#METABOLOMICS WORKBENCH jgooding_20170217_113747 DATATRACK_ID:846 STUDY_ID:ST000559 ANALYSIS_ID:AN000858
VERSION                          	1
CREATED_ON                       	02-08-2024
#PROJECT
PR:PROJECT_TITLE                 	Urine metabolomic profiling of diabetic nephropathy in the steptozotocin induced
PR:PROJECT_TITLE                 	type-1 diabetes mouse model.
PR:PROJECT_SUMMARY               	Diabetic nephropathy (DN) is the leading cause of end stage renal disease, and
PR:PROJECT_SUMMARY               	is associated with high morbidity and mortality rates. The pathophysiology of DN
PR:PROJECT_SUMMARY               	includes both glomerular and tubulointerstitial damage. Meprins are
PR:PROJECT_SUMMARY               	metalloproteinases which are most abundantly expressed in the brush border
PR:PROJECT_SUMMARY               	membranes of proximal kidney tubules. Meprins are also expressed in leukocytes
PR:PROJECT_SUMMARY               	(monocytes and macrophages) and podocytes. Meprins have been implicated in the
PR:PROJECT_SUMMARY               	pathology of acute and chronic kidney injury. Single nucleotide polymorphisms
PR:PROJECT_SUMMARY               	(SNPs) in the meprin β gene were associated in human DN in the Pima Indians,
PR:PROJECT_SUMMARY               	suggesting a role for meprins in the pathophysiology of DN. The current study
PR:PROJECT_SUMMARY               	was done to determine the mechanisms by which meprins modulate the progression
PR:PROJECT_SUMMARY               	of DN in mice.
PR:INSTITUTE                     	North Carolina A&T State University
PR:DEPARTMENT                    	Department of Biology
PR:LAST_NAME                     	Ongeri
PR:FIRST_NAME                    	Elimelda Moige
PR:ADDRESS                       	1601 E Market Street, Greensboro, NC 27411
PR:EMAIL                         	eongeri@ncat.edu
PR:PHONE                         	336-285-2182
PR:FUNDING_SOURCE                	NIH/NIGMS Grant # SC3102049 To Elimelda Moige Ongeri; NIH Center Grant #
PR:FUNDING_SOURCE                	U24DK097193 to Susan Sumner; NIH/NCATS award # UL1TR001111 to John Buse, UNC-CH,
PR:FUNDING_SOURCE                	PI, Sumner- Director, Metabolomics core; and NIH/NIGMS Grant # K01GM109320 to
PR:FUNDING_SOURCE                	Jessica Gooding.
PR:DOI                           	http://dx.doi.org/10.21228/M82889
#STUDY
ST:STUDY_TITLE                   	Urine metabolomic profiling of diabetic nephropathy in the steptozotocin induced
ST:STUDY_TITLE                   	type-1 diabetes mouse model.
ST:STUDY_SUMMARY                 	This metabolomics study evaluated urine from wild-type and meprin β knockout
ST:STUDY_SUMMARY                 	mice after induction of diabetes with streptozotocin or treatment with sodium
ST:STUDY_SUMMARY                 	citrate control to understand how these factors influence the metabotype.
