#METABOLOMICS WORKBENCH mdm_20250120_012241 DATATRACK_ID:5543 STUDY_ID:ST003680 ANALYSIS_ID:AN006041 PROJECT_ID:PR002283 VERSION 1 CREATED_ON January 21, 2025, 1:16 pm #PROJECT PR:PROJECT_TITLE Ketogenic diet suppresses colorectal cancer through the gut microbiome long PR:PROJECT_TITLE chain fatty acid stearate PR:PROJECT_SUMMARY Our manuscript entitled “Ketogenic diet suppresses colorectal cancer through PR:PROJECT_SUMMARY the gut microbiome long chain fatty acid stearate” describes a reduced colonic PR:PROJECT_SUMMARY tumor burden upon ketogenic diet (KD) consumption in a CRC mouse model with a PR:PROJECT_SUMMARY humanized microbiome. Importantly, we demonstrate a causal relationship through PR:PROJECT_SUMMARY microbiome transplantation into germ-free mice, whereby alterations in the gut PR:PROJECT_SUMMARY microbiota were maintained in the absence of continued selective pressure from PR:PROJECT_SUMMARY the KD. Specifically, we identify a shift toward bacterial species that produce PR:PROJECT_SUMMARY stearic acid in ketogenic conditions, whereas consumers were depleted, resulting PR:PROJECT_SUMMARY in elevated levels of free stearate in the gut lumen. This microbial product PR:PROJECT_SUMMARY demonstrates tumor-suppressing properties by inducing apoptosis in cancer cells PR:PROJECT_SUMMARY and decreasing colonic Th17 immune cell populations. As part of this study, we PR:PROJECT_SUMMARY used different metabolomics workflows to study metabolites in mouse fecal and PR:PROJECT_SUMMARY plasma samples as well as the used rodent diet. PR:INSTITUTE University of Luxembourg PR:LAST_NAME Letellier PR:FIRST_NAME Elisabeth PR:ADDRESS 6, avenue du Swing, Belval, Esch, 4367, Luxembourg PR:EMAIL madita.brauer@uni.lu PR:PHONE (+352) 46 66 44 6954 #STUDY ST:STUDY_TITLE Ketogenic diet suppresses colorectal cancer through the gut microbiome long ST:STUDY_TITLE chain fatty acid stearate - untargeted LCMS data from CMT experiment ST:STUDY_SUMMARY Germ-free mice received cecal content from mice having received a ketogenic diet ST:STUDY_SUMMARY (KC) or from mice having received a standard diet (SC) and were subjected to ST:STUDY_SUMMARY AOM/DSS treatment as described in Tsenkova et al. (2025). Mouse fecal pellets ST:STUDY_SUMMARY were analyzed by a LC-MS approach to characterize the fecal metabolome of mice ST:STUDY_SUMMARY from different treatment groups. Metabolites were extracted from mouse fecal ST:STUDY_SUMMARY samples as described in the method file. Resulting samples were used for LC-MS ST:STUDY_SUMMARY untargeted metabolomics screen using a Vanquish UHPLC (ThermoFisher Scientific), ST:STUDY_SUMMARY coupled to a Q Exactive HF mass spectrometer (ThermoFisher Scientific). ST:INSTITUTE University of Luxembourg ST:LAST_NAME Letellier ST:FIRST_NAME Elisabeth ST:ADDRESS 6, avenue du Swing, Belval, Esch, 4367, Luxembourg ST:EMAIL madita.brauer@uni.lu ST:PHONE (+352) 46 66 44 6954 #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 16 KC_16 Sample source:mouse feces | Condition:KC RAW_FILE_NAME(raw_file_name)=MT011_LC_GlasgowMA_Mouse_feces_KC_16_1.mzML SUBJECT_SAMPLE_FACTORS 18 KC_18 Sample source:mouse feces | Condition:KC RAW_FILE_NAME(raw_file_name)=MT011_LC_GlasgowMA_Mouse_feces_KC_18_1.mzML SUBJECT_SAMPLE_FACTORS 23 KC_23 Sample source:mouse feces | Condition:KC RAW_FILE_NAME(raw_file_name)=MT011_LC_GlasgowMA_Mouse_feces_KC_23_1.mzML SUBJECT_SAMPLE_FACTORS 28 KC_28 Sample source:mouse feces | Condition:KC RAW_FILE_NAME(raw_file_name)=MT011_LC_GlasgowMA_Mouse_feces_KC_28_1.mzML SUBJECT_SAMPLE_FACTORS 34 KC_34 Sample source:mouse feces | Condition:KC RAW_FILE_NAME(raw_file_name)=MT011_LC_GlasgowMA_Mouse_feces_KC_34_1.mzML SUBJECT_SAMPLE_FACTORS 38 KC_38 Sample source:mouse feces | Condition:KC RAW_FILE_NAME(raw_file_name)=MT011_LC_GlasgowMA_Mouse_feces_KC_38_1.mzML