#METABOLOMICS WORKBENCH blfitz_20180926_213803_mwtab.txt DATATRACK_ID:1518 STUDY_ID:ST001104 ANALYSIS_ID:AN001797 PROJECT_ID:PR000716
VERSION             	1
CREATED_ON             	December 4, 2018, 11:25 am
PR:PROJECT_TITLE                 	Urine Metabolite Identification
PR:PROJECT_TYPE                  	MS and MS/MS analyses
PR:PROJECT_SUMMARY               	Structural identification of unknown metabolites in human urine
PR:INSTITUTE                     	Colorado State University
PR:DEPARTMENT                    	MIP
PR:LABORATORY                    	Belisle
PR:LAST_NAME                     	Fitzgerald
PR:FIRST_NAME                    	Bryna
PR:ADDRESS                       	3185 Rampart Rd, Fort Collins, CO, 80521, USA
PR:EMAIL                         	blfitz@colostate.edu
PR:PHONE                         	9704918905
PR:FUNDING_SOURCE                	NIH U01 AI-115619
ST:STUDY_TITLE                   	Seryl-leucine core 1 O-glycosylated peptide (SLC1G) identification
ST:STUDY_SUMMARY                 	An untargeted metabolomics approach was utilized to determine urinary
ST:STUDY_SUMMARY                 	metabolites that could serve as small molecule biomarkers for treatment response
ST:STUDY_SUMMARY                 	to standard tuberculosis treatment. However, the majority of metabolites that
ST:STUDY_SUMMARY                 	most accurately distinguished patient samples at time of diagnosis from those at
ST:STUDY_SUMMARY                 	one month after the start of therapy lacked structural identification. The
ST:STUDY_SUMMARY                 	detection of unknown metabolite structures is a well-known limitation of
ST:STUDY_SUMMARY                 	untargeted metabolomics, and underscores a need for continued elucidation of
ST:STUDY_SUMMARY                 	novel metabolite structures. In this study, we sought to define the structure of
ST:STUDY_SUMMARY                 	a urine metabolite with an experimentally determined mass of 202.1326 Da,
ST:STUDY_SUMMARY                 	classified as molecular feature (MF) 874.3547. Using mass spectrometry combined
ST:STUDY_SUMMARY                 	with enzymatic digestions, the metabolite was structurally characterized as a
ST:STUDY_SUMMARY                 	seryl-leucine core 1 O-glycosylated peptide (SLC1G) of human origin.
ST:INSTITUTE                     	Colorado State University
ST:LAST_NAME                     	Fitzgerald
ST:FIRST_NAME                    	Bryna
ST:ADDRESS                       	3185 Rampart Rd
ST:EMAIL                         	blfitz@colostate.edu
ST:PHONE                         	9704918905
SU:SUBJECT_TYPE                  	Human
SU:SUBJECT_SPECIES               	Homo sapiens
SU:TAXONOMY_ID                   	9606
#SUBJECT_SAMPLE_FACTORS:         	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data
SUBJECT_SAMPLE_FACTORS           	-	7728_CE20.d	Sample Type:Patient urine	
SUBJECT_SAMPLE_FACTORS           	-	SL_10.d	Sample Type:Seryl-leucine Standard	
SUBJECT_SAMPLE_FACTORS           	-	SLC1G_only.d	Sample Type:Enriched SLC1G	
SUBJECT_SAMPLE_FACTORS           	-	SLC1G+a2-3.d	Sample Type:Enriched SLC1G after treatment with a2-3 neuraminidase	
SUBJECT_SAMPLE_FACTORS           	-	SLC1G+a2-368.d	Sample Type:Enriched SLC1G after treatment with a2-3,6,8 neuraminidase	
SUBJECT_SAMPLE_FACTORS           	-	SLC1G+both.d	Sample Type:Enriched SLC1G after treatment with a2-3,6,8 neuraminidase and O-glycosidase	
SUBJECT_SAMPLE_FACTORS           	-	SLC1G+both_10.d	Sample Type:Enriched SLC1G after treatment with a2-3,6,8 neuraminidase and O-glycosidase for comparison with seryl-leucine standard	
SUBJECT_SAMPLE_FACTORS           	-	SLC1G+both-r002.d	Sample Type:Enriched SLC1G after treatment with a2-3,6,8 neuraminidase and O-glycosidase for comparison with seryl-leucine standard	
SUBJECT_SAMPLE_FACTORS           	-	SL-r002.d	Sample Type:Seryl-leucine Standard	
CO:COLLECTION_SUMMARY            	Urine was collected and frozen at -20 for storage. Prior to analysis, urine was
CO:COLLECTION_SUMMARY            	thawed and centrifuged to remove particulates.
CO:SAMPLE_TYPE                   	Urine
TR:TREATMENT_SUMMARY             	Urine, SLC1G enriched from urine, and seryl-leucine synthetic standard were
TR:TREATMENT_SUMMARY             	analyzed by LC-MS and LC-MS/MS
SP:SAMPLEPREP_SUMMARY            	Urine was thawed on ice and centrifuged to remove particulates prior to
SP:SAMPLEPREP_SUMMARY            	analysis.
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Agilent 1200
CH:COLUMN_NAME                   	Atlantis T3 reverse-phase C18 3.5µm column (2.1 by 150mm
CH:FLOW_GRADIENT                 	100% Solvent A to 90% Solvent B
CH:FLOW_RATE                     	0.25 ml/min
CH:SOLVENT_A                     	0.1% Formic Acid in Water
CH:SOLVENT_B                     	0.1% Formic Acid in Methanol
AN:ANALYSIS_TYPE                 	MS
AN:ANALYSIS_PROTOCOL_FILE        	SLC1G_Identification_Methods
AN:DATA_FORMAT                   	.d
MS:MS_COMMENTS                   	-
MS:INSTRUMENT_NAME               	Agilent 6520 QTOF
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	POSITIVE
MS:CAPILLARY_VOLTAGE             	2000 V
MS:COLLISION_ENERGY              	10 V
MS:DRY_GAS_FLOW                  	8 L/min
MS:FRAGMENT_VOLTAGE              	120 V
MS:NEBULIZER                     	45 psi
MS:OCTPOLE_VOLTAGE               	750 V
MS:SCANNING_CYCLE                	1.2 spectra/sec
MS:SCANNING_RANGE                	100-1700 m/z
MS:SKIMMER_VOLTAGE               	60 V
MS:MS_RESULTS_FILE               	ST001104_AN001797_Results.txt	UNITS:m/z	Has m/z:Yes	Has RT:Yes	RT units:Minutes