Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA008926

Local Sample ID:	MKRP4
Subject ID:	SU000183
Subject Type:	Animal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Genotype Strain:	FVB (wild type)\|MKR (FVB background)
Age Or Age Range:	12 week old\|12 week old
Gender:	male\|male
Animal Animal Supplier:	JAX lab\|LeRoith laboratory
Species Group:	Mammals

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Subject:

Subject ID:	SU000183
Subject Type:	Animal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Genotype Strain:	FVB (wild type)\|MKR (FVB background)
Age Or Age Range:	12 week old\|12 week old
Gender:	male\|male
Animal Animal Supplier:	JAX lab\|LeRoith laboratory
Species Group:	Mammals

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
MKRP4	SA008926	FL002130	MKR	Mice health condition
MKRP4	SA008926	FL002130	PBS buffer	Treatment

Collection:

Collection ID:	CO000169
Collection Summary:	-
Collection Protocol Filename:	Protocol_Methods_metformin_Li.pdf
Sample Type:	total bone marrow flush (cells)
Storage Conditions:	-20°C

Treatment:

Treatment ID:	TR000189
Treatment:	abiotic
Treatment Compound:	metformin
Treatment Route:	Intraperitoneal injection
Treatment Dose:	200 mg/kg body weight
Treatment Dosevolume:	50 microliters
Treatment Doseduration:	daily for 14 days
Treatment Vehicle:	phosphate buffered solution

Sample Preparation:

Sampleprep ID:	SP000183
Sampleprep Summary:	Frozen cells were mixed with dry ice-cool 80% methanol (mass-spec grade) at a ratio of 30 ?L/million cells, and then quickly thawed on heat block set at 50 °C for 5 min. The suspension was then processed by three rounds of 1 min vortex at max speed, chilled briefly on dry ice. The mixture was incubated at 4°C for 1 hour before centrifuge at 20,000 x g for 20 min at 4 °C. The supernatant was stored at -20 °C and used as metabolite extract for LC-MS analysis.
Sampleprep Protocol Filename:	Protocol_Methods_metformin_Li.pdf
Processing Method:	Homogenization and Centrifugation
Processing Storage Conditions:	On ice
Extraction Method:	80% Methanol
Extract Concentration Dilution:	30 microliters solvent per million cells
Extract Storage:	-20°C
Cell Type:	bone marrow

Combined analysis:

Chromatography:

Chromatography ID:	CH000181
Chromatography Summary:	Untarged mix-mode method
Instrument Name:	Thermo Exactive
Column Name:	Shodex ODP2 HP-4B 5um polymer (50 x 4.6mm)
Column Pressure:	150 Bar (Max)
Column Temperature:	45
Flow Gradient:	0-15 min -1% B, 15-19 min- gradient 1 to 99% B, 18-24 min- 99% B, 24-25 min-gradient 99% to 1% B, 25-30 min-1% B
Flow Rate:	0.1 ml/min
Injection Temperature:	4 C
Sample Injection:	5 uL
Solvent A:	10% acetonitrile/90% water; 10 mM ammonium acetate ; 10 mM acetic acid (positive) or 10 mM NH3.H2O (negative)
Solvent B:	90% acetonitrile/10% water; 10 mM ammonium acetate; 10 mM acetic acid (positive) or 10 mM ammonia (negative)
Analytical Time:	30 min
Capillary Voltage:	3.5 kv (positive), 3.1 kv (negative)
Transferline Temperature:	275C
Washing Buffer:	10% Acetonitrile
Weak Wash Solvent Name:	10% Acetonitrile
Weak Wash Volume:	600 uL
Strong Wash Solvent Name:	90% Acetonitrile
Strong Wash Volume:	200 uL
Sample Loop Size:	20 uL
Chromatography Type:	Reversed phase

MS: