#METABOLOMICS WORKBENCH armbrusm_20240215_193601 DATATRACK_ID:4647 STUDY_ID:ST003282 ANALYSIS_ID:AN005377 PROJECT_ID:PR002034 VERSION 1 CREATED_ON February 15, 2024, 9:10 pm #PROJECT PR:PROJECT_TITLE High Throughput Metabolomics using 96-plex Isotope Tagging PR:PROJECT_SUMMARY Chemical tags with 96 unique masses are reported which alleviate the metabolomic PR:PROJECT_SUMMARY workflow bottleneck and allow for absolute quantitation. A metabolic screen for PR:PROJECT_SUMMARY carboxylic acids was performed on mammalian cells deprived of various nutrients PR:PROJECT_SUMMARY and showed 24% RSD and analysis of 288 samples in 2 hours. PR:INSTITUTE Saint Louis University PR:DEPARTMENT Chemistry PR:LAST_NAME Armbruster PR:FIRST_NAME Michael PR:ADDRESS 3501 Laclede Ave, St Louis, MO 63103 PR:EMAIL armbrustermr@gmail.com PR:PHONE 3145501620 PR:FUNDING_SOURCE 5R01GM134081 #STUDY ST:STUDY_TITLE 96-plex metabolomics studies on nutrient-deprived endothelial cells ST:STUDY_SUMMARY A NeuCode tag which allows for cost efficient incorporation of isotopes is used ST:STUDY_SUMMARY to enable a 96-plex assay for small molecule metabolomics. A 96-well plate of ST:STUDY_SUMMARY cell lysates is analyzed simultaneously, providing absolute quantitation of 38 ST:STUDY_SUMMARY metabolites across 96 samples in a single LC-MS injection. ST:INSTITUTE Saint Louis University ST:LAST_NAME Armbruster ST:FIRST_NAME Michael ST:ADDRESS 4448 Swiss Stone Ln E Apt 3C, Ypsilanti, Michigan, 48197, USA ST:EMAIL armbrustermr@gmail.com ST:PHONE 3145501620 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Bos taurus SU:TAXONOMY_ID 9913 #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 - ctrl_1_1hr Treatment:ctrl_1hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_A.mzML SUBJECT_SAMPLE_FACTORS - ctrl_2_1hr Treatment:ctrl_1hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_B.mzML SUBJECT_SAMPLE_FACTORS - ctrl_3_1hr Treatment:ctrl_1hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_C.mzML SUBJECT_SAMPLE_FACTORS - gln_1_1hr Treatment:gln_1hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_A.mzML SUBJECT_SAMPLE_FACTORS - gln_2_1hr Treatment:gln_1hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_B.mzML SUBJECT_SAMPLE_FACTORS - gln_3_1hr Treatment:gln_1hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_C.mzML SUBJECT_SAMPLE_FACTORS - gluc_1_1hr Treatment:gluc_1hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_A.mzML SUBJECT_SAMPLE_FACTORS - gluc_2_1hr Treatment:gluc_1hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_B.mzML SUBJECT_SAMPLE_FACTORS - gluc_3_1hr Treatment:gluc_1hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_C.mzML SUBJECT_SAMPLE_FACTORS - pyr_1_1hr Treatment:pyr_1hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_A.mzML SUBJECT_SAMPLE_FACTORS - pyr_2_1hr Treatment:pyr_1hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_B.mzML SUBJECT_SAMPLE_FACTORS - pyr_3_1hr Treatment:pyr_1hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_C.mzML SUBJECT_SAMPLE_FACTORS - gln_gluc_1_1hr Treatment:gln_gluc_1hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_A.mzML SUBJECT_SAMPLE_FACTORS - gln_gluc_2_1hr Treatment:gln_gluc_1hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_B.mzML SUBJECT_SAMPLE_FACTORS - gln_gluc_3_1hr Treatment:gln_gluc_1hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_C.mzML SUBJECT_SAMPLE_FACTORS - gln_pyr_1_1hr Treatment:gln_pyr_1hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_A.mzML SUBJECT_SAMPLE_FACTORS - gln_pyr_2_1hr Treatment:gln_pyr_1hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_B.mzML SUBJECT_SAMPLE_FACTORS - gln_pyr_3_1hr Treatment:gln_pyr_1hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_C.mzML SUBJECT_SAMPLE_FACTORS - gluc_pyr_1_1hr Treatment:gluc_pyr_1hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_A.mzML SUBJECT_SAMPLE_FACTORS - gluc_pyr_2_1hr Treatment:gluc_pyr_1hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_B.mzML SUBJECT_SAMPLE_FACTORS - gluc_pyr_3_1hr Treatment:gluc_pyr_1hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_C.mzML SUBJECT_SAMPLE_FACTORS - ctrl_1_4hr Treatment:ctrl_4hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_A.mzML SUBJECT_SAMPLE_FACTORS - ctrl_2_4hr Treatment:ctrl_4hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_B.mzML SUBJECT_SAMPLE_FACTORS - ctrl_3_4hr Treatment:ctrl_4hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_C.mzML SUBJECT_SAMPLE_FACTORS - gln_1_4hr Treatment:gln_4hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_A.mzML SUBJECT_SAMPLE_FACTORS - gln_2_4hr Treatment:gln_4hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_B.mzML SUBJECT_SAMPLE_FACTORS - gln_3_4hr Treatment:gln_4hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_C.mzML SUBJECT_SAMPLE_FACTORS - gluc_1_4hr Treatment:gluc_4hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_A.mzML SUBJECT_SAMPLE_FACTORS - gluc_2_4hr Treatment:gluc_4hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_B.mzML SUBJECT_SAMPLE_FACTORS - gluc_3_4hr Treatment:gluc_4hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_C.mzML SUBJECT_SAMPLE_FACTORS - pyr_1_4hr Treatment:pyr_4hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_A.mzML SUBJECT_SAMPLE_FACTORS - pyr_2_4hr Treatment:pyr_4hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_B.mzML SUBJECT_SAMPLE_FACTORS - pyr_3_4hr Treatment:pyr_4hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_C.mzML SUBJECT_SAMPLE_FACTORS - gln_gluc_1_4hr Treatment:gln_gluc_4hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_A.mzML SUBJECT_SAMPLE_FACTORS - gln_gluc_2_4hr Treatment:gln_gluc_4hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_B.mzML SUBJECT_SAMPLE_FACTORS - gln_gluc_3_4hr Treatment:gln_gluc_4hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_C.mzML SUBJECT_SAMPLE_FACTORS - gln_pyr_1_4hr Treatment:gln_pyr_4hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_A.mzML