#METABOLOMICS WORKBENCH engelmab_20231008_003851 DATATRACK_ID:4382 STUDY_ID:ST002920 ANALYSIS_ID:AN004790 PROJECT_ID:PR001814 VERSION 1 CREATED_ON October 10, 2023, 2:18 am #PROJECT PR:PROJECT_TITLE MINCH causes metabolic rewiring towards lipid accumulation and adipogenesis PR:PROJECT_SUMMARY Humans are ubiquitously exposed to plastic additives, including plasticizers. PR:PROJECT_SUMMARY There is growing evidence that exposure to certain plasticizers is associated PR:PROJECT_SUMMARY with the development of obesity due to their metabolism-disrupting properties. PR:PROJECT_SUMMARY Following the restriction of the use of the phthalate plasticizer PR:PROJECT_SUMMARY di-(2-ethylhexyl) phthalate (DEHP) due to its adverse health effects, it has PR:PROJECT_SUMMARY been replaced by new substitutes such as the plasticizer PR:PROJECT_SUMMARY diisononylcyclohexane-1,2-dicarboxylate (DINCH). Despite recent studies PR:PROJECT_SUMMARY suggesting that the primary metabolite monoisononylcyclohexane-1,2-dicarboxylic PR:PROJECT_SUMMARY acid ester (MINCH) promotes human adipocyte differentiation, the adipogenic PR:PROJECT_SUMMARY properties of MINCH remain controversial. Because the metabolome largely PR:PROJECT_SUMMARY reflects the molecular phenotype and is sensitive to perturbation by external PR:PROJECT_SUMMARY factors, we used targeted metabolomics to investigate the effects of DINCH and PR:PROJECT_SUMMARY MINCH on key metabolic pathways of adipocytes. Analysis of central carbon PR:PROJECT_SUMMARY metabolism is particularly relevant because it provides cellular energy through PR:PROJECT_SUMMARY the degradation of organic compounds and metabolic precursors for anabolic PR:PROJECT_SUMMARY functions that are critical for adipocyte function, such as de novo lipogenesis. PR:PROJECT_SUMMARY The project consists of three main studies: analysis of the effects of DINCH and PR:PROJECT_SUMMARY MINCH on central carbon metabolism of human SGSB cells, analysis of the insulin PR:PROJECT_SUMMARY response of DINCH- and MINCH-treated SGSB cells, and analysis of the effects of PR:PROJECT_SUMMARY DINCH and MINCH on central carbon metabolism of human SGSB cells in the presence PR:PROJECT_SUMMARY of the PPARG inhibitor GW9662. PR:INSTITUTE Helmholtz Centre for Environmental Research PR:LAST_NAME Engelmann PR:FIRST_NAME Beatrice PR:ADDRESS Permoserstr. 