#METABOLOMICS WORKBENCH michaelsa93_20160414_130539_mwtab.txt DATATRACK_ID:594 VERSION 1 CREATED_ON November 16, 2016, 9:34 am #PROJECT PR:PROJECT_TITLE Temporal metabolomic responses of cultured HepG2 liver cells to high fructose PR:PROJECT_TITLE and high glucose exposures PR:PROJECT_TYPE Time-course Study PR:PROJECT_SUMMARY High fructose consumption has been implicated with deleterious effects on human PR:PROJECT_SUMMARY health, including hyperlipidemia elicited through de novo lipogenesis. However, PR:PROJECT_SUMMARY more global effects of fructose on cellular metabolism have not been elucidated. PR:PROJECT_SUMMARY In order to explore the metabolic impact of fructose-containing nutrients, we PR:PROJECT_SUMMARY applied both GC-TOF and HILIC-QTOF mass spectrometry metabolomic strategies PR:PROJECT_SUMMARY using extracts from cultured HepG2 cells exposed to fructose, glucose, or PR:PROJECT_SUMMARY fructose + glucose. Cellular responses were analyzed in a time-dependent manner, PR:PROJECT_SUMMARY incubated in media containing 5.5 mM glucose + 5.0 mM fructose in comparison to PR:PROJECT_SUMMARY controls incubated in media containing either 5.5 mM glucose or 10.5 mM glucose. PR:PROJECT_SUMMARY Mass spectrometry identified 156 unique known metabolites and a large number of PR:PROJECT_SUMMARY unknown compounds, which revealed metabolite changes due to both utilization of PR:PROJECT_SUMMARY fructose and high-carbohydrate loads independent of hexose structure. Fructose PR:PROJECT_SUMMARY was shown to be partially converted to sorbitol, and generated higher levels of PR:PROJECT_SUMMARY fructose-1-phosphate as a precursor for glycolytic intermediates. Differentially PR:PROJECT_SUMMARY regulated ratios of 3-phosphoglycerate to serine pathway intermediates in high PR:PROJECT_SUMMARY fructose media indicated a diversion of carbon backbones away from energy PR:PROJECT_SUMMARY metabolism. Additionally, high fructoseconditions changed levels of complex PR:PROJECT_SUMMARY lipids toward phosphatidylethanolamines. Patterns of acylcarnitines in response PR:PROJECT_SUMMARY to high hexose exposure (10.5 mM glucose or glucose/fructose combination) PR:PROJECT_SUMMARY suggested a reduction in mitochondrial beta-oxidation. PR:INSTITUTE University of California, Davis PR:DEPARTMENT Genome and Biomedical Sciences Facility PR:LABORATORY WCMC Metabolomics Core PR:LAST_NAME Fiehn PR:FIRST_NAME Oliver PR:ADDRESS 1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, PR:ADDRESS CA 95616 PR:EMAIL ofiehn@ucdavis.edu PR:PHONE (530) 754-8258 PR:FUNDING_SOURCE NIH U24DK097154 #STUDY ST:STUDY_TITLE Temporal metabolomic responses of cultured HepG2 liver cells to high fructose ST:STUDY_TITLE and high glucose exposures ST:STUDY_SUMMARY High fructose consumption has been implicated with deleterious effects on human ST:STUDY_SUMMARY health, including hyperlipidemia elicited through de novo lipogenesis. However, ST:STUDY_SUMMARY more global effects of fructose on cellular metabolism have not been elucidated. ST:STUDY_SUMMARY In order to explore the metabolic impact of fructose-containing nutrients, we ST:STUDY_SUMMARY applied both GC-TOF and HILIC-QTOF mass spectrometry metabolomic strategies ST:STUDY_SUMMARY using extracts from cultured HepG2 cells exposed to fructose, glucose, or ST:STUDY_SUMMARY fructose + glucose. Cellular responses were analyzed in a time-dependent manner, ST:STUDY_SUMMARY incubated in media containing 5.5 mM glucose + 5.0 mM fructose in comparison to ST:STUDY_SUMMARY controls incubated in media containing either 5.5 mM glucose or 10.5 mM glucose. ST:STUDY_SUMMARY Mass spectrometry identified 156 unique known metabolites and a large number of ST:STUDY_SUMMARY unknown compounds, which revealed metabolite changes due to both utilization of ST:STUDY_SUMMARY fructose and high-carbohydrate loads independent of hexose structure. Fructose ST:STUDY_SUMMARY was shown to be partially converted to sorbitol, and generated higher levels of ST:STUDY_SUMMARY fructose-1-phosphate as a precursor for glycolytic intermediates. Differentially ST:STUDY_SUMMARY regulated ratios of 3-phosphoglycerate to serine pathway intermediates in high ST:STUDY_SUMMARY fructose media indicated a diversion of carbon backbones away from energy ST:STUDY_SUMMARY metabolism. Additionally, high fructoseconditions changed levels of complex ST:STUDY_SUMMARY lipids toward phosphatidylethanolamines. Patterns of acylcarnitines in response ST:STUDY_SUMMARY to high hexose exposure (10.5 mM glucose or glucose/fructose combination) ST:STUDY_SUMMARY suggested a reduction in mitochondrial beta-oxidation. ST:INSTITUTE University of California, Davis ST:DEPARTMENT Genome and Biomedical Sciences Facility ST:LABORATORY WCMC Metabolomics Core ST:LAST_NAME Fiehn ST:FIRST_NAME Oliver ST:ADDRESS 1315 Genome and Biomedical Sciences Facility, 451 Health Sciences Drive, Davis, ST:ADDRESS CA 95616 ST:EMAIL ofiehn@ucdavis.edu ST:PHONE (530) 754-8258 #SUBJECT SU:SUBJECT_TYPE Cells SU:SUBJECT_SPECIES Homo sapiens #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS GlcN-10min-R1 10Glucosamine10minReplicate1_0047.d Treatment:Glucosamine | Time:10min SUBJECT_SAMPLE_FACTORS GlcN-10min-R2 10Glucosamine10minReplicate2_0093.d Treatment:Glucosamine | Time:10min SUBJECT_SAMPLE_FACTORS GlcN-10min-R3 10Glucosamine10minReplicate3_0026.d Treatment:Glucosamine | Time:10min SUBJECT_SAMPLE_FACTORS GlcN-10min-R4 10Glucosamine10minReplicate4_0073.d Treatment:Glucosamine | Time:10min SUBJECT_SAMPLE_FACTORS GlcN-10min-R5 10Glucosamine10minReplicate5_0021.d Treatment:Glucosamine | Time:10min SUBJECT_SAMPLE_FACTORS GlcN-10min-R6 10Glucosamine10minReplicate6_0091.d Treatment:Glucosamine | Time:10min SUBJECT_SAMPLE_FACTORS 10.5mM Glu-10min-R1 10Glucose10minReplicate1_0042.d Treatment:10.5mM Glu | Time:10min SUBJECT_SAMPLE_FACTORS 10.5mM Glu-10min-R2 10Glucose10minReplicate2_0077.d Treatment:10.5mM Glu | Time:10min SUBJECT_SAMPLE_FACTORS 10.5mM Glu-10min-R3 10Glucose10minReplicate3_0022.d Treatment:10.5mM Glu | Time:10min SUBJECT_SAMPLE_FACTORS 10.5mM Glu-10min-R4 10Glucose10minReplicate4_0008.d Treatment:10.5mM Glu | Time:10min SUBJECT_SAMPLE_FACTORS 10.5mM Glu-10min-R5 10Glucose10minReplicate5_0094.d Treatment:10.5mM Glu | Time:10min SUBJECT_SAMPLE_FACTORS 10.5mM Glu-10min-R6 10Glucose10minReplicate6_0024.d Treatment:10.5mM Glu | Time:10min SUBJECT_SAMPLE_FACTORS 10.5mM Glu-1hr-R1 10Glucose1hrReplicate1_0005.d Treatment:10.5mM Glu | Time:1hr SUBJECT_SAMPLE_FACTORS 10.5mM Glu-1hr-R2 10Glucose1hrReplicate2_0034.d Treatment:10.5mM Glu | Time:1hr SUBJECT_SAMPLE_FACTORS 10.5mM Glu-1hr-R3 10Glucose1hrReplicate3_0083.d Treatment:10.5mM