#METABOLOMICS WORKBENCH csclendinen_20140910_9105751_mwtab.txt DATATRACK_ID:149 STUDY_ID:ST000098 ANALYSIS_ID:AN000159 PROJECT_ID:PR000090
VERSION             	1
CREATED_ON          	2016-09-17
#PROJECT
PR:PROJECT_TITLE                 	13C Metabolomics: NMR and IROA for Unknown Identification
PR:PROJECT_SUMMARY               	Demonstrate a method using high-sensitivity NMR to identify an unknown
PR:PROJECT_SUMMARY               	from a fraction isolated from an IROA LC-MS experiment.
PR:INSTITUTE                     	University of Florida
PR:DEPARTMENT                    	Department of Biochemistry and Molecular Biology / SECIM
PR:LABORATORY                    	Edison
PR:LAST_NAME                     	Edison
PR:FIRST_NAME                    	Arthur
PR:ADDRESS                       	-
PR:EMAIL                         	aedison@ufl.edu
PR:PHONE                         	352-392-4535
#STUDY
ST:STUDY_TITLE                   	Heatshock response of C. elegans using IROA (II)
ST:STUDY_SUMMARY                 	-
ST:INSTITUTE                     	University of Florida
ST:DEPARTMENT                    	Department of Biochemistry and Molecular Biology / SECIM
ST:LABORATORY                    	Arthur Edison
ST:LAST_NAME                     	Edison
ST:FIRST_NAME                    	Arthur
ST:ADDRESS                       	R3-226 Academic Research Building, Department of Biochemistry & Molecular
ST:ADDRESS                       	PO Box 100245, Gainesville, FL 32610-0245
ST:EMAIL                         	aedison@ufl.edu
ST:PHONE                         	352-392-4535
ST:NUM_GROUPS                    	1
ST:TOTAL_SUBJECTS                	1
#SUBJECT
SU:SUBJECT_TYPE                  	Animal
SU:SUBJECT_SPECIES               	Caenorhabditis elegans
SU:GENOTYPE_STRAIN               	N2
SU:SPECIES_GROUP                 	Invertebrate
#SUBJECT_SAMPLE_FACTORS:         	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data
SUBJECT_SAMPLE_FACTORS           	HSP	HSP_F9_ms2_neg.raw	Fractionation:fraction 4-4.5 min	
SUBJECT_SAMPLE_FACTORS           	HSP	HSP_F9_pos.raw	Fractionation:fraction 4-4.5 min	
SUBJECT_SAMPLE_FACTORS           	HSP	HSP_F10_ms2_neg.raw	Fractionation:fraction 4.5-5 min	
SUBJECT_SAMPLE_FACTORS           	HSP	HSP_F10_pos.raw	Fractionation:fraction 4.5-5 min	
SUBJECT_SAMPLE_FACTORS           	HSP	HSP_ms2_pos.raw	Fractionation:Unfractionated	
SUBJECT_SAMPLE_FACTORS           	HSP	HSP_pos.raw	Fractionation:Unfractionated	
#COLLECTION
CO:COLLECTION_SUMMARY            	Samples were already collected.
CO:COLLECTION_PROTOCOL_FILENAME  	NMR_Methods.docx;LCMSMethod.docx
#TREATMENT
TR:TREATMENT_SUMMARY             	No further treatment
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Samples were mixed together and resuspended in 120 ?L of water.
SP:SAMPLEPREP_PROTOCOL_FILENAME  	NMR_Methods.docx;LCMSMethod.docx
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	Samples were injected onto an Waters Acquity UPLC HSS T3 (2.1x100 mm, 1.8
CH:CHROMATOGRAPHY_SUMMARY        	column using a column temperature of 35 C and a flow rate of 300 L/min with
CH:CHROMATOGRAPHY_SUMMARY        	A (0.1% formic acid in water (Fisher LC-MS Optima)) held at 100% A for 1 min,
CH:CHROMATOGRAPHY_SUMMARY        	by linear increase to 80% B (0.1% formic acid in acetonitrile) over 10 min,
CH:CHROMATOGRAPHY_SUMMARY        	constant for 2 min, followed by a return to initial conditions over 0.5 min and
CH:CHROMATOGRAPHY_SUMMARY        	for 3 min.
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
MS:INSTRUMENT_NAME               	Thermo Q Exactive Orbitrap
CH:COLUMN_NAME                   	Waters Acquity HSS T3 (100 x 2.1mm, 1.8um)
CH:FLOW_RATE                     	300 L/min
CH:INJECTION_TEMPERATURE         	4 C
CH:SOLVENT_A                     	0.1% formic acid in water (Fisher LC-MS Optima)
CH:SOLVENT_B                     	0.1% formic acid in acetonitrile
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
AN:LABORATORY_NAME               	Timothy Garrett
AN:OPERATOR_NAME                 	Timothy Garrett
AN:DETECTOR_TYPE                 	Thermo Scientific Q-Exactive Orbitrap/MS2 Negative
#MS
MS:INSTRUMENT_NAME               	Thermo Q Exactive Orbitrap
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	NEGATIVE
MS:MS_COMMENTS                   	HSP_F10_ms2_neg.raw, HSP_F9_ms2_neg.raw
MS:FRAGMENTATION_METHOD          	Full MS/DD-MS2 (TOPN)
MS:RESOLUTION_SETTING            	70,000, 17,500
MS:SCAN_RANGE_MOVERZ             	70-1000
MS:MS_COMMENTS                   	Samples were analyzed using a mass range of 70-1000 in positive and negative
MS:MS_COMMENTS                   	modes, externally calibrated, using a Thermo Scientific Q-Exactive Orbitrap
MS:MS_COMMENTS                   	spectrometer equipped with a Dionex UltiMate 3000 RS autosampler and pump. The
MS:MS_COMMENTS                   	was equipped with a Heated Electrospray Ionization (HESI) source which operated
MS:MS_COMMENTS                   	a spray temperature of 350 ?C, a spray voltage of 3.5 kV, and sheath and
MS:MS_COMMENTS                   	gas flow rates of 50 and 10 arbitrary units, respectively. The capillary
MS:MS_COMMENTS                   	was held at 325 ?C, and the S-lens RF Level was set to 40%. The FR resolution
MS:MS_COMMENTS                   	set to 70,000 at m/z 200. The accuracy achieved was routinely less than 1.5
MS:MS_COMMENTS                   	In addition to full MS, data-dependent MS/MS was performed on the 10 most
MS:MS_COMMENTS                   	peaks with an exclusion period of 10 seconds (40 normalized collision energy).
MS:MS_COMMENTS                   	was performed using a Dionex UltiMate 3000 fraction collector by triggering
MS:MS_COMMENTS                   	30 seconds.
#END