#METABOLOMICS WORKBENCH agoldman_20250918_123454 DATATRACK_ID:6450 STUDY_ID:ST004228 ANALYSIS_ID:AN007038 PROJECT_ID:PR002668 VERSION 1 CREATED_ON September 26, 2025, 4:27 pm #PROJECT PR:PROJECT_TITLE Aromatic Microbial Metabolite Hippuric Acid Potentiates Pro-Inflammatory PR:PROJECT_TITLE Responses in Macrophages through TLR-MyD88 Signaling and Lipid Remodeling PR:PROJECT_SUMMARY The gut microbiome generates a diverse array of metabolites that actively shape PR:PROJECT_SUMMARY host immunity, yet the pro-inflammatory potential of microbial metabolites PR:PROJECT_SUMMARY remains incompletely understood. Using a non-targeted, liquid PR:PROJECT_SUMMARY chromatography-tandem mass spectrometry (LC-MS/MS)-based metabolomics, we PR:PROJECT_SUMMARY identified hippuric acid, an aromatic gut microbe-derived metabolite, as a PR:PROJECT_SUMMARY potent enhancer of pro-inflammatory responses in Escherichia coli infection PR:PROJECT_SUMMARY model. Intraperitoneal administration of hippuric acid significantly heightened PR:PROJECT_SUMMARY pro-inflammatory responses, promoted innate immune cell activation, and reduced PR:PROJECT_SUMMARY survival in infected mice. Similar pro-inflammatory effects were observed in an PR:PROJECT_SUMMARY LPS-induced inflammation model. In vitro, hippuric acid selectively potentiated PR:PROJECT_SUMMARY M1-like macrophage polarization (LPS + IFNγ) but had no effect on M2-like PR:PROJECT_SUMMARY polarization (IL-4). Hippuric acid further augmented responses to multiple PR:PROJECT_SUMMARY myeloid differentiation primary response 88 (MyD88)-dependent toll-like receptor PR:PROJECT_SUMMARY (TLR) ligands, but not to TRIF-dependent TLR3, or to cytosolic innate immune PR:PROJECT_SUMMARY stimuli such as STING and NOD2 agonists, implicating TLR-MyD88 signaling as a PR:PROJECT_SUMMARY likely mechanism of action. Genetic deletion of MyD88 abrogated the PR:PROJECT_SUMMARY pro-inflammatory effects of hippuric acid both in vitro and in vivo, confirming PR:PROJECT_SUMMARY its dependence on the MyD88 pathway. Transcriptomic and lipidomic analyses PR:PROJECT_SUMMARY revealed that hippuric acid upregulated cholesterol biosynthesis and induced PR:PROJECT_SUMMARY lipid accumulation. Pharmacological reduction of cellular cholesterol using PR:PROJECT_SUMMARY fluvastatin or 25-hydroxycholesterol attenuated its pro-inflammatory effects. PR:PROJECT_SUMMARY Notably, hippuric acid also enhanced pro-inflammatory responses in human PR:PROJECT_SUMMARY macrophages, and its elevated levels correlated with increased sepsis mortality, PR:PROJECT_SUMMARY underscoring its clinical relevance. Together, these findings identify hippuric PR:PROJECT_SUMMARY acid as a previously unrecognized microbial-derived pro-inflammatory modulator PR:PROJECT_SUMMARY that links gut microbial metabolism, lipid remodeling, and innate immune PR:PROJECT_SUMMARY signaling, and offer new insights into its role in infection and inflammation. PR:INSTITUTE The Wistar Institute PR:LAST_NAME Shinde PR:FIRST_NAME Rahul PR:ADDRESS 3601 Spruce Street, Philadelphia, PA, 19104, USA PR:EMAIL rshinde@wistar.org PR:PHONE 215-898-3717 #STUDY ST:STUDY_TITLE Aromatic Microbial Metabolite Hippuric Acid Potentiates Pro-Inflammatory ST:STUDY_TITLE Responses in Macrophages through TLR-MyD88 Signaling and Lipid Remodeling - ST:STUDY_TITLE Non-targeted metabolomics of C57BL/6 mice infected with Escherichia coli ST:STUDY_SUMMARY Authors have investigated changes in microbial metabolites during bacterial ST:STUDY_SUMMARY infection. In this study, C57BL/6 mice were infected intraperitoneally with ST:STUDY_SUMMARY Escherichia coli (2 × 10⁷ CFU), and non-targeted metabolomics using liquid ST:STUDY_SUMMARY chromatography-tandem mass spectrometry (LC-MS/MS) was performed on sera ST:STUDY_SUMMARY collected 48 hours post-infection. The metabolomics screen revealed significant ST:STUDY_SUMMARY shifts in metabolic profiles, including altered levels of 2/3-hydroxybutyrate (a