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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:Mammal

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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:Mammal

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
MKRP4SA008926FL002130MKRMice health condition
MKRP4SA008926FL002130PBS bufferTreatment

Collection:

Collection ID:CO000169
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:

Analysis ID AN000256 AN000257
Analysis type MS MS
Chromatography type Reversed phase Reversed phase
Chromatography system Thermo Exactive Thermo Exactive
Column Shodex ODP2 HP-4B 5um polymer (50 x 4.6mm) Shodex ODP2 HP-4B 5um polymer (50 x 4.6mm)
MS Type ESI ESI
MS instrument type Orbitrap Orbitrap
MS instrument name Thermo Q Exactive Orbitrap Thermo Q Exactive Orbitrap
Ion Mode POSITIVE NEGATIVE
Units Peak area Peak area

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:

MS ID:MS000206
Analysis ID:AN000256
Instrument Name:Thermo Q Exactive Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
Ion Mode:POSITIVE
Capillary Temperature:30V
Collision Gas:40 units
Ion Source Temperature:3.5 kv
Ion Spray Voltage:Electrospray Ionization
Ionization Potential:< 2ppm
Atom Gun Current:3000000
Sample Dripping:75-1200
Scanning Range:20 v
Skimmer Voltage:120 v
  
MS ID:MS000207
Analysis ID:AN000257
Instrument Name:Thermo Q Exactive Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
Ion Mode:NEGATIVE
Capillary Temperature:30V
Collision Gas:40 units
Ion Source Temperature:3.1 kv
Ion Spray Voltage:Electrospray Ionization
Ionization Potential:< 4 ppm
Atom Gun Current:3000000
Sample Dripping:75-1200
Scanning Range:20 v
Skimmer Voltage:120 v
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