15 PR:EMAIL beatrice.engelmann@ufz.de PR:PHONE 00493412351099 #STUDY ST:STUDY_TITLE Possible PPARG-independent effects of DINCH and MINCH on central carbon ST:STUDY_TITLE metabolism ST:STUDY_SUMMARY In the third part of the project, we investigated the PPARG-independent effects ST:STUDY_SUMMARY of DINCH and MINCH on the central carbon metabolism of human SGBS cells. SGBS ST:STUDY_SUMMARY preadipocytes were treated for 12 days with 10 µM MINCH, 10 µM DINCH, or ST:STUDY_SUMMARY rosiglitazone in the presence of the PPARG inhibitor GW9662. Irreversible ST:STUDY_SUMMARY blocking of PPARG was achieved by incubating cells with 10 µM GW9662 1 hour ST:STUDY_SUMMARY before treatment and maintaining a regular treatment interval of 2 days. In ST:STUDY_SUMMARY conclusion, although the PPARG inhibitor GW9662 greatly reduced the effects of ST:STUDY_SUMMARY MINCH and rosiglitazone, slightly increased lipid accumulation along with ST:STUDY_SUMMARY increased lactate secretion and increased concentrations of pyruvate cycle ST:STUDY_SUMMARY metabolites were consistently induced by MINCH treatment even after PPARG ST:STUDY_SUMMARY inhibition. This suggests that MINCH exerts its effects on lipid accumulation ST:STUDY_SUMMARY and central carbon metabolism at least in part via a PPARG-independent ST:STUDY_SUMMARY mechanism. ST:INSTITUTE Helmholtz Centre for Environmental Research ST:LAST_NAME Engelmann ST:FIRST_NAME Beatrice ST:ADDRESS Permoserstr. 15 ST:EMAIL beatrice.engelmann@ufz.de ST:PHONE 00493412351099 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 SU:GENDER Male #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_GW_1 Treatment:Control GW9662 treatment Replicate=1; Species=Homo sapiens; RAW_FILE_NAME=230418_SGBS_DINCH_MINCH_GW_intracellular_CG.wiff SUBJECT_SAMPLE_FACTORS - Ctrl_GW_2 Treatment:Control GW9662 treatment Replicate=2; Species=Homo sapiens; RAW_FILE_NAME=230418_SGBS_DINCH_MINCH_GW_intracellular_CG.wiff SUBJECT_SAMPLE_FACTORS - Ctrl_GW_3 Treatment:Control GW9662 treatment Replicate=3; Species=Homo sapiens; RAW_FILE_NAME=230418_SGBS_DINCH_MINCH_GW_intracellular_CG.wiff SUBJECT_SAMPLE_FACTORS - Ctrl_GW_4 Treatment:Control GW9662 treatment Replicate=4; Species=Homo sapiens; RAW_FILE_NAME=230418_SGBS_DINCH_MINCH_GW_intracellular_CG.wiff SUBJECT_SAMPLE_FACTORS - DINCH_GW_1 Treatment:DINCH 10µM + GW9662 treatment Replicate=1; Species=Homo sapiens; RAW_FILE_NAME=230418_SGBS_DINCH_MINCH_GW_intracellular_CG.wiff SUBJECT_SAMPLE_FACTORS - DINCH_GW_2 Treatment:DINCH 10µM + GW9662 treatment Replicate=2; Species=Homo sapiens; RAW_FILE_NAME=230418_SGBS_DINCH_MINCH_GW_intracellular_CG.wiff SUBJECT_SAMPLE_FACTORS - DINCH_GW_3 Treatment:DINCH 10µM + GW9662 treatment Replicate=3; Species=Homo sapiens; RAW_FILE_NAME=230418_SGBS_DINCH_MINCH_GW_intracellular_CG.wiff SUBJECT_SAMPLE_FACTORS - DINCH_GW_4 Treatment:DINCH 10µM + GW9662 treatment Replicate=4; Species=Homo sapiens; RAW_FILE_NAME=230418_SGBS_DINCH_MINCH_GW_intracellular_CG.wiff SUBJECT_SAMPLE_FACTORS - MINCH_GW_1 Treatment:MINCH 10µM + GW9662 treatment Replicate=1; Species=Homo sapiens; RAW_FILE_NAME=230418_SGBS_DINCH_MINCH_GW_intracellular_CG.wiff SUBJECT_SAMPLE_FACTORS - MINCH_GW_2 Treatment:MINCH 10µM + GW9662 treatment Replicate=2; Species=Homo sapiens; RAW_FILE_NAME=230418_SGBS_DINCH_MINCH_GW_intracellular_CG.wiff SUBJECT_SAMPLE_FACTORS - MINCH_GW_3 Treatment:MINCH 10µM + GW9662 treatment Replicate=3; Species=Homo sapiens; RAW_FILE_NAME=230418_SGBS_DINCH_MINCH_GW_intracellular_CG.wiff SUBJECT_SAMPLE_FACTORS - MINCH_GW_4 Treatment:MINCH 10µM + GW9662 treatment Replicate=4; Species=Homo sapiens; RAW_FILE_NAME=230418_SGBS_DINCH_MINCH_GW_intracellular_CG.wiff SUBJECT_SAMPLE_FACTORS - Rosi_GW_1 Treatment:Rosi (d0-d4) + GW9662 treatment Replicate=1; Species=Homo sapiens; RAW_FILE_NAME=230418_SGBS_DINCH_MINCH_GW_intracellular_CG.wiff SUBJECT_SAMPLE_FACTORS - Rosi_GW_2 Treatment:Rosi (d0-d4) + GW9662 treatment Replicate=2; Species=Homo sapiens; RAW_FILE_NAME=230418_SGBS_DINCH_MINCH_GW_intracellular_CG.wiff SUBJECT_SAMPLE_FACTORS - Rosi_GW_3 Treatment:Rosi (d0-d4) + GW9662 treatment Replicate=3; Species=Homo sapiens; RAW_FILE_NAME=230418_SGBS_DINCH_MINCH_GW_intracellular_CG.wiff SUBJECT_SAMPLE_FACTORS - Rosi_GW_4 Treatment:Rosi (d0-d4) + GW9662 treatment Replicate=4; Species=Homo sapiens; RAW_FILE_NAME=230418_SGBS_DINCH_MINCH_GW_intracellular_CG.wiff SUBJECT_SAMPLE_FACTORS - Ctrl_GW_SN1 Treatment:Control GW9662 treatment supernatant Replicate=1; Species=Homo sapiens; RAW_FILE_NAME=230420_SGBS_DINCH_MINCH_GW_SN_CG.wiff SUBJECT_SAMPLE_FACTORS - Ctrl_GW_SN2 Treatment:Control GW9662 treatment supernatant Replicate=2; Species=Homo sapiens; RAW_FILE_NAME=230420_SGBS_DINCH_MINCH_GW_SN_CG.wiff SUBJECT_SAMPLE_FACTORS - Ctrl_GW_SN3 Treatment:Control GW9662 treatment supernatant Replicate=3; Species=Homo sapiens; RAW_FILE_NAME=230420_SGBS_DINCH_MINCH_GW_SN_CG.wiff SUBJECT_SAMPLE_FACTORS - Ctrl_GW_SN4 Treatment:Control GW9662 treatment supernatant Replicate=4; Species=Homo sapiens; RAW_FILE_NAME=230420_SGBS_DINCH_MINCH_GW_SN_CG.wiff SUBJECT_SAMPLE_FACTORS - DINCH_GW_SN1 Treatment:DINCH 10µM + GW9662 treatment supernatant Replicate=1; Species=Homo sapiens; RAW_FILE_NAME=230420_SGBS_DINCH_MINCH_GW_SN_CG.wiff SUBJECT_SAMPLE_FACTORS - DINCH_GW_SN2 Treatment:DINCH 10µM + GW9662 treatment supernatant Replicate=2; Species=Homo sapiens; RAW_FILE_NAME=230420_SGBS_DINCH_MINCH_GW_SN_CG.wiff SUBJECT_SAMPLE_FACTORS - DINCH_GW_SN3 Treatment:DINCH 10µM + GW9662 treatment supernatant Replicate=3; Species=Homo sapiens; RAW_FILE_NAME=230420_SGBS_DINCH_MINCH_GW_SN_CG.wiff SUBJECT_SAMPLE_FACTORS - DINCH_GW_SN4 Treatment:DINCH 10µM + GW9662 treatment supernatant Replicate=4; Species=Homo