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MB Sample ID: SA097868
Local Sample ID: | AR124_2-2 |
Subject ID: | SU001421 |
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: | SU001421 |
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 ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
AR124_2-2 | SA097868 | FL013727 | Control (normal protein diet) D12450B | Diet |
Collection:
Collection ID: | CO001416 |
Collection Summary: | Mice were fasted overnight and then re-fed for 5 to 6 hours before blood was collected from the hepatic portal vein. |
Sample Type: | Blood (plasma) |
Storage Conditions: | -80℃ |
Treatment:
Treatment ID: | TR001436 |
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 sample collections and being humanely euthanised for tissue collection. |
Sample Preparation:
Sampleprep ID: | SP001429 |
Sampleprep Summary: | Plasma (25 µL) was extracted by addition of 200 µL of 1:1 acetonitrile:MeOH v/v with 1 µM CHAPS, CAPS, PIPES and TRIS as internal standards at 0°C. Samples were then mixed on a vortex mixer for 30 minutes at 4°C after which they were centrifuged (20,000 rcf, 4℃, 15 min) and the supernatant then transferred to a mass spectrometry vial for LC-MS analysis. |
Processing Storage Conditions: | Described in summary |
Extract Storage: | -80℃ |
Combined analysis:
Analysis ID | AN002241 |
---|---|
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: | CH001647 |
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: | MS002087 |
Analysis ID: | AN002241 |
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 |