ST:STUDY_SUMMARY marker of fatty acid metabolism), hypoxanthine (a marker of purine metabolism), ST:STUDY_SUMMARY and N-acetylglutamine (involved in nitrogen metabolism), indicating broad ST:STUDY_SUMMARY metabolic reprogramming during infection. Among the most significantly altered ST:STUDY_SUMMARY metabolites, hippuric acid—an aromatic compound produced by microbial ST:STUDY_SUMMARY metabolism of dietary polyphenols and amino acids was markedly depleted, showing ST:STUDY_SUMMARY a 24-fold reduction in infected mice compared to uninfected controls (p < ST:STUDY_SUMMARY 0.001). These findings establish an association between bacterial infection and ST:STUDY_SUMMARY the depletion of hippuric acid, indicating its role in modulating immune ST:STUDY_SUMMARY responses during infection. ST:INSTITUTE The Wistar Institute ST:LAST_NAME Shinde ST:FIRST_NAME Rahul ST:ADDRESS 3601 Spruce Street, Philadelphia, PA, 19104, USA ST:EMAIL rshinde@wistar.org ST:PHONE 215-898-3717 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 #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 - QC-6 Treatment:n/a | Sample source:Serum RAW_FILE_NAME(raw_data)=m23-L034-QC-FullMS-6.raw SUBJECT_SAMPLE_FACTORS - QC-7 Treatment:n/a | Sample source:Serum RAW_FILE_NAME(raw_data)=m23-L034-QC-FullMS-7.raw SUBJECT_SAMPLE_FACTORS - QC-8 Treatment:n/a | Sample source:Serum RAW_FILE_NAME(raw_data)=m23-L034-QC-FullMS-8.raw SUBJECT_SAMPLE_FACTORS - QC-9 Treatment:n/a | Sample source:Serum RAW_FILE_NAME(raw_data)=m23-L034-QC-FullMS-9.raw SUBJECT_SAMPLE_FACTORS - C1 Treatment:Control | Sample source:Serum RAW_FILE_NAME(raw_data)=m23-L034-C1.raw SUBJECT_SAMPLE_FACTORS - C2 Treatment:Control | Sample source:Serum RAW_FILE_NAME(raw_data)=m23-L034-C2.raw SUBJECT_SAMPLE_FACTORS - C3 Treatment:Control | Sample source:Serum RAW_FILE_NAME(raw_data)=m23-L034-C3.raw SUBJECT_SAMPLE_FACTORS - C4 Treatment:Control | Sample source:Serum RAW_FILE_NAME(raw_data)=m23-L034-C4.raw SUBJECT_SAMPLE_FACTORS - C5 Treatment:Control | Sample source:Serum RAW_FILE_NAME(raw_data)=m23-L034-C5.raw SUBJECT_SAMPLE_FACTORS - E1 Treatment:E. coli | Sample source:Serum RAW_FILE_NAME(raw_data)=m23-L034-E1.raw SUBJECT_SAMPLE_FACTORS - E2 Treatment:E. coli | Sample source:Serum RAW_FILE_NAME(raw_data)=m23-L034-E2.raw SUBJECT_SAMPLE_FACTORS - E3 Treatment:E. coli | Sample source:Serum RAW_FILE_NAME(raw_data)=m23-L034-E3.raw SUBJECT_SAMPLE_FACTORS - E4 Treatment:E. coli | Sample source:Serum RAW_FILE_NAME(raw_data)=m23-L034-E4.raw SUBJECT_SAMPLE_FACTORS - E5 Treatment:E. coli | Sample source:Serum RAW_FILE_NAME(raw_data)=m23-L034-E5.raw #COLLECTION CO:COLLECTION_SUMMARY Serum samples were collected 48 hours post-infection CO:SAMPLE_TYPE Blood (serum) #TREATMENT TR:TREATMENT_SUMMARY Escherichia coli peritoneal infection was performed as previously described. TR:TREATMENT_SUMMARY Briefly, E. coli (ATCC 25922) was cultured in LB medium with 100 µg/mL TR:TREATMENT_SUMMARY ampicillin overnight at 37 °C with gentle shaking to an optical density at TR:TREATMENT_SUMMARY wavelength 600 nm of 0.5−0.6 to achieve a logarithmic growth phase. C57BL/6J TR:TREATMENT_SUMMARY or MyD88 KO mice of 8–12 weeks of age received an intraperitoneal (i.p.) TR:TREATMENT_SUMMARY injection of 100 μl PBS with 2 × 10^7 E. coli. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Polar metabolites were extracted from serum samples with 10-times the volume of SP:SAMPLEPREP_SUMMARY ice-cold 80% methanol spiked with 1 μM HA-13C6 as an internal standard for HA SP:SAMPLEPREP_SUMMARY in isotope dilution MS. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Thermo Vanquish CH:COLUMN_NAME SeQuant ZIC- pHILIC (150 x 2.1mm,5um) CH:SOLVENT_A 100% water; 20 mM ammonium carbonate; 5 µM medronic acid; 0.1% ammonium CH:SOLVENT_A hydroxide; pH 9.2 CH:SOLVENT_B 100% Acetonitrile CH:FLOW_GRADIENT 85% B for 2 min, 85% B to 20% B over 15 min, 20% B to 85% B over 0.1 min, CH:FLOW_GRADIENT and 85% B for 8.9 min CH:FLOW_RATE 0.2 ml/min CH:COLUMN_TEMPERATURE 45 #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive HF-X Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS All samples were analyzed by full MS with polarity switching. A quality control MS:MS_COMMENTS (QC) sample pool was also analyzed by data-dependent MS/MS with separate runs MS:MS_COMMENTS for positive and negative ion modes. Full MS scans were acquired at 120,000 MS:MS_COMMENTS resolution with a scan range of 50-750 m/z for positive ion mode and 65-975 m/z MS:MS_COMMENTS for negative ion mode. Data-dependent MS/MS scans were acquired for the top 10 MS:MS_COMMENTS highest intensity ions at 15,000 resolution with 1.0 m/z isolation width and MS:MS_COMMENTS stepped normalized collision energy (NCE) of 20/40/60. Global metabolite MS:MS_COMMENTS profiling was performed using Compound Discoverer 3.3 SP1 (Thermo Fisher MS:MS_COMMENTS Scientific). Periodic technical injections of the QC pool were used to correct MS:MS_COMMENTS metabolite quantifications over the course of a batch analysis. Serum data were MS:MS_COMMENTS normalized per volume serum injected, which was equivalent for all samples. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Peak Area MS_METABOLITE_DATA_START Samples QC-6 QC-7 QC-8 QC-9 C1 C2 C3 C4 C5 E1 E2 E3 E4 E5 Factors Treatment:n/a | Sample source:Serum Treatment:n/a | Sample source:Serum Treatment:n/a | Sample source:Serum Treatment:n/a | Sample source:Serum Treatment:Control | Sample source:Serum Treatment:Control | Sample source:Serum Treatment:Control | Sample source:Serum Treatment:Control | Sample source:Serum Treatment:Control | Sample source:Serum Treatment:E. coli | Sample source:Serum Treatment:E. coli | Sample source:Serum Treatment:E. coli | Sample source:Serum Treatment:E. coli | Sample source:Serum Treatment:E. coli | Sample source:Serum (+/-)11(12)-EET 330334129.6 392074383.1 396472094.9 420063312.6 383149323.7 313673400.4 396125661 305845100.7 473558621.4 663007984.3 515042555.9 655517730.7 324674596.7 317959679.8 (±)10(11)-EpDPA 118148573.7 118064763.8 118565621.9 116823556 113980423.3 101834244.8 101550080.4 108641859.4 121182248.6 238777963.6 205903430.3 320463860.1 92516296.83 89532629.29 (1S,3R,4R,5R)-1,3,4-trihydroxy-5-{[(2E)-3-(4-hydroxy-3-methoxyphenyl)prop-2-enoyl]oxy}cyclohexane-1-carboxylic acid 620733213.5 640703661.7 635011379.8 641598312.1 483245457.9 624242032.1 469421808.4 621377440.7 582849388.6 105866525.4 18882586.97 287634938.7 306108026.2 447503054 Linolenic Acid 696210604.1 701997053.5 699782628.6 704208075.5 662604659.7 424251548.5 863949764.7 432582392.7 907174787.1 1054203959 1266340698 1056623761 692978656.7 901727072.8 beta-Muricholic acid 197309630.7 201138050 201595023.2 198928842.2 230535155.6 790595826.5 310381320.8 183783913.4 430761459.2 20679730.55 104837448.9 95635396.46 57714517.84 26122493.91 1,5-Anhydro-D-glucitol 318686480.7 318637433.9 319222644.9 321278427.5 468746009.2 178794381.5 425017137.1 450440172.6 420026663.7 214777635.3 129186336.3 92457739.71 104976584.7 375399712.5 11(E)-Eicosenoic Acid 134660553.6 135853927.1 135808482.9 136879529.6 136707638.1 178024121.1 167893219.6 132333843.3 167486717.3 104429373.6 98376462.16 132526705 105632208.1 126657252.9 11(Z),14(Z)-Eicosadienoic acid 98074048.09 97625818.3 97693415.6 97369200.45 109279897.5 106441372.6 116091098.2 90770065.51 114130934.9 124995333.9 131567509.9 130221989.9 98517157.6 113542560.7 16-Hydroxyhexadecanoic acid 49114232.54 41473754.6 43781939.75 47050012.49 45962284.77 64346863.23 60117315.21 50525765.85 78021325.14 67696605.17 47355635.74 56348626.74 81018205.83 54707061.92 2,3-Dihydroxybenzoic acid 4838597.796 4702096.853 4870168.073 4870769.424 243365.1675 323027.8244 223884.9898 297811.3103 249574.6759 8727104.72 35220505 19412342.2 255289.0123 238735.6351 2/3-Hydroxybutyrate 780949887.2 791957446.7 781473153 788992691 168158437.1 200094018.9 201132472.5 325508771.5 256261396.1 1903826670 1024499441 1057908785 