Glu | Time:1hr SUBJECT_SAMPLE_FACTORS 10.5mM Glu-1hr-R4 10Glucose1hrReplicate4_0037.d Treatment:10.5mM Glu | Time:1hr SUBJECT_SAMPLE_FACTORS 10.5mM Glu-1hr-R5 10Glucose1hrReplicate5_0092.d Treatment:10.5mM Glu | Time:1hr SUBJECT_SAMPLE_FACTORS 10.5mM Glu-1hr-R6 10Glucose1hrReplicate6_0030.d Treatment:10.5mM Glu | Time:1hr SUBJECT_SAMPLE_FACTORS 10.5mM Glu-6hr-R1 10Glucose6hrReplicate1_0071.d Treatment:10.5mM Glu | Time:6hr SUBJECT_SAMPLE_FACTORS 10.5mM Glu-6hr-R2 10Glucose6hrReplicate2_0070.d Treatment:10.5mM Glu | Time:6hr SUBJECT_SAMPLE_FACTORS 10.5mM Glu-6hr-R3 10Glucose6hrReplicate3_0006.d Treatment:10.5mM Glu | Time:6hr SUBJECT_SAMPLE_FACTORS 10.5mM Glu-6hr-R4 10Glucose6hrReplicate4_0009.d Treatment:10.5mM Glu | Time:6hr SUBJECT_SAMPLE_FACTORS 10.5mM Glu-6hr-R5 10Glucose6hrReplicate5_0046.d Treatment:10.5mM Glu | Time:6hr SUBJECT_SAMPLE_FACTORS 10.5mM Glu-6hr-R6 10Glucose6hrReplicate6_0031.d Treatment:10.5mM Glu | Time:6hr SUBJECT_SAMPLE_FACTORS 10.5mM Glu-24hr-R1 10Glucose24hrReplicate1_0043.d Treatment:10.5mM Glu | Time:24hr SUBJECT_SAMPLE_FACTORS 10.5mM Glu-24hr-R2 10Glucose24hrReplicate2_0090.d Treatment:10.5mM Glu | Time:24hr SUBJECT_SAMPLE_FACTORS 10.5mM Glu-24hr-R3 10Glucose24hrReplicate3_0044.d Treatment:10.5mM Glu | Time:24hr SUBJECT_SAMPLE_FACTORS 10.5mM Glu-24hr-R4 10Glucose24hrReplicate4_0061.d Treatment:10.5mM Glu | Time:24hr SUBJECT_SAMPLE_FACTORS 10.5mM Glu-24hr-R5 10Glucose24hrReplicate5_0062.d Treatment:10.5mM Glu | Time:24hr SUBJECT_SAMPLE_FACTORS 10.5mM Glu-24hr-R6 10Glucose24hrReplicate6_0053.d Treatment:10.5mM Glu | Time:24hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu 5mM Fru-10min-R1 5Glucose5Fructose10minReplicate1_0036.d Treatment:5.5mM Glu 5mM Fru | Time:10min SUBJECT_SAMPLE_FACTORS 5.5mM Glu 5mM Fru-10min-R2 5Glucose5Fructose10minReplicate2_0065.d Treatment:5.5mM Glu 5mM Fru | Time:10min SUBJECT_SAMPLE_FACTORS 5.5mM Glu 5mM Fru-10min-R3 5Glucose5Fructose10minReplicate3_0013.d Treatment:5.5mM Glu 5mM Fru | Time:10min SUBJECT_SAMPLE_FACTORS 5.5mM Glu 5mM Fru-10min-R4 5Glucose5Fructose10minReplicate4_0023.d Treatment:5.5mM Glu 5mM Fru | Time:10min SUBJECT_SAMPLE_FACTORS 5.5mM Glu 5mM Fru-10min-R5 5Glucose5Fructose10minReplicate5_0029.d Treatment:5.5mM Glu 5mM Fru | Time:10min SUBJECT_SAMPLE_FACTORS 5.5mM Glu 5mM Fru-10min-R6 5Glucose5Fructose10minReplicate6_0082.d Treatment:5.5mM Glu 5mM Fru | Time:10min SUBJECT_SAMPLE_FACTORS 5.5mM Glu 5mM Fru-1hr-R1 5Glucose5Fructose1hrReplicate1_0038.d Treatment:5.5mM Glu 5mM Fru | Time:1hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu 5mM Fru-1hr-R2 5Glucose5Fructose1hrReplicate2_0081.d Treatment:5.5mM Glu 5mM Fru | Time:1hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu 5mM Fru-1hr-R3 5Glucose5Fructose1hrReplicate3_0067.d Treatment:5.5mM Glu 5mM Fru | Time:1hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu 5mM Fru-1hr-R4 5Glucose5Fructose1hrReplicate4_0048.d Treatment:5.5mM Glu 5mM Fru | Time:1hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu 5mM Fru-1hr-R5 5Glucose5Fructose1hrReplicate5_0059.d Treatment:5.5mM Glu 5mM Fru | Time:1hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu 5mM Fru-1hr-R6 5Glucose5Fructose1hrReplicate6_0017.d Treatment:5.5mM Glu 5mM Fru | Time:1hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu 5mM Fru-6hr-R1 5Glucose5Fructose6hrReplicate1_0014.d Treatment:5.5mM Glu 5mM Fru | Time:6hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu 5mM Fru-6hr-R2 5Glucose5Fructose6hrReplicate2_0012.d Treatment:5.5mM Glu 5mM Fru | Time:6hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu 5mM Fru-6hr-R3 5Glucose5Fructose6hrReplicate3_0018.d Treatment:5.5mM Glu 5mM Fru | Time:6hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu 5mM Fru-6hr-R4 5Glucose5Fructose6hrReplicate4_0041.d Treatment:5.5mM Glu 5mM Fru | Time:6hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu 5mM Fru-6hr-R6 5Glucose5Fructose6hrReplicate6_0045.d Treatment:5.5mM Glu 5mM Fru | Time:6hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu 5mM Fru-24hr-R1 5Glucose5Fructose24hrReplicate1_0056.d Treatment:5.5mM Glu 5mM Fru | Time:24hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu 5mM Fru-24hr-R2 5Glucose5Fructose24hrReplicate2_0054.d Treatment:5.5mM Glu 5mM Fru | Time:24hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu 5mM Fru-24hr-R3 5Glucose5Fructose24hrReplicate3_0033.d Treatment:5.5mM Glu 5mM Fru | Time:24hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu 5mM Fru-24hr-R4 5Glucose5Fructose24hrReplicate4_0007.d Treatment:5.5mM Glu 5mM Fru | Time:24hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu 5mM Fru-24hr-R5 5Glucose5Fructose24hrReplicate5_0074.d Treatment:5.5mM Glu 5mM Fru | Time:24hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu 5mM Fru-24hr-R6 5Glucose5Fructose24hrReplicate6_0035.d Treatment:5.5mM Glu 5mM Fru | Time:24hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu-10min-R1 5Glucose10minReplicate1_0068.d Treatment:5.5mM Glu | Time:10min SUBJECT_SAMPLE_FACTORS 5.5mM Glu-10min-R2 5Glucose10minReplicate2_0020.d Treatment:5.5mM Glu | Time:10min SUBJECT_SAMPLE_FACTORS 5.5mM Glu-10min-R3 5Glucose10minReplicate3_0086.d Treatment:5.5mM Glu | Time:10min SUBJECT_SAMPLE_FACTORS 5.5mM Glu-10min-R4 5Glucose10minReplicate4_0032.d Treatment:5.5mM Glu | Time:10min SUBJECT_SAMPLE_FACTORS 5.5mM Glu-10min-R5 5Glucose10minReplicate5_0089.d Treatment:5.5mM Glu | Time:10min SUBJECT_SAMPLE_FACTORS 5.5mM Glu-10min-R6 5Glucose10minReplicate6_0019.d Treatment:5.5mM Glu | Time:10min SUBJECT_SAMPLE_FACTORS 5.5mM Glu-1hr-R1 5Glucose1hrReplicate1_0069.d Treatment:5.5mM Glu | Time:1hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu-1hr-R2 5Glucose1hrReplicate2_0060.d Treatment:5.5mM Glu | Time:1hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu-1hr-R3 5Glucose1hrReplicate3_0057.d Treatment:5.5mM Glu | Time:1hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu-1hr-R4 5Glucose1hrReplicate4_0095.d Treatment:5.5mM Glu | Time:1hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu-1hr-R5 5Glucose1hrReplicate5_0080.d Treatment:5.5mM Glu | Time:1hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu-1hr-R6 5Glucose1hrReplicate6_0085.d Treatment:5.5mM Glu | Time:1hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu-6hr-R1 5Glucose6hrReplicate1_0010.d Treatment:5.5mM Glu | Time:6hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu-6hr-R2 5Glucose6hrReplicate2_0049.d Treatment:5.5mM Glu | Time:6hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu-6hr-R3 5Glucose6hrReplicate3_0079.d Treatment:5.5mM Glu | Time:6hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu-6hr-R4 5Glucose6hrReplicate4_0011.d Treatment:5.5mM Glu | Time:6hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu-6hr-R5 5Glucose6hrReplicate5_0072.d Treatment:5.5mM Glu | Time:6hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu-6hr-R6 5Glucose6hrReplicate6_0055.d Treatment:5.5mM Glu | Time:6hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu-24hr-R1 5Glucose24hrReplicate1_0050.d Treatment:5.5mM Glu | Time:24hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu-24hr-R2 5Glucose24hrReplicate2_0066.d Treatment:5.5mM Glu | Time:24hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu-24hr-R3 5Glucose24hrReplicate3_0078.d Treatment:5.5mM Glu | Time:24hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu-24hr-R4 5Glucose24hrReplicate4_0084.d Treatment:5.5mM Glu | Time:24hr SUBJECT_SAMPLE_FACTORS 5.5mM Glu-24hr-R5 5Glucose24hrReplicate5_0025.d Treatment:5.5mM Glu | Time:24hr #COLLECTION CO:COLLECTION_SUMMARY HepG2 cells (ATCC HB-8065) were cultured in MEM containing 10 % (v:v) FBS, 100 CO:COLLECTION_SUMMARY U/mL penicillin and 100 mg/mL streptomycin (Invitrogen, Carlsbad, CA), 1 x MEM CO:COLLECTION_SUMMARY non-essential amino acids, and 5.5 mM glucose at 37 °C in a 5 % CO2 CO:COLLECTION_SUMMARY environment. Cells were grown for four to six passages in 10 cm tissue culture CO:COLLECTION_SUMMARY dishes with 14 mL MEM, and transferred to MULTIWELL™ 12 well culture dishes CO:COLLECTION_SUMMARY for incubation in 2 mL of the treatment medium. Upon reaching 80 % confluency, CO:COLLECTION_SUMMARY the cell culture medium was changed to media representative of experimental CO:COLLECTION_SUMMARY condition: 5.5 mM glucose MEM, 5.5 mM glucose MEM + 5 mM glucose, or 5.5 mM CO:COLLECTION_SUMMARY glucose MEM + 5 mM fructose. Media was replenished after 24 and 48 h. Cell CO:COLLECTION_SUMMARY material was collected at 10 min, 1, 6, and 24 h time points following the 48 h CO:COLLECTION_SUMMARY media replacement. Culture dishes were placed on ice and each well was washed CO:COLLECTION_SUMMARY twice with 1 mL ice-cold PBS. 4 mL ice-cold 3:1 methanol/H2O extraction solvent CO:COLLECTION_SUMMARY was added to each well. Cell material was manually scraped from each well, and CO:COLLECTION_SUMMARY the extraction solvent cell material suspension was transferred to collection CO:COLLECTION_SUMMARY tubes and frozen at −80 °C prior to further processing. CO:COLLECTION_PROTOCOL_FILENAME metabolomic_responses_of_cultured_HepG2_liver_cells.pdf CO:SAMPLE_TYPE Cell CO:COLLECTION_LOCATION Invitrogen, Carlsbad, CA CO:COLLECTION_FREQUENCY Cell material was collected at 10 min, 1, 6, and 24 h time points following the CO:COLLECTION_FREQUENCY 48 h media replacement. CO:TISSUE_CELL_IDENTIFICATION HepG2 #TREATMENT TR:TREATMENT_SUMMARY 3 Treatments: A) Control group of HepG2 cells incubated in media with 5.5 mM TR:TREATMENT_SUMMARY glucose (Glc5) B) HepG2 cells incubated in media containing 5.5 mM glucose + 5.0 TR:TREATMENT_SUMMARY mM fructose (Glc5Fru5) C) HepG2 cells incubated in media containing 10.5 mM TR:TREATMENT_SUMMARY glucose (Glc10) TR:TREATMENT_PROTOCOL_FILENAME metabolomic_responses_of_cultured_HepG2_liver_cells.pdf TR:TREATMENT_PROTOCOL_COMMENTS HepG2 cells incubated in media containing 10.5 mM glucose (Glc10) was included TR:TREATMENT_PROTOCOL_COMMENTS to enable differentiation of metabolic effects caused by fructose from metabolic TR:TREATMENT_PROTOCOL_COMMENTS effects caused by increased hexose resources TR:CELL_MEDIA Cells were acclimated to their respective media condition for 48 h prior to TR:CELL_MEDIA collection of sample material to obtain a metabolite profile representative of TR:CELL_MEDIA continued exposure instead of response to a sudden change in carbohydrate TR:CELL_MEDIA resources. Media was replenished after 24 and 48 h #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Sample material was thawed on ice, vortexed for 20 s, sonicated for 5 min with a SP:SAMPLEPREP_SUMMARY VWR 50HT Ultrasonic Bath (VWR International Inc., Bridgeport, NJ), and separated SP:SAMPLEPREP_SUMMARY into 500 μL aliquots. Each aliquot was centrifuged for 5 min @ 14,000 rcf, and SP:SAMPLEPREP_SUMMARY supernatant was collected and lyophilized to dryness. Samples was kept on ice SP:SAMPLEPREP_SUMMARY and removed only for sonication, centrifugation, and lyophilization steps. SP:SAMPLEPREP_SUMMARY Lyophilized material was used for HILIC-QTOF metabolite profiling without SP:SAMPLEPREP_SUMMARY additional clean-up steps. Lyophilized material for GC-TOF analysis was SP:SAMPLEPREP_SUMMARY redissolved in 1:1 acetonitrile/H2O, vortexed for 10 s, and centrifuged for 5 SP:SAMPLEPREP_SUMMARY min @ 14,000 rcf. Supernatant was collected and lyophilized to dryness. SP:SAMPLEPREP_PROTOCOL_FILENAME metabolomic_responses_of_cultured_HepG2_liver_cells.pdf SP:PROCESSING_METHOD Lysophilization #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Agilent 6530 CH:COLUMN_NAME Waters Acquity BEH Amide (150 x 2.1mm, 1.7um) CH:COLUMN_NAME Waters Acquity CH:FLOW_GRADIENT 100% B to 30% B CH:FLOW_RATE 0.4 mL/min CH:COLUMN_TEMPERATURE 45 C CH:METHODS_FILENAME Data_Dictionary_Fiehn_laboratory_HILIC_QTOF_MS.pdf CH:SOLVENT_A Ultrapure water with 10 mM ammonium formiate + 0.125% formic acid, pH 3 CH:SOLVENT_B Ultrapure water with 10 mM ammonium formiate + 0.125% formic acid, pH 3 CH:COLUMN_PRESSURE 200-700 bar CH:INJECTION_TEMPERATURE 4 C CH:INTERNAL_STANDARD See data dictionary CH:RETENTION_TIME See data dictionary CH:SAMPLE_INJECTION 3uL CH:ANALYTICAL_TIME 14 min CH:CAPILLARY_VOLTAGE 4500 V CH:TIME_PROGRAM 16.75 min CH:WEAK_WASH_SOLVENT_NAME 1:1 ACN:H2O CH:STRONG_WASH_SOLVENT_NAME 1:1 ACN:H2O CH:TARGET_SAMPLE_TEMPERATURE Autosampler temp 4 C CH:RANDOMIZATION_ORDER Excel generated #ANALYSIS AN:ANALYSIS_TYPE MS AN:LABORATORY_NAME WCMC Metabolomics Core AN:SOFTWARE_VERSION ChromaTOF AN:DATA_FORMAT .peg, .txt, .cdf #MS MS:MS_COMMENTS - MS:INSTRUMENT_NAME Agilent 6530 QTOF MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:CAPILLARY_VOLTAGE 3500 MS:COLLISION_GAS Nitrogen MS:DRY_GAS_FLOW 8 L/min MS:DRY_GAS_TEMP 325 C MS:FRAGMENT_VOLTAGE 120 V MS:FRAGMENTATION_METHOD Auto MS/MS MS:ION_SOURCE_TEMPERATURE 325 MS:ION_SPRAY_VOLTAGE 1000 MS:IONIZATION Pos MS:PRECURSOR_TYPE Intact Molecule MS:REAGENT_GAS Nitrogen MS:SOURCE_TEMPERATURE 325 C MS:DATAFORMAT .d MS:DESOLVATION_GAS_FLOW 11 L/min MS:DESOLVATION_TEMPERATURE 350 C MS:NEBULIZER 35 psig MS:OCTPOLE_VOLTAGE 750 MS:RESOLUTION_SETTING extended dynamic range MS:SCAN_RANGE_MOVERZ 60-1700 Da MS:SCANNING_CYCLE 2 Hz MS:SCANNING_RANGE 60-1700 Da MS:SKIMMER_VOLTAGE 1850 V #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS counts MS_METABOLITE_DATA_START