SUBJECT_SAMPLE_FACTORS - gln_pyr_2_4hr Treatment:gln_pyr_4hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_B.mzML SUBJECT_SAMPLE_FACTORS - gln_pyr_3_4hr Treatment:gln_pyr_4hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_C.mzML SUBJECT_SAMPLE_FACTORS - gluc_pyr_1_4hr Treatment:gluc_pyr_4hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_A.mzML SUBJECT_SAMPLE_FACTORS - gluc_pyr_2_4hr Treatment:gluc_pyr_4hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_B.mzML SUBJECT_SAMPLE_FACTORS - gluc_pyr_3_4hr Treatment:gluc_pyr_4hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_C.mzML SUBJECT_SAMPLE_FACTORS - ctrl_1_8hr Treatment:ctrl_8hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_A.mzML SUBJECT_SAMPLE_FACTORS - ctrl_2_8hr Treatment:ctrl_8hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_B.mzML SUBJECT_SAMPLE_FACTORS - ctrl_3_8hr Treatment:ctrl_8hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_C.mzML SUBJECT_SAMPLE_FACTORS - gln_1_8hr Treatment:gln_8hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_A.mzML SUBJECT_SAMPLE_FACTORS - gln_2_8hr Treatment:gln_8hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_B.mzML SUBJECT_SAMPLE_FACTORS - gln_3_8hr Treatment:gln_8hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_C.mzML SUBJECT_SAMPLE_FACTORS - gluc_1_8hr Treatment:gluc_8hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_A.mzML SUBJECT_SAMPLE_FACTORS - gluc_2_8hr Treatment:gluc_8hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_B.mzML SUBJECT_SAMPLE_FACTORS - gluc_3_8hr Treatment:gluc_8hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_C.mzML SUBJECT_SAMPLE_FACTORS - pyr_1_8hr Treatment:pyr_8hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_A.mzML SUBJECT_SAMPLE_FACTORS - pyr_2_8hr Treatment:pyr_8hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_B.mzML SUBJECT_SAMPLE_FACTORS - pyr_3_8hr Treatment:pyr_8hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_C.mzML SUBJECT_SAMPLE_FACTORS - gln_gluc_1_8hr Treatment:gln_gluc_8hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_A.mzML SUBJECT_SAMPLE_FACTORS - gln_gluc_2_8hr Treatment:gln_gluc_8hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_B.mzML SUBJECT_SAMPLE_FACTORS - gln_gluc_3_8hr Treatment:gln_gluc_8hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_C.mzML SUBJECT_SAMPLE_FACTORS - gln_pyr_1_8hr Treatment:gln_pyr_8hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_A.mzML SUBJECT_SAMPLE_FACTORS - gln_pyr_2_8hr Treatment:gln_pyr_8hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_B.mzML SUBJECT_SAMPLE_FACTORS - gln_pyr_3_8hr Treatment:gln_pyr_8hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_C.mzML SUBJECT_SAMPLE_FACTORS - gluc_pyr_1_8hr Treatment:gluc_pyr_8hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_A.mzML SUBJECT_SAMPLE_FACTORS - gluc_pyr_2_8hr Treatment:gluc_pyr_8hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_B.mzML SUBJECT_SAMPLE_FACTORS - gluc_pyr_3_8hr Treatment:gluc_pyr_8hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_C.mzML SUBJECT_SAMPLE_FACTORS - ctrl_1_24hr Treatment:ctrl_24hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_A.mzML SUBJECT_SAMPLE_FACTORS - ctrl_2_24hr Treatment:ctrl_24hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_B.mzML SUBJECT_SAMPLE_FACTORS - ctrl_3_24hr Treatment:ctrl_24hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_C.mzML SUBJECT_SAMPLE_FACTORS - gln_1_24hr Treatment:gln_24hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_A.mzML SUBJECT_SAMPLE_FACTORS - gln_2_24hr Treatment:gln_24hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_B.mzML SUBJECT_SAMPLE_FACTORS - gln_3_24hr Treatment:gln_24hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_C.mzML SUBJECT_SAMPLE_FACTORS - gluc_1_24hr Treatment:gluc_24hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_A.mzML SUBJECT_SAMPLE_FACTORS - gluc_2_24hr Treatment:gluc_24hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_B.mzML SUBJECT_SAMPLE_FACTORS - gluc_3_24hr Treatment:gluc_24hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_C.mzML SUBJECT_SAMPLE_FACTORS - pyr_1_24hr Treatment:pyr_24hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_A.mzML SUBJECT_SAMPLE_FACTORS - pyr_2_24hr Treatment:pyr_24hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_B.mzML SUBJECT_SAMPLE_FACTORS - pyr_3_24hr Treatment:pyr_24hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_C.mzML SUBJECT_SAMPLE_FACTORS - gln_gluc_1_24hr Treatment:gln_gluc_24hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_A.mzML SUBJECT_SAMPLE_FACTORS - gln_gluc_2_24hr Treatment:gln_gluc_24hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_B.mzML SUBJECT_SAMPLE_FACTORS - gln_gluc_3_24hr Treatment:gln_gluc_24hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_C.mzML SUBJECT_SAMPLE_FACTORS - gln_pyr_1_24hr Treatment:gln_pyr_24hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_A.mzML SUBJECT_SAMPLE_FACTORS - gln_pyr_2_24hr Treatment:gln_pyr_24hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_B.mzML SUBJECT_SAMPLE_FACTORS - gln_pyr_3_24hr Treatment:gln_pyr_24hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_C.mzML SUBJECT_SAMPLE_FACTORS - gluc_pyr_1_24hr Treatment:gluc_pyr_24hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_A.mzML SUBJECT_SAMPLE_FACTORS - gluc_pyr_2_24hr Treatment:gluc_pyr_24hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_B.mzML SUBJECT_SAMPLE_FACTORS - gluc_pyr_3_24hr Treatment:gluc_pyr_24hr | Sample source:BAEC RAW_FILE_NAME=nDeP_96Plex_C.mzML #COLLECTION CO:COLLECTION_SUMMARY Cell culture media was removed and the cells were rinsed 3x with PBS using an CO:COLLECTION_SUMMARY 8-channel pipette. Lysing solution (100µL of 80/20 ACN/H2O)) was added and the CO:COLLECTION_SUMMARY cells were put over ice. The wells were flush-mixed 10 times, sonicated for 5 CO:COLLECTION_SUMMARY minutes, then