#METABOLOMICS WORKBENCH stuppie_20140203_9436091_mwtab.txt DATATRACK_ID:32 stuppie_20140203_9544301_mwtab.txt DATATRACK_ID:31 TEXT OUTPUT STUDY_ID:ST000019 ANALYSIS_ID:AN000037 PROJECT_ID:PR000018 VERSION 1 CREATED_ON 10-02-2015 #PROJECT PR:PROJECT_TITLE Isotopic Ratio Outlier Analysis Global Metabolomics ofCaenorhabditis elegans PR:PROJECT_TYPE IROA labeling study PR:PROJECT_SUMMARY We demonstrate the global metabolic analysis ofCaenorhabditis elegansstress PR:PROJECT_SUMMARY responses using a mass-spectrometry-based technique called isotopic ratio PR:PROJECT_SUMMARY outlier analysis (IROA). In an IROA protocol, control and experimental samples PR:PROJECT_SUMMARY are isotopically labeled with 95 and 5%13C, and the two sample populations are PR:PROJECT_SUMMARY mixed together for uniform extraction, sample preparation, and LC-MS analysis.To PR:PROJECT_SUMMARY illustrate the utility of IROA for global metabolomics, we exposed wild-type PR:PROJECT_SUMMARY (N2) worms to a heat shock (30 min heat shock at 33 C), which causes PR:PROJECT_SUMMARY significant, widespread changes in metabolism. We collected and analyzed PR:PROJECT_SUMMARY material from the exometabolome (all material that worms release in the PR:PROJECT_SUMMARY supernatant) and the endometabolome (homogenized total extracts from the worm PR:PROJECT_SUMMARY bodies). PR:INSTITUTE University of Florida PR:DEPARTMENT Biochemistry & Molecular Biology PR:LABORATORY Edison PR:LAST_NAME Edison PR:FIRST_NAME Arthur PR:ADDRESS R3-226 Academic Research Building, Department of Biochemistry & Molecular PR:ADDRESS Biology, PO Box 100245, Gainesville, FL 32610-0245 PR:EMAIL aedison@ufl.edu PR:PHONE (352) 392-4535 #STUDY ST:STUDY_TITLE Determine purity and quality of IROA labelled glucose ST:STUDY_TYPE NMR and MS analysis ST:STUDY_SUMMARY We demonstrate the global metabolic analysis ofCaenorhabditis elegansstress ST:STUDY_SUMMARY responses using a mass-spectrometry-based technique called isotopic ratio ST:STUDY_SUMMARY outlier analysis (IROA). In an IROA protocol, control and experimental samples ST:STUDY_SUMMARY are isotopically labeled with 95 and 5%13C, and the two sample populations are ST:STUDY_SUMMARY mixed together for uniform extraction, sample preparation, and LC-MS analysis.To ST:STUDY_SUMMARY illustrate the utility of IROA for global metabolomics, we exposed wild-type ST:STUDY_SUMMARY (N2) worms to a heat shock (30 min heat shock at 33 C), which causes ST:STUDY_SUMMARY significant, widespread changes in metabolism. We collected and analyzed ST:STUDY_SUMMARY material from the exometabolome (all material that worms release in the ST:STUDY_SUMMARY supernatant) and the endometabolome (homogenized total extracts from the worm ST:STUDY_SUMMARY bodies). ST:INSTITUTE University of Florida ST:DEPARTMENT Biochemistry & Molecular Biology ST:LAST_NAME Stupp ST:FIRST_NAME Gregory ST:EMAIL stuppie@ufl.edu ST:STUDY_COMMENTS Determine purity and quality of IROA labelled glucose ST:PHONE - ST:ADDRESS - #SUBJECT SU:SUBJECT_SPECIES N/A SU:SUBJECT_TYPE - #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - 5p %13C:0.05 SUBJECT_SAMPLE_FACTORS - 95p %13C:0.95 SUBJECT_SAMPLE_FACTORS - Blank %13C:none #COLLECTION CO:COLLECTION_SUMMARY none #TREATMENT TR:TREATMENT_SUMMARY none #SAMPLEPREP SP:SAMPLEPREP_PROTOCOL_FILENAME IROAGlucoseNMR_Feb11_2014.txt SP:SAMPLEPREP_PROTOCOL_COMMENTS 5%: 10.7 mg dissolved in 594 L of 99% D2O and 6 L of 111.11 mM d4-TSP in D2O SP:SAMPLEPREP_PROTOCOL_COMMENTS 95%: 2.8 mg dissolved in 594 L of 99% D2O and 6 L of 111.11 mM d4-TSP in D2O SP:SAMPLEPREP_PROTOCOL_COMMENTS Blank: 594 L of 99% D2O and 6 L of 111.11 mM d4-TSP in D2O SP:SAMPLEPREP_SUMMARY - #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE - MS:INSTRUMENT_NAME Thermo Accela 1250 CH:COLUMN_NAME - #ANALYSIS AN:ANALYSIS_TYPE NMR AN:ACQUISITION_PARAMETERS_FILE NMR_Methods.docx AN:PROCESSING_PARAMETERS_FILE NMR_Methods.docx #NMR NM:INSTRUMENT_TYPE Bruker Avance III NM:NMR_EXPERIMENT_TYPE 1D 1H NM:STANDARD_CONCENTRATION 10 mM NM:SPECTROMETER_FREQUENCY 500 MHz NM:NMR_PROBE 5mm FBBO-Z Probe NM:NMR_SOLVENT D2O NM:NMR_TUBE_SIZE 5mm NM:PULSE_SEQUENCE zg NM:PULSE_WIDTH 4 NM:RECEIVER_GAIN 152 NM:CHEMICAL_SHIFT_REF_CPD TSP NM:TEMPERATURE 27 NM:NUMBER_OF_SCANS 16 NM:ACQUISITION_TIME 4 s NM:RELAXATION_DELAY 3 s NM:SPECTRAL_WIDTH 5498.5 NM:NUM_DATA_POINTS_ACQUIRED 65536 NM:REAL_DATA_POINTS 16384 NM:LINE_BROADENING 0.33 Hz NM:ZERO_FILLING Yes NM:APODIZATION Exponential NM:BASELINE_CORRECTION_METHOD Whittaker Smoother NM:CHEMICAL_SHIFT_REF_STD TSP NM:INSTRUMENT_NAME Bruker Avance III #NMR_METABOLITE_DATA NMR_METABOLITE_DATA:UNITS N/A NMR_METABOLITE_DATA_START Samples 5p 95p Blank Factors %13C:0.05 %13C:0.95 %13C:none no data NMR_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name no data METABOLITES_END #END