Samples 10Glucosamine10minReplicate1_0047.d 10Glucosamine10minReplicate2_0093.d 10Glucosamine10minReplicate3_0026.d 10Glucosamine10minReplicate4_0073.d 10Glucosamine10minReplicate5_0021.d 10Glucosamine10minReplicate6_0091.d 10Glucose10minReplicate1_0042.d 10Glucose10minReplicate2_0077.d 10Glucose10minReplicate3_0022.d 10Glucose10minReplicate4_0008.d 10Glucose10minReplicate5_0094.d 10Glucose10minReplicate6_0024.d 10Glucose1hrReplicate1_0005.d 10Glucose1hrReplicate2_0034.d 10Glucose1hrReplicate3_0083.d 10Glucose1hrReplicate4_0037.d 10Glucose1hrReplicate5_0092.d 10Glucose1hrReplicate6_0030.d 10Glucose6hrReplicate1_0071.d 10Glucose6hrReplicate2_0070.d 10Glucose6hrReplicate3_0006.d 10Glucose6hrReplicate4_0009.d 10Glucose6hrReplicate5_0046.d 10Glucose6hrReplicate6_0031.d 10Glucose24hrReplicate1_0043.d 10Glucose24hrReplicate2_0090.d 10Glucose24hrReplicate3_0044.d 10Glucose24hrReplicate4_0061.d 10Glucose24hrReplicate5_0062.d 10Glucose24hrReplicate6_0053.d 5Glucose5Fructose10minReplicate1_0036.d 5Glucose5Fructose10minReplicate2_0065.d 5Glucose5Fructose10minReplicate3_0013.d 5Glucose5Fructose10minReplicate4_0023.d 5Glucose5Fructose10minReplicate5_0029.d 5Glucose5Fructose10minReplicate6_0082.d 5Glucose5Fructose1hrReplicate1_0038.d 5Glucose5Fructose1hrReplicate2_0081.d 5Glucose5Fructose1hrReplicate3_0067.d 5Glucose5Fructose1hrReplicate4_0048.d 5Glucose5Fructose1hrReplicate5_0059.d 5Glucose5Fructose1hrReplicate6_0017.d 5Glucose5Fructose6hrReplicate1_0014.d 5Glucose5Fructose6hrReplicate2_0012.d 5Glucose5Fructose6hrReplicate3_0018.d 5Glucose5Fructose6hrReplicate4_0041.d 5Glucose5Fructose6hrReplicate6_0045.d 5Glucose5Fructose24hrReplicate1_0056.d 5Glucose5Fructose24hrReplicate2_0054.d 5Glucose5Fructose24hrReplicate3_0033.d 5Glucose5Fructose24hrReplicate4_0007.d 5Glucose5Fructose24hrReplicate5_0074.d 5Glucose5Fructose24hrReplicate6_0035.d 5Glucose10minReplicate1_0068.d 5Glucose10minReplicate2_0020.d 5Glucose10minReplicate3_0086.d 5Glucose10minReplicate4_0032.d 5Glucose10minReplicate5_0089.d 5Glucose10minReplicate6_0019.d 5Glucose1hrReplicate1_0069.d 5Glucose1hrReplicate2_0060.d 5Glucose1hrReplicate3_0057.d 5Glucose1hrReplicate4_0095.d 5Glucose1hrReplicate5_0080.d 5Glucose1hrReplicate6_0085.d 5Glucose6hrReplicate1_0010.d 5Glucose6hrReplicate2_0049.d 5Glucose6hrReplicate3_0079.d 5Glucose6hrReplicate4_0011.d 5Glucose6hrReplicate5_0072.d 5Glucose6hrReplicate6_0055.d 5Glucose24hrReplicate1_0050.d 5Glucose24hrReplicate2_0066.d 5Glucose24hrReplicate3_0078.d 5Glucose24hrReplicate4_0084.d 5Glucose24hrReplicate5_0025.d Factors Treatment:Glucosamine | Time:10min Treatment:Glucosamine | Time:10min Treatment:Glucosamine | Time:10min Treatment:Glucosamine | Time:10min Treatment:Glucosamine | Time:10min Treatment:Glucosamine | Time:10min Treatment:10.5mM Glu | Time:10min Treatment:10.5mM Glu | Time:10min Treatment:10.5mM Glu | Time:10min Treatment:10.5mM Glu | Time:10min Treatment:10.5mM Glu | Time:10min Treatment:10.5mM Glu | Time:10min Treatment:10.5mM Glu | Time:1hr Treatment:10.5mM Glu | Time:1hr Treatment:10.5mM Glu | Time:1hr Treatment:10.5mM Glu | Time:1hr Treatment:10.5mM Glu | Time:1hr Treatment:10.5mM Glu | Time:1hr Treatment:10.5mM Glu | Time:6hr Treatment:10.5mM Glu | Time:6hr Treatment:10.5mM Glu | Time:6hr Treatment:10.5mM Glu | Time:6hr Treatment:10.5mM Glu | Time:6hr Treatment:10.5mM Glu | Time:6hr Treatment:10.5mM Glu | Time:24hr Treatment:10.5mM Glu | Time:24hr Treatment:10.5mM Glu | Time:24hr Treatment:10.5mM Glu | Time:24hr Treatment:10.5mM Glu | Time:24hr Treatment:10.5mM Glu | Time:24hr Treatment:5.5mM Glu 5mM Fru | Time:10min Treatment:5.5mM Glu 5mM Fru | Time:10min Treatment:5.5mM Glu 5mM Fru | Time:10min Treatment:5.5mM Glu 5mM Fru | Time:10min Treatment:5.5mM Glu 5mM Fru | Time:10min Treatment:5.5mM Glu 5mM Fru | Time:10min Treatment:5.5mM Glu 5mM Fru | Time:1hr Treatment:5.5mM Glu 5mM Fru | Time:1hr Treatment:5.5mM Glu 5mM Fru | Time:1hr Treatment:5.5mM Glu 5mM Fru | Time:1hr Treatment:5.5mM Glu 5mM Fru | Time:1hr Treatment:5.5mM Glu 5mM Fru | Time:1hr Treatment:5.5mM Glu 5mM Fru | Time:6hr Treatment:5.5mM Glu 5mM Fru | Time:6hr Treatment:5.5mM Glu 5mM Fru | Time:6hr Treatment:5.5mM Glu 5mM Fru | Time:6hr Treatment:5.5mM Glu 5mM Fru | Time:6hr Treatment:5.5mM Glu 5mM Fru | Time:24hr Treatment:5.5mM Glu 5mM Fru | Time:24hr Treatment:5.5mM Glu 5mM Fru | Time:24hr Treatment:5.5mM Glu 5mM Fru | Time:24hr Treatment:5.5mM Glu 5mM Fru | Time:24hr Treatment:5.5mM Glu 5mM Fru | Time:24hr Treatment:5.5mM Glu | Time:10min Treatment:5.5mM Glu | Time:10min Treatment:5.5mM Glu | Time:10min Treatment:5.5mM Glu | Time:10min Treatment:5.5mM Glu | Time:10min Treatment:5.5mM Glu | Time:10min Treatment:5.5mM Glu | Time:1hr Treatment:5.5mM Glu | Time:1hr Treatment:5.5mM Glu | Time:1hr Treatment:5.5mM Glu | Time:1hr Treatment:5.5mM Glu | Time:1hr Treatment:5.5mM Glu | Time:1hr Treatment:5.5mM Glu | Time:6hr Treatment:5.5mM Glu | Time:6hr Treatment:5.5mM Glu | Time:6hr Treatment:5.5mM Glu | Time:6hr Treatment:5.5mM Glu | Time:6hr Treatment:5.5mM Glu | Time:6hr Treatment:5.5mM Glu | Time:24hr Treatment:5.5mM Glu | Time:24hr Treatment:5.5mM Glu | Time:24hr Treatment:5.5mM Glu | Time:24hr Treatment:5.5mM Glu | Time:24hr Choline 4825 5245 3613 4150 4855 4385 3172 3996 3804 3353 3426 3745 3558 3456 3216 3396 2713 2654 4066 2875 3211 3894 3023 3081 5164 4219 4575 4307 5147 5510 3113 2108 2983 3828 2291 3118 3104 3660 3402 3393 3439 2795 3191 3403 3481 3073 2377 4214 3111 4260 4488 4454 4350 1935 1884 2147 1899 1760 2047 1584 1732 1711 1729 1751 1668 2503 2632 2213 2894 2426 2736 1112 1694 2261 1761 1328 Creatinine 5730 7836 4978 6575 5759 6141 12343 19029 16851 13868 21400 16159 12982 16273 17503 10655 18200 14294 19015 15532 16552 24800 13834 12652 16341 10712 8819 11019 13132 27800 11758 16830 14818 16323 11269 15830 14125 20145 16683 17714 21695 14717 21645 15970 19965 13542 21821 13827 15909 11640 13334 13346 14938 16575 17479 15007 14300 17292 18474 16809 21571 22715 26001 23190 20160 24755 19121 26511 35401 18342 18056 10476 8687 13389 11636 9148 Proline 36792 43553 27581 35314 34577 33244 41821 42428 51102 46225 40587 44437 51259 49818 44789 43505 44918 42999 67357 63128 59484 72108 54558 53589 86475 63952 69646 62879 63336 70904 45317 41907 34243 47960 35976 44077 46618 47909 49924 48811 55339 48848 50773 56381 61714 54128 54963 63824 53052 69374 67021 71592 72651 43735 39607 33460 41006 40620 43513 40841 59519 51525 44348 45923 46519 65070 61281 69953 72246 62782 54705 26823 40242 48227 42645 34366 Betaine 151346 164714 119065 163192 139880 157260 144963 160701 127919 131050 136443 129804 142946 157465 151458 123990 142714 136489 203228 