ST:INSTITUTE                     	RTI International
ST:LABORATORY                    	NIH Eastern Regional Comphrehensive Metabolomics Resource Core at UNC Chapel
ST:LABORATORY                    	Hill (ERCMRC)
ST:LAST_NAME                     	Sumner
ST:FIRST_NAME                    	Susan
ST:ADDRESS                       	3040 E. Cornwallis Road, Research Triangle Park, NC 27709
ST:EMAIL                         	susan_sumner@unc.edu
ST:PHONE                         	704-250-5000
ST:SUBMIT_DATE                   	2017-02-17
#SUBJECT
SU:SUBJECT_TYPE                  	Animal
SU:SUBJECT_SPECIES               	Mus musculus
SU:TAXONOMY_ID                   	10090
SU:GENOTYPE_STRAIN               	C57BL/6
SU:GENDER                        	Male
SU:ANIMAL_ANIMAL_SUPPLIER        	KO: bred and housed at the laboratory animal resource unit (LARU) of North
SU:ANIMAL_ANIMAL_SUPPLIER        	Carolina A&T State University; WT: Charles River Laboratories
SU:ANIMAL_HOUSING                	group cages of up to 5 mice per cage
SU:ANIMAL_LIGHT_CYCLE            	12:12h light:dark cycle
SU:ANIMAL_FEED                   	rat chow
SU:ANIMAL_WATER                  	water ad libitum
SU:SPECIES_GROUP                 	Mammal
#SUBJECT_SAMPLE_FACTORS:         	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data
SUBJECT_SAMPLE_FACTORS           	Equilibrium_6	Equilibrium_6	Treatment:- | genotype:- | Week:-	"Subject Identifer Instrument Run Name (POS)"=MO_U_Equil6.raw; "Subject Identifier Instrument Run Name (NEG)"=-; Gender=-
SUBJECT_SAMPLE_FACTORS           	Total Pool_1	Total Pool_1	Treatment:- | genotype:- | Week:-	"Subject Identifer Instrument Run Name (POS)"=MO_U_Pool_1.raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U_Pool_1_NEG.raw; Gender=-
SUBJECT_SAMPLE_FACTORS           	Total Pool_2	Total Pool_2	Treatment:- | genotype:- | Week:-	"Subject Identifer Instrument Run Name (POS)"=MO_U_Pool_2.raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U_Pool_2_NEG.raw; Gender=-
SUBJECT_SAMPLE_FACTORS           	Total Pool_3	Total Pool_3	Treatment:- | genotype:- | Week:-	"Subject Identifer Instrument Run Name (POS)"=MO_U_Pool_3.raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U_Pool_3_NEG.raw; Gender=-
SUBJECT_SAMPLE_FACTORS           	Total Pool_4	Total Pool_4	Treatment:- | genotype:- | Week:-	"Subject Identifer Instrument Run Name (POS)"=MO_U_Pool_4.raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U_Pool_4_NEG.raw; Gender=-
SUBJECT_SAMPLE_FACTORS           	Total Pool_5	Total Pool_5	Treatment:- | genotype:- | Week:-	"Subject Identifer Instrument Run Name (POS)"=MO_U_Pool_5.raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U_Pool_5_NEG.raw; Gender=-
SUBJECT_SAMPLE_FACTORS           	383	383-4	Treatment:NaC | genotype:Meprin bKO | Week:4	"Subject Identifer Instrument Run Name (POS)"=MO_U4_KO_NaC_383 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U4_KO_NaC_383 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	387	387-4	Treatment:NaC | genotype:Meprin bKO | Week:4	"Subject Identifer Instrument Run Name (POS)"=MO_U4_KO_NaC_387 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U4_KO_NaC_387 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	391	391-4	Treatment:NaC | genotype:Meprin bKO | Week:4	"Subject Identifer Instrument Run Name (POS)"=MO_U4_KO_NaC_391 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U4_KO_NaC_391 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	383	383-8	Treatment:NaC | genotype:Meprin bKO | Week:8	"Subject Identifer Instrument Run Name (POS)"=MO_U8_KO_NaC_383 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U8_KO_NaC_383 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	387	387-8	Treatment:NaC | genotype:Meprin bKO | Week:8	"Subject Identifer Instrument Run Name (POS)"=MO_U8_KO_NaC_387 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U8_KO_NaC_387 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	390	390-8	Treatment:NaC | genotype:Meprin bKO | Week:8	"Subject Identifer Instrument Run Name (POS)"=MO_U8_KO_NaC_390 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U8_KO_NaC_390 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	391	391-8	Treatment:NaC | genotype:Meprin bKO | Week:8	"Subject Identifer Instrument Run Name (POS)"=MO_U8_KO_NaC_391 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U8_KO_NaC_391 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	399	399-8	Treatment:NaC | genotype:Meprin bKO | Week:8	"Subject Identifer Instrument Run Name (POS)"=MO_U8_KO_NaC_399 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U8_KO_NaC_399 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	519	519-4	Treatment:NaC | genotype:wild-type | Week:4	"Subject Identifer Instrument Run Name (POS)"=MO_U4_WT_NaC_519 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U4_WT_NaC_519 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	520	520-4	Treatment:NaC | genotype:wild-type | Week:4	"Subject Identifer Instrument Run Name (POS)"=MO_U4_WT_NaC_520 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U4_WT_NaC_520 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	521	521-4	Treatment:NaC | genotype:wild-type | Week:4	"Subject Identifer Instrument