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MB Sample ID: SA028883
Local Sample ID: | 532-4 |
Subject ID: | SU000581 |
Subject Type: | Animal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Genotype Strain: | C57BL/6 |
Gender: | Male |
Animal Animal Supplier: | KO: bred and housed at the laboratory animal resource unit (LARU) of North Carolina A&T State University; WT: Charles River Laboratories |
Animal Housing: | group cages of up to 5 mice per cage |
Animal Light Cycle: | 12:12h light:dark cycle |
Animal Feed: | rat chow |
Animal Water: | water ad libitum |
Species Group: | Mammal |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU000581 |
Subject Type: | Animal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Genotype Strain: | C57BL/6 |
Gender: | Male |
Animal Animal Supplier: | KO: bred and housed at the laboratory animal resource unit (LARU) of North Carolina A&T State University; WT: Charles River Laboratories |
Animal Housing: | group cages of up to 5 mice per cage |
Animal Light Cycle: | 12:12h light:dark cycle |
Animal Feed: | rat chow |
Animal Water: | water ad libitum |
Species Group: | Mammal |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
532-4 | SA028883 | FL007039 | STZ | Treatment |
532-4 | SA028883 | FL007039 | wild-type | genotype |
532-4 | SA028883 | FL007039 | 4 | Week |
Collection:
Collection ID: | CO000575 |
Collection Summary: | Urine samples were collected at 4 and 8 weeks post-STZ injection. Urine samples were obtained by bladder massage unto sterile petri dishes and stored at -80oC until used. |
Sample Type: | Urine |
Storage Conditions: | -80 C |
Treatment:
Treatment ID: | TR000595 |
Treatment Summary: | Type 1 diabetes was induced at age 8 weeks. The mice were fasted for 6 hours prior to being injected with low dose STZ (50 mg/kg body weight) in sodium citrate buffer (10 mmol/L, pH 4.5) daily for 5 days following the protocols described by Tesch and Allen and recommended by the Animal Models of Diabetic Complications Consortium (AMDCC; amdcc.org)1. Control mice were injected with equivalent volumes of the sodium citrate buffer. Diabetes was confirmed by measuring fasting blood glucose levels using a standard glucose meter at 10 days post-STZ injection. Mice with a fasting glucose level ≥280 mg/dL were considered diabetic. STZ-injected mice with a fasting blood glucose of <280 mg/dL (15mmol/L) were culled and eliminated from the study. |
Treatment Compound: | streptozotocin |
Treatment Dose: | 50 mg/kg body weight |
Treatment Doseduration: | daily; 5 days |
Treatment Vehicle: | sodium citrate |
Animal Endp Tissue Coll List: | plasma, urine, kidney tissue |
Sample Preparation:
Sampleprep ID: | SP000588 |
Sampleprep Summary: | Urine samples were thawed on ice for 30–60 min and vortexed on a multi-tube vortexer for 2 mins at 5,000 rpm, followed by centrifugation at room temperature at 16,000 rcf for 4 min. An aliquot of the supernatant was transferred to pre-labeled 1.5 mL low protein-binding microcentrifuge tube to make the individual study sample (40 µL). Another aliquot of supernatant (11 µL) was combined with those from all other urine samples in a 1.5 mL low protein-binding microcentrifuge tube to make the total pool quality control sample. The total pool quality control sample was aliquoted (40 µL) to make 5 Total Pool Samples and 4 Column Equilibration samples. 132 µL of the total pool quality control sample was added to the total study sample pool to make a pool of kidney, urine, and plasma samples. The total study pool sample was used to prepare 9 Total Study Samples (40 µL) to aid in alignment of the three studies. Of these, 3 total study samples were included in the urine analysis. Acetonitrile containing the internal standard Tryptophan-d5 (120 µL; 0.0167 mg/ml) was added to all samples, and samples were vortexed on a multiple tube vortexer for 2 min at 5000 rpm followed by centrifugation at 16,000 rcf for 4 min. The supernatants (420 µL) were transferred into new, pre-labeled 2.0 mL low protein-binding microcentrifuge tubes and capped with disposable rubber stoppers. Samples were placed at -80 °C for 1.5 h and lyophilized for 18 h. Acetonitrile:Water (150 µL; 95:5) was added to each tube and vortex mixed for 10 min at 5000 rpm, followed by centrifugation at room temperature at 16,000 rcf for 4 min. The supernatants were transferred to new, pre-labeled autosampler vials and 3 uL was injected into SYNAPT G2-Si. |
Combined analysis:
Analysis ID | AN000858 | AN000859 |
---|---|---|
Analysis type | MS | MS |
Chromatography type | HILIC | HILIC |
Chromatography system | Waters Acquity I-Class | Waters Acquity I-Class |
Column | Waters Acquity BEH Amide (150 x 2.1mm,1.7um) | Waters Acquity BEH Amide (150 x 2.1mm,1.7um) |
MS Type | ESI | ESI |
MS instrument type | QTOF | QTOF |
MS instrument name | Waters Synapt G2 Si QTOF | Waters Synapt G2 Si QTOF |
Ion Mode | POSITIVE | NEGATIVE |
Units | normalized ion counts | normalized ion counts |
Chromatography:
Chromatography ID: | CH000611 |
Chromatography Summary: | Hydrophilic Interaction Liquid Chromatography (HILIC) Gradient Seperation |
Instrument Name: | Waters Acquity I-Class |
Column Name: | Waters Acquity BEH Amide (150 x 2.1mm,1.7um) |
Chromatography Type: | HILIC |
MS:
MS ID: | MS000759 |
Analysis ID: | AN000858 |
Instrument Name: | Waters Synapt G2 Si QTOF |
Instrument Type: | QTOF |
MS Type: | ESI |
Ion Mode: | POSITIVE |
MS ID: | MS000760 |
Analysis ID: | AN000859 |
Instrument Name: | Waters Synapt G2 Si QTOF |
Instrument Type: | QTOF |
MS Type: | ESI |
Ion Mode: | NEGATIVE |