146139 144219 168794 137533 120729 191807 127570 139220 143361 171282 223047 167737 170993 130456 158432 137009 204267 172855 190881 181427 170078 174897 164964 151771 150880 157044 164574 143696 177528 149702 203937 173842 179806 189372 164398 139770 160018 137874 176950 151596 179168 161420 171735 162452 169575 168701 144768 153863 187961 158294 184257 156056 134193 151678 205499 177299 123386 Taurine 16984 23931 14127 22156 20713 19984 63262 70295 73990 64681 66829 68390 70944 61185 68233 55834 66618 63805 83947 69264 66565 76478 54616 67253 76255 66434 59138 68283 65489 71918 76273 74272 66754 91940 74163 89606 74222 85158 84823 78711 94460 88124 60223 71560 64736 62491 54538 90458 76693 100632 92055 114375 99332 88663 90065 85488 88083 120670 99959 86094 97192 94713 87905 92563 91572 56692 64590 73812 69848 67936 58780 67791 108280 136772 122597 91832 Creatine 154610 200504 108258 173606 157148 171324 263061 294395 251514 278861 264963 263188 218171 263553 228279 194900 228164 229386 339769 322765 283622 335388 287449 242645 326019 240659 264640 277535 246874 239623 339571 262389 257016 352695 274439 399456 312475 336470 350853 335520 384539 306397 296322 353239 348579 361144 279281 349279 312722 421424 325258 453012 421782 271976 246695 251485 225413 302106 265839 222679 278931 252100 242278 249152 249368 270391 294882 376211 312520 318917 330642 319943 452162 571321 500226 359262 Adenine 1609 2746 1504 2709 1836 2586 3125 4334 4501 3118 3094 4332 3790 3465 3768 3062 3251 3657 2308 2660 1942 1442 1701 1995 5061 6042 4978 6108 4979 6508 3597 3611 3029 3436 3645 3528 2928 3052 2941 2693 2996 2992 2318 2019 1849 1858 1471 4651 4338 4186 4333 5954 6589 4320 3548 5121 3893 4872 4182 2838 4153 4312 3767 3981 4850 2140 1697 2209 2069 1977 1993 3156 4846 7337 6743 6264 Glutamine 21787 33478 13988 15140 20473 15018 24425 16303 36192 43369 21213 24655 66111 41458 26848 37516 23427 27105 22280 39314 48732 57459 22572 25048 9062 4078 5392 8084 5575 10897 25984 18243 23093 32350 16958 13706 37865 32333 51487 35061 93382 33766 22707 38379 39462 22964 26420 5105 3636 4964 9448 7153 6557 32124 29666 11872 25661 19362 32132 24340 87766 44111 30482 32555 28535 48071 30988 31044 58754 22128 29108 3212 4266 3184 2505 1241 Carnitine 18511 25722 11695 18784 15711 16380 33618 35220 46546 35354 41320 33065 57233 46424 53860 45933 40704 33436 52505 55863 45620 56790 30006 41961 58205 46618 41536 51802 50005 49921 44650 45411 38703 53691 34348 45237 58363 64647 79365 59796 93608 55483 33233 45069 51738 39797 44818 53240 41196 48288 56618 67073 51408 61234 51504 45788 42930 57060 46221 54543 92656 64472 59682 60341 59917 61126 57758 66028 62021 58005 45873 19807 36273 38890 34427 22451 Phenylalanine 11880 12732 11701 9992 14638 8827 14544 11867 20173 20648 7560 18755 27043 22722 11942 16337 11533 23330 16744 13380 22537 32251 14156 15919 27368 9030 14269 11308 12344 29763 20070 17975 18573 25292 17175 12893 24228 15234 20218 23272 27957 26357 19873 21742 27310 17782 14073 16151 12345 19218 22735 15645 26240 15157 17847 8536 22954 13101 25426 12875 24510 19088 12238 13589 12437 24144 15043 13397 29900 13154 18223 9922 9493 14092 9175 14085 Acylcarnitine C2:0 15584 22066 10911 16432 13301 15503 50556 55723 68519 58856 68738 54234 50412 45184 46686 42977 40195 30770 40863 81856 51817 42431 36962 28518 36791 28875 29750 35535 36743 37681 51760 36881 41704 61354 40304 56659 36315 37333 50981 38825 66061 29636 34748 50323 39389 47704 45101 44806 33216 40972 48364 52912 37739 53292 43502 41882 35060 53970 44264 32536 67774 41050 36382 40836 38241 34748 54125 57406 51655 40561 34695 27637 42166 49930 47985 34434 Acylcarnitine C3:0 5701 6842 4124 6377 6259 5058 3922 3419 4722 4017 3606 3896 6056 4609 4354 5420 4441 4854 3094 3592 3599 5748 3048 2951 4504 3286 4521 4614 4563 5413 4090 4841 3486 5627 3633 4841 6167 6624 5918 4768 8414 5572 3172 3841 4370 3767 2935 3844 3311 5003 4792 4323 4824 5178 5143 4134 4814 5783 4681 4827 6371 5307 4370 4708 5364 3705 2990 2634 4000 3346 3228 3086 2892 4051 4246 2558 Pantothenic Acid 33311 40823 27194 43724 36932 37091 46621 51552 58523 52525 49983 54920 78909 75532 75921 65435 75585 81028 89036 71670 73624 96421 59573 71275 29344 23075 23920 23291 24539 27589 44621 41079 39461 53938 45204 52146 78574 74614 86950 78767 92003 82402 69541 78316 87752 68060 77980 27479 22276 28526 33050 30240 26695 45642 48208 40242 46295 53496 51938 76875 97973 84549 77213 85939 80043 93403 79765 100476 97884 89304 70390 11993 17508 20383 17450 15978 Acylcarnitine C4:0 7913 16579 4598 7941 7086 6893 14862 15953 20345 16670 19691 15840 32214 24315 31513 26098 22868 17066 20968 33603 21370 22865 14408 14737 13387 10135 11164 12939 12083 12863 18170 18787 15529 21137 15866 21211 29121 35610 51905 29087 89597 26798 16591 23483 20206 19298 21283 14424 11092 13648 17423 16297 12215 24617 17315 14029 15001 23001 17414 24851 80898 31666 27968 32453 32232 21437 25582 30741 24471 22147 19388 4766 6738 8433 7907 5859 Taurine Dimer 776 1257 413 1176 1210 699 5641 6001 9525 6617 6139 8139 7193 5866 6313 5738 6256 6706 6626 7315 7556 7357 5094 7823 8158 7679 6063 8002 5660 5097 8698 7558 7256 10913 8286 8721 8002 9415 9691 8076 12450 9942 5040 8741 7001 5338 5782 10952 8764 11610 11913 17017 12814 11954 12738 9095 11395 15874 13153 9230 13964 12392 9898 10274 10826 4939 7039 7261 7999 6133 5890 5948 16544 16644 16337 13496 Thiamine 32220 48893 26937 37843 33354 33817 34049 41254 40695 36023 27285 38074 59061 57866 57698 46753 45809 54434 46936 40427 41509 56752 34194 35221 56330 36799 32720 34851 36668 71507 40608 55334 35030 47727 38551 45073 58049 47056 61289 53407 71374 56530 41198 45700 51105 40730 34511 44069 34847 47622 47061 54107 56899 49953 41819 40857 49962 55436 53675 45761 60907 55137 45609 45581 43545 45637 35793 37403 54969 37761 42377 41431 45467 61724 53905 42437 Adenosine 2431 2911 2004 3219 1790 3071 2899 3865 4642 4413 3620 4221 3985 2610 3388 2447 2375 2557 4071 5855 6215 4935 4554 1556 3589 3914 3769 4208 3453 4421 3751 4366 3916 4092 3209 4715 3936 4414 4427 3749 6273 5075 5815 6169 4730 5151 3042 4671 4299 4758 5964 5115 5112 2685 2022 2558 2516 3240 3479 1586 3386 2244 1768 2699 2871 1905 3518 2696 3010 3868 4156 2037 2749 3580 4061 3701 5-Methylthioadenosine 35475 55710 29000 51308 39244 48072 68214 94103 97162 78724 74539 84358 91842 72063 72799 64913 60138 92376 38174 42228 41259 36181 39012 46015 118898 137701 119733 138907 120364 155157 90344 70959 64224 68138 75460 88357 51173 60412 64942 58264 71194 67389 46100 44489 40344 35853 41667 109672 98032 95852 96280 131707 141519 83974 75835 101216 86422 98071 93946 