centrifuged at 3,400 rpm for 10 minutes at 10 ºC. The supernatant CO:COLLECTION_SUMMARY was then transferred to a 96-well plate containing 200µg of pre-aliquoted tag CO:COLLECTION_SUMMARY in each well. CO:SAMPLE_TYPE Endothelial cells #TREATMENT TR:TREATMENT_SUMMARY Bulk bovine aortic endothelial cells were cultured in complete media. These bulk TR:TREATMENT_SUMMARY plates were used to seed 96-well plates at 40,000 cells per well in 200µL of TR:TREATMENT_SUMMARY media. These cultures were maintained for 24 hours in complete media TR:TREATMENT_SUMMARY supplemented with 10% FBS before nutrient deprivation. Triplicate controls were TR:TREATMENT_SUMMARY included at every time point and were media swapped to complete media + 10% TR:TREATMENT_SUMMARY dialyzed FBS. Triplicate 96-well plates were used for nutrient deprivation TR:TREATMENT_SUMMARY experiments, and a separate plate was cultured for cell counting using a TR:TREATMENT_SUMMARY hemocytometer following manufacturer instructions. Tags were allocated to TR:TREATMENT_SUMMARY different wells to reduce bias caused by tag interactions. A single row (B) was TR:TREATMENT_SUMMARY left empty during culture. These wells were used to create a calibration curve TR:TREATMENT_SUMMARY for each analyte centered around the approximate biological concentration in TR:TREATMENT_SUMMARY controls. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY DMTMM mediated coupling of amine tags to biological acids is well studied and SP:SAMPLEPREP_SUMMARY previous reports were used as a basis for acid tagging. Briefly, the coupling of SP:SAMPLEPREP_SUMMARY biological acids to the amine tag was initiated with the addition of 25µL of a SP:SAMPLEPREP_SUMMARY DMTMM/NMM solution (75mM DMTMM, 100mM NMM in 95/5 ACN/H2O)). The 96-well plate SP:SAMPLEPREP_SUMMARY was allowed to react for 30 minutes at room temperature before another addition SP:SAMPLEPREP_SUMMARY (12.5µL) of DMTMM/NMM to ensure reaction completion and account for SP:SAMPLEPREP_SUMMARY water-mediated DMTMM degradation. After 4 hours total reaction time, 50µL of SP:SAMPLEPREP_SUMMARY 50mM myristic acid was added and allowed to react for an additional 30 minutes SP:SAMPLEPREP_SUMMARY to quench the reaction. All 96 well lysates are mixed into a 15mL falcon tube SP:SAMPLEPREP_SUMMARY after the reaction is quenched. This reaction solution is mixed 1:1 with SP:SAMPLEPREP_SUMMARY chloroform, vortexed for 30s, then centrifuged at 2,000 rpm for 5 minutes to SP:SAMPLEPREP_SUMMARY ensure phase separation. The top aqueous layer is then taken for WCX enrichment. SP:SAMPLEPREP_SUMMARY Supelclean 500 mg WCX columns were used to selectively enrich quaternary SP:SAMPLEPREP_SUMMARY amine-tagged metabolites. Columns were conditioned with 4 mL of 90/10 (MeOH/H2O) SP:SAMPLEPREP_SUMMARY and then equilibrated with 4 mL of loading buffer (10% MeOH in 20mM ammonium SP:SAMPLEPREP_SUMMARY formate pH 8.5). The extracted reaction solution was mixed 1:1 with loading SP:SAMPLEPREP_SUMMARY buffer before injection. The column was then rinsed with 4 mL of loading buffer, SP:SAMPLEPREP_SUMMARY followed by elution with 8 mL of 90/10 (MeOH/H2O) + 1% formic acid. While singly SP:SAMPLEPREP_SUMMARY tagged analytes readily elute at mL 2-3 under these conditions, doubly tagged SP:SAMPLEPREP_SUMMARY analytes require more fractions to be collected. The first 1mL of this elution SP:SAMPLEPREP_SUMMARY was discarded and the rest were pooled, dried, and reconstituted into 50µL of SP:SAMPLEPREP_SUMMARY 25/25/50 (MeOH/H2O/ACN) to ensure solubility of all tagged analytes. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Thermo Vanquish CH:COLUMN_NAME Waters ACQUITY UPLC BEH Amide (150 x 2.1mm,1.7um) CH:SOLVENT_A 10/90 ACN/H2) CH:SOLVENT_B 95/5 ACN/H2O CH:FLOW_GRADIENT 0–1 min, 100% B; 6 min, 45% B; 8.5 min, 45% B; 8.6 – 18 min, 0% B CH:FLOW_RATE 300µL/min CH:COLUMN_TEMPERATURE 50 #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Orbitrap ID-X tribrid MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS The IDX was operated with an isolation width of 12 m/z, maximum injection time MS:MS_COMMENTS of 502 ms, AGC of 1e6, positive spray voltage of 3.5 kV, sheath gas of 50 MS:MS_COMMENTS arbitrary units (Arb), aux gas of 10 Arb, sweep gas of 1 Arb, a vaporizer MS:MS_COMMENTS temperature of 350 ºC, and a capillary temperature of 325 ºC. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS nM MS_METABOLITE_DATA_START Samples ctrl_1_1hr ctrl_2_1hr ctrl_3_1hr gln_1_1hr gln_2_1hr gln_3_1hr gln_gluc_1_1hr gln_gluc_2_1hr gln_gluc_3_1hr gln_pyr_1_1hr gln_pyr_2_1hr gln_pyr_3_1hr gluc_1_1hr gluc_2_1hr gluc_3_1hr gluc_pyr_1_1hr gluc_pyr_2_1hr gluc_pyr_3_1hr pyr_1_1hr pyr_2_1hr pyr_3_1hr ctrl_1_4hr ctrl_2_4hr ctrl_3_4hr gln_1_4hr gln_2_4hr gln_3_4hr gln_gluc_1_4hr gln_gluc_2_4hr gln_gluc_3_4hr gln_pyr_1_4hr gln_pyr_2_4hr gln_pyr_3_4hr gluc_1_4hr gluc_2_4hr gluc_3_4hr gluc_pyr_1_4hr gluc_pyr_2_4hr gluc_pyr_3_4hr pyr_1_4hr pyr_2_4hr pyr_3_4hr ctrl_1_8hr ctrl_2_8hr ctrl_3_8hr gln_1_8hr gln_2_8hr gln_3_8hr gln_gluc_1_8hr gln_gluc_2_8hr gln_gluc_3_8hr gln_pyr_1_8hr gln_pyr_2_8hr gln_pyr_3_8hr gluc_1_8hr gluc_2_8hr gluc_3_8hr gluc_pyr_1_8hr gluc_pyr_2_8hr gluc_pyr_3_8hr pyr_1_8hr pyr_2_8hr pyr_3_8hr ctrl_1_24hr ctrl_2_24hr ctrl_3_24hr gln_1_24hr gln_2_24hr gln_3_24hr gln_gluc_1_24hr gln_gluc_2_24hr gln_gluc_3_24hr gln_pyr_1_24hr gln_pyr_2_24hr gln_pyr_3_24hr gluc_1_24hr gluc_2_24hr gluc_3_24hr gluc_pyr_1_24hr gluc_pyr_2_24hr gluc_pyr_3_24hr pyr_1_24hr pyr_2_24hr pyr_3_24hr Factors Treatment:ctrl_1hr | Sample source:BAEC Treatment:ctrl_1hr | Sample source:BAEC Treatment:ctrl_1hr | Sample source:BAEC Treatment:gln_1hr | Sample source:BAEC Treatment:gln_1hr | Sample source:BAEC Treatment:gln_1hr | Sample source:BAEC Treatment:gln_gluc_1hr | Sample source:BAEC Treatment:gln_gluc_1hr | Sample source:BAEC Treatment:gln_gluc_1hr | Sample source:BAEC Treatment:gln_pyr_1hr | Sample source:BAEC Treatment:gln_pyr_1hr | Sample source:BAEC Treatment:gln_pyr_1hr | Sample source:BAEC Treatment:gluc_1hr | Sample source:BAEC Treatment:gluc_1hr | Sample source:BAEC Treatment:gluc_1hr | Sample source:BAEC Treatment:gluc_pyr_1hr | Sample source:BAEC Treatment:gluc_pyr_1hr | Sample source:BAEC Treatment:gluc_pyr_1hr | Sample source:BAEC Treatment:pyr_1hr | Sample source:BAEC Treatment:pyr_1hr | Sample source:BAEC Treatment:pyr_1hr | Sample source:BAEC Treatment:ctrl_4hr | Sample source:BAEC Treatment:ctrl_4hr | Sample source:BAEC Treatment:ctrl_4hr | Sample source:BAEC Treatment:gln_4hr | Sample source:BAEC Treatment:gln_4hr | Sample source:BAEC Treatment:gln_4hr | Sample source:BAEC Treatment:gln_gluc_4hr | Sample source:BAEC Treatment:gln_gluc_4hr | Sample source:BAEC Treatment:gln_gluc_4hr | Sample source:BAEC Treatment:gln_pyr_4hr | Sample source:BAEC Treatment:gln_pyr_4hr | Sample source:BAEC Treatment:gln_pyr_4hr | Sample source:BAEC Treatment:gluc_4hr | Sample source:BAEC Treatment:gluc_4hr | Sample source:BAEC Treatment:gluc_4hr | Sample source:BAEC Treatment:gluc_pyr_4hr | Sample source:BAEC Treatment:gluc_pyr_4hr | Sample source:BAEC Treatment:gluc_pyr_4hr | Sample source:BAEC Treatment:pyr_4hr | Sample source:BAEC Treatment:pyr_4hr | Sample source:BAEC Treatment:pyr_4hr | Sample source:BAEC Treatment:ctrl_8hr | Sample source:BAEC Treatment:ctrl_8hr | Sample source:BAEC Treatment:ctrl_8hr | Sample source:BAEC Treatment:gln_8hr | Sample source:BAEC Treatment:gln_8hr | Sample source:BAEC Treatment:gln_8hr | Sample source:BAEC Treatment:gln_gluc_8hr | Sample source:BAEC Treatment:gln_gluc_8hr | Sample source:BAEC Treatment:gln_gluc_8hr | Sample source:BAEC Treatment:gln_pyr_8hr | Sample source:BAEC Treatment:gln_pyr_8hr | Sample source:BAEC Treatment:gln_pyr_8hr | Sample source:BAEC Treatment:gluc_8hr | Sample source:BAEC Treatment:gluc_8hr | Sample source:BAEC Treatment:gluc_8hr | Sample source:BAEC Treatment:gluc_pyr_8hr | Sample source:BAEC Treatment:gluc_pyr_8hr | Sample source:BAEC Treatment:gluc_pyr_8hr | Sample source:BAEC Treatment:pyr_8hr | Sample source:BAEC Treatment:pyr_8hr | Sample source:BAEC Treatment:pyr_8hr | Sample source:BAEC Treatment:ctrl_24hr | Sample source:BAEC Treatment:ctrl_24hr | Sample source:BAEC Treatment:ctrl_24hr | Sample source:BAEC Treatment:gln_24hr | Sample source:BAEC Treatment:gln_24hr | Sample source:BAEC Treatment:gln_24hr | Sample source:BAEC Treatment:gln_gluc_24hr | Sample source:BAEC Treatment:gln_gluc_24hr | Sample source:BAEC Treatment:gln_gluc_24hr | Sample source:BAEC Treatment:gln_pyr_24hr | Sample source:BAEC Treatment:gln_pyr_24hr | Sample source:BAEC Treatment:gln_pyr_24hr | Sample source:BAEC Treatment:gluc_24hr | Sample source:BAEC Treatment:gluc_24hr | Sample source:BAEC Treatment:gluc_24hr | Sample source:BAEC Treatment:gluc_pyr_24hr | Sample source:BAEC Treatment:gluc_pyr_24hr | Sample source:BAEC Treatment:gluc_pyr_24hr | Sample source:BAEC Treatment:pyr_24hr | Sample source:BAEC Treatment:pyr_24hr | Sample source:BAEC Treatment:pyr_24hr | Sample source:BAEC (+/-)-3-methyl-2-oxovaleric acid sodium salt 398.3135632 623.6422268 494.8065929 324.5948383 251.6179188 433.2801276 175.3380382 293.0201579 705.7208838 NA 117.4341478 13.97164483 189.1614987 291.0413765 281.6431893 49.74883982 69.76773931 3.19770888 NA 110.8714308 8.891433269 NA 49.41255142 91.72820493 273.1255439 469.2264831 291.9952329 50.2286743 46.90348783 248.3975944 165.0078027 479.6797516 134.4699716 236.2541785 658.0949479 343.6894984 33.36235977 84.4315923 4.871202039 309.9010696 303.3575744 319.0737451 138.7853907 425.0779506 194.5247159 43.66724368 127.4688971 54.27433381 48.38563101 269.7199672 98.1928323 59.63079527 197.6188806 141.7379592 NA 26.250627 100.6523659 119.9340693 189.0579398 117.2520118 219.8129969 322.0896738 32.99250244 235.4187489 198.002329 290.7782124 15.07753596 108.1650684 78.12910279 280.0981612 703.726107 475.3416516 172.9073798 413.5406985 99.19101933 54.66723777 268.9554763 162.9971458 NA 9.833379121 NA 161.3378078 171.9162176 308.326018 2-oxobutyric acid 75.43991147 97.63316588 147.0412149 51.75211894 67.20567947 125.9213634 66.70754694 83.53126964 178.9190501 249.9991261 291.5300845 440.403729 51.32725803 55.24840823 112.2951925 111.7168451 121.5181025 341.5162631 104.2993886 164.6220803 338.2122272 74.11313322 90.08193831 174.5975525 56.55437126 71.39543013 125.4837968 73.54679242 96.41540217 154.6483183 73.44973795 87.31986302 120.0295424 64.91998349 100.7954307 160.6974828 64.70868162 117.443096 102.4070176 250.3007752 366.086486 562.6950755 52.44174309 62.07195283 139.0317728 82.43082955 84.51425028 164.6355489 70.36331361 72.6589399 139.3183629 63.69159368 67.89705935 146.0312016 73.70106421 74.34290914 173.3482374 58.19807846 58.83389227 136.3206652 57.03067474 56.64399383 127.0698622 293.1441713 335.5174864 812.3383168 51.62808919 58.81171109 90.2877406 345.0380494 375.972165 900.5170853 478.6939562 484.4460347 1144.881912 83.06393089 73.69913256 221.2032936 57.96304752 91.71943078 124.3298873 114.5219342 132.6850724 154.1677221 4-acetamidobutanoate NA NA NA NA NA NA NA NA 1.432122586 NA NA NA NA NA NA NA NA NA NA NA NA 0.64412494 3.869681876 19.45814801 NA NA NA NA NA NA 119.5429982 49.2600355 27.0624196 NA 10.40353559 24.64126611 NA NA 18.59575025 78.71340011 76.37454848 57.84642876 NA NA NA NA NA NA NA NA NA 58.35179666 47.4795804 49.69302455 53.76374521 33.71358766 53.70577484 NA NA NA NA NA NA 58.69645943 60.33284638 64.82220156 NA NA 16.0465842 NA NA NA 81.44367156 85.24458921 72.18645272 NA NA NA NA NA NA 184.011883 122.4297095 64.50436527 4-imidazoleacetate 148.7134836 94.94564401 62.39682964 38.24476375 31.40204741 39.42369544 1369.467154 343.4115375 1550.412396 142.3741567 88.7187539 100.0233445 252.6706292 249.932293 96.47963546 34.92238523 131.6923955 51.38815193 36.98479401 27.71966762 68.02760923 33.6095079 