sapiens; RAW_FILE_NAME=230420_SGBS_DINCH_MINCH_GW_SN_CG.wiff SUBJECT_SAMPLE_FACTORS - MINCH_GW__SN1 Treatment:MINCH 10µM + GW9662 treatment supernatant Replicate=1; Species=Homo sapiens; RAW_FILE_NAME=230420_SGBS_DINCH_MINCH_GW_SN_CG.wiff SUBJECT_SAMPLE_FACTORS - MINCH_GW_SN2 Treatment:MINCH 10µM + GW9662 treatment supernatant Replicate=2; Species=Homo sapiens; RAW_FILE_NAME=230420_SGBS_DINCH_MINCH_GW_SN_CG.wiff SUBJECT_SAMPLE_FACTORS - MINCH_GW_SN3 Treatment:MINCH 10µM + GW9662 treatment supernatant Replicate=3; Species=Homo sapiens; RAW_FILE_NAME=230420_SGBS_DINCH_MINCH_GW_SN_CG.wiff SUBJECT_SAMPLE_FACTORS - MINCH_GW_SN4 Treatment:MINCH 10µM + GW9662 treatment supernatant Replicate=4; Species=Homo sapiens; RAW_FILE_NAME=230420_SGBS_DINCH_MINCH_GW_SN_CG.wiff SUBJECT_SAMPLE_FACTORS - Rosi_GW_SN1 Treatment:Rosi (d0-d4) + GW9662 treatment supernatant Replicate=1; Species=Homo sapiens; RAW_FILE_NAME=230420_SGBS_DINCH_MINCH_GW_SN_CG.wiff SUBJECT_SAMPLE_FACTORS - Rosi_GW_SN2 Treatment:Rosi (d0-d4) + GW9662 treatment supernatant Replicate=2; Species=Homo sapiens; RAW_FILE_NAME=230420_SGBS_DINCH_MINCH_GW_SN_CG.wiff SUBJECT_SAMPLE_FACTORS - Rosi_GW_SN3 Treatment:Rosi (d0-d4) + GW9662 treatment supernatant Replicate=3; Species=Homo sapiens; RAW_FILE_NAME=230420_SGBS_DINCH_MINCH_GW_SN_CG.wiff SUBJECT_SAMPLE_FACTORS - Rosi_GW_SN4 Treatment:Rosi (d0-d4) + GW9662 treatment supernatant Replicate=4; Species=Homo sapiens; RAW_FILE_NAME=230420_SGBS_DINCH_MINCH_GW_SN_CG.wiff #COLLECTION CO:COLLECTION_SUMMARY The SGBS cells were obtained from Prof. Martin Wabitsch laboratory at the CO:COLLECTION_SUMMARY University Clinic Ulm. SGBS preadipocytes were differentiated according to the CO:COLLECTION_SUMMARY standard protocol described previously (Wabitsch et al., 2001). CO:SAMPLE_TYPE Adipose tissue CO:STORAGE_CONDITIONS -80℃ #TREATMENT TR:TREATMENT_SUMMARY SGSB preadipocytes were maintained at 37°C and 5% CO2 in 95% humidity. To TR:TREATMENT_SUMMARY assess the PPARG-independent effects of DINCH and MINCH on central carbon TR:TREATMENT_SUMMARY metabolism, SGBS preadipocytes were exposed to differentiation media containing TR:TREATMENT_SUMMARY DINCH (10 µM DINCH+GW) or MINCH (10 µM MINCH+GW) supplemented with the PPARG TR:TREATMENT_SUMMARY antagonist GW9662 for 12 days. Irreversible blocking of PPARG prior to treatment TR:TREATMENT_SUMMARY was achieved by adding the antagonist 1 hour before the addition of the TR:TREATMENT_SUMMARY respective chemical. For comparison, SGBS cells were treated with rosiglitazone TR:TREATMENT_SUMMARY (d0-d4) as in the standard protocol but in the presence of GW9662 (Rosi+GW), and TR:TREATMENT_SUMMARY SGBS cells were treated with GW9662 only for 12 days (Ctrl+GW). During TR:TREATMENT_SUMMARY differentiation, a final concentration of 0.01% (v/v) MeOH and 0.02% (v/v) DMSO TR:TREATMENT_SUMMARY was added to all differentiation media. Continuous exposure was mimicked by TR:TREATMENT_SUMMARY replacing the cell culture medium every other day. Each treatment was performed TR:TREATMENT_SUMMARY in four biological replicates (n=4). #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Extraction of intracellular and extracellular metabolites was performed by a SP:SAMPLEPREP_SUMMARY 1:1:1 methanol:water:chloroform extraction protocol. For the extraction of SP:SAMPLEPREP_SUMMARY intracellular metabolites, the culture medium was removed and the cells were SP:SAMPLEPREP_SUMMARY rinsed twice with 1 ml of 0.9% ice-cold NaCl. The rinsing solution was removed, SP:SAMPLEPREP_SUMMARY and the metabolism of the cells was stopped by adding 400 µL of MeOH (-20 °C) SP:SAMPLEPREP_SUMMARY followed by 400 µL of ice-cold H2O containing 10 µM d6-glutarate. Cells were SP:SAMPLEPREP_SUMMARY collected using a cell lifter and 400 µL of chloroform was added. After shaking SP:SAMPLEPREP_SUMMARY at 1,400 rpm and 4 °C for 20 min, the extraction mixture was centrifuged at SP:SAMPLEPREP_SUMMARY 18,000 g and 4 °C for 5 min. Subsequently, 300 µL volume of the polar upper SP:SAMPLEPREP_SUMMARY phase was collected and evaporated to complete dryness. For the extraction of SP:SAMPLEPREP_SUMMARY extracellular metabolites, 300 µL of the supernatant was extracted by adding SP:SAMPLEPREP_SUMMARY 400 µL MeOH (-20 °C) containing 100 nM MEHP, 100 µL ice-cold H2O containing SP:SAMPLEPREP_SUMMARY 40 µM d6-glutarate, and 400 µL chloroform (-20 °C). Subsequent sample SP:SAMPLEPREP_SUMMARY preparation was identical to the extraction of intracellular metabolites. Note: SP:SAMPLEPREP_SUMMARY After measurement of the samples by LC-MS, the raw AUC values uploaded here were SP:SAMPLEPREP_SUMMARY normalized to the internal standard (d6-glutarate, if applicable) and DNA SP:SAMPLEPREP_SUMMARY content per well (measured by DAPI fluorescence). After normalization, log2 fold SP:SAMPLEPREP_SUMMARY changes were calculated by dividing the normalized peak area from each replicate SP:SAMPLEPREP_SUMMARY of each treatment by the normalized peak area from each control. Insulin data SP:SAMPLEPREP_SUMMARY were not normalized to DAPI because fold changes were calculated by dividing the SP:SAMPLEPREP_SUMMARY intensities of the insulin-stimulated cells by the noninsulin-stimulated cells SP:SAMPLEPREP_SUMMARY from each treatment. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Agilent 1290 Infinity II CH:COLUMN_NAME Waters Xselect XP HSS T3 (150 x 2.1mm, 2.5µm) CH:SOLVENT_A 10mM TBA, 10mM acetic acid, 5% MeOH, 2% IPA in water CH:SOLVENT_B 100% IPA CH:FLOW_GRADIENT 0-5 min 0% B, 5-9 min 0%- 2% B, 9-9.5 min 2-6% B, 9.5-11.5 min 6% B, 11.5-12 min CH:FLOW_GRADIENT 6-11% B, 12-13.5 min 11% B, 13.5-15.5 min 11-28% B, 15.5-16.5 min 28-53% B, CH:FLOW_GRADIENT 16.5-22.5 53% B, 22.5-23 min 53-0% B, 23-33 min 0% B CH:FLOW_RATE 0-15.5 min 0.4 mL/min, 15.5-16.5 min 0.4-0.15 mL/min, 16.5-23 min 0.15 mL/min, CH:FLOW_RATE 23-27 min 0.15-0.4 mL/min, 27-33 min 0.4 mL/min CH:COLUMN_TEMPERATURE 40 #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME ABI Sciex 6500+ MS:INSTRUMENT_TYPE QTRAP MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS For identification and quantitation, a scheduled MRM method was used, with MS:MS_COMMENTS specific transitions for every metabolite. Data acquisition and peak integration MS:MS_COMMENTS were performed in Analyst® software (Version 1.7.1). #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Peak AUC MS_METABOLITE_DATA_START Samples Ctrl_GW_1 Ctrl_GW_2 Ctrl_GW_3 Ctrl_GW_4 DINCH_GW_1 DINCH_GW_2 DINCH_GW_3 DINCH_GW_4 MINCH_GW_1 MINCH_GW_2 MINCH_GW_3 MINCH_GW_4 Rosi_GW_1 Rosi_GW_2 Rosi_GW_3 Rosi_GW_4 Ctrl_GW_SN1 Ctrl_GW_SN2 Ctrl_GW_SN3 Ctrl_GW_SN4 DINCH_GW_SN1 DINCH_GW_SN2 DINCH_GW_SN3 DINCH_GW_SN4 MINCH_GW__SN1 MINCH_GW_SN2 MINCH_GW_SN3 MINCH_GW_SN4 Rosi_GW_SN1 Rosi_GW_SN2 Rosi_GW_SN3 Rosi_GW_SN4 Factors Treatment:Control GW9662 treatment Treatment:Control GW9662 treatment Treatment:Control GW9662 treatment Treatment:Control GW9662 treatment Treatment:DINCH 10µM + GW9662 treatment Treatment:DINCH 10µM + GW9662 treatment Treatment:DINCH 10µM + GW9662 treatment Treatment:DINCH 10µM + GW9662 treatment Treatment:MINCH 10µM + GW9662 treatment Treatment:MINCH 10µM + GW9662 treatment Treatment:MINCH 10µM + GW9662 treatment Treatment:MINCH 10µM + GW9662 treatment Treatment:Rosi (d0-d4) + GW9662 treatment Treatment:Rosi (d0-d4) + GW9662 treatment Treatment:Rosi (d0-d4) + GW9662 treatment Treatment:Rosi (d0-d4) + GW9662 treatment Treatment:Control GW9662 treatment supernatant Treatment:Control GW9662 treatment supernatant Treatment:Control GW9662 treatment supernatant Treatment:Control GW9662 treatment supernatant Treatment:DINCH 10µM + GW9662 treatment supernatant Treatment:DINCH 10µM + GW9662 treatment supernatant Treatment:DINCH 10µM + GW9662 treatment supernatant Treatment:DINCH 10µM + GW9662 treatment supernatant Treatment:MINCH 10µM + GW9662 treatment supernatant Treatment:MINCH 10µM + GW9662 treatment supernatant Treatment:MINCH 10µM + GW9662 treatment supernatant Treatment:MINCH 10µM + GW9662 treatment supernatant Treatment:Rosi (d0-d4) + GW9662 treatment