62852 95061 83495 73569 86536 86591 42411 35172 40098 41839 42745 42113 72065 101977 170192 174432 167519 Acylcarnitine C12:0 354 86 360 491 243 106 1352 1432 1957 1360 1269 1472 938 827 622 853 798 1022 1617 1477 1635 537 1119 540 3395 1462 1994 1705 1709 1739 3623 2146 2341 4020 2773 3388 2098 2142 2306 3263 2804 2032 2764 2834 2610 3302 1672 7284 5071 6310 7835 9435 6151 3428 3687 3119 3404 3524 4119 2164 2704 2417 2179 2340 2434 925 2322 1463 1977 1280 1874 6357 9376 11970 8870 9749 Acylcarnitine C14:1 525 612 275 699 560 679 1504 1594 1043 1414 1125 1204 865 1064 937 700 973 915 1455 1684 1169 822 1124 639 2483 1423 1370 1809 1792 1993 3619 2576 2071 2996 2503 3942 2973 2677 3161 3235 2800 2169 2848 2571 1984 3388 2028 6228 4873 6167 5800 7682 5986 4688 4374 4440 4601 5141 5108 2927 3307 4265 3174 3786 3931 1131 2021 1697 2337 1655 2284 9911 12099 14519 12291 10372 Acylcarnitine C14:0 3227 3036 2249 3613 3714 2627 6590 7289 6419 6772 6094 7120 5123 6147 4117 3832 4903 4353 6924 8456 6757 5138 7987 3316 9530 5361 5774 6452 6277 6578 15437 9816 9874 13058 11793 14498 9925 9922 9618 10965 10258 8457 10834 10518 8532 12895 9575 19156 17406 21094 18761 22204 20459 17598 17478 18200 15357 20622 19904 11945 11474 13447 11896 12885 12562 3817 9671 7714 7465 7949 8270 24646 31061 41570 33083 28480 Acylcarnitine C16:1 1748 1602 1320 1797 1802 1861 2837 3590 2660 4033 2687 3356 2302 3085 2021 1820 2458 2599 3413 4207 3517 3254 2941 1773 2983 1720 1990 1742 1941 2458 5080 3374 3911 5209 4479 5279 4256 4488 4851 4621 4039 4745 5447 4670 3884 6824 4572 6078 4962 6211 5349 6830 5820 8004 7838 8663 7627 10020 10322 6277 6259 7517 6233 6581 6162 2083 4099 4017 3511 3290 3833 7400 10664 12933 9817 9372 Acylcarnitine C16:0 9547 9256 6849 8593 8976 9129 14913 15480 15577 15601 13025 16457 10324 11167 9298 8286 9800 9764 12942 14920 12061 8527 10992 5821 14085 8760 8713 9690 10096 12201 23379 16772 17112 25291 19520 27330 15382 15610 14627 16746 15463 15067 16145 15465 14127 17908 12996 26316 24431 30203 26572 28810 30758 25119 27222 29760 26613 31132 32054 19594 18007 22868 17872 17170 18397 6303 14698 10209 12313 11850 12262 35240 44466 61608 45090 43675 Acylcarnitine C18:1 4538 3071 3189 3324 3895 3231 6962 6152 6119 8002 5753 6906 4453 4865 4232 3815 4112 4747 6480 6195 5686 4352 5903 3063 7832 4291 4421 4880 4889 6820 9228 6138 7119 10464 7901 10512 6805 6942 7242 7714 6003 7218 8503 8219 6507 9933 6557 12404 10173 14862 12083 12590 13114 12599 15101 14189 13982 17232 17845 9833 8835 12895 10228 10051 10869 3481 6823 4871 5837 4731 5885 18531 20311 29640 20403 20915 Acylcarnitine C18:0 2008 1664 1406 1619 2580 2152 4348 3409 3063 4273 2999 4173 2248 2562 1495 2031 2328 2633 4662 3673 2793 2262 2979 1871 7557 4242 4848 4585 4511 6734 7329 3890 5089 7297 4596 6382 3585 3073 2964 4266 2972 3765 4857 4037 3380 5257 3394 14287 12483 16554 14844 12582 14896 7292 9201 8156 8006 7813 9977 5199 4476 6083 4727 5050 4731 2511 4255 2589 3434 2560 3303 20938 22446 28410 19677 23456 lysoPE 16:0 5959 6083 4356 5532 5431 6337 7135 6857 6025 6579 6328 6847 6426 6212 5664 4320 6468 6143 6214 5431 4913 5381 5010 3583 7304 5458 6030 5616 6093 6341 7621 6547 6129 8230 6394 9223 7719 7166 7487 7816 6961 6726 4469 4952 5148 5937 4783 7438 6812 7549 7563 6852 6587 7242 7677 7479 7094 9559 7912 7687 7922 7690 7157 7574 8220 5161 5441 5225 4895 6118 4173 5527 6889 10706 8948 6348 lysoPE 18:1 5420 6268 3534 5819 6254 5788 8720 8370 8718 9052 8378 8833 9717 8716 7538 6852 8213 8689 8125 7248 7169 7495 6755 6237 11326 9626 9652 8106 9200 11234 9203 7821 8228 9340 7905 10839 9168 8575 8778 8973 8496 9242 6097 6972 7191 7733 6378 10624 9938 10678 11955 10949 10104 8097 8920 7712 7997 9527 8486 8872 9617 9113 7914 8017 9046 5937 6922 6315 7080 6447 6010 7827 8426 11408 11489 9584 lysoPE 18:0 8770 9141 6694 8395 9256 8207 12872 13204 13657 13834 13687 13333 12068 11142 10743 9326 10305 10827 9921 10289 10504 10006 8983 8538 11259 10554 10366 9758 9068 9045 12043 9986 9984 13665 10032 13569 11094 10616 11370 11098 11678 11732 7412 8329 8590 9609 6788 10291 9685 11168 12088 9924 10242 9754 10932 8632 8551 11076 10850 8962 9208 9648 9121 9242 8638 7286 7662 7885 8831 7112 7103 7111 8577 8413 8135 8484 lysoPC 16:1 4892 4650 3218 4978 4481 4593 5221 5615 5326 4587 5790 4801 7119 6498 6300 6952 6494 5578 4893 7605 5742 5565 5968 4719 5709 11036 6920 7236 5009 5502 5596 4828 3959 4172 4850 4713 7137 8096 6852 6940 7771 5304 5233 4983 5747 6499 6329 8082 7982 6707 6509 9179 9940 7206 5308 5843 5390 5067 5246 5618 8235 6385 7068 7304 6003 5226 6694 6953 5477 5858 5936 5374 6450 7107 7872 6595 lysoPC 16:0 89271 97427 63560 88637 88536 87298 103571 106396 115535 103740 98834 105344 119140 128330 117538 106612 117450 115492 106971 106661 88721 104644 88936 79166 122266 116007 100725 103931 104483 110853 94072 80458 69474 98573 78409 96196 113369 103585 111895 104125 121402 102297 65345 70551 79922 77826 75522 86835 79805 93883 86236 98625 94608 90709 93079 84490 85164 98336 91666 92665 109025 107044 100707 93520 92377 77078 84727 82162 85140 77674 70270 54482 72292 89277 85620 69656 lysoPC 18:1 31516 33224 23231 32888 30955 35248 38358 42590 44154 37737 45691 35728 48083 45817 46795 47052 44946 40238 39752 49589 41249 38247 42972 35045 59018 67526 53742 55816 48147 51639 37717 33308 27183 32197 30214 34546 44074 46534 40765 41572 45927 37492 27920 31220 31712 36727 37036 43350 42380 41444 42151 51651 50067 38545 34849 33233 33211 36554 32934 33598 44678 39080 39412 40704 34597 32389 38468 35342 35312 31862 33771 31885 36500 41889 42564 38271 lysoPC 18:0 64617 72974 48462 63576 66153 65200 92780 93323 98014 95474 86198 94678 100825 110826 98575 98458 99499 101216 93605 88575 78742 95340 76763 70336 84974 86715 73832 72288 82183 76131 72827 64988 57842 81635 62404 77428 90217 82106 89018 85754 98161 86039 52552 58675 65763 59025 62115 50239 45388 55787 53424 54891 57697 80495 77577 71104 74674 86040 80228 88473 97806 97729 87906 81470 79125 62150 72441 68502 75890 62227 56345 37831 50003 61296 51383 46541 SM 32:1 35832 35220 26941 30989 34084 32422 41783 39095 37480 42512 40716 39221 39618 43697 35529 35985 39202 35122 36414 41060 44055 45445 38934 36532 35834 37575 41430 42996 33951 29007 53261 38391 40998 43428 35635 44634 44944 47843 50306 52976 49017 45667 43178 46346 52439 51756 42175 47209 52657 49968 48627 51046 53060 47206 46384 40569 37742 44531 46451 46028 51813 46928 42979 44565 42259 49113 53125 50699 45121 50088 44222 45163 46009 42674 44139 47263 PE 32:2 11633 13120 11877 15024 13786 10986 7913 8026 8511 9013 7213 8561 7895 9874 7772 9541 8915 7551 6850 