SUBJECT_SAMPLE_FACTORS 42 KC_42 Sample source:mouse feces | Condition:KC RAW_FILE_NAME(raw_file_name)=MT011_LC_GlasgowMA_Mouse_feces_KC_42_1.mzML SUBJECT_SAMPLE_FACTORS 47 KC_47 Sample source:mouse feces | Condition:KC RAW_FILE_NAME(raw_file_name)=MT011_LC_GlasgowMA_Mouse_feces_KC_47_1.mzML SUBJECT_SAMPLE_FACTORS 11 SC_11 Sample source:mouse feces | Condition:SC RAW_FILE_NAME(raw_file_name)=MT011_LC_GlasgowMA_Mouse_feces_SC_11_1.mzML SUBJECT_SAMPLE_FACTORS 14 SC_14 Sample source:mouse feces | Condition:SC RAW_FILE_NAME(raw_file_name)=MT011_LC_GlasgowMA_Mouse_feces_SC_14_1.mzML SUBJECT_SAMPLE_FACTORS 20 SC_20 Sample source:mouse feces | Condition:SC RAW_FILE_NAME(raw_file_name)=MT011_LC_GlasgowMA_Mouse_feces_SC_20_1.mzML SUBJECT_SAMPLE_FACTORS 21 SC_21 Sample source:mouse feces | Condition:SC RAW_FILE_NAME(raw_file_name)=MT011_LC_GlasgowMA_Mouse_feces_SC_21_1.mzML SUBJECT_SAMPLE_FACTORS 30 SC_30 Sample source:mouse feces | Condition:SC RAW_FILE_NAME(raw_file_name)=MT011_LC_GlasgowMA_Mouse_feces_SC_30_1.mzML SUBJECT_SAMPLE_FACTORS 32 SC_32 Sample source:mouse feces | Condition:SC RAW_FILE_NAME(raw_file_name)=MT011_LC_GlasgowMA_Mouse_feces_SC_32_1.mzML SUBJECT_SAMPLE_FACTORS 35 SC_35 Sample source:mouse feces | Condition:SC RAW_FILE_NAME(raw_file_name)=MT011_LC_GlasgowMA_Mouse_feces_SC_35_1.mzML SUBJECT_SAMPLE_FACTORS 41 SC_41 Sample source:mouse feces | Condition:SC RAW_FILE_NAME(raw_file_name)=MT011_LC_GlasgowMA_Mouse_feces_SC_41_1.mzML #COLLECTION CO:COLLECTION_SUMMARY Germ-free mice received cecal content from mice having received a ketogenic diet CO:COLLECTION_SUMMARY (KC) or from mice having received a standard diet (SC) and were subjected to CO:COLLECTION_SUMMARY AOM/DSS treatment as described in Tsenkova et al. (2025). Feces samples were CO:COLLECTION_SUMMARY collected and flesh frozen in liquid nitrogen and kept at -80 degree until CO:COLLECTION_SUMMARY metabolite extraction. CO:SAMPLE_TYPE Feces CO:STORAGE_CONDITIONS -80℃ #TREATMENT TR:TREATMENT_SUMMARY Germ-free mice received cecal content from mice having received a ketogenic diet TR:TREATMENT_SUMMARY (KC) or from mice having received a standard diet (SC) and were subjected to TR:TREATMENT_SUMMARY AOM/DSS treatment as described in Tsenkova et al. (2025). Briefly, mice were TR:TREATMENT_SUMMARY administered a single intraperitoneal injection of sterile-filtered AOM (10mg/kg TR:TREATMENT_SUMMARY mouse body weight). Microbiomes were replaced with donor microbiomes via oral TR:TREATMENT_SUMMARY gavage. Mice were administered a total of three cycles of DSS, with a TR:TREATMENT_SUMMARY two-week-long recovery period between each cycle. DSS was refreshed mid-cycle. TR:TREATMENT_SUMMARY Half of the mice were randomly allocated to each dietary condition at the end of TR:TREATMENT_SUMMARY the last cycle of DSS. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Mouse fecal pellets were placed in 0.5mL Precellys® tubes (VWR) containing five SP:SAMPLEPREP_SUMMARY ceramic beads each and MilliQ® water was added to each sample at a 1:16 dry SP:SAMPLEPREP_SUMMARY weight to water ratio. The samples were homogenized at 6000rpm, for two SP:SAMPLEPREP_SUMMARY 30-second-long cycles at 4°C, in a Precellys®24 Homogenizer (Bertin Corp.). SP:SAMPLEPREP_SUMMARY Samples were then incubated at 4°C for ten minutes, then centrifuged at maximum SP:SAMPLEPREP_SUMMARY speed for 10 minutes at 4°C. Samples were maintained on ice and in the dark. SP:SAMPLEPREP_SUMMARY The supernatant was used for further downstream processing. An ISM was prepared SP:SAMPLEPREP_SUMMARY (2μg/mL of ribitol, pentanedioic-d6 acid and d-mannose and 10μg/mL SP:SAMPLEPREP_SUMMARY tridecanoic-d25 acid, 6-chloropurine riboside, 4-chloro-DL-phenylalanine, SP:SAMPLEPREP_SUMMARY Nε-trifluoroacetyl-L-lysine and thionicotinamide adenine dinucleotide SP:SAMPLEPREP_SUMMARY (Sigma-Aldrich) in MilliQ® water). 