#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