9724 9904 8380 9219 9506 7100 11711 8540 8734 7258 4852 13229 10520 12691 9353 12754 8500 13132 12639 13018 12373 14421 14146 13073 13302 13428 12370 12708 14367 14087 14618 14771 18587 16705 13657 13054 12976 14832 11918 19219 13226 17434 14411 12326 14907 14329 12879 11866 13124 13184 13692 12641 17226 16448 12251 18378 21474 PE 32:1 71338 76182 74340 80307 68985 66346 43484 44408 57709 47795 48787 49542 44583 49601 46988 58424 47230 43960 32986 62580 53660 39463 48371 48528 36625 59228 46553 53076 35250 21799 63192 51268 58473 44429 65628 32893 65893 65643 69799 59037 83614 65737 60225 61658 63595 63976 70040 63034 68424 59348 66277 88047 74748 78146 70947 61765 75184 53777 81653 62288 89186 68404 67143 70538 68313 65576 58920 70707 68625 69207 67643 70005 91365 59076 74119 89960 SM 34:1 479592 403674 388304 353325 381001 389282 520017 467719 364106 486282 470304 453809 529138 487651 430057 436386 503200 422908 435022 464546 501364 443883 463783 415298 463420 271099 425714 451514 406670 397979 572487 419045 439435 399467 400486 463614 513412 545668 515739 608254 434781 475632 480381 383792 465209 559394 454077 452464 480967 455992 470347 343111 411593 497604 452384 410041 366592 494529 478905 471705 467516 508529 476432 570999 446625 445738 543673 489705 431258 480217 418488 504775 449319 413675 379103 434902 PE 34:3 13503 15493 14203 15767 14729 13197 11117 11331 13289 13252 10632 12591 11530 12530 10971 14405 11741 10897 8951 13370 13652 10919 12919 12848 8573 13805 10286 10361 8625 5536 14334 12285 13503 11630 14376 9533 14520 14605 15232 13997 16960 16429 15380 15385 16453 16194 15882 13982 14372 13791 16684 17053 16597 14571 14609 14233 15864 11864 16696 13660 16698 14716 13289 13664 15993 16035 14126 17244 16096 16046 16966 13066 15070 10834 13513 16312 PE 34:2 187513 196415 194670 209483 187340 176922 138767 143506 185152 150077 145998 161568 150721 157024 160018 187439 150549 147450 104470 187492 177807 129429 157014 163219 106432 183793 137697 143225 105229 66070 168612 151145 169102 130582 181613 95070 194179 186468 197278 167972 209366 202113 183473 202250 194884 187570 212360 169307 176634 158592 181928 209364 192113 208051 196547 180519 215920 150570 216293 178331 209114 195476 172302 190797 191430 200971 186750 210182 197611 205012 202789 174291 214137 141124 176382 220039 PE 34:1 144601 158666 159880 156447 134405 146192 118187 105277 129968 111393 118173 115461 108255 122466 120416 130874 119559 120255 89514 126400 111005 87215 101336 98742 106386 122472 111919 126333 98853 59857 131973 114947 125174 106172 139543 81793 154242 133942 157521 125546 173065 144980 135246 131767 138431 124387 131267 144548 140648 135191 138855 167706 148614 191876 149750 146970 162977 126309 173739 143814 172548 165120 140317 147374 147261 133915 127732 139782 120621 137412 142989 148574 207264 148456 190405 192457 PC 32:2 211265 192964 192949 197159 229599 227190 202975 188350 140429 190972 152779 191219 213824 226053 171383 153946 203549 194460 325410 182244 252486 305060 209314 210407 223790 161799 184739 162209 212946 272728 238466 186126 214962 251182 179339 334002 256618 213569 227105 274169 181033 255327 281795 256911 268217 280748 202522 263305 253632 324493 272425 206615 265932 198449 227110 244111 193798 312687 263964 269814 201814 269837 269828 229942 268722 271803 260328 245363 261256 283222 232008 276343 212407 333579 316613 241909 PE 36:4 142525 150852 142374 156176 144105 132037 129256 125654 172557 134680 130534 143257 138010 146272 146962 188339 134825 137601 100308 164674 161820 117984 148144 155678 99311 177217 132811 136074 96625 57879 141441 125456 147226 108388 157099 75154 171368 161125 159402 138571 193219 171827 168963 182129 163833 165768 191936 142026 149683 136407 156792 179109 171031 170928 161419 148037 178027 118071 172809 148541 181315 170476 139266 148994 149222 168790 160972 182417 169399 176596 188704 133334 172026 100783 132546 174497 PE 36:3 177487 194853 190713 198768 184557 173125 166269 160992 213403 171391 169667 185548 182064 191367 185942 229861 172058 187730 130335 214209 211866 145730 185328 196640 130745 206990 170917 181507 126943 78120 191360 176596 209285 152778 215985 107517 239080 219052 231569 192736 247478 246529 237348 248096 234468 227702 236946 205619 215403 194419 225817 234964 233352 226827 217607 199923 239269 154529 230316 201457 217354 224830 184440 188063 198661 228896 213303 238497 222096 233157 254204 169307 215782 140986 184253 231425 PE 36:2 290198 317541 332336 312949 289592 297180 302955 270897 334370 273591 277475 292806 283431 307916 299430 357245 285168 307135 234499 304585 289919 234978 264632 273239 251566 323894 284360 292018 242502 151972 284675 269697 313807 265831 323297 188228 373811 307447 354576 293958 380624 358111 336536 358665 331058 298314 321823 313124 314785 328973 332938 376694 338573 384156 336872 327895 374047 281179 364321 309737 344646 354425 289256 291663 311289 332230 305508 328308 294814 327605 378477 298596 378328 281651 328556 365376 PC 34:4 356365 372698 439195 509884 451935 350750 367955 381496 429769 448568 352960 466215 432364 501580 372075 396387 388805 477210 433446 464083 455441 526799 416125 531865 384536 392693 396982 385075 359440 417771 444166 331671 435125 445106 433473 402772 451584 411012 492796 429242 515628 510236 468929 512169 568593 426144 423452 369515 361879 533666 484776 477202 438704 389980 467171 365287 449861 446541 478083 374035 425024 367030 369174 426744 405118 510454 437789 426899 540158 464468 413578 394729 354023 393923 375930 455487 PC 34:2 1038508 977338 1031028 940897 1055997 1069071 1117963 1102488 921201 1075273 1032520 1084102 1077930 1093882 1044109 943376 1133823 1114608 1392119 1065741 1134528 1345052 1181459 1169057 1039087 884179 971038 918107 1070790 1273352 893253 1006205 965793 1033433 899611 1236928 1116486 1097916 1089729 1242478 968938 1126261 1197230 1172263 1177034 1210188 1114900 1082077 1087876 1085558 1044499 952368 950083 970904 1047318 1123777 987726 1225198 1012672 1169689 1007097 1056086 1163194 1093448 1170443 1304828 1294398 1215286 1224630 1275182 1179237 1054925 880725 1064402 1028387 913166 PC 34:1 2963360 2600160 3173826 2828846 3047953 3008582 3451673 3206858 2891887 3248843 2999763 3258242 3268501 3312256 3007859 3005648 3200719 3292044 3594286 2779762 3179917 3312156 3196417 3166857 3292943 2548038 3037615 2583346 3063717 3550507 2913508 2771235 3014954 2868101 2801883 3082409 3442532 3205961 2976105 3306618 2589629 3388809 3414935 3142297 3110152 3355657 3207330 3004220 3027118 3416300 3139480 2523733 3059581 2732575 3145098 3183245 2973116 3165954 3087952 3215574 