supernatant Treatment:Rosi (d0-d4) + GW9662 treatment supernatant Treatment:Rosi (d0-d4) + GW9662 treatment supernatant Treatment:Rosi (d0-d4) + GW9662 treatment supernatant Glucose 6-phosphate 6.36E+04 2.40E+05 9.86E+04 3.00E+05 6.77E+04 4.43E+05 8.45E+04 4.51E+05 2.81E+04 3.30E+05 4.34E+04 3.40E+05 8.46E+04 2.58E+05 9.29E+04 3.01E+05 Citrate 1.73E+07 2.34E+07 2.07E+07 3.00E+07 1.66E+07 4.03E+07 1.78E+07 3.81E+07 1.20E+07 2.60E+07 1.48E+07 2.64E+07 1.88E+07 2.29E+07 2.26E+07 2.50E+07 a-Ketoglutarate 5.94E+05 1.41E+06 7.30E+05 1.36E+06 8.47E+05 2.28E+06 9.07E+05 2.42E+06 8.10E+05 1.57E+06 8.95E+05 1.72E+06 7.64E+05 1.00E+06 7.80E+05 1.04E+06 Succinate 2.17E+06 1.53E+06 1.71E+06 1.79E+06 1.76E+06 1.70E+06 1.58E+06 1.50E+06 1.92E+06 1.31E+06 1.63E+06 1.63E+06 1.18E+06 1.78E+06 1.75E+06 1.42E+06 Glutamate 6.80E+07 7.35E+07 7.87E+07 7.87E+07 7.96E+07 7.91E+07 8.14E+07 7.62E+07 7.44E+07 8.20E+07 8.28E+07 8.67E+07 5.66E+07 5.76E+07 5.90E+07 5.84E+07 Aspartate 2.18E+07 1.47E+07 1.91E+07 1.52E+07 1.66E+07 8.68E+06 1.69E+07 8.09E+06 1.23E+07 1.05E+07 1.37E+07 1.15E+07 1.93E+07 1.68E+07 1.96E+07 1.68E+07 Lactate 1.02E+07 8.58E+06 9.36E+06 1.08E+07 1.11E+07 1.03E+07 8.48E+06 7.81E+06 1.35E+07 1.57E+07 1.41E+07 1.42E+07 1.35E+07 1.16E+07 1.33E+07 1.17E+07 1.46E+08 1.32E+08 1.62E+08 1.44E+08 1.51E+08 1.44E+08 1.41E+08 1.46E+08 2.06E+08 1.99E+08 1.93E+08 1.89E+08 2.02E+08 2.06E+08 2.22E+08 1.80E+08 Alanine 2.85E+04 3.10E+04 2.96E+04 3.26E+04 2.83E+04 2.77E+04 2.99E+04 2.84E+04 3.26E+04 3.18E+04 3.39E+04 3.57E+04 3.26E+04 3.63E+04 3.22E+04 3.48E+04 Phosphoenolpyruvate 3.89E+05 2.77E+05 4.46E+05 3.59E+05 4.32E+05 2.28E+05 3.90E+05 2.37E+05 2.23E+05 2.80E+05 2.40E+05 3.60E+05 1.03E+06 6.26E+05 1.16E+06 5.77E+05 Pyruvate 1.04E+05 1.18E+05 1.16E+05 9.01E+04 7.11E+04 7.15E+04 6.82E+04 5.97E+04 1.14E+05 9.86E+04 1.26E+05 1.05E+05 1.68E+05 1.17E+05 1.97E+05 1.22E+05 Fructose 6-phosphate 9.69E+04 3.60E+05 1.52E+05 3.91E+05 1.11E+05 5.41E+05 1.30E+05 5.75E+05 4.98E+04 3.96E+05 7.34E+04 3.85E+05 1.65E+05 2.90E+05 1.53E+05 3.58E+05 Glyceraldehyde 3-phosphate 2.62E+04 4.54E+04 2.95E+04 4.50E+04 3.52E+04 6.91E+04 2.90E+04 6.15E+04 2.40E+04 6.40E+04 2.62E+04 7.37E+04 9.37E+04 7.61E+04 1.03E+05 7.72E+04 Ribulose 5-phosphate 2.36E+05 4.11E+05 2.72E+05 4.61E+05 3.28E+05 5.26E+05 3.09E+05 5.16E+05 2.42E+05 4.67E+05 2.43E+05 4.77E+05 5.41E+05 5.50E+05 5.77E+05 5.10E+05 Ribose 5-phosphate 9.84E+04 2.21E+05 1.60E+05 2.10E+05 1.46E+05 2.54E+05 1.54E+05 2.69E+05 9.30E+04 2.38E+05 1.19E+05 2.58E+05 2.90E+05 2.82E+05 2.92E+05 2.97E+05 Fumarate 7.26E+04 7.82E+04 6.90E+04 1.03E+05 7.97E+04 9.18E+04 7.93E+04 7.30E+04 7.59E+04 6.07E+04 5.99E+04 6.52E+04 2.09E+05 1.10E+05 