40μL of the ISM was added to 100μL of the SP:SAMPLEPREP_SUMMARY fecal supernatant fluid. 80μL of this mixture was added to 320μL of methanol, SP:SAMPLEPREP_SUMMARY vortexed thoroughly, incubated for five minutes at 4°C at maximum speed in an SP:SAMPLEPREP_SUMMARY Eppendorf ThermoMixer, and then centrifugated for five minutes at 4°C at SP:SAMPLEPREP_SUMMARY maximum speed. 350μL of supernatant were added to 280μL of chloroform. 180μL SP:SAMPLEPREP_SUMMARY of MilliQ® water were added, the samples were vortexed thoroughly, incubated SP:SAMPLEPREP_SUMMARY for ten minutes at 4°C at maximum speed in an ThermoMixer (Eppendorf), then SP:SAMPLEPREP_SUMMARY centrifugated for five minutes at 4°C at maximum speed. The extract was then SP:SAMPLEPREP_SUMMARY split – 200μL of the upper (polar) phase were aliquoted into a GC vial with a SP:SAMPLEPREP_SUMMARY micro-insert and the rest of the upper (polar) phase was filtered through a SP:SAMPLEPREP_SUMMARY PHENEX-RC syringe filter (Phenomenex), and 200μL were retained in Eppendorf SP:SAMPLEPREP_SUMMARY tubes for further processing. Then, the temperature of the SpeedVac® was SP:SAMPLEPREP_SUMMARY increased to 25°C for 25 minutes to avoid water condensation on the surface of SP:SAMPLEPREP_SUMMARY the glass vial. Samples were protected from light exposure and stored at -80°C. SP:SAMPLEPREP_SUMMARY Samples for LC-MS analysis (polar) were reconstituted in 80μL 50% ACN in water, SP:SAMPLEPREP_SUMMARY transferred into LC vials for LC-MS analysis and analyzed on a Vanquish UHPLC SP:SAMPLEPREP_SUMMARY (ThermoFisher Scientific), coupled to a Q Exactive HF mass spectrometer SP:SAMPLEPREP_SUMMARY (ThermoFisher Scientific). #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, 0.01% formic acid CH:SOLVENT_B 90% acetonitrile, 0.1% formic acid, 20 mM ammonium acetate CH:FLOW_GRADIENT 90 % B for 1.5 min, followed by a decrease to 20% B within 15 min, then 20% B CH:FLOW_GRADIENT for 2 min, increase to 90% B within 2 min, then 90% B for 13 min CH:FLOW_RATE 200 µl/ml CH:COLUMN_TEMPERATURE 50°C #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 UNSPECIFIED MS:MS_COMMENTS Resolution - 120,000, m/z range - 60-900, AGC target - 1e6, maximum injection MS:MS_COMMENTS time – 70 ms. ddMS2 was applied using the following settings: Resolution - MS:MS_COMMENTS 30,000, AGC target - 5e5, maximum injection time – 70, topN - 5, Normalized MS:MS_COMMENTS collision energy – 20. Samples were acquired in positive and negative MS:MS_COMMENTS ionization mode simultaneously (polarity switching). #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS normalized area MS_METABOLITE_DATA_START Samples KC_16 KC_18 KC_23 KC_28 KC_34 KC_38 KC_42 KC_47 SC_11 SC_14 SC_20 SC_21 SC_30 SC_32 SC_35 SC_41 Factors Sample source:mouse feces | Condition:KC Sample source:mouse feces | Condition:KC Sample source:mouse feces | Condition:KC Sample source:mouse feces | Condition:KC Sample source:mouse feces | Condition:KC Sample source:mouse feces | Condition:KC Sample source:mouse feces | Condition:KC Sample source:mouse feces | Condition:KC Sample source:mouse feces | Condition:SC Sample source:mouse feces | Condition:SC Sample source:mouse feces | Condition:SC Sample source:mouse feces | Condition:SC Sample source:mouse feces | Condition:SC Sample source:mouse feces | Condition:SC Sample source:mouse feces | Condition:SC Sample source:mouse feces | Condition:SC 2-Aminobenzoic acid N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A 2-Hydroxyadipic acid 0.004 0.004 0.002 0.003 0.002 0.002 0.004 0.002 0.004 0.002 0.001 0.002 0.015 0.004 0.002 0.014 2-Hydroxybutyric acid N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A 2-Ketobutyric acid N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A 2-Methylglutaric acid