2511466 3172505 3124161 3163019 3063347 3238896 3124923 3155952 3290915 3400722 3107439 3186864 2554963 2969125 2796588 2703407 PE 38:6 52709 53076 53488 56993 55625 49976 49155 50419 61600 51711 48741 55718 55518 56576 56680 73332 51797 56020 42952 62002 66047 50026 59970 63454 38361 61238 48354 47979 37922 25568 52304 48011 57713 44145 61393 32299 69368 62045 62126 55373 69960 73657 72965 76599 66862 71197 74309 57034 59958 56998 61969 62690 67469 61246 61589 56816 66552 45668 64794 57744 63904 66114 52081 54215 58582 65355 65221 71448 64206 68532 77525 47763 57562 39549 48415 61015 PC 36:3 217069 260322 181668 219228 180990 221656 228231 277353 277934 244936 354077 231967 235420 203776 333987 282316 287938 274204 251821 371455 258483 273091 345097 339843 177967 330228 253941 358445 302108 206636 164705 286047 201209 227531 236752 198795 250216 335773 340931 299590 373773 251220 278867 291946 283489 279978 334983 323809 339188 160926 240084 355436 233567 309847 238162 285313 275905 257916 207883 294756 352845 259195 314842 269394 303959 330943 402318 352746 317725 296961 417343 243379 295809 195948 235820 235880 PC 36:2 904033 1061883 853414 883082 767367 917655 1094034 1264895 1434364 1186373 1643840 1116024 1139015 987999 1602957 1445826 1370809 1234695 1034794 1631344 1290258 1136469 1580936 1552675 823803 1434657 1157657 1497926 1354919 872598 757624 1278706 899297 957136 1103590 768095 1115944 1463677 1410432 1303042 1530191 1133714 1187552 1303563 1249734 1167780 1544528 1309702 1378670 716395 988317 1376802 957038 1376390 1105842 1244914 1329037 1032913 908075 1275785 1449032 1052039 1394016 1220256 1311503 1490884 1640980 1448711 1416103 1276445 1679464 992998 1169028 738996 886597 968139 PC 38:5 247308 228191 263860 269399 261402 241600 300121 313492 254614 280636 249075 285931 290915 283037 268379 270854 284668 291654 356184 253113 289331 302789 295837 296491 332647 227469 277071 209567 261217 260582 248870 272776 260058 264858 244945 254632 297874 305245 268589 292940 273364 297251 297373 281899 283459 303045 293424 262349 262031 284686 271935 206436 281127 235242 274348 282019 259700 258276 270257 284301 205550 277804 252745 279245 268834 280269 268545 310103 294448 328222 296193 243394 208230 218379 217727 229114 PC 38:4 93621 118764 95732 114293 107507 88659 113863 118642 189364 131302 119647 142147 126023 140985 122077 198724 118559 119902 76252 164877 166788 107373 146530 149275 90152 212229 144621 147212 84687 46047 121605 100278 129173 90418 131330 52694 134954 120610 142344 109463 169193 134889 135877 156353 150095 130433 144612 109491 119823 111690 136331 168117 167624 156264 145102 120819 164681 92177 156193 101678 155519 127028 102435 107364 115980 157585 129908 137157 144029 142279 147847 89536 131337 67737 91517 151006 PC 38:3 51811 64448 59610 64226 63352 51497 78080 84960 140229 93292 81216 99920 92655 95746 87797 140578 85268 90132 50530 105452 113651 68166 87557 101474 61170 163064 94761 102362 55248 29434 82856 70533 96083 62941 97636 36024 98606 90862 99632 70221 135139 106739 101915 112905 108190 83751 88484 75244 77662 72730 99286 108237 109111 106875 96038 79879 114612 53161 106550 69658 107242 81848 65350 72988 70450 90966 70118 89373 90141 89961 95842 55314 83023 41957 56607 93725 PC 38:2 46411 61818 54909 57278 57126 47744 93060 103287 169714 104957 102956 127984 119133 120217 123524 184535 104113 111610 66256 141255 144358 73369 113119 132602 79455 187167 120816 137280 74366 37646 89005 88370 105665 64620 113547 37242 118416 101619 108703 75200 155752 127541 130702 135053 125561 104084 115167 88286 88221 84257 108198 134160 121960 117018 106701 88946 123713 54307 112492 77384 125295 92934 76406 78194 76086 102278 92945 106412 99840 112154 138125 56750 93816 42269 56613 101002 PC 40:7 32120 40368 32971 38561 39640 28801 40720 40957 63199 48165 40473 53544 41956 50210 42482 61997 40685 43527 26174 54090 61106 40095 50763 55687 32020 68892 52347 48402 27500 13572 42757 35066 50252 34019 49096 16454 46857 40738 48865 39122 50400 53073 52821 60401 58396 48899 51023 40141 44008 38130 52174 53848 61612 51000 55059 40991 60697 30616 59092 36590 50153 45659 34667 36187 40965 61920 48116 50531 52091 50063 52194 32160 47054 22922 34160 58551 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name row ID row m/z retention time Pubchem ID Choline 2655 104.1071794 13.753 305 Creatinine 124 114.0662848 4.167 588 Proline 4854 116.0711927 11.194 145742 Betaine 5018 118.0864932 11.424 247 Taurine 5248 126.0220355 4.995 1123 Creatine 8522 132.0768601 12.209 586 Adenine 2970 136.0639019 2.491 190 Glutamine 8626 147.0804022 12.372 5961 Carnitine 359 162.1115757 13.482 10917 Phenylalanine 3523 166.0864615 9.57 6140 Acylcarnitine C2:0 8099 204.1229547 13.103 7045767 Acylcarnitine C3:0 3752 218.1357026 12.818 188824 Pantothenic Acid 6783 220.1179959 4.898 6613 Acylcarnitine C4:0 1941 232.1535555 12.493 213144 Taurine Dimer 5105 251.0358709 4.968 1123 Thiamine 8570 265.1122176 13.319 1130 Adenosine 397 268.1055386 3.152 60961 5-Methylthioadenosine 5256 298.0973149 2.475 439176 Acylcarnitine C12:0 26324 344.2781712 11.147 168381 Acylcarnitine C14:1 4029 370.2942133 10.991 53634646 Acylcarnitine C14:0 5601 372.3105627 10.977 53477791 Acylcarnitine C16:1 5522 398.3270304 10.869 53951914 Acylcarnitine C16:0 4964 400.3424124 10.842 11953816 Acylcarnitine C18:1 4944 426.3576773 10.747 46907933 Acylcarnitine C18:0 1130 428.3735135 10.734 52922056 lysoPE 16:0 6707 454.292605 8.888 9547069 lysoPE 18:1 66 480.308566 8.575 9547071 lysoPE 18:0 7817 482.324123 8.454 9547068 lysoPC 16:1 2704 494.324484 11.422 24779461 lysoPC 16:0 4219 496.3398435 11.381 460602 lysoPC 18:1 1499 522.3557595 11.178 16081932 lysoPC 18:0 7408 524.3711797 11.286 497299 SM 32:1 6033 675.543565 11.01 11433862 PE 32:2 728 688.4914041 4.274 9546809 PE 32:1 3140 690.5067966 4.372 52924223 SM 34:1 7335 703.5742272 10.946 9939941 PE 34:3 8574 714.5064345 4.212 52924360 PE 34:2 5001 716.522987 4.252 52924899 PE 34:1 1482 718.5373503 4.37 9546802 PC 32:2 2391 730.5386662 9.945 24778764 PE 36:4 1357 740.5220477 4.049 52924875 PE 36:3 599 742.5377229 4.144 52924364 PE 36:2 8487 744.5537567 4.198 9546757 PC 34:4 6780 754.5366323 9.985 52922835 PC 34:2 7085 758.5702511 9.907 24778935 PC 34:1 4681 760.5845333 9.907 24778933 PE 38:6 996 764.5225557 3.989 52924873 PC 36:3 7627 784.5854303 9.62 52922727 PC 36:2 230 786.6014517 9.75 10350317 PC 38:5 8240 808.5842462 9.5 52923293 PC 38:4 8607 810.599117 8.5 53478957 PC 38:3 6357 812.6143887 9.15 52923173 PC 38:2 491 814.6301964 9.247 52923113 PC 40:7 6434 832.5834274 8.43 52923693 METABOLITES_END #END