2551719105 1517006199 2'-Deoxycytidine 7282450.044 7297868.337 7314275.037 7153033.331 8354199.134 11939301.87 8157253.688 8722622.944 9212490.6 216062.1996 874449.3962 7315228.007 4954059.852 10317387.08 3-Hydroxy-3-methylglutaric acid 185010852.8 184696386.4 184918912.6 183883887.9 53972140.2 41759527.72 49037252.64 52151997.46 66247340.23 249933129.5 497337430.7 214614245.4 374879680.8 386091534 3-Hydroxybutyric acid 4182311934 3584399486 3882458354 4138206912 1427175333 2213103056 1165235369 989214420 2950233579 10231079295 3729991595 5939158559 11522386892 19507969000 3-Indoxyl sulphate 1352820582 1381275523 1377998814 1368492052 3331194659 1388056372 3096675729 2385283564 2337094076 507259734.3 304757950.1 526158682.8 283777887.8 368336516 3-methylphenylacetic acid 16702579.26 17159640.43 16697381.46 16565182.91 13536242.97 81345541.3 42145047.13 10695162.02 8679843.913 1837694.424 2561473.016 1758176.508 3691243.786 1416541.17 3-Phosphoglyceric acid 48245840.53 48083569.99 48303187.1 46737739.04 27365137.63 17864069.01 23726523.47 20985091.71 18639294.46 194902406.9 205939518.4 58519820.4 27066460.17 17206687.54 4-Methylphenol 11309088.25 11237364.51 11227753.58 10769915.77 15469224.99 12746115.79 11200573.27 12767318.13 13240499.57 2918620.692 10668697 5156352.24 8178019.027 6891815.25 5-Hydroxyindole-3-acetic acid 10832219.09 11065950.75 11131682.95 11028365.02 16267524.5 8761330.495 18633576.73 17720783.59 26347943.33 1729927.029 4988972.613 12106143.29 229286.3199 405235.4473 Adenosine diphosphate ribose 2571035.582 2519963.507 2489080.29 2528997.353 802586.3339 4211515.014 629538.7681 5989091.172 4016614.641 2417901.323 4762875.999 5828757.631 1133980.237 7892122.057 Allantoin 263574475.6 259781888.4 263623237.9 261237352.6 266331212.9 262680085.8 181730344.9 233877539.2 258185137.2 401044226 328675240.7 512739300.3 146794383 229960769.7 Arachidonic acid 562821032.7 662632793.2 663372342.1 714216548.2 862007311.4 274086571.4 709663709.4 443595951.6 762460840.3 819104160.5 1229674605 767089576.2 444427103.6 597792767.5 Cinnamoylglycine 5701677.679 5612458.805 5405428.858 5657528.334 5297963.68 4915509.504 17488546.47 11082070.07 13779626.04 403658.4147 578916.0022 1041036.982 320139.6951 350648.7775 cis-Aconitic acid 183952141.1 187088016.8 194670906.2 186462938.3 193159426.6 244868371.7 220720264.8 242830092.7 265148867.8 154068068.2 216515076.3 319359866.2 129391066.4 105625767.7 Citraconic Acid 37701417.81 41678078.08 41415191.89 41383158.87 32129846.94 17202313.3 29260658.56 35408142.52 30239825.31 35936335 23189735.44 29957343.23 33123308.61 40967170.34 Citric acid 2492237971 2448005520 2443388981 2467357062 2452875423 3172144778 2500382125 3155880042 2917746267 2798800711 2637985902 4187234727 1824704692 1666495216 D-2-Hydroxyglutaric acid 403820048.4 397574368.6 400902316.5 395931132.6 163212583.4 122699706.9 142703893.2 189979461.3 241529893.5 218491519.7 1587650689 171520621.9 648776511 482664193.6 Deoxycholic Acid 85367068.55 85765395.32 85398099.54 84448591.99 156848249.7 242546851.7 135683102.4 74055380.83 227094006.1 3923126.024 28911589.56 14029161.5 37174682.04 18451710.31 Deoxyuridine 6515496.062 6616661.41 6575437.512 6560615.906 9104699.367 12822637.51 7821385.741 8647386.822 10230379 554961.0287 378982.7599 4283401.011 10849644.34 10778977.15 DHAP/GAP 15272860.92 15283057.83 14846760.16 15004906.6 17188312.92 12704211.56 12602475.67 14397745.38 13892912.33 7328466.153 48824704.49 4162811.968 22570376.18 14279246.33 Dihomo-gamma-Linolenic Acid 124512876 144290519.3 142643200.7 146113402.5 188197274.4 95246648.73 170697641.8 147441841.6 188370198.6 201848024.2 157286016.8 212926156.3 83191659.82 123960530.7 DL-4-Hydroxyphenyllactic acid 156751235.3 157706967.6 157079459.5 157026752.1 131933251.6 180962754.9 110600113.5 135581583.7 141049185.7 110799530.5 376605054.4 201558335.2 95776330.13 96625012.65 Docosahexaenoic acid 1430211672 1422842629 1426729877 1421875917 