2050.0000 16852.5000 41961.5000 7427.0000 8563.0000 8289.5000 62292.0000 8050.0000 2026.5000 6687.5000 2424891.0000 2840281.0000 903258.5000 2913762.5000 2257966.0000 2661470.5000 951.0000 14874.5000 3772711.5000 3436779.0000 2955622.5000 1834552.0000 L-glutamic acid 1322183.0000 4634850.0000 3367625.0000 1208713.0000 2801329.0000 4975321.0000 4505804.0000 3449898.0000 282390.0000 2967113.0000 3378785.0000 3258184.0000 4103602.0000 3115084.0000 2107178.0000 2286819.0000 1309087.0000 3994508.0000 4379188.0000 4313190.0000 778971.0000 1282452.0000 3871326.0000 1166823.0000 422372.0000 3705203.0000 2862261.0000 2539128.0000 3170576.0000 2115712.0000 L-histidine 3428741.9460 5653506.2990 3954397.4950 4593253.3080 5172192.0390 9678382.5060 6255364.5600 5361014.5480 7090810.9320 8772390.8390 4663944.4520 8767443.3850 4522412.1070 L-homoserine 113414.3322 178550.2423 229332.9648 133933.6261 98847.1740 173863.2873 36790.1303 38125.0000 190869.4666 205157.2922 150696.5890 223919.2067 200374.4454 157996.2109 52327.0000 212814.9525 219274.6752 202648.6340 186949.8208 82080.5237 47661.7970 47591.4010 59539.0000 57762.6536 104057.3354 65309.1520 57659.0842 94183.6397 69351.0000 98716.9186 melibiose 14088.5000 12466.0000 4268.5000 6666.0000 990.0000 4510.0000 4411.5000 3907.0000 2242869.5000 6659044.0000 987308.5000 4572262.0000 1369913.0000 3056349.5000 10672.5000 3745028.5000 2488504.0000 2658464.0000 1348533.0000 methionine sulfoxide 303089.4348 246238.0000 196945.8237 266569.5000 162743.5000 158751.9618 201292.1750 55756.6238 254890.6424 113922.6876 167251.1190 368582.0194 157880.2775 336798.0239 80666.3874 207358.6877 272065.4594 254522.5154 217531.6399 259247.1800 76508.9949 169278.2770 432968.0511 147841.7152 351318.5634 240420.6594 395650.4386 271433.1225 418650.8645 145543.3483 raffinose 30470.0000 8379.0000 5998.0000 7676.0000 9168.0000 14403.0000 9182.0000 9921.0000 6033.0000 6012.0000 3039.0000 6080.0000 2078.0000 4118.0000 8572.0000 5803107.0000 11731037.0000 2252028.0000 9491301.0000 3003515.0000 7115220.0000 5050.0000 26303.0000 8136145.0000 7267019.0000 6545848.0000 2828983.0000 shikimic acid 1138789.2770 888332.8260 643769.1444 424332.9996 843085.5664 669868.2063 416722.7231 966912.9809 962065.3568 810530.7703 urea 83551.4829 97233.4965 83174.2909 79953.0236 69364.7798 75814.7769 93050.9337 25414.0000 116265.5154 51193.1456 53368.7012 71670.9352 72359.6726 54666.1879 27128.7316 111191.8088 58084.2329 71764.4081 77580.6242 167422.4875 173669.8480 656186.3348 42078.8615 75955.5673 248055.5500 337172.7049 1091191.8150 618727.4347 2325193.4160 662968.5050 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 2-ketohexanoic acid 159664 CHEBI:17308 PNNL_ID Galactinol 439451 CID45109775 PNNL_ID gluconic acid lactone 7027 CHEBI:16217 PNNL_ID hypoxanthine CHEBI:17368 PNNL_ID inositol 892 CHEBI:17268 PNNL_ID lactose 440995 CHEBI:17716 PNNL_ID L-glutamic acid 33032 CHEBI:16015 PNNL_ID L-histidine 6274 CHEBI:15971 PNNL_ID L-homoserine CHEBI:15699 PNNL_ID melibiose 440658 CID46905267 PNNL_ID methionine sulfoxide 847 CHEBI:49033 PNNL_ID raffinose 10542 CHEBI:16634 PNNL_ID shikimic acid 8742 CHEBI:16119 PNNL_ID urea CHEBI:16199 PNNL_ID METABOLITES_END #END