Run Name (POS)"=MO_U4_WT_NaC_521 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U4_WT_NaC_521 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	523	523-4	Treatment:NaC | genotype:wild-type | Week:4	"Subject Identifer Instrument Run Name (POS)"=MO_U4_WT_NaC_523 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U4_WT_NaC_523 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	519	519-8	Treatment:NaC | genotype:wild-type | Week:8	"Subject Identifer Instrument Run Name (POS)"=MO_U8_WT_NaC_519 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U8_WT_NaC_519 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	520	520-8	Treatment:NaC | genotype:wild-type | Week:8	"Subject Identifer Instrument Run Name (POS)"=MO_U8_WT_NaC_520 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U8_WT_NaC_520 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	521	521-8	Treatment:NaC | genotype:wild-type | Week:8	"Subject Identifer Instrument Run Name (POS)"=MO_U8_WT_NaC_521 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U8_WT_NaC_521 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	522	522-8	Treatment:NaC | genotype:wild-type | Week:8	"Subject Identifer Instrument Run Name (POS)"=MO_U8_WT_NaC_522 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U8_WT_NaC_522 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	523	523-8	Treatment:NaC | genotype:wild-type | Week:8	"Subject Identifer Instrument Run Name (POS)"=MO_U8_WT_NaC_523 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U8_WT_NaC_523 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	395	395-4	Treatment:STZ | genotype:Meprin bKO | Week:4	"Subject Identifer Instrument Run Name (POS)"=MO_U4_KO_STZ_395 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U4_KO_STZ_395 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	396	396-4	Treatment:STZ | genotype:Meprin bKO | Week:4	"Subject Identifer Instrument Run Name (POS)"=MO_U4_KO_STZ_396 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U4_KO_STZ_396 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	397	397-4	Treatment:STZ | genotype:Meprin bKO | Week:4	"Subject Identifer Instrument Run Name (POS)"=MO_U4_KO_STZ_397 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U4_KO_STZ_397 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	400	400-4	Treatment:STZ | genotype:Meprin bKO | Week:4	"Subject Identifer Instrument Run Name (POS)"=MO_U4_KO_STZ_400 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U4_KO_STZ_400 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	514	514-4	Treatment:STZ | genotype:Meprin bKO | Week:4	"Subject Identifer Instrument Run Name (POS)"=MO_U4_KO_STZ_514 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U4_KO_STZ_514 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	515	515-4	Treatment:STZ | genotype:Meprin bKO | Week:4	"Subject Identifer Instrument Run Name (POS)"=MO_U4_KO_STZ_515 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U4_KO_STZ_515 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	516	516-4	Treatment:STZ | genotype:Meprin bKO | Week:4	"Subject Identifer Instrument Run Name (POS)"=MO_U4_KO_STZ_516 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U4_KO_STZ_516 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	518	518-4	Treatment:STZ | genotype:Meprin bKO | Week:4	"Subject Identifer Instrument Run Name (POS)"=MO_U4_KO_STZ_518 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U4_KO_STZ_518 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	395	395-8	Treatment:STZ | genotype:Meprin bKO | Week:8	"Subject Identifer Instrument Run Name (POS)"=MO_U8_KO_STZ_395 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U8_KO_STZ_395 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	396	396-8	Treatment:STZ | genotype:Meprin bKO | Week:8	"Subject Identifer Instrument Run Name (POS)"=MO_U8_KO_STZ_396 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U8_KO_STZ_396 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	400	400-8	Treatment:STZ | genotype:Meprin bKO | Week:8	"Subject Identifer Instrument Run Name (POS)"=MO_U8_KO_STZ_400 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U8_KO_STZ_400 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	515	515-8	Treatment:STZ | genotype:Meprin bKO | Week:8	"Subject Identifer Instrument Run Name (POS)"=MO_U8_KO_STZ_515 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U8_KO_STZ_515 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	516	516-8	Treatment:STZ | genotype:Meprin bKO | Week:8	"Subject Identifer Instrument Run Name (POS)"=MO_U8_KO_STZ_516 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U8_KO_STZ_516 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	518	518-8	Treatment:STZ | genotype:Meprin bKO | Week:8	"Subject Identifer Instrument Run Name (POS)"=MO_U8_KO_STZ_518 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U8_KO_STZ_518 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	525	525-4	Treatment:STZ | genotype:wild-type | Week:4	"Subject Identifer Instrument Run Name (POS)"=MO_U4_WT_STZ_525 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U4_WT_STZ_525 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	526	526-4	Treatment:STZ | genotype:wild-type | Week:4	"Subject Identifer Instrument Run Name (POS)"=MO_U4_WT_STZ_526 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U4_WT_STZ_526 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	527	527-4	Treatment:STZ | genotype:wild-type | Week:4	"Subject Identifer Instrument Run Name (POS)"=MO_U4_WT_STZ_527 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U4_WT_STZ_527 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	528	528-4	Treatment:STZ | genotype:wild-type | Week:4	"Subject Identifer Instrument Run Name (POS)"=MO_U4_WT_STZ_528 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U4_WT_STZ_528 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	529	529-4	Treatment:STZ | genotype:wild-type | Week:4	"Subject Identifer Instrument Run Name (POS)"=MO_U4_WT_STZ_529 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U4_WT_STZ_529 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	530	530-4	Treatment:STZ | genotype:wild-type | Week:4	"Subject Identifer Instrument Run Name (POS)"=MO_U4_WT_STZ_530 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U4_WT_STZ_530 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	531	531-4	Treatment:STZ | genotype:wild-type | Week:4	"Subject Identifer Instrument Run Name (POS)"=MO_U4_WT_STZ_531 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U4_WT_STZ_531 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	532	532-4	Treatment:STZ | genotype:wild-type | Week:4	"Subject Identifer Instrument Run Name (POS)"=MO_U4_WT_STZ_532 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U4_WT_STZ_532 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	533	533-4	Treatment:STZ | genotype:wild-type | Week:4	"Subject Identifer Instrument Run Name (POS)"=MO_U4_WT_STZ_533 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U4_WT_STZ_533 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	525	525-8	Treatment:STZ | genotype:wild-type | Week:8	"Subject Identifer Instrument Run Name (POS)"=MO_U8_WT_STZ_525 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U8_WT_STZ_525 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	526	526-8	Treatment:STZ | genotype:wild-type | Week:8	"Subject Identifer Instrument Run Name (POS)"=MO_U8_WT_STZ_526 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U8_WT_STZ_526 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	528	528-8	Treatment:STZ | genotype:wild-type | Week:8	"Subject Identifer Instrument Run Name (POS)"=MO_U8_WT_STZ_528 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U8_WT_STZ_528 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	529	529-8	Treatment:STZ | genotype:wild-type | Week:8	"Subject Identifer Instrument Run Name (POS)"=MO_U8_WT_STZ_529 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U8_WT_STZ_529 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	530	530-8	Treatment:STZ | genotype:wild-type | Week:8	"Subject Identifer Instrument Run Name (POS)"=MO_U8_WT_STZ_530 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U8_WT_STZ_530 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	531	531-8	Treatment:STZ | genotype:wild-type | Week:8	"Subject Identifer Instrument Run Name (POS)"=MO_U8_WT_STZ_531 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U8_WT_STZ_531 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	532	532-8	Treatment:STZ | genotype:wild-type | Week:8	"Subject Identifer Instrument Run Name (POS)"=MO_U8_WT_STZ_532 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U8_WT_STZ_532 _NEG.raw; Gender=Male
SUBJECT_SAMPLE_FACTORS           	533	533-8	Treatment:STZ | genotype:wild-type | Week:8	"Subject Identifer Instrument Run Name (POS)"=MO_U8_WT_STZ_533 .raw; "Subject Identifier Instrument Run Name (NEG)"=MO_U8_WT_STZ_533 _NEG.raw; Gender=Male
#COLLECTION
CO:COLLECTION_SUMMARY            	Urine samples were collected at 4 and 8 weeks post-STZ injection. Urine samples
CO:COLLECTION_SUMMARY            	were obtained by bladder massage unto sterile petri dishes and stored at -80oC
CO:COLLECTION_SUMMARY            	until used.
CO:SAMPLE_TYPE                   	Urine
CO:STORAGE_CONDITIONS            	-80 C
#TREATMENT
TR:TREATMENT_SUMMARY             	Type 1 diabetes was induced at age 8 weeks. The mice were fasted for 6 hours
TR:TREATMENT_SUMMARY             	prior to being injected with low dose STZ (50 mg/kg body weight) in sodium
TR:TREATMENT_SUMMARY             	citrate buffer (10 mmol/L, pH 4.5) daily for 5 days following the protocols
TR:TREATMENT_SUMMARY             	described by Tesch and Allen and recommended by the Animal Models of Diabetic
TR:TREATMENT_SUMMARY             	Complications Consortium (AMDCC; amdcc.org)1. Control mice were injected with
TR:TREATMENT_SUMMARY             	equivalent volumes of the sodium citrate buffer. Diabetes was confirmed by
TR:TREATMENT_SUMMARY             	measuring fasting blood glucose levels using a standard glucose meter at 10 days
TR:TREATMENT_SUMMARY             	post-STZ injection. Mice with a fasting glucose level ≥280 mg/dL were
TR:TREATMENT_SUMMARY             	considered diabetic. STZ-injected mice with a fasting blood glucose of <280
TR:TREATMENT_SUMMARY             	mg/dL (15mmol/L) were culled and eliminated from the study.