N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A 3,4-Dihydroxybenzeneacetic acid N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A 3-Hydroxyanthranilic acid N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A 3-Hydroxyglutaric acid 0.037 0.01 0.027 0.026 0.019 0.038 0.016 0.019 0.013 0.008 0.014 0.025 0.042 0.014 0.011 0.051 3-Indolebutyric acid N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A 3-Indoleglyoxylic acid N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A 4-Hydroxy-2-oxoglutaric acid 0.001 0.002 0 0.002 0.002 0.002 0.002 0.002 0 0.001 0.001 0.001 0.001 0.002 0.001 0.001 4-Hydroxy-L-glutamic acid N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A 4-Hydroxyproline 0.019 0.011 0.02 0.021 0.043 0.037 0.033 0.028 0.017 0.011 0.014 0.044 0.035 0.023 0.017 0.029 5-Hydroxyindoleacetic acid N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A 5-Hydroxy-L-tryptophan N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A 5-Methyl-DL-Tryptophan N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A 6-Phosphogluconic acid N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Acetyl-CoA N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Acetylcysteine N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Adenine 0.022 0.007 0.035 0.011 0.008 0.014 0.014 0.008 0.062 0.084 0.013 0.08 0.025 0.166 0.15 0.023 Adenosine monophosphate 0.002 0.002 0.089 0 0.001 0.002 0.002 0.002 0.002 0.002 0.002 0.001 0.002 0.009 0.002 0.002 Adenosine triphosphate N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A ADP N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A AICAR N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Betaine 0.923 0.906 0.966 0.411 1.243 0.732 1.069 1.02 1.09 0.41 0.919 0.345 0.114 1.157 0.415 0.472 Biotin N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A CDP N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Choline 0.201 0.312 0.168 0.222 0.207 0.217 0.296 0.31 0.526 0.238 0.343 0.211 0.2 0.268 0.454 0.15 cis-Aconitic acid N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Citramalic acid 0.015 0.013 0.006 0.013 0.021 0.014 0.022 0.021 0.007 0.006 0.006 0.004 0.005 0.011 0.007 0.006 Citric acid 0.234 0.188 0.223 0.161 0.181 0.306 0.235 0.172 0.164 0.135 0.196 0.117 0.153 0.169 0.092 0.217 Creatine 0.161 0.158 0.382 0.023 0.104 0.253 0.085 0.096 0.625 0.367 0.642 0.195 0.06 0.072 0.637 0.047 Creatinine 0.048 0.031 0.045 0.018 0.024 0.038 0.023 0.023 0.08 0.039 0.1 0.024 0.026 0.023 0.047 0.015 Cytidine monophosphate 0 0.001 0.004 0 0.001 0 0.001 0.001 0.001 0.002 0.002 0.001 0.001 0.002 0.001 0.001 Cytidine triphosphate N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Cytosine 0.005 0.012 0.016 0.008 0.013 0.009 0.013 0.014 0.005 0.005 0.007 0.01 0.004 0.008 0.008 0.007 D-2-Hydroxyglutaric acid N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Deoxyadenosine triphosphate N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A D-Erythrose 4-phosphate N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Desaminotyrosine N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A D-Fructose N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A D-Galactose N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A D-Glucose 0.032 0.058 0.018 0.051 0.052 0.026 0.048 0.065 0.012 0.029 0.02 0.025 0.029 0.073 0.018 0.017 D-Glucuronic acid N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A D-Glycerate 3-phosphate N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A D-Mannose N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A D-Mannose 1-phosphate 0.011 0.004 0.005 0.021 0.014 0.012 0.018 0.006 0.001 0.001 0.007 0.005 0.012 0.001 0 0.011 D-Ribose 5-phosphate 0.003 0.002 0.004 0.019 0.011 0.002 0.013 0.004 0.001 0.001 0.004 0.002 0.008 0.001 0 0.013 D-Sedoheptulose 7-phosphate 0.01 0.003 0.007 0.039 0.021 0.012 