Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA330395

Local Sample ID:	Serum3DMN4negative
Subject ID:	SU003158
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Gender:	Female

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Subject:

Subject ID:	SU003158
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Gender:	Female

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
Serum3DMN4negative	SA330395	FL039091	Mutant	Genotype
Serum3DMN4negative	SA330395	FL039091	tamoxifen, 3day	Treatment

Collection:

Collection ID:	CO003151
Collection Summary:	serum and kidney were obtained from GCERRARaD females 3 days or 3 months post-tam (n=5/group) and age-matched wild-type females (n=4)
Sample Type:	Kidney; Serum

Treatment:

Treatment ID:	TR003167
Treatment Summary:	Mutant and wild-type female mice were treated with tamoxifen for 3 days or 3 months.

Sample Preparation:

Sampleprep ID:	SP003164
Sampleprep Summary:	To extract metabolites, frozen kidney cortices were weighed, and ca 20 mg of each cortex was added to a tube containing 650 μL of 80% methanol (diluted in dH2O). Sample tubes were placed in a bead homogenizer for 1 minute at a frequency of 30/s. Samples were then placed in a -20℃ freezer for 10 minutes, centrifuged for 10 minutes at 15,000 RPM in 4℃, and the supernatant was moved to a new tube. The remaining pellets were then re-extracted with 400 μL of 80% methanol, following the above procedure of homogenization and centrifugation. The supernatant was then added to the supernatant pool obtained from the first extraction. The pellet was re-extracted for a third time using 350 μL of 80% MeOH and supernatant was again added to the pool from the first two extractions. The supernatant pool was placed at -20℃ for 15 minutes, centrifuged for 20 minutes at 15,000 RPM, and transferred to a new tube. The methanolic extract samples were speed-vacuumed and reconstituted in 70% acetonitrile (diluted in dH2O) containing 0.025% acetic acid at a protein concentration of 3μg/μL. For serum preparation, samples were diluted 10-fold with 70% acetonitrile (diluted in dH2O) containing 0.025% acetic acid, and centrifuged for 20 minutes at 20,000 g.

Sampleprep ID:

SP003164

Sampleprep Summary:

To extract metabolites, frozen kidney cortices were weighed, and ca 20 mg of each cortex was added to a tube containing 650 μL of 80% methanol (diluted in dH2O). Sample tubes were placed in a bead homogenizer for 1 minute at a frequency of 30/s. Samples were then placed in a -20℃ freezer for 10 minutes, centrifuged for 10 minutes at 15,000 RPM in 4℃, and the supernatant was moved to a new tube. The remaining pellets were then re-extracted with 400 μL of 80% methanol, following the above procedure of homogenization and centrifugation. The supernatant was then added to the supernatant pool obtained from the first extraction. The pellet was re-extracted for a third time using 350 μL of 80% MeOH and supernatant was again added to the pool from the first two extractions. The supernatant pool was placed at -20℃ for 15 minutes, centrifuged for 20 minutes at 15,000 RPM, and transferred to a new tube. The methanolic extract samples were speed-vacuumed and reconstituted in 70% acetonitrile (diluted in dH2O) containing 0.025% acetic acid at a protein concentration of 3μg/μL. For serum preparation, samples were diluted 10-fold with 70% acetonitrile (diluted in dH2O) containing 0.025% acetic acid, and centrifuged for 20 minutes at 20,000 g.

Combined analysis:

Analysis ID	AN004992
Analysis type	MS
Chromatography type	HILIC
Chromatography system	Agilent 1290 Infinity II
Column	MicroSolv Cogent Diamond Hydride (150 × 2.1 mm, 4.0 um)
MS Type	ESI
MS instrument type	QTOF
MS instrument name	Agilent 6550 QTOF
Ion Mode	UNSPECIFIED
Units	abundance

Chromatography:

Chromatography ID:	CH003771
Instrument Name:	Agilent 1290 Infinity II
Column Name:	MicroSolv Cogent Diamond Hydride (150 × 2.1 mm, 4.0 um)
Column Temperature:	25
Flow Gradient:	0-1 min, 99% B; 1-15 min, a linear gradient to 20% B; 15.1-29 min, 0% B; 29.1-37 min, 99% B
Flow Rate:	0.4mL/min
Solvent A:	6 uM edta and 0.025% acetic acid in isopropanol:H2O (50:50)
Solvent B:	6 uM edta and 5mM ammonium acetate in CH3CN: H2O (90:10)
Chromatography Type:	HILIC

MS:

MS ID:	MS004732
Analysis ID:	AN004992
Instrument Name:	Agilent 6550 QTOF
Instrument Type:	QTOF
MS Type:	ESI
MS Comments:	The serum samples have negative and positive mode data. The kidney samples have negative mode data. MassHunter B10 MassHunter B10 MassHunter Profinder (B10)
Ion Mode:	UNSPECIFIED