TR:TREATMENT_COMPOUND            	streptozotocin
TR:TREATMENT_DOSE                	50 mg/kg body weight
TR:TREATMENT_DOSEDURATION        	daily; 5 days
TR:TREATMENT_VEHICLE             	sodium citrate
TR:ANIMAL_ENDP_TISSUE_COLL_LIST  	plasma, urine, kidney tissue
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Urine samples were thawed on ice for 30–60 min and vortexed on a multi-tube
SP:SAMPLEPREP_SUMMARY            	vortexer for 2 mins at 5,000 rpm, followed by centrifugation at room temperature
SP:SAMPLEPREP_SUMMARY            	at 16,000 rcf for 4 min. An aliquot of the supernatant was transferred to
SP:SAMPLEPREP_SUMMARY            	pre-labeled 1.5 mL low protein-binding microcentrifuge tube to make the
SP:SAMPLEPREP_SUMMARY            	individual study sample (40 µL). Another aliquot of supernatant (11 µL) was
SP:SAMPLEPREP_SUMMARY            	combined with those from all other urine samples in a 1.5 mL low protein-binding
SP:SAMPLEPREP_SUMMARY            	microcentrifuge tube to make the total pool quality control sample. The total
SP:SAMPLEPREP_SUMMARY            	pool quality control sample was aliquoted (40 µL) to make 5 Total Pool Samples
SP:SAMPLEPREP_SUMMARY            	and 4 Column Equilibration samples. 132 µL of the total pool quality control
SP:SAMPLEPREP_SUMMARY            	sample was added to the total study sample pool to make a pool of kidney, urine,
SP:SAMPLEPREP_SUMMARY            	and plasma samples. The total study pool sample was used to prepare 9 Total
SP:SAMPLEPREP_SUMMARY            	Study Samples (40 µL) to aid in alignment of the three studies. Of these, 3
SP:SAMPLEPREP_SUMMARY            	total study samples were included in the urine analysis. Acetonitrile containing
SP:SAMPLEPREP_SUMMARY            	the internal standard Tryptophan-d5 (120 µL; 0.0167 mg/ml) was added to all
SP:SAMPLEPREP_SUMMARY            	samples, and samples were vortexed on a multiple tube vortexer for 2 min at 5000
SP:SAMPLEPREP_SUMMARY            	rpm followed by centrifugation at 16,000 rcf for 4 min. The supernatants (420
SP:SAMPLEPREP_SUMMARY            	µL) were transferred into new, pre-labeled 2.0 mL low protein-binding
SP:SAMPLEPREP_SUMMARY            	microcentrifuge tubes and capped with disposable rubber stoppers. Samples were
SP:SAMPLEPREP_SUMMARY            	placed at -80 °C for 1.5 h and lyophilized for 18 h. Acetonitrile:Water (150
SP:SAMPLEPREP_SUMMARY            	µL; 95:5) was added to each tube and vortex mixed for 10 min at 5000 rpm,
SP:SAMPLEPREP_SUMMARY            	followed by centrifugation at room temperature at 16,000 rcf for 4 min. The
SP:SAMPLEPREP_SUMMARY            	supernatants were transferred to new, pre-labeled autosampler vials and 3 uL was
SP:SAMPLEPREP_SUMMARY            	injected into SYNAPT G2-Si.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	Hydrophilic Interaction Liquid Chromatography (HILIC) Gradient Seperation
CH:INSTRUMENT_NAME               	Waters Acquity I-Class
CH:COLUMN_NAME                   	Waters Acquity BEH Amide (150 x 2.1mm,1.7um)
CH:CHROMATOGRAPHY_TYPE           	HILIC
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Waters Synapt G2 Si QTOF
MS:INSTRUMENT_TYPE               	QTOF
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	POSITIVE
#END