#METABOLOMICS WORKBENCH flingrus_20200403_125632 DATATRACK_ID:1969 STUDY_ID:ST001379 ANALYSIS_ID:AN002299 PROJECT_ID:000000 VERSION 1 CREATED_ON May 14, 2020, 11:08 am #PROJECT PR:PROJECT_TITLE 2,3,7,8 -Tetrachlorodibenzo-p-dioxin Mediated Effects on Metabolite Levels PR:PROJECT_TYPE Dioxin PR:PROJECT_SUMMARY Experiment to study TCDD-elicited effects on hepatic coenzyme A (CoA) and PR:PROJECT_SUMMARY Acetyl-CoA levels in liver of male mice treated with sesame oil vehicle or PR:PROJECT_SUMMARY 0.01-30 ug/kg TCDD every 4 days for 28 days. PR:INSTITUTE Michigan State University PR:DEPARTMENT Biochemistry and Molecular Biology PR:LABORATORY Zacharewski Lab PR:LAST_NAME Fling PR:FIRST_NAME Russell PR:ADDRESS 1129 Farm Ln, East Lansing, MI, USA PR:EMAIL flingrus@msu.edu PR:PHONE (517) 884-2054 PR:FUNDING_SOURCE NIH/NIEHS ES029541 PR:PROJECT_COMMENTS FUNDING SOURCE: NIH/NIEHS; PR:PUBLICATIONS - PR:CONTRIBUTORS Russ Fling, Kelly A Fader, Claire M Doskey, Rance Nault, Tim Zacharewski #STUDY ST:STUDY_TITLE 2,3,7,8 -Tetrachlorodibenzo-p-dioxin Mediated Effects on Hepatic Coenzyme A ST:STUDY_TITLE (CoA) and Acetyl-CoA Levels (part-II) ST:STUDY_TYPE Dioxin ST:STUDY_SUMMARY Experiment to study TCDD-elicited effects on hepatic Coenzyme A (CoA) and ST:STUDY_SUMMARY Acetyl-CoA levels in liver of male mice treated with sesame oil vehicle or ST:STUDY_SUMMARY 0.01-30 ug/kg TCDD every 4 days for 28 days. ST:INSTITUTE Michigan State University ST:DEPARTMENT Biochemistry and Molecular Biology ST:LABORATORY Zacharewski Lab ST:LAST_NAME Fling ST:FIRST_NAME Russell ST:ADDRESS 1129 Farm Ln, East Lansing, MI, USA ST:EMAIL flingrus@msu.edu ST:PHONE (517) 884-2054 ST:NUM_GROUPS 9 ST:TOTAL_SUBJECTS 80 ST:NUM_MALES 29 ST:STUDY_COMMENTS Fax: (517) 353-9334; ORCID: 0000-0002-6822-4962; ROLE: Graduate Student; Pubmed ST:STUDY_COMMENTS ID: -;Publication DOI: -; ST:PUBLICATIONS - #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:GENOTYPE_STRAIN C57BL/6 SU:AGE_OR_AGE_RANGE postnatal day 56 SU:GENDER Male SU:ANIMAL_HOUSING Cage Type: Innovive, Catalog#M-BTM, San Diego, CA; Bedding Type: cellulose fiber SU:ANIMAL_HOUSING chips; SU:ANIMAL_LIGHT_CYCLE 12:12 SU:ANIMAL_FEED 22/5 Rodent Diet SU:ANIMAL_WATER Aquavive, Catalog#M-WB-300A, INNOVIVE, San Diego, CA #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 - mouse_1 TCDD treatment(ug/kg):vehicle Experiment Length=28 days; RAW_FILE_NAME=XS1_081417_002.raw SUBJECT_SAMPLE_FACTORS - mouse_2 TCDD treatment(ug/kg):vehicle Experiment Length=28 days; RAW_FILE_NAME=XS1_081417_003.raw SUBJECT_SAMPLE_FACTORS - mouse_6 TCDD treatment(ug/kg):vehicle Experiment Length=28 days; RAW_FILE_NAME=XS1_081417_004.raw SUBJECT_SAMPLE_FACTORS - mouse_7 TCDD treatment(ug/kg):vehicle