25.1741149 27.17410242 149.4327327 79.2053114 78.93877499 327.3477893 123.2787034 185.8727866 119.5699581 117.7188377 NA 722.5671934 237.2583704 1051.427999 185.7918992 7.573527048 28.85408772 116.711863 94.28432309 99.45789446 NA 55.06866065 NA 59.56144536 14.30657329 2.436579032 208.3904193 115.4608622 43.53785081 156.282217 77.95078543 75.38134115 195.1619387 136.3602107 432.7334073 76.06703649 10.63658325 34.74763199 57.32097976 4.439743847 4.34896667 68.79434298 72.09439407 37.05476963 226.2504678 101.4036235 94.95903608 82.93038972 15.19307349 56.17274103 78.97734349 37.48935206 16.18243029 6163.476863 947.9187664 9295.049252 74.1101678 23.08240868 46.16049834 229.7490652 94.62821517 58.06059528 adipic acid 40.85726057 66.2492806 46.6225524 562.3027013 314.9078375 415.019492 40.947582 48.05835922 18.07928529 146.9597498 168.9327873 223.9021999 68.87492536 56.23862667 60.46452058 71.39074897 60.87499202 49.53044065 120.4837907 80.99919238 83.58483064 138.4323059 59.27442116 24.26167721 49.18597036 52.81705626 27.75626971 52.94016279 45.28957691 30.40540881 38.58336323 42.88888516 25.94212608 60.18403179 80.07674311 158.3213423 315.7350024 121.7310259 163.0424491 91.11740175 63.46718869 86.34971272 32.5102986 44.46108834 53.71251721 105.9091412 61.84304394 36.30712353 49.35730051 60.40770543 26.84460364 55.31417814 50.93569303 59.98138901 51.74899572 53.56845952 47.98968958 57.29761272 51.03703898 22.55009417 32.42635477 39.0304293 45.07791549 70.93932017 60.37712679 56.00765173 86.07923498 72.29806639 85.14428276 126.9012886 119.2611266 173.9976459 122.6737824 84.20645565 118.3025165 125.4319392 123.6880417 154.5603686 152.0444884 124.9754318 139.8518384 41.48705723 46.80726589 43.43694792 akg 4976.440436 5031.109807 988.2467018 1866.849422 1734.77525 NA 1817.821936 2661.437527 6495.95925 1132.527652 637.4515001 NA 1229.685411 980.5936257 NA 2615.950831 2392.405586 14174.90115 5257.089748 5136.584113 20602.02706 993.4156229 590.3076975 NA 1440.970262 1171.944103 NA 5303.748671 5316.029041 10994.34308 1429.269237 1074.999986 NA 5503.98263 6242.754177 21014.43929 1527.787843 1086.184225 NA 1252.29204 936.7031623 NA 3339.3357 3593.99089 9016.988889 2115.897442 2371.450659 NA 4644.733976 7041.842232 26332.20459 2134.290099 1616.340413 NA 6304.851944 9781.209746 37122.03625 6669.482463 8866.227948 26659.63594 1236.606851 659.3552035 NA 3790.436653 4744.463657 3965.434291 3594.960004 3772.941598 1550.5281 1416.062255 735.339156 NA 1404.543469 1695.974922 NA 4849.431897 5552.188219 6624.443449 1114.339796 934.3076406 NA 1281.322597 747.1767264 NA azelate 92.85879514 42.78664713 206.2358882 97.87150991 127.326012 234.9355964 2093.536379 4631.774086 4763.913454 102.5564092 134.7707555 267.073094 187.4089983 390.9160257 430.1443196 97.27146135 64.43178976 221.6284484 96.37701933 48.2816919 217.5597996 76.08331686 70.00480115 219.9667686 76.87639604 NA 165.6556652 97.13596657 58.27393044 215.0913756 295.6377515 1677.329348 559.3445675 124.2828055 394.8668711 251.1552842 129.9488992 144.3915942 268.0239261 148.4677162 229.6021392 322.8023645 98.41844113 75.54958552 222.5691196 96.64351782 36.28354879 217.1080466 98.65346504 79.25069521 218.3944537 97.12433773 55.44558189 222.7003641 94.16226247 64.33919997 200.8024995 97.54480601 34.78777796 218.204822 94.32633906 14.83921744 215.4853442 136.9666567 NA NA 458.0703788 1105.468342 1002.615716 101.7311854 50.94159697 207.4133488 1891.136507 4613.557876 4783.690523 131.9152839 111.0020976 288.193867 101.0926355 82.26166453 228.5751729 100.5325869 55.44878476 212.6925599 benzoate 18358.88153 8222.060609 3813.522149 19645.35927 16157.99041 5172.022639 19796.82763 15670.72387 5858.420163 10777.97017 9492.590344 4190.426719 22952.04365 17595.49698 6363.564446 19355.24205 15859.16482 5913.979738 17207.78431 15506.83823 6851.871135 21095.00257 15854.55343 6905.535808 13520.51069 15813.47046 6945.952372 3776.323847 6438.957682 7926.02315 2867.041329 7028.435293 4294.374528 11648.13181 14098.78926 6517.17796 22541.12451 17542.13773 6350.14815 NA NA NA 33069.2937 16536.86185 6785.025515 32444.73032 17528.76904 8013.428871 17658.02772 14632.95841 5853.451492 25067.5857 15986.74198 6531.130945 16642.75696 11960.4638 5805.133681 12540.58304 6979.63993 2947.839271 15190.17034 14319.9592 6638.543419 26517.81463 15107.87547 5223.93515 25895.16174 18600.11116 7112.685535 18100.95197 15277.72816 6616.935056 NA 2574.862404 2337.400241 25996.34556 17445.9768 5133.167273 12221.96951 6346.591862 2741.408562 1070.355744 9558.655066 4376.15542 butyric acid 80.32690482 182.5020645 118.4064833 26.36099387 NA 22.33731342 10.95493058 NA 125.1404731 NA NA 11.09399888 6.834597846 55.34228807 67.60652158 7.204652714 NA 72.35000196 NA NA 25.43965942 NA NA 41.29050113 NA 46.43868551 34.10382665 NA 66.1056706 80.9743593 NA 66.43930543 31.83653543 6.589781895 22.8352618 94.76590849 27.62245212 NA 69.94298807 38.03836103 132.9562705 169.1065877 NA NA 10.18104858 11.20584028 192.3604167 77.24362988 12.50765589 344.2337777 20.67848877 67.1062491 14.69871031 122.0083443 NA 9.857356678 44.17320943 NA NA 19.69237719 NA 97.09634841 31.43935995 109.0409574 44.11050906 154.0552811 13.14417103 140.0848681 32.86410131 57.71821012 113.6476325 107.4025888 93.23862827 168.5685685 100.5288846 NA 61.33978287 36.73366638 33.10613439 8.163299608 54.32653044 11.43531912 56.10891108 126.3627864 fumarate 162.9088833 124.6069076 89.29719996 146.1825026 163.9754208 163.3742451 47.49625612 20.97598319 NA 59.88971819 25.13479245 NA 299.9919497 178.8112363 238.5544904 42.97270977 9.34428675 NA 261.8926133 289.2743445 289.2993986 153.8145361 148.0651525 170.6634585 163.5443785 65.3826388 104.126992 480.0534487 604.2813359 719.7586004 153.9298805 153.0074253 203.4360973 35.73064815 6.7878031 NA 48.76723729 15.47173963 NA 83.83460455 69.6258812 NA 161.3684701 135.2565908 284.1559481 159.5160695 166.5541234 153.3223724 544.5234349 