1652754676 1051261870 1318839160 1233494195 1471002032 2145814239 2403665049 2166838076 1107758126 1526750179 D-Ribose 5-phosphate 22881780.34 23001572.57 22671932.11 22904715.15 29672702.77 23769558.95 31056199.59 32108735.27 27606911.43 6978616.274 20162654.26 7660470.635 34337740.84 29538067.99 D-Ribulose 5-phosphate 12836312.33 13087379.77 12811683.05 12963056.98 9499051.503 10501691.91 6206211.805 6394823.911 11840417.14 7416360.783 3396329.985 3463916.383 50192659.94 32674573.14 D-Saccharic acid 24871322.07 22279851.74 22692861.91 22685897.33 24905277.62 27274284.95 18548763.74 17170443.48 16860082 44781718.27 20730414.62 12425096.71 14959678.17 16207614.37 D-Sedoheptulose 7-phosphate 4837194.504 4692329.59 4652666.903 4641005.876 4026077.442 3219960.802 3630428.377 3837327.565 3523502.933 7574629.093 8750878.222 6886839.208 7094343.54 6501309.367 Ethyl myristate 8700200837 8769758161 8749847375 8773423409 6786370579 9284180542 9284970717 4253312083 9751014171 8193893259 12274847401 10679653517 8057051423 7796920450 Gluconic acid 194946200.3 217629879.7 193312498 192405375.3 142517243.6 108400322.8 164590802 139836742.8 122792650.1 259317222.3 220135509.7 266670915.4 109960989.4 177917525.6 Glucose 6-phosphate 34412690.74 34970504.66 33788400.83 33810079.18 29080868.1 21501474.17 23915626.86 22710983.3 24288024.97 78200306.66 69511042.16 57137065.55 41514888.69 20492557.01 Glycerol 250962180.7 246595317.9 245617480.3 256767962.7 211920678.1 235451632.5 258250059.8 291419167.9 398053267.8 210427556.6 391068054.8 273649122.8 335110902.2 272736098.2 Glycerol 3-phosphate 118171561.9 118068575.6 116841248.5 116720816.9 148863309.2 86654187.34 151212256 135603778.8 123312242.5 118394995.3 78107597.55 78323477.12 163753150.6 105008553.9 Glycerophospho-N-palmitoyl ethanolamine 45622765.51 45841289.67 45925721.91 45078063.24 45214793.22 55532153.1 44985427.55 42822492.81 51084152.87 40050873.61 15664775.84 29399188.59 49880991.74 71856681.01 Glycyl-L-leucine 5347221.862 5497666.119 5449460.456 5511109.657 9010078.45 6402854.758 8656984.185 5694209.403 8996942.765 6329007.427 4403920.718 6339019.1 3457268.083 4975147.443 Hippuric Acid 177928419.4 191055861.8 195360226.1 190597020.8 185028048.7 226071810.4 392164584.8 388776894.7 354073262 14146021.24 27020427.77 37419621.07 6309820.782 4286490.37 Indole-3-pyruvic acid 129493586.6 130122342.1 129575925.7 127553205.1 176101464.8 366348750.9 207043244.2 332872073.9 327057935.5 17695216.06 15752123.93 6143063.167 74717578.29 135246380.7 Inosine 7229107.327 7122196.714 7204995.47 7204474.487 15235808.46 3456915.493 8117538.89 27251083.01 27437312.55 195863.1969 153238.129 178028.3224 6606741.323 3950073.842 Jasmonic acid 7082157.91 8020312.293 7264885.359 7080812.141 13453845.95 23489111.88 21050165.71 13819860.55 19455093.51 2889010.654 2257382.261 3001493.722 25898434.45 17100271.57 L-Anserine 16410045.82 16236807.84 16367028.41 16783880.53 3762408.325 4461360.588 3446771.993 4370853.218 4696299.414 28211462.93 29207030.97 67352436.45 3672283.342 3852077.142 L-Ascorbic acid 2-sulfate 219413964.3 220860968.3 222688060.9 219858148.8 170302310.6 77779503.54 147770657.6 157057086.7 182299979.8 395388131.4 528769393.9 632850502.9 128106639.5 243478949.5 L-Aspartic acid 233960253.1 230178196.1 231255702.3 229898128.6 331224767.7 253510340.3 334789231.3 294569146.5 238118001.8 204319365.7 75113493.15 131278482.5 164511910.3 72878974.63 L-Cysteine-S-sulfate 68591767.15 68529352.71 70772483.85 72098768.48 28754983.26 147095905.1 39008169.32 32303792.47 40613875.87 11779452.36 867964.4048 18319547.28 267701251.8 63507129.77 L-Cysteinesulfinic acid 22717717.05 22964965.29 22919333.36 23099867.57 14267268.83 13135205.78 23829078.76 17834368.75 21977093.46 30633498.58 31425433.87 46103456.6 13460590.87 32404162.71 L-Glutathione oxidized 20034208.81 18818145.96 18577526.51 19306798.85 17175255.65 8060998.304 10421654.6 17342591.14 13928867.53 16002415.04 55109079.71 