0.024 0.006 0.001 0.001 0.008 0.006 0.028 0.001 0 0.027 dUMP N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A FAD 0.001 0 0.007 0.003 0.002 0.002 0.001 0.001 0 0 0 0.001 0.001 0.001 0 0.001 Flavin Mononucleotide 0.002 N/F 0.002 0.009 0.003 0.004 0.003 N/F N/F N/F 0.001 0.001 0.002 N/F N/F 0.003 Fructose 1,6-bisphosphate N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Fructose 6-phosphate 0.019 0.006 0.01 0.032 0.021 0.015 0.028 0.008 0.001 0.001 0.008 0.005 0.017 0.002 0 0.012 Fumaric acid 0.004 0.005 0.004 0.008 0.006 0.005 0.008 0.007 0.013 0.008 0.018 0.012 0.011 0.012 0.008 0.009 Galactose 1-phosphate N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Galacturonic acid 0.012 0.008 0.007 0.007 0.007 0.012 0.009 0.006 0.006 0.007 0.01 0.008 0.008 0.009 0.004 0.009 Gamma Glutamylglutamic acid 0.006 0.002 0.003 0.002 0.003 0.007 0.002 0.001 0 0.001 0.001 0.001 0.001 0.001 0 0.001 Gamma-Glutamylcysteine N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Glucaric acid N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Gluconic acid 0.066 0.008 0.026 0.007 0.01 0.091 0.012 0.021 0.008 0.013 0.028 0.009 0.016 0.018 0.005 0.015 Glucose 1-phosphate N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Glucose 6-phosphate 0.045 0.017 0.025 0.076 0.052 0.037 0.068 0.024 0.003 0.003 0.02 0.017 0.047 0.005 0 0.032 Glutaric acid 0.001 0.001 0.002 0.004 0.002 0.002 0.001 0.001 0.008 0.002 0.001 0.002 0.006 0.005 0.001 0.005 Glutathione N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Glyceric acid 0.059 0.041 0.052 0.005 0.011 0.07 0.043 0.025 0.021 0.041 0.04 0.029 0.022 0.034 0.015 0.025 Glycerol 3-phosphate 0.003 0.002 0.009 0.005 0.002 0.002 0.005 0.005 0.004 0.001 0.006 0.003 0.003 0.002 0.002 0.003 Glycine 0.001 0.001 0.001 0.001 0.001 0.001 0.001 0 0.001 0 0 0.001 0.001 0 0 0.001 Guanine 0.017 0.051 0.014 0.044 0.045 0.044 0.074 0.036 0.005 0.026 0.01 0.017 0.028 0.022 0.012 0.046 Guanosine diphosphate N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Guanosine diphosphate mannose N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Guanosine monophosphate 0 0 0.002 0 0 0 0.001 0 0 0.001 0.001 0 0.001 0.001 0 0.001 Guanosine triphosphate N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Homocysteine N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Hydroxykynurenine N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Hydroxypropanedioic acid 0.001 0 0 0 0 0 0.001 0 0 0 0.001 0 0 0 0 0 Indole N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Indole-3-acetamide N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Indole-3-carboxaldehyde N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Indole-3-carboxylic acid N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Indole-3-propionic acid 0.006 0.005 0.002 0.005 0.01 0.005 0.005 0.009 0.005 0.007 0.006 0.016 0.031 0.008 0.006 0.015 Indoleacetaldehyde N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Indoleacetic acid 0.047 0.047 0.02 0.069 0.067 0.081 0.057 0.041 0.025 0.051 0.045 0.053 0.05 0.05 0.033 0.029 Indoleacrylic acid N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Indolelactic acid 0.003 0.004 N/F N/F N/F N/F N/F 0.014 0.002 0.011 0.013 0.014 N/F 0.027 0.013 N/F Indolepyruvate N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Inosine 0.149 0.068 0.065 0.263 0.227 0.199 0.13 0.118 0.013 0.025 0.08 0.037 0.147 0.02 0.009 0.215 Inosinic acid N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Isocitric acid N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Itaconic acid N/F N/F N/F N/F N/F N/F N/F N/F N/F N/F N/F N/F N/F N/F N/F N/F Ketoleucine 0.103 0.047 0.017 0.058 0.024 0.053 0.032 0.039 0.023 0.024 0.041 0.021 0.06 0.022 0.017 0.03 Kynurenic acid 0.017 0.032 0.007 0.03 0.045 0.011 0.059 0.032 0.023 0.034 0.023 0.013 0.01 0.028 0.029 