642.2186199 1156.250265 121.7463593 90.20091516 63.10481144 92.50948976 67.65158786 NA 54.80159153 27.57445905 NA 219.8433344 238.4290516 315.109041 92.6807132 84.73339743 NA 304.2909943 439.1508829 800.3450988 36.5463054 NA NA 472.1733094 522.4385083 655.7226773 49.72252926 35.41465263 NA 36.20046067 NA NA 90.13401308 71.16735399 NA glutarate 378.205127 717.5209514 336.3563392 132.1164454 142.3416309 184.9591649 51.14245029 97.40439374 104.0464168 630.8972691 980.9127857 633.1264476 150.0796144 181.2702453 151.9103864 150.9945705 163.8277629 142.0080658 570.3300239 775.0354294 526.7054503 317.3620693 395.5446903 288.0674888 430.4560014 647.3268949 379.1316541 225.0428734 454.3902598 209.5837442 69.33824262 76.71449522 107.8286345 255.4328638 402.2410859 257.4625282 357.6438138 425.6042883 347.1006312 236.693238 275.582533 229.3033506 147.6397483 291.6388429 149.083884 NA NA 12.22445626 96.62514153 132.500093 68.01533371 NA NA NA 21.21606175 27.18563515 71.16587786 46.70121182 40.80063501 39.54983386 186.7164613 230.6761089 192.9959891 NA NA NA 371.5090336 522.8608325 307.5609441 268.0214017 464.5977531 288.387442 312.3698697 453.2589184 248.0675562 43.91205441 NA 80.58806098 291.4587173 429.273422 338.7824757 135.0060241 159.9599568 122.2022616 hexanoate 51.45161621 NA 1073.311567 291.1163111 102.6704814 254.8600865 NA NA 872.766045 NA NA 253.6666807 NA NA 618.8065135 NA 64.11942042 407.2856246 787.5805168 300.8236253 NA NA NA 313.0044648 56.38353093 62.57674539 862.3520291 1203.788943 578.4558747 66.70630951 188.7195004 83.01803589 101.8992338 295.8907566 126.3857218 44.29204946 NA NA 198.6674057 120.6306941 NA 258.3591373 NA 105.7532766 123.9325787 NA NA 130.7920922 163.9128034 89.87748907 413.0643717 121.2639105 134.3585952 54.40631439 NA NA 951.1333767 NA 66.17591017 1489.786947 207.4801605 20.34855733 199.7024945 120.3799697 191.8300219 NA NA NA 133.6739787 NA NA 378.256487 NA NA 250.0150722 6246.134066 2940.058109 NA NA 110.9330709 296.9114892 NA NA 463.7166428 lactate 83.50395664 64597.59519 1486421.031 NA 29539.48502 746994.3304 7657.521248 190111.7853 1992142.75 NA NA NA 2376.26016 114431.1276 1077011.08 1905.978352 118984.4661 1539905.733 7871.104331 184151.6464 NA 3207.528696 116704.9215 1597811.43 4759.281033 161913.1137 1493686.021 9466.948717 241252.5808 1751321.46 NA 28788.14664 138736.9043 1705.782949 99354.50943 1105623.541 4577.025747 162170.2988 1639238.478 8507.059433 250903.0389 2901617.83 NA NA NA NA NA NA NA NA 75454.98234 NA NA NA 6214.344304 181800.2144 2112327.607 NA NA NA NA NA NA 1607.872516 36345.53243 536337.4374 45049.0602 724392.3268 3272832.243 10386.53667 266211.6613 2633227.14 3961.421993 120532.0196 7801.642763 NA NA NA 6423.551286 173032.6325 1743846.409 2350.147749 124093.4283 1576991.816 levulinic acid 21.65564088 410.6565323 213.7960355 NA NA NA NA NA 2.615925991 NA NA NA NA 152.4355136 148.8150471 NA 125.9825606 220.3897239 NA NA NA NA NA NA NA 48.51737934 59.00038198 NA 152.582744 91.05535938 NA NA NA NA NA NA NA 54.47071566 97.9104815 NA 191.0570777 271.2701 NA 447.5868132 NA 1.161975118 486.9902616 318.987699 59.380963 651.021842 533.4726023 NA NA NA 225.4820249 1225.006656 306.5106956 NA 473.4089995 789.4616817 33.65744063 589.60935 641.3843606 NA NA NA 3.903849383 559.8887671 668.7160927 NA 156.0339332 167.1764206 NA 58.00994284 NA NA NA NA NA 86.87128032 100.9549598 NA NA NA malate 6625.936431 20659.87792 539191.6234 1363.97051 7423.038949 NA 8877.467194 26267.8423 743591.1725 94.66195953 NA NA 2496.464979 5331.978775 243740.3969 10787.58114 32517.57171 1055290.23 2349.675794 3236.845495 175237.9786 240.5160146 NA 5268.265085 6931.048085 18792.73262 666150.9892 6117.552056 11387.00084 504384.1011 1214.980716 2230.979907 107710.5146 6943.626859 19790.15207 602861.0958 NA NA NA 585.3034697 109.30292 330.3106168 10651.63464 30974.35298 908242.8921 NA NA NA 6498.554978 16582.90784 631247.1355 53.45243488 NA NA 7948.195453 22471.53479 796683.2776 6942.867081 14688.58045 655334.6316 1031.668205 2468.939744 93878.56532 4677.577974 13720.46099 389722.5364 6049.527727 17016.85137 598750.3603 1568.804274 4967.216048 109233.4113 185.7404995 NA 5526.565086 8379.443924 21335.94746 738704.1527 2384.89403 4431.148465 199643.2436 226.6079026 NA NA malonate NA NA 189.6007697 150.941617 NA 405.6570411 NA NA NA NA NA 372.4117065 12119.68766 147.4251983 NA 12031.32136 2866.388669 451.7931767 NA 228.3819637 156.0685532 NA NA NA NA NA NA NA NA NA NA 749.8816079 122.4362423 21.33879458 NA 146.3193544 12573.12062 1294.466234 NA 7042.242522 1454.377247 31.71044476 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA 1636.252398 NA NA 3089.920543 153.9741494 106.5490344 19421.06908 3520.391272 128.7970334 135252.4361 38974.31221 7014.986225 204460.6017 55980.80319 9912.782295 50894.7145 15121.3509 2766.287591 2692.225395 1238.368111 NA 1598.032871 NA 180.118473 methylmalonate 37.27896629 73.98739358 51.84101953 98.9908711 83.02692613 84.47550848 59.20920681 68.75087002 111.7971228 202.6300917 186.181218 60.70157551 1008.047518 1184.83789 135.6549358 168.5402667 190.3652818 54.82926908 181.6935062 87.86989123 38.64529686 272.0910439 438.4327209 243.2594945 99.35330801 90.20744275 16.111131 19.24794514 33.65493439 154.1526336 60.23255322 99.07213873 43.67953307 160.2768517 151.7731371 68.74606689 436.1085025 828.847393 133.5736651 227.46261 202.3534572 106.9424342 173.7159747 82.51583651 112.9453266 175.0739531 136.0485211 123.8975498 73.0684547 115.4425679 31.20437375 406.9456991 220.9069585 157.918204 93.36163906 22.61840884 58.37831332 10.46998404 NA 51.92663229 196.5643996 186.7480139 10.88147533 31.58995304 48.24492046 29.40325102 144.7852711 193.1432259 75.88413839 1754.217903 1659.812299 1011.265364 785.2506741 1077.286752 125.2830041 185.0619013 190.4959705 91.16542753 331.3267735 477.3940464 37.54893976 120.0689839 187.5053408 133.8066902 n-acetyl-beta-alanine NA