1.71E+05 1.09E+05 Oxaloacetate 7.40E+04 6.14E+04 8.00E+04 6.90E+04 6.66E+04 6.35E+04 6.69E+04 6.05E+04 1.44E+05 1.19E+05 1.36E+05 1.21E+05 1.27E+05 1.05E+05 1.22E+05 1.03E+05 Glutamine 4.03E+07 3.83E+07 4.36E+07 4.06E+07 4.03E+07 3.33E+07 4.04E+07 3.36E+07 3.94E+07 3.64E+07 3.96E+07 3.89E+07 3.94E+07 3.83E+07 3.97E+07 3.59E+07 Fructose 1,6-bisphosphate 1.51E+06 4.29E+06 1.64E+06 5.92E+06 1.58E+06 9.11E+06 1.68E+06 9.21E+06 6.65E+05 5.11E+06 8.23E+05 5.65E+06 3.11E+06 3.28E+06 3.53E+06 3.07E+06 cis-Aconitate 5.16E+05 7.80E+05 6.62E+05 1.11E+06 6.06E+05 1.54E+06 6.29E+05 1.58E+06 3.64E+05 9.09E+05 3.71E+05 9.71E+05 5.92E+05 9.27E+05 6.48E+05 8.56E+05 Malate 2.19E+07 2.24E+07 2.28E+07 2.50E+07 2.31E+07 1.08E+08 2.17E+07 2.13E+07 2.01E+07 2.00E+07 2.06E+07 1.98E+07 3.99E+07 2.99E+07 4.16E+07 2.96E+07 Acetyl-CoA 3.33E+05 1.71E+05 3.99E+05 2.05E+05 4.73E+05 2.97E+05 4.50E+05 3.42E+05 6.60E+05 4.41E+05 6.16E+05 4.22E+05 8.14E+05 2.75E+05 1.09E+06 2.67E+05 Asparagine 5.39E+05 5.34E+05 5.80E+05 5.93E+05 5.39E+05 6.85E+05 5.28E+05 4.79E+05 5.59E+05 5.63E+05 5.63E+05 5.94E+05 5.27E+05 5.80E+05 5.45E+05 5.38E+05 Sedoheptulose 7-phosphate 4.05E+04 9.10E+04 5.45E+04 1.32E+05 5.28E+04 1.39E+05 6.22E+04 1.67E+05 3.31E+04 1.42E+05 3.80E+04 1.38E+05 8.34E+04 1.48E+05 7.87E+04 1.76E+05 Glycerol 3-phosphate 9.85E+06 1.16E+07 1.08E+07 1.16E+07 1.21E+07 1.59E+07 1.14E+07 1.61E+07 1.07E+07 1.79E+07 9.88E+06 1.96E+07 4.04E+07 2.88E+07 3.92E+07 2.36E+07 3-Phosphoglycerate 5.31E+05 3.53E+05 5.23E+05 4.52E+05 5.75E+05 3.36E+05 5.17E+05 3.42E+05 3.38E+05 4.24E+05 3.17E+05 4.63E+05 1.34E+06 7.88E+05 1.55E+06 7.52E+05 Malonyl-CoA 1.89E+04 1.24E+04 2.03E+04 1.62E+04 5.69E+04 4.57E+04 6.25E+04 5.28E+04 9.63E+04 9.29E+04 9.79E+04 9.16E+04 6.41E+04 4.68E+04 7.00E+04 3.92E+04 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name m/z PubChem ID KEGG_ID Glucose 6-phosphate 259 5958 C00668 Fructose 6-phosphate 259 69507 C00085 Fructose 1,6-bisphosphate 339 10267 C05378 Glycerol 3-phosphate 171 439162 C00093 Glyceraldehyde 3-phosphate 169 439168 C00118 3-Phosphoglycerate 185 724 C00197 Phosphoenolpyruvate 167 1005 C00074 Pyruvate 87 107735 C00022 Lactate 89 91435 C00186 Alanine 88 5950 C00041 Acetyl-CoA 808 444493 C00024 Malonyl-CoA 852 644066 C00083 Citrate 191 311 C00158 cis-Aconitate 173 643757 C00417 a-Ketoglutarate 145 51 C00026 Aspartate 132 5960 C00049 Asparagine 131 6267 C00152 Succinate 117 1110 C00042 Fumarate 115 444972 C00122 Malate 133 525 C00711 Oxaloacetate 131 970 C00036 Glutamate 146 33032 C00025 Glutamine 145 5961 C00064 Ribulose 5-phosphate 229 439184 C00199 Ribose 5-phosphate 229 439167 C00117 Sedoheptulose 7-phosphate 289 65007 C05382 METABOLITES_END #END