0.017 L-Acetylcarnitine 0.024 0.037 0.028 0.002 0.178 0.052 0.202 0.058 0.236 0.128 0.16 0.019 0.004 0.022 0.064 0.004 L-Alanine 0.109 0.033 0.076 0.108 0.043 0.109 0.04 0.028 0.033 0.028 0.037 0.036 0.077 0.02 0.031 0.035 L-Arginine 0.157 0.034 0.447 0.32 0.023 0.064 0.04 0.019 0.007 0.015 0.022 0.013 0.072 0.012 0.003 0.039 L-Asparagine 0.017 0.008 0.023 0.033 0.013 0.018 0.011 0.005 N/F N/F N/F N/F N/F N/F N/F N/F L-Aspartic acid 0.067 0.014 0.09 0.057 0.026 0.075 0.028 0.015 0.035 0.018 0.023 0.036 0.046 0.014 0.025 0.035 L-Cystathionine N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A L-Cystine N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A L-Glutamic acid 0.196 0.046 0.238 0.257 0.137 0.273 0.143 0.074 0.054 0.026 0.055 0.1 0.178 0.05 0.036 0.171 L-Glutamine 0.141 0.066 0.149 0.178 0.082 0.12 0.083 0.056 0.069 0.064 0.064 0.041 0.083 0.042 0.023 0.051 L-Histidine N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A L-Isoleucine 0.452 0.092 0.232 0.545 0.086 0.317 0.096 0.058 0.045 0.058 0.069 0.083 0.224 0.038 0.03 0.079 L-Kynurenine N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A L-Lactic acid 31.191 20.868 8.668 4.627 43.462 7.044 5.784 49.681 35.935 45.752 52.905 4.231 2.464 84.727 47.699 2.958 L-Leucine 0.697 0.09 0.359 0.681 0.102 0.406 0.115 0.053 0.073 0.051 0.091 0.082 0.301 0.03 0.031 0.074 L-Lysine 0.039 0.007 0.033 0.102 0.013 0.032 0.011 0.006 0.006 0.004 0.007 0.008 0.018 0.004 0.003 0.01 L-Malic acid 0.028 0.034 0.03 0.058 0.042 0.036 0.058 0.048 0.085 0.056 0.124 0.079 0.067 0.083 0.058 0.058 L-Methionine 0.408 0.075 0.041 0.484 0.099 0.344 0.1 0.054 0.048 0.048 0.061 0.074 0.143 0.045 0.034 0.082 L-Phenylalanine 0.652 0.126 0.327 0.762 0.111 0.353 0.125 0.068 0.08 0.09 0.095 0.081 0.282 0.061 0.06 0.074 L-Proline 0.856 0.59 0.572 0.705 0.768 0.999 0.737 0.581 0.299 0.516 0.584 0.512 0.637 0.595 0.539 0.59 L-Serine 0.013 0.007 0.008 0.014 0.006 0.014 0.007 0.005 0.004 0.005 0.006 0.007 0.007 0.004 0.003 0.006 L-Threonine 0.079 0.028 0.059 0.101 0.052 0.087 0.04 0.029 0.019 0.024 0.036 0.037 0.053 0.022 0.015 0.04 L-Tryptophan 0.083 0.027 0.063 0.097 0.031 0.067 0.031 0.016 0.011 0.028 0.026 0.023 0.046 0.024 0.053 0.025 L-Tyrosine 0.331 0.064 0.256 0.501 0.092 0.242 0.077 0.035 0.045 0.044 0.058 0.052 0.154 0.037 0.049 0.061 L-Valine 0.569 0.106 0.266 0.653 0.109 0.324 0.112 0.068 0.08 0.082 0.1 0.092 0.301 0.053 0.064 0.069 Maleic acid 0.185 0.24 0.358 0.183 0.187 0.227 0.218 0.263 0.241 0.204 0.392 0.207 0.156 0.184 0.223 0.262 Malonic acid 0.004 0.003 0.008 0.002 0.003 0.003 0.004 0.003 0.002 0.002 0.003 0.003 0.003 0.003 0.002 0.003 Melatonin N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Methylmalonic acid 0 0.001 0.001 0.001 0.001 N/F 0.001 0.001 0.001 N/F N/F 0.001 0.001 N/F N/F N/F Myo-inositol 1-phosphate N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N-Acetylglutamic acid 0.12 0.09 0.139 0.154 0.157 0.161 0.124 0.106 0.06 0.06 0.075 0.122 0.143 0.123 0.092 0.147 N-Acetyl-L-aspartic acid 0.093 0.049 0.088 0.132 0.098 0.119 0.078 0.09 0.061 0.047 0.072 0.089 0.074 0.118 0.066 0.088 N-Acetylserotonin N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A NAD N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A NADH N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A NADP N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A NADPH N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N-Formyl-L-methionine N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Niacinamide 0.015 0.008 0.989 0.045 0.105 0.018 0.006 0.01 0.054 0.039 0.007 0.009 0.01 0.038 0.078 0.008 Nicotinic acid 0.267 0.091 0.02 0.523 0.314 0.341 0.279 0.139 0.037 0.033 0.076 0.174 0.34 0.077 0.026 0.397 O-Acetylserine 1.019 1.299 0.53 1.322 1.879 1.334 1.571 1.084 1.602 1.313 1.199 2.051 1.151 1.999 2.869 1.567 