NA NA NA NA 6.987492846 NA NA NA NA 18.11943472 19.16177368 NA NA NA NA NA 8.837436841 NA NA 5.640046147 1.220182235 NA 21.17955091 NA NA 9.318068058 NA NA NA NA NA 13.14490286 24.12234194 39.0554293 27.54721008 7.818590072 4.289410615 20.53688682 NA NA 17.82167426 NA NA NA NA NA NA NA NA NA NA NA 4.088518966 49.09669388 76.85312959 36.98423767 NA NA NA NA NA 15.25057558 NA NA NA 21.56394642 NA 28.86017712 NA NA 2.888820736 NA NA NA NA NA NA 24.28899394 63.4325425 25.25987431 15.30094593 23.27534253 21.70535344 n-acetyl-isoleucine 73.89895631 109.9040438 146.7848315 54.15750957 49.82999686 427.7042663 52.35363144 74.20134797 61.96758664 87.49532045 54.84589943 238.0219369 284.4773364 118.354023 350.6357357 98.54693762 76.95083754 177.9634273 293.8864592 299.2931356 634.090277 68.62479947 88.99365256 74.87299854 58.62570653 61.16370505 101.5059838 31.37350325 2.034085838 9.897200449 43.35959699 50.68875436 287.3058627 33.47688949 104.762783 40.44183803 41.15991759 49.88832708 172.4660866 59.43958388 148.8786953 149.1319667 110.5947892 60.90397172 NA 237.2800046 126.6827045 625.8479609 207.70139 48.59579391 389.5339854 596.0343782 69.62358457 637.3168579 68.7489174 132.906017 86.08176075 15.93739923 107.8521825 NA 18.90111907 29.87733133 291.5348092 87.27195255 108.7119845 NA 93.51932722 217.7107197 149.0580554 108.4491509 71.94628008 299.9755918 97.69355086 12.83426365 367.7293255 93.2235334 32.68627926 234.874617 28.35400248 72.93375629 NA 161.7713479 207.0079618 156.2346566 n-acetylalanine 28.75456086 91.49437247 2414.993282 84.13631054 87.53293598 1255.930619 NA 27.34550426 901.3651026 NA 17.46524794 548.3120072 NA 23.48808631 NA 31.1507051 72.8270816 365.8144108 5.7954396 64.18782759 2936.798406 137.6765264 128.6834856 547.5478075 43.55476089 78.93057836 2430.560272 45.21320899 94.42875471 293.314653 NA 21.03404019 166.5908664 76.53556013 78.60931618 544.0029835 NA 30.76056568 216.7003606 NA 47.40749239 497.7579991 NA 29.58845178 4872.747167 NA 59.57894075 2795.427363 NA 68.14688256 542.2292338 NA 18.43284259 NA 98.15553514 52.05172735 245.3569794 NA 55.95827106 NA 215.3450182 39.15324056 NA NA 33.28933936 NA 190.75912 101.4499257 1143.555288 13.73917281 81.13966641 2245.354827 NA 32.42363994 1032.913386 NA 11.53048544 NA 145.8271557 92.98640322 1681.901401 NA 47.34818507 321.5347128 n-acetylasparagine 1.521889539 0.079174666 2.976969257 30.75698996 14.90344033 18.20406023 5.021583632 8.242768691 4.086344079 4.697164949 10.82305733 8.205678843 22.43022185 16.87911628 14.71077143 1.153944818 NA 1.415619932 1.17561442 NA 1.118436038 5.079693391 2.362966048 5.411740713 4.200711768 3.299650022 4.327822337 2.394627379 NA 2.560640745 31.00377119 47.29745551 40.42623667 4.328647895 4.839234711 5.889541816 5.714458404 4.100788279 3.954798495 4.499636252 NA NA 4.584197067 3.56692511 4.100851778 57.37909876 76.48412919 63.89310742 31.9910074 42.20685266 34.78688506 10.24892002 8.557245742 5.778782408 5.659235704 7.489231164 5.440625508 2.617205158 4.90198881 3.530870661 NA NA NA 20.58573458 20.81805169 14.00590655 196.9103432 238.592194 185.9676845 49.42789082 50.54465021 50.04451337 41.80194842 49.16653221 40.32506626 5.435146257 3.346179309 2.563714105 NA NA NA 29.42420188 17.22084588 25.81946791 n-acetylglycine 60.60920778 236.6238539 347.6084501 NA 11.90402037 103.4372547 95.86897657 53.85996361 98.63024312 NA 12.34250712 20.86334501 4.370749032 19.20449675 22.77797954 NA 17.88192571 46.20140213 116.4856349 21.97987664 0.585359835 9.97322038 18.74668621 16.37015201 35.16870041 28.22206418 NA 23.13381232 34.08872964 106.8055753 NA 150.5653947 254.8183022 NA 22.4597011 12.12655062 323.8772005 29.36834396 40.88699592 46.7297899 12.40672113 19.94191431 22.79962023 23.13087889 16.68185203 20.05483532 17.12203178 150.8007583 26.09481882 55.52691835 228.4755167 34.47614609 20.02414749 81.9259085 NA 16.38453769 13.99208019 NA NA 55.05242711 5.598487107 60.89984502 941.3260242 NA 15.76337051 NA 25.74835577 23.48158329 8.778334114 222.5147955 57.28680751 21.93968021 43.76896706 40.88280239 67.35280351 213.5064671 33.85567928 NA 62.74739258 16.34933115 37.33853109 3.875046018 53.28168358 421.5215062 n-acetylproline NA 136.4313636 47.81702713 1.512880602 0.69306454 46.55514091 13.65329292 NA 15.45162696 16.39344366 NA 14.55261112 NA 99.2088175 72.94902555 14.58754729 23.03810425 22.27142771 NA 217.5268906 37.61314876 9.602586255 57.88379049 24.12925095 NA 804.1535284 99.60149159 15.81393279 NA 15.27692145 NA 159.5844017 58.38315116 0.553942124 NA 6.878650545 6.140989964 102.0192999 34.06976309 12.62127357 58.80251181 13.61259013 NA 160.1792763 37.9650231 6.862895212 108.7473607 17.18748106 25.58792621 NA 4.224131874 7.305285827 55.43282497 37.07114411 21.39152798 NA 9.042477648 18.87272604 NA 8.24061623 20.21082786 NA 3.711511836 16.76644561 NA 10.29739662 13.01047713 NA 17.89246258 6.327539492 NA 21.59947558 3.739039004 68.42249465 17.62402339 16.5035884 NA 15.78780416 20.80368965 NA NA 13.46417215 NA 9.336260891 pantothenate 2224.948951 1551.211082 256.0725718 214.3481481 258.929487 158.6088795 70.25368237 28.97558424 149.5276181 172.1073447 42.94397233 211.508144 802.5367245 780.1439235 537.1481658 14.20140944 NA 126.8975601 228.8303094 441.6991126 718.2821861 3481.743212 6055.826156 908.3512245 195.4178233 68.17991139 167.948985 314.1850512 68.13676411 469.7539293 NA NA 252.4385144 NA NA 88.28477318 1776.350466 2456.055209 441.8641733 122.1431495 NA 150.5076337 2639.730213 2259.836364 98.1132505 NA NA 182.6803664 588.4692409 426.7175762 376.1714108 2869.952037 4820.133457 637.5149087 395.8854313 176.1617286 220.4039713 682.4709567 541.1843498 314.5323154 2283.280012 2717.26834 1221.764188 NA NA 163.0560929 67.6832671 61.22629672 251.0748547 NA NA 48.48817097 836.4704633 975.7993552 600.9756171 84.84148502 NA 239.3664026 3326.830996 4627.77221 1696.115236 16.48562433 NA 154.9253673 pyroglutamate 30117.14837 