5025979.702 7827325.873 15274493.48 L-Iditol 48017555 47973131.69 48525934.4 48148059.77 64582419.5 53506581.32 63387126.7 65200097.09 51394891.63 17442904.37 31811297.47 28212109.57 22300020.34 24546134.86 Linoleic acid 7830000962 7892453521 7864830851 7912116363 8220210680 7422812702 8401181240 6887940525 8654367426 12283065599 13906405451 11186148642 7074516353 9479573042 L-Lactic acid 51252363710 51571926443 51607540932 51801803018 48473438126 39243241373 49585591172 59856523600 51220904949 29487101070 37327870613 31304790138 60868875557 57270670942 Malic acid 1881074024 1860465131 1874721471 1846811909 1514794608 1102725614 1927781949 2237810762 2361043569 1443070956 2736783884 1260538049 2978457766 1790977380 Malonic acid 56017422.02 55536242.65 55655356.69 55742973.31 12101354.98 12514119.11 10435267.94 12666978.33 14322229.73 164874401.4 118978652.9 232418244.2 115710967.8 81600293.38 Methyl indole-3-acetate 83665520.99 84774847.36 83684877.06 83475735.99 173811169.2 57132805.52 158767390.9 210115265.5 153031210.1 1289499.553 1253011.199 2235416.883 2963178.219 11126286.04 Methylsuccinic acid 50820775.22 51655436.89 51512890.5 51793214.58 52286893.6 45791749.79 57524074.09 67524401.55 67593405.5 11115657.86 9650794.92 13791721.11 23426586.84 43763754.77 Myo-inositol 33509586.44 33700625.88 34214816.81 34360621.87 24131209.67 28039929.1 34976528.58 37384342.44 32300329.28 43848679.04 38074653.61 59174432.85 15819406.28 21861603.47 Myristic acid 658167185.6 810462296.5 819092471.7 835514797.6 552763922 937131000.8 938615419.8 839591781.6 1522050829 907749081.9 1429177719 1096807020 1227122345 1034621743 N-alpha-Acetyl-L-asparagine 84226390.29 84411334.18 85485395.54 84892362.26 25662581.73 42978181.51 32813000.77 30197970.74 32858734.49 193238365.1 175054890.8 247155985.8 20713986.24 13364505.61 N-Acetyl-alpha-D-glucosamine 1-phosphate 4402762.931 4324214.637 4435413.42 4344539.339 4920602.621 4412325.936 3941885.393 4749453.86 3892201.199 7573270.928 4888391.876 4110988.114 7367023.419 5782907.471 N-Acetyl-1-aspartylglutamic acid 3925314.518 3947371.46 3934582.166 3886381.591 3251843.17 2881807.671 2509649.372 2719931.896 2738103.748 2852138.229 3630868.809 3882937.111 7155672.447 4520679.243 N-Acetyl-DL-tryptophan 38694194.94 39249610.08 39134128.3 38121785.13 45072450.67 81007490.12 39438245.66 72063527.11 78131064.49 4253130.436 20119193.44 11800876.91 31267399.28 21462890.73 N-Acetylglutamic acid 61080914.83 61008268.56 62321012.07 61518321.56 17985278.82 16874536.7 21974061.56 27975958.69 30168093.47 135443158 177783048.4 156732282 49446360.61 40338383.22 N-Acetyl-L-methionine 9679206.557 9568204.451 9644176.943 9660851.561 8482767.918 11055006.53 12303948.33 15634031.51 15089721.01 1826934.064 6733813.741 3123714.021 10699365.01 4918944.972 N-Acetyl-L-phenylalanine 34229160.4 34947625.99 34238009.39 34508564.58 21518876.85 45391996.56 34000208.3 38119424.87 36516805.79 22788190.16 55254869.14 30023942.99 25959555.81 27485024.76 N-Acetylneuraminic acid 22419712.27 22127930.87 22221425.81 22149665.5 8657656.201 12227341.83 9811083.091 9767091.896 9145450.043 34526251.33 39145555.69 128376006.2 15753620.5 11724647.84 N-Glycolylneuraminic acid 43385974.65 43343908.75 43441643.44 43307791.73 5174847.479 5220136.384 4909872.479 6186564.673 5093136.67 90779616.35 110284881.4 275407301.3 22349180.97 17391957.43 N-Isovalerylglycine 21116508.59 21000447.48 20757248.8 20098714.02 11787995.32 4323063.419 16830837.82 22387100.79 29039352.44 9728570.525 51637902.36 26277480.18 9565880.284 36312915.78 N-Tigloylglycine 13926474 14091435.1 14341216.03 14014698.65 7856176.152 1639642.017 6305036.68 15178171.16 13409485.69 6047666.961 36921233.62 35323856.71 4859438.288 15966627.34 O-Acetylserine 139693892.6 140226390 139851078 138430114.7 87287022.78 98219537.64 102260184.7 107261509.1 113962163.7 193064701.3 268117194.2 290440706.7 101795911.4 96051306.14 Oleic acid 9534627924 9646748806 9539202874 