Experiment Length=28 days; RAW_FILE_NAME=XS1_081417_005.raw SUBJECT_SAMPLE_FACTORS - mouse_8 TCDD treatment(ug/kg):vehicle Experiment Length=28 days; RAW_FILE_NAME=XS1_081417_006.raw SUBJECT_SAMPLE_FACTORS - mouse_33 TCDD treatment(ug/kg):0.3 Experiment Length=28 days; RAW_FILE_NAME=XS1_081417_007.raw SUBJECT_SAMPLE_FACTORS - mouse_34 TCDD treatment(ug/kg):0.3 Experiment Length=28 days; RAW_FILE_NAME=XS1_081417_008.raw SUBJECT_SAMPLE_FACTORS - mouse_35 TCDD treatment(ug/kg):0.3 Experiment Length=28 days; RAW_FILE_NAME=XS1_081417_009.raw SUBJECT_SAMPLE_FACTORS - mouse_37 TCDD treatment(ug/kg):0.3 Experiment Length=28 days; RAW_FILE_NAME=XS1_081417_010.raw SUBJECT_SAMPLE_FACTORS - mouse_38 TCDD treatment(ug/kg):0.3 Experiment Length=28 days; RAW_FILE_NAME=XS1_081417_011.raw SUBJECT_SAMPLE_FACTORS - mouse_42 TCDD treatment(ug/kg):1 Experiment Length=28 days; RAW_FILE_NAME=XS1_081417_012.raw SUBJECT_SAMPLE_FACTORS - mouse_43 TCDD treatment(ug/kg):1 Experiment Length=28 days; RAW_FILE_NAME=XS1_081417_013.raw SUBJECT_SAMPLE_FACTORS - mouse_44 TCDD treatment(ug/kg):1 Experiment Length=28 days; RAW_FILE_NAME=XS1_081417_014.raw SUBJECT_SAMPLE_FACTORS - mouse_46 TCDD treatment(ug/kg):1 Experiment Length=28 days; RAW_FILE_NAME=XS1_081417_015.raw SUBJECT_SAMPLE_FACTORS - mouse_48 TCDD treatment(ug/kg):1 Experiment Length=28 days; RAW_FILE_NAME=XS1_081417_016.raw SUBJECT_SAMPLE_FACTORS - mouse_49 TCDD treatment(ug/kg):3 Experiment Length=28 days; RAW_FILE_NAME=XS1_081417_017.raw SUBJECT_SAMPLE_FACTORS - mouse_50 TCDD treatment(ug/kg):3 Experiment Length=28 days; RAW_FILE_NAME=XS1_081417_018.raw SUBJECT_SAMPLE_FACTORS - mouse_53 TCDD treatment(ug/kg):3 Experiment Length=28 days; RAW_FILE_NAME=XS1_081417_019.raw SUBJECT_SAMPLE_FACTORS - mouse_56 TCDD treatment(ug/kg):3 Experiment Length=28 days; RAW_FILE_NAME=XS1_081417_020.raw SUBJECT_SAMPLE_FACTORS - mouse_59 TCDD treatment(ug/kg):3 Experiment Length=28 days; RAW_FILE_NAME=XS1_081417_021.raw SUBJECT_SAMPLE_FACTORS - mouse_60 TCDD treatment(ug/kg):10 Experiment Length=28 days; RAW_FILE_NAME=XS1_081417_022.raw SUBJECT_SAMPLE_FACTORS - mouse_61 TCDD treatment(ug/kg):10 Experiment Length=28 days; RAW_FILE_NAME=XS1_081417_023.raw SUBJECT_SAMPLE_FACTORS - mouse_63 TCDD treatment(ug/kg):10 Experiment Length=28 days; RAW_FILE_NAME=XS1_081417_024.raw SUBJECT_SAMPLE_FACTORS - mouse_64 TCDD treatment(ug/kg):10 Experiment Length=28 days; RAW_FILE_NAME=XS1_081417_025.raw SUBJECT_SAMPLE_FACTORS - mouse_65 TCDD treatment(ug/kg):30 Experiment Length=28 days; RAW_FILE_NAME=XS1_081417_026.raw SUBJECT_SAMPLE_FACTORS - mouse_66 TCDD treatment(ug/kg):30 Experiment Length=28 days; RAW_FILE_NAME=XS1_081417_027.raw SUBJECT_SAMPLE_FACTORS - mouse_67 TCDD treatment(ug/kg):30 