30853.14518 NA 21810.95487 17987.74985 3211.332206 19038.66098 15839.5855 29731.63151 25620.22287 15868.53894 11647.84149 24990.08635 29205.57259 39343.28886 21130.9475 16191.10119 3548.701367 25004.85996 30772.44398 NA 61756.58665 28070.03979 37095.48314 14714.92938 10040.54774 12361.13565 11119.30767 9605.955151 68045.3022 11150.24562 12526.37902 25029.05891 22863.71476 27625.29105 23792.18402 18022.13642 21350.45581 NA 37110.82817 31632.99955 16891.86452 32214.43228 26894.10783 35337.29341 8336.38116 10343.4379 NA 7083.144601 7642.392834 NA 7632.771914 6193.957479 31054.26754 22401.3438 26457.36491 180.744822 22304.11358 21644.91034 12144.41573 28253.22341 31203.70621 944.9998019 41017.86148 16268.63138 6560.960361 4864.396475 4507.476906 16952.89365 3412.349855 3229.175622 19623.04232 8257.659624 1791.165507 1664.677051 20812.68087 23540.57816 26909.28265 37239.96236 16371.61493 1908.612681 16644.33605 28471.37677 45696.14028 pyruvate 63489.83579 63046.92738 51730.35793 32504.3893 15214.52596 11227.80996 34206.83722 24533.58737 16701.40033 8951.432073 1796.280909 2456.042927 47482.78906 41465.60982 26321.47127 39940.93655 24928.78704 18752.50172 93731.02534 47152.36118 45761.67205 8327.585707 5917.480702 3035.052069 34284.96835 24768.57607 13349.07261 19970.62942 19766.34775 10358.99702 31877.538 13103.95352 5912.87703 14628.4695 15418.27409 7204.429894 NA 788.6778859 2185.193537 27363.14686 31947.4622 16878.94432 19081.08319 9902.374289 7192.322328 14505.98593 11459.31652 8746.406468 267410.8655 143525.0629 117384.4232 NA 743.9100498 2243.197838 18410.69489 15894.74543 11344.95528 32896.24748 17913.60599 14137.13195 39173.47594 29335.99286 7301.136887 6405.737355 2926.273394 4973.817335 40637.38256 24946.51431 18818.39581 9329.299994 6606.45008 6609.103365 98913.59376 50863.79229 21512.28934 23242.54044 12710.41027 4751.250966 7013.294295 4854.166654 4061.182868 435.3008359 441.6295375 3868.986519 sebacic acid 46.77275264 66.34838895 131.0756893 67.97949906 49.34163487 157.4549001 42.83969269 37.91491751 58.14789977 3.936091943 36.28173256 61.0625455 30.43235025 118.4185018 98.92391541 18.65567046 39.11483208 19.34011051 5.464663245 57.53157027 NA 8.477169576 28.7572025 32.82487694 16.51883084 32.70218817 NA 24.30750334 27.1700312 98.9801099 32.21860048 80.33838684 65.28698169 45.77264612 46.45941949 NA 20.09174913 20.6515806 19.36782487 27.3958546 44.32508919 211.3470276 30.21403652 50.48731499 135.4816572 36.68301378 49.63942382 99.3941276 9.17255651 91.78876319 38.02147171 137.6726798 75.0659538 197.4222139 0.445669593 165.9066176 91.90100265 35.9931411 42.97734243 31.3256969 21.07409466 33.41070799 134.9804409 40.74533199 42.6055868 6.529922 6.269647766 43.52307594 40.95858173 23.20171697 33.26156291 NA 14.4216593 39.18191143 NA 52.21810903 42.2162263 NA 3.440421735 49.03390732 NA 20.66809951 54.85841747 NA suberate 48.42102327 NA NA 241.6614499 NA 2432.130738 NA NA NA 1755.114595 30.68976578 1225.421665 1271.993075 NA 532.5887213 NA NA NA 774.3967498 0.469337394 526.293948 406.3961163 NA 860.4850002 NA NA NA NA NA 80.23066381 NA NA 1118.694056 293.6329099 NA 148.748723 4450.565792 68.75669418 5297.803156 953.4868792 NA 1793.86819 NA NA NA 638.9266767 NA NA NA NA NA NA NA NA NA NA NA NA NA NA 1153.736408 3.016434172 392.383648 NA NA 153.8584923 1006.239465 NA 1373.124748 NA NA NA 160.4102388 NA 1171.135161 393.0713286 NA 31.94585161 2273.177931 NA NA 2217.403312 11.8344158 3790.712769 succinate 167.7382605 1798.04314 NA 588.7101968 589.5968477 402.1168871 252.2794215 836.0716916 529.6074031 705.7180808 3822.404135 495.587927 4549.988312 32823.75216 764.6791689 773.7144249 5842.090433 408.2801693 405.3481632 3940.198087 86.81905276 993.790979 8456.757757 1826.780338 483.2748854 1093.31969 NA 163.9970402 NA 1127.710645 245.2520358 982.7777482 83.77631296 536.0471763 5314.591668 675.5701057 1692.724531 18903.36041 641.524818 867.5812298 4468.631256 809.3319456 805.5751764 1807.144107 455.1712063 858.8834787 4136.979143 685.8631862 374.6860311 1603.121701 NA 1810.582239 5906.314507 1095.823866 736.8826464 NA 233.7064268 63.89767917 NA 50.25475036 637.0884713 4242.176898 NA 247.392577 153.9032 NA 828.3637119 3296.272234 278.153181 10260.85872 37246.96828 10486.29946 3584.802843 24613.39241 668.0786804 1336.12895 1735.025648 753.2614219 1397.784216 10570.88876 369.0328609 538.7186579 3303.564563 966.3094069 ureidopropionate 84.97377675 84.61168904 53.85006537 286.1142231 222.9675848 88.73614791 90.21832394 84.81688215 56.37069815 593.6096507 656.6665315 616.5217872 91.77555548 90.85782784 59.90216137 86.67884527 84.81810693 53.58958224 219.6545761 216.4487971 142.0198131 340.75244 333.9986052 296.0712475 87.3148468 82.04358656 53.94519077 280.0165165 256.4955447 188.9645358 167.6237528 195.785129 138.6024181 615.8770779 443.7445093 384.3864987 420.2649733 306.9829463 332.3092297 231.0571418 225.208742 223.2471453 89.54319644 85.02526464 52.41866342 90.07991385 85.69799483 54.68147694 85.50989321 82.01483842 48.39807273 135.3862632 122.8448315 123.1449783 277.8992223 293.9086433 351.1575806 109.1901249 90.7618053 55.94312644 89.33186872 84.70759266 54.87307019 100.7614706 91.33490567 61.36696422 465.8007388 490.5064645 227.9090252 95.42399344 88.55142449 53.73513626 250.4975515 246.7851718 243.6668306 244.125595 253.7572559 151.3253235 370.6846308 252.4071859 283.2891947 307.9617736 281.9169546 324.8143084 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name (+/-)-3-methyl-2-oxovaleric acid sodium salt 2-oxobutyric acid 4-acetamidobutanoate 4-imidazoleacetate adipic acid akg azelate benzoate butyric acid fumarate glutarate hexanoate lactate levulinic acid malate malonate methylmalonate n-acetyl-beta-alanine n-acetyl-isoleucine n-acetylalanine n-acetylasparagine n-acetylglycine n-acetylproline pantothenate pyroglutamate pyruvate sebacic acid suberate succinate ureidopropionate METABOLITES_END #END