O-Phosphoethanolamine N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Ornithine 0.008 0.002 0.002 0.009 0.003 0.002 0.002 0.002 0.002 0.001 0.001 0.002 0.003 0.002 0.002 0.002 Orotic acid 0.408 0.423 0.16 0.322 0.29 0.336 0.209 0.281 0.302 0.249 0.254 0.125 0.276 0.301 0.187 0.192 Oxalacetic acid N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Oxalic acid 0.026 0.023 0.026 0.021 0.023 0.034 0.026 0.023 0.02 0.02 0.022 0.02 0.025 0.019 0.021 0.026 Oxidized glutathione N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Oxoglutaric acid 0.022 0.015 0.002 0.013 0.013 0.023 0.017 0.028 0.005 0.006 0.017 0.009 0.023 0.015 0.003 0.03 Pantothenic acid 0.098 0.022 0.027 0.061 0.035 0.089 0.037 0.029 0.025 0.032 0.042 0.066 0.097 0.029 0.018 0.097 Phenylpyruvic acid N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Phosphoenolpyruvic acid N/F 0 0 0.002 0.001 N/F 0.002 0 0 N/F 0 0 0.001 0 0 0.002 Phosphoserine N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Picolinic acid 0.27 0.087 0.016 0.533 0.319 0.341 0.282 0.137 0.034 0.028 0.073 0.174 0.346 0.075 0.022 0.403 Pyridoxal N/F N/F N/F N/F N/F N/F N/F N/F N/F N/F N/F N/F N/F N/F N/F N/F Pyridoxal 5'-phosphate N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Pyridoxine 0.03 0.056 0.015 0.023 0.015 0.036 0.027 0.057 0.019 0.075 0.053 0.073 0.055 0.086 0.055 0.043 Pyroglutamic acid 1.852 1.508 1.718 1.202 1.246 1.806 2.093 1.039 1.348 0.907 1.93 2.016 1.745 1.091 1.333 1.668 Pyrophosphate N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Pyruvic acid N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Quinaldic acid N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Quinolinic acid N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Riboflavin 0.013 N/F 0.005 0.012 0.004 0.021 N/F N/F N/F 0.005 0.003 0.003 0.005 0.003 N/F 0.006 Ribonic acid 0.002 0.008 0.005 0.009 0.008 0.01 0.011 0.005 0.003 N/F N/F N/F N/F N/F N/F N/F Sarcosine 0.033 0.018 0.029 0.006 0.009 0.024 0.01 0.014 0.016 0.022 0.053 0.042 0.027 0.017 0.028 0.008 Serotonin N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Succinic acid 0.013 0.008 0.02 0.009 0.005 0.008 0.006 0.015 0.405 0.064 0.036 0.018 0.028 0.188 0.104 0.029 Taurine 0.096 0.032 0.051 0.005 0.027 0.039 0.009 0.042 0.085 0.089 0.05 0.03 0.02 0.061 0.051 0.013 Thiamine 0.003 0 0.001 0.002 0.001 0.002 0.001 0.001 0.001 0.001 0.001 0.002 0.005 0 0 0.005 Thiamine monophosphate N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Thiamine pyrophosphate N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Thiosulfate 0.037 0.021 0.009 0.021 0.023 0.043 0.018 0.027 0.032 0.011 0.021 0.032 0.051 0.01 0.015 0.036 Threonic acid 0.001 0.001 0.003 0 0.001 0.001 0.001 0.001 0.004 0.002 0.003 0.001 0 0.001 0.002 0 Thymidine 5'-triphosphate N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Thymine 0.05 0.021 0.006 0.076 0.042 0.045 0.034 0.021 0.008 0.013 0.019 0.023 0.035 0.009 0.009 0.042 Tryptamine N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Tryptophol N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Uracil 0.215 0.058 0.052 0.777 0.247 0.227 0.155 0.082 0.038 0.028 0.094 0.132 0.191 0.032 0.015 0.2 Uric acid 0.011 0.006 0.009 0.018 0.009 0.009 0.006 0.008 0.02 0.009 0.028 0.014 0.011 0.006 0.008 0.007 Uridine 3.608 1.322 1.117 6.142 3.619 5.689 2.42 2.16 1.11 2.247 2.177 2.015 2.967 1.289 0.669 3.426 Uridine 5'-diphosphate N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Uridine 5'-monophosphate 0.001 0.001 0.025 0 0 0.001 0.002 0.001 0.002 0.002 0.003 0.002 0.003 0.007 0.002 0.003 Uridine diphosphategalactose N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Uridine triphosphate N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A N/A Xanthurenic acid 0.002 0.003 0 0.002 0.004 0.001 0.008 0.003 0.007 0.004 0.002 0.001 0.001 0.003 