Summary of Study ST000540

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR000395. The data can be accessed directly via it's Project DOI: 10.21228/M80G60 This work is supported by NIH grant, U2C- DK119886.

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Study ID	ST000540
Study Title	Kidney tissue metabolomic profiling of diabetic nephropathy in the steptozotocin induced type-1 diabetes mouse model.
Study Type	Metabolomics
Study Summary	This metabolomics study evaluated kidney tissue from wild-type and meprin β knockout mice after induction of diabetes with streptozotocin or treatment with sodium citrate control to understand how these factors influence the metabotype.
Institute	RTI International
Department	Discovery-Science-Technology
Laboratory	NIH Eastern Regional Comphrehensive Metabolomics Resource Core at UNC Chapel Hill (ERCMRC)
Last Name	Sumner
First Name	Susan
Address	3040 E Cornwallis Road, Research Triangle Park, NC 27709
Email	susan_sumner@unc.edu
Phone	704-250-5000
Submit Date	2017-01-20
Raw Data Available	Yes
Raw Data File Type(s)	raw(Waters)
Analysis Type Detail	LC-MS
Release Date	2018-02-07
Release Version	1

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Sample Preparation:

Sampleprep ID:	SP000569
Sampleprep Summary:	Frozen kidney tissue samples on dry ice were transferred to pre-chilled, pre-labeled tubes, and their weights recorded. For every 1 mg of tissue sample, 2 µL of 50:50 Acetonitrile:Water was added to the tube. Ceramic beads (2.3 mm; ~15-20 prewashed & dried) were added to the tubes, and the samples were homogenized on the MagNA Lyser system using a 30 s pulse at 4,000 rpm. The samples were centrifuged at room temperature at 16,000 rcf for 4 min. An aliquot of the supernatant was transferred to pre-labeled 2.0 mL low protein-binding microcentrifuge tube to make the individual study sample (40 µL). Another aliquot of supernatant (18 µL) was combined with those from all other kidney tissue 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 4 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 kidney tissue analysis. Acetonitrile containing the internal standard Tryptophan-d5 (460 µL; 0.0125 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 pre-labeled autosampler vials and 3 µL was injected into SYNAPT G2 Si.

Sampleprep ID:

SP000569

Sampleprep Summary:

Frozen kidney tissue samples on dry ice were transferred to pre-chilled, pre-labeled tubes, and their weights recorded. For every 1 mg of tissue sample, 2 µL of 50:50 Acetonitrile:Water was added to the tube. Ceramic beads (2.3 mm; ~15-20 prewashed & dried) were added to the tubes, and the samples were homogenized on the MagNA Lyser system using a 30 s pulse at 4,000 rpm. The samples were centrifuged at room temperature at 16,000 rcf for 4 min. An aliquot of the supernatant was transferred to pre-labeled 2.0 mL low protein-binding microcentrifuge tube to make the individual study sample (40 µL). Another aliquot of supernatant (18 µL) was combined with those from all other kidney tissue 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 4 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 kidney tissue analysis. Acetonitrile containing the internal standard Tryptophan-d5 (460 µL; 0.0125 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 pre-labeled autosampler vials and 3 µL was injected into SYNAPT G2 Si.