9590000822 10329736316 9081635889 10143031416 8495914634 11887248000 15885885265 15448253106 12584813856 8772338680 11998141648 Oleoyl-L-alpha-lysophosphatidic acid 463268437.7 464618536.6 465336978.7 464178047.8 536767902.5 377985858.4 535121268.1 514098979 579539224.1 352179434.6 164025406.9 306695535.2 322056334.1 441647927.1 Orotic acid 88223490.77 88371200.58 88059524.12 87969596.72 90423734.37 113735532.6 90555016.7 96011261.1 125510547.7 46489958.94 151192590.7 47912395.31 55662955.89 64014332.46 Orotidine 13069685.48 13106591.07 13108005.4 12935723.98 8014475.736 12519750.28 8724037.879 11200009.26 8891999.083 24276664.48 22573707.66 25998695.43 4725697.172 5246462.343 Palmitoleic acid 2278684737 2286347434 2315880036 2301246028 1478228011 1541270306 2386913103 1060227573 3896468766 4049218792 3292752092 4147815464 2594111978 3331450903 Pentadecanoic acid 110167055.4 90859141.03 110471977.1 111659509.3 111636836.8 110828519.3 92846110.4 58586975.35 141400786 105572502.4 250770311.9 103147804 92741311.84 111135994.8 Phenylacetylglycine 47351687.13 47368775.4 46991702.52 46622074.33 83768464.51 16489067.25 64239734.63 87766753.56 119464714.3 18388479.13 41043315.93 24057596.21 8963331.202 22651435.03 Phenyllactic acid 10808892.68 10727170.47 10555956.35 10112018.97 6436452.482 10300290.08 13442410.15 10667305.66 12364534.15 5750701.585 23552117.08 11518042.66 6639898.119 5766909.7 Pseudouridine 124668216.7 124469340 124857399.6 124609577.9 147990132.7 144676469.6 137609488.8 156253388.5 151338725.4 154385940.8 89024395.67 102220108.6 80164699.69 104026024.5 Pyroglutamic acid 1246673731 1238969017 1235184120 1235494333 634794132.5 1179360324 546846357.9 1064958552 1218337416 2475881145 1317331845 1262497493 1672524458 1273496864 Pyruvic acid 1713551505 1724206793 1748171415 1734363099 2083894516 2017745909 2096449678 2196258560 2010035232 899392053.6 321598299.2 1136591974 1811132170 1567998120 Quinolinic acid 3719799.463 3850807.346 3709005.396 3748030.59 1762963.112 1871729.376 1810718.483 3665414.446 3733415.003 6416427.547 5506412.146 4323188.428 1047799.649 4005330.273 Stearic acid 2551894004 2592014536 2571572375 2585110428 1604855628 3106305967 2343052754 1600218834 1757966510 1749522962 4219245947 4029256855 1950074596 1324667540 Succinic acid 810067008.6 809034988.7 810875057.6 802229272.6 503682062 300350234 606177954.8 904351627.2 698165149.9 185054931.6 1330806518 179129425.4 1112472747 1854540294 Taurine 7021929887 7066796754 7099758776 7108321898 6657195848 6642098881 6861921220 7523974593 6856225777 5996366342 4554331851 8653217694 5403221260 6148513016 Taurochenodeoxycholic Acid (sodium salt) 104406350.7 105030716 103418266.7 105485727.6 19177367.32 969586183.4 13004512.43 13156324.99 22196021.86 4715885.612 61638030.65 17472673.59 454891507.3 6624103.553 Tetradecanedioic acid 8388336.554 7873921.143 7922526.379 7995168.041 10228174.87 14014007.9 11620777.19 6497629.76 9821131.599 14734173 16972786.09 9951831.638 14760521.36 5048760.774 Theophylline 37396816.41 36123738.99 37175105.46 37412386.18 21982327.86 22252258.63 25305980.52 34450702.62 30306839.22 78699176.89 51749983.52 77316282.69 19989491.1 29522842.34 Thymine 99313297.52 99786574.01 100796155.7 100461693.6 111008756.6 210166922.6 86301265.17 110733330 148022879.6 52671437.8 52508064.33 146385258.6 150471205.7 164627347.1 trans-10-Heptadecenoic acid 115144668.4 101545920.1 102804348.6 114318805.4 141076472 124801931.1 159895408 89584863.87 185589133.3 183158579.4 146833243.6 172776444.6 108154566.8 151526470.6 trans-Cinnamic acid 20397416.89 20843496.36 20841591.21 20984326.67 20519132.77 24715786.99 21075462.77 21431892.66 22942046.63 20466665.96 18782349.34 15062266.13 26580349.99 25549614.17 Uric acid 1642690334 1651942444 1656431209 1663618237 1418679623 1512820075 1558885095 1558339960 1325999559 857263639.4 1963910419 947519994.7 1994292641 2177269233 Uridine 131303404.8 129870366.7 126863607.7 125682841.5 189544567.3 128868733.3 174872122.1 221514243.1 179396299.5 12904157.43 10441886.46 7940167.653 195497390.8 268033940.3 Xanthosine 26505823.4 26271338.44 26351098.56 25946570.46 22107019.2 12453587.68 22106707.48 41531195.48 22236571.94 418984.7939 587471.7715 71440.34084 33214132.47 31803781.89 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name m/z RT [min] (+/-)11(12)-EET 319.22796 1.314 (±)10(11)-EpDPA 343.228 1.298 (1S,3R,4R,5R)-1,3,4-trihydroxy-5-{[(2E)-3-(4-hydroxy-3-methoxyphenyl)prop-2-enoyl]oxy}cyclohexane-1-carboxylic acid 367.10562 8.425 Linolenic Acid 277.21765 1.308 beta-Muricholic acid 407.28055 2.773 1,5-Anhydro-D-glucitol 163.06124 6.515 11(E)-Eicosenoic Acid 309.28007 1.279 11(Z),14(Z)-Eicosadienoic acid 307.26445 1.285 16-Hydroxyhexadecanoic acid 271.22816 1.351 2,3-Dihydroxybenzoic acid 153.01937 11.833 2/3-Hydroxybutyrate 103.04004 2.794 2'-Deoxycytidine 226.08347 5.701 3-Hydroxy-3-methylglutaric acid 161.04554 9.481 3-Hydroxybutyric acid 103.04003 3.416 3-Indoxyl sulphate 212.0024 2.812 3-methylphenylacetic acid 149.06071 1.861 3-Phosphoglyceric acid 184.9857 10.922 4-Methylphenol 107.05018 3.181 5-Hydroxyindole-3-acetic acid 190.05104 3.104 Adenosine diphosphate ribose 558.06531 10.713 Allantoin 157.03669 7.954 Arachidonic acid 303.23309 1.29 Cinnamoylglycine 204.06666 1.915 cis-Aconitic acid 173.00926 11.12 Citraconic Acid 129.01928 3.183 Citric acid 191.01978 11.216 D-2-Hydroxyglutaric acid 147.02987 9.768 Deoxycholic Acid 391.28577 1.733 Deoxyuridine 227.0675 3.295 DHAP/GAP 168.99079 9.74 Dihomo-gamma-Linolenic Acid 305.24867 1.288 DL-4-Hydroxyphenyllactic acid 181.05064 3.842 Docosahexaenoic acid 327.23313 1.286 D-Ribose 5-phosphate 229.01199 9.952 D-Ribulose 5-phosphate 229.01195 9.667 D-Saccharic acid 209.03044 10.748 D-Sedoheptulose 7-phosphate 289.03324 10.39 Ethyl myristate 255.23314 1.311 Gluconic acid 195.05104 8.788 Glucose 6-phosphate 259.02258 10.73 Glycerol 91.03998 4.413 Glycerol 3-phosphate 171.00646 9.567 Glycerophospho-N-palmitoyl ethanolamine 452.27874 1.8 Glycyl-L-leucine 187.10883 7.68 Hippuric Acid 178.05101 2.764 Indole-3-pyruvic acid 202.0511 2.786 Inosine 267.07367 6.299 Jasmonic acid 209.11844 1.357 L-Anserine 239.11509 8.495 L-Ascorbic acid 2-sulfate 254.98183 8.992 L-Aspartic acid 132.03018 9.56 L-Cysteine-S-sulfate 199.96941 10.086 L-Cysteinesulfinic acid 152.00223 4.231 L-Glutathione oxidized 611.14577 10.944 L-Iditol 181.07176 8.062 Linoleic acid 279.23322 1.305 L-Lactic acid 89.02437 3.735 Malic acid 133.01417 10.228 Malonic acid 103.00367 9.557 Methyl indole-3-acetate 188.07174 2.792 Methylsuccinic acid 131.03487 8.769 Myo-inositol 179.05616 10.089 Myristic acid 227.20183 1.344 N-alpha-Acetyl-L-asparagine 173.05678 6.9 N-Acetyl-alpha-D-glucosamine 1-phosphate 300.0491 9.46 N-Acetyl-1-aspartylglutamic acid 303.08362 10.846 N-Acetyl-DL-tryptophan 245.09327 2.79 N-Acetylglutamic acid 188.05647 9.118 N-Acetyl-L-methionine 190.05443 1.965 N-Acetyl-L-phenylalanine 206.08231 1.88 N-Acetylneuraminic acid 308.09891 8.176 N-Glycolylneuraminic acid 324.09376 9.05 N-Isovalerylglycine 158.08224 1.942 N-Tigloylglycine 156.06654 2.775 O-Acetylserine 146.04585 5.367 Oleic acid 281.2486 1.297 Oleoyl-L-alpha-lysophosphatidic acid 435.25189 1.862 Orotic acid 155.00982 4.838 Orotidine 287.05223 7.46 Palmitoleic acid 253.21762 1.323 Pentadecanoic acid 241.2175 1.321 Phenylacetylglycine 192.06672 1.961 Phenyllactic acid 165.05565 1.911 Pseudouridine 243.06235 7.008 Pyroglutamic acid 128.03528 4.865 Pyruvic acid 87.00873 2.898 Quinolinic acid 166.01458 9.904 Stearic acid 283.26411 1.289 Succinic acid 117.01928 9.666 Taurine 124.00727 8.622 Taurochenodeoxycholic Acid (sodium salt) 498.29006 1.438 Tetradecanedioic acid 257.17595 2.763 Theophylline 179.05614 7.114 Thymine 125.03558 2.819 trans-10-Heptadecenoic acid 267.23316 1.303 trans-Cinnamic acid 147.04513 5.426 Uric acid 167.021 7.71 Uridine 243.06235 4.774 Xanthosine 283.06855 7.52 METABOLITES_END #END