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MB Sample ID: SA097858

Local Sample ID:AR104_1-4
Subject ID:SU001420
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090
Genotype Strain:C57Bl/6NCrl
Age Or Age Range:7 to 10 weeks
Gender:Male
Animal Animal Supplier:Charles River Laboratories, DEU
Animal Light Cycle:12-12h light-dark cycle

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Subject:

Subject ID:SU001420
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090
Genotype Strain:C57Bl/6NCrl
Age Or Age Range:7 to 10 weeks
Gender:Male
Animal Animal Supplier:Charles River Laboratories, DEU
Animal Light Cycle:12-12h light-dark cycle

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
AR104_1-4SA097858FL013725Control (normal protein diet) D12450BDiet

Collection:

Collection ID:CO001415
Collection Summary:Mice were fasted overnight and then re-fed for 5 to 6 hours before being humanely euthanised for tissue collection.
Sample Type:Liver
Storage Conditions:-80℃

Treatment:

Treatment ID:TR001435
Treatment Summary:Male mice aged 7 weeks upon arrival, were acclimated to the local housing facility (12-12h light-dark cycle, 22-24°C) for one week prior to experimentation and were fed standard rodent chow (3437, Provimi Kliba, DEU). Following acclimation, mice were placed on either a normal protein diet (D12450B, Research Diets, New Jersey, USA) or a reduced protein diet (D10062201, Research Diets, New Jersey, USA) for 3 weeks prior to being humanely euthanised for tissue collection.

Sample Preparation:

Sampleprep ID:SP001428
Sampleprep Summary:Livers were cryogenically pulverized (cryopulverization) using a 12-well biopulverizer (BioSpec Products, OK USA Part number 59012MS) according to the manufacturer’s instructions. The frozen tissue powder was then weighed and extracted in 20 µL of extraction solvent (0°C) per mg of tissue. The mixture was then briefly vortexed before sonication in an ice-water bath for 10 minutes followed by centrifugation (20,000 rcf, 4℃, 10 minutes). The supernatant was then transferred to a mass spectrometry vial for LC-MS analysis. The extraction solvent consisted of 2:6:1 CHCl3:MeOH:H2O v/v/v with 2 µM CHAPS, CAPS, PIPES and TRIS as internal standards.
Processing Storage Conditions:Described in summary
Extract Storage:-80℃

Combined analysis:

Analysis ID AN002240
Analysis type MS
Chromatography type HILIC
Chromatography system Thermo Dionex Ultimate 3000 RS
Column SeQuant ZIC-pHILIC (150 x 4.6mm,5um)
MS Type ESI
MS instrument type Orbitrap
MS instrument name Thermo Q Exactive Orbitrap
Ion Mode UNSPECIFIED
Units Peak Area

Chromatography:

Chromatography ID:CH001646
Chromatography Summary:Chromatography utilized a ZIC-pHILIC column (column temperature 25 °C) with a gradient elution of 20 mM ammonium carbonate (A) and acetonitrile (B) (linear gradient time-%B as follows: 0 min-80%, 15 min-50%, 18 min-5%, 21 min-5%, 24 min-80%, 32 min-80%) on a Dionex RSLC3000 UHPLC (Thermo). The flow rate was maintained at 300 μL/min. Samples were kept at 4 °C in the autosampler and 10 μL injected for analysis.
Instrument Name:Thermo Dionex Ultimate 3000 RS
Column Name:SeQuant ZIC-pHILIC (150 x 4.6mm,5um)
Column Temperature:25
Flow Gradient:linear gradient time-%B as follows: 0 min-80%, 15 min-50%, 18 min-5%, 21 min-5%, 24 min-80%, 32 min-80%
Flow Rate:300 µL/min
Solvent A:100% water; 20 mM ammonium carbonate
Solvent B:100% water; acetonitrile
Chromatography Type:HILIC

MS:

MS ID:MS002086
Analysis ID:AN002240
Instrument Name:Thermo Q Exactive Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:Mass spectrometry was performed at 35 000 resolution (accuracy calibrated to <1 ppm) on a Q-Exactive Orbitrap MS (Thermo) operating in rapid switching positive (4 kV) and negative (−3.5 kV) mode electrospray ionization (capillary temperature 300 °C; sheath gas 50; Aux gas 20; sweep gas 2; probe temp 120 °C).
Ion Mode:UNSPECIFIED
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