0.005 0.001 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name RT 2-Aminobenzoic acid 2.69 2-Hydroxyadipic acid 10.27 2-Hydroxybutyric acid 2.52 2-Ketobutyric acid 2.26 2-Methylglutaric acid 2.14 3,4-Dihydroxybenzeneacetic acid 7.4 3-Hydroxyanthranilic acid 5.44 3-Hydroxyglutaric acid 10.63 3-Indolebutyric acid 1.92 3-Indoleglyoxylic acid 3.03 4-Hydroxy-2-oxoglutaric acid 11.3 4-Hydroxy-L-glutamic acid 10.23 4-Hydroxyproline 8.99 5-Hydroxyindoleacetic acid 6.99 5-Hydroxy-L-tryptophan 9.07 5-Methyl-DL-Tryptophan 4.06 6-Phosphogluconic acid 12.63 Acetyl-CoA 9.44 Acetylcysteine 2.57 Adenine 3.33 Adenosine monophosphate 9.54 Adenosine triphosphate 11.96 ADP 10.99 AICAR 4.53 Betaine 4.51 Biotin 3.08 CDP 12.16 Choline 12.63 cis-Aconitic acid 12.28 Citramalic acid 10.13 Citric acid 13 Creatine 9.14 Creatinine 3.6 Cytidine monophosphate 11.05 Cytidine triphosphate 13.07 Cytosine 4.7 D-2-Hydroxyglutaric acid 10.44 Deoxyadenosine triphosphate 11.41 D-Erythrose 4-phosphate 11.04 Desaminotyrosine 2.84 D-Fructose 7.2 D-Galactose 8.71 D-Glucose 8.75 D-Glucuronic acid 10.35 D-Glycerate 3-phosphate 12.19 D-Mannose 8.04 D-Mannose 1-phosphate 11.46 D-Ribose 5-phosphate 11.13 D-Sedoheptulose 7-phosphate 11.43 dUMP 9.4 FAD 7.75 Flavin Mononucleotide 7.27 Fructose 1,6-bisphosphate 13.19 Fructose 6-phosphate 11.15 Fumaric acid 11.15 Galactose 1-phosphate 11.09 Galacturonic acid 10.6 Gamma Glutamylglutamic acid 12.24 Gamma-Glutamylcysteine 9.41 Glucaric acid 11.64 Gluconic acid 8.88 Glucose 1-phosphate 11.05 Glucose 6-phosphate 11.8 Glutaric acid 10.29 Glutathione 9.63 Glyceric acid 5.69 Glycerol 3-phosphate 10.41 Glycine 9.99 Guanine 6.07 Guanosine diphosphate 12.71 Guanosine diphosphate mannose 12.72 Guanosine monophosphate 11.75 Guanosine triphosphate 13.56 Homocysteine 7.12 Hydroxykynurenine 7.46 Hydroxypropanedioic acid 11.85 Indole 1.83 Indole-3-acetamide 2.03 Indole-3-carboxaldehyde 1.85 Indole-3-carboxylic acid 3.51 Indole-3-propionic acid 2.15 Indoleacetaldehyde 1.95 Indoleacetic acid 3.1 Indoleacrylic acid 2.6 Indolelactic acid 2.74 Indolepyruvate 2.24 Inosine 4.84 Inosinic acid 10.85 Isocitric acid 13.09 Itaconic acid 9.94 Ketoleucine 1.69 Kynurenic acid 1.86 L-Acetylcarnitine 4.3 L-Alanine 8.99 L-Arginine 15.7 L-Asparagine 9.68 L-Aspartic acid 10.08 L-Cystathionine 11.26 L-Cystine 10.6 L-Glutamic acid 9.56 L-Glutamine 9.61 L-Histidine 9.24 L-Isoleucine 4.83 L-Kynurenine 4.56 L-Lactic acid 3.38 L-Leucine 4.32 L-Lysine 15.2 L-Malic acid 11.2 L-Methionine 5.16 L-Phenylalanine 3.84 L-Proline 6.65 L-Serine 10 L-Threonine 8.7 L-Tryptophan 5.51 L-Tyrosine 7.31 L-Valine 5.99 Maleic acid 3.53 Malonic acid 10.36 Melatonin 1.65 Methylmalonic acid 9.27 Myo-inositol 1-phosphate 12.18 N-Acetylglutamic acid 9.85 N-Acetyl-L-aspartic acid 10.22 N-Acetylserotonin 1.91 NAD 9.7 NADH 9.12 NADP 11.96 NADPH 11.96 N-Formyl-L-methionine 2.1 Niacinamide 2.3 Nicotinic acid 2.45 O-Acetylserine 4.22 O-Phosphoethanolamine 11 Ornithine 14.34 Orotic acid 4.15 Oxalacetic acid 11.8 Oxalic acid 12.22 Oxidized glutathione 12.18 Oxoglutaric acid 10.8 Pantothenic acid 2.7 Phenylpyruvic acid 1.7 Phosphoenolpyruvic acid 12.54 Phosphoserine 11.63 Picolinic acid 2.81 Pyridoxal 2.5 Pyridoxal 5'-phosphate 9.6 Pyridoxine 2.64 Pyroglutamic acid 4.06 Pyrophosphate 10.12 Pyruvic acid 2.84 Quinaldic acid 2.05 Quinolinic acid 10.73 Riboflavin 2.8 Ribonic acid 7.59 Sarcosine 8.27 Serotonin 16.01 Succinic acid 10.38 Taurine 8.94 Thiamine 12.7 Thiamine monophosphate 9.61 Thiamine pyrophosphate 10.14 Thiosulfate 14.39 Threonic acid 7.26 Thymidine 5'-triphosphate 11.6 Thymine 2.3 Tryptamine 13.61 Tryptophol 1.84 Uracil 2.96 Uric acid 7.71 Uridine 3.78 Uridine 5'-diphosphate 11.78 Uridine 5'-monophosphate 10.47 Uridine diphosphategalactose 11.56 Uridine triphosphate 12.71 Xanthurenic acid 4.82 METABOLITES_END #END