Experiment Length=28 days; RAW_FILE_NAME=XS1_081417_028.raw SUBJECT_SAMPLE_FACTORS - mouse_68 TCDD treatment(ug/kg):30 Experiment Length=28 days; RAW_FILE_NAME=XS1_081417_029.raw SUBJECT_SAMPLE_FACTORS - mouse_69 TCDD treatment(ug/kg):30 Experiment Length=28 days; RAW_FILE_NAME=XS1_081417_030.raw #COLLECTION CO:COLLECTION_SUMMARY Mice were taken down with CO2. Livers were excised and immediately frozen in CO:COLLECTION_SUMMARY liquid nitrogen. CO:COLLECTION_PROTOCOL_COMMENTS Time-point: 28; Time-point Unit: DAYS; CO:SAMPLE_TYPE Liver #TREATMENT TR:TREATMENT_SUMMARY oral gavage every 4 days for 28 days. 7 total exposures with sesame oil vehicle TR:TREATMENT_SUMMARY or 0.3-30 ug/kg TCDD. TR:TREATMENT_PROTOCOL_COMMENTS Administration Interval: 4; Administration Interval Unit: DAYS; Dose Unit: TR:TREATMENT_PROTOCOL_COMMENTS ug/kg; Number of Administrations: 7; Chemical Soucre: AccuStandard; Chemical TR:TREATMENT_PROTOCOL_COMMENTS Catalog Number: S-70982; Chemical Purity: -; Vehicle Substance: CHEBI 28119 TR:TREATMENT_ROUTE GAVAGE TR:TREATMENT_DOSE 0.3,1,3,10,30 TR:TREATMENT_DOSEVOLUME 0.1 ml TR:TREATMENT_VEHICLE Sesame oil TR:ANIMAL_FASTING 2 hrs before end-point TR:ANIMAL_ENDP_EUTHANASIA CO2 TR:TREATMENT_COMPOUND 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Frozen liver tissue was homegenized in 4.6 ml h20:methanol (5:3) mixture and SP:SAMPLEPREP_SUMMARY homegenized, 2.9 ml chloroform was added,vortexed and spun down at 5,000G. Top SP:SAMPLEPREP_SUMMARY layer was removed and blown down with nitrogen and reconstituted in 400 l 10mM SP:SAMPLEPREP_SUMMARY tetrabutylammonium (TBA)+10mM acetate (pH 5). A further 1:16 dilution was SP:SAMPLEPREP_SUMMARY performed for analysis of acetyl-CoA. Coenzyme A (CoA) was measured by standard SP:SAMPLEPREP_SUMMARY addition method by diluting with TBA containing CoA standard resulting in 1 uM SP:SAMPLEPREP_SUMMARY CoA standard addition. 1 uM CoA standard addition samples are denoted in the SP:SAMPLEPREP_SUMMARY subject sample factor section. SP:PROCESSING_METHOD Liquid extraction SP:PROCESSING_STORAGE_CONDITIONS -80℃ #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Waters Acquity CH:COLUMN_NAME Waters Ascentis Express C18 (2.1 X 5 cm) #ANALYSIS AN:ANALYSIS_TYPE MS AN:OPERATOR_NAME Russ Fling #MS MS:INSTRUMENT_NAME Waters Xevo G2 XS QTof MS:INSTRUMENT_TYPE TOF MS:MS_TYPE ESI MS:ION_MODE Negative MS:MS_COMMENTS Samples were run in MS^e mode. MassLynx V4.2 was used to measure area under the MS:MS_COMMENTS curve. Accurate mass and retention were used for identification of signal for MS:MS_COMMENTS respective metabolites. MS:MS_RESULTS_FILE ST001379_AN002299_Results.txt UNITS:normalized signal area Has m/z:Yes Has RT:Yes RT units:Minutes #END