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MB Sample ID: SA262769
Local Sample ID: | 90430015508 |
Subject ID: | SU002744 |
Subject Type: | Mammal |
Subject Species: | Rattus norvegicus |
Taxonomy ID: | 10116 |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU002744 |
Subject Type: | Mammal |
Subject Species: | Rattus norvegicus |
Taxonomy ID: | 10116 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
90430015508 | SA262769 | FL034253 | Training | Group |
90430015508 | SA262769 | FL034253 | 1 week of training or control time | Timepoint |
90430015508 | SA262769 | FL034253 | Male | Sex |
Collection:
Collection ID: | CO002737 |
Collection Summary: | - |
Sample Type: | Gastrocnemius |
Treatment:
Treatment ID: | TR002753 |
Treatment Summary: | - |
Sample Preparation:
Sampleprep ID: | SP002750 |
Sampleprep Summary: | Tissue samples (10 mg) were homogenized in 300 µL of 10/67.4/22.4/0.18 (v/v/v/v) water/acetonitrile/methanol/formic acid containing stable isotope-labeled internal standards (0.18 ng/µL valine-d8, Isotec; 0.18 ng/µL phenylalanine-d8, Cambridge Isotope Laboratories) using a TissueLyserII bead mill (20 Hz, 2 X 2min; QIAGEN). The samples were place in a -80?C freezer for one hour and then centrifuged for 10 min (9000g, 4?C). 100uL of each supernatent was transferred to a de-activated inserts within an autsampler vial (Waters). |
Sampleprep Protocol Filename: | pass1b_experimental_design_metabolomics.pdf |
Combined analysis:
Analysis ID | AN004314 |
---|---|
Analysis type | MS |
Chromatography type | HILIC |
Chromatography system | Shimadzu Nexera X2 |
Column | Waters Atlantis HILIC (150 x 2.1mm,3um) |
MS Type | ESI |
MS instrument type | Orbitrap |
MS instrument name | Thermo Q Exactive Orbitrap |
Ion Mode | POSITIVE |
Units | peak area |
Chromatography:
Chromatography ID: | CH003220 |
Chromatography Summary: | Extracts (10 µL) were injected onto a 150 x 2.1 mm Atlantis HILIC column (Waters). The column was eluted isocratically at a flow rate of 250 µL/min with 5% mobile phase A (10 mM ammonium formate and 0.1% formic acid in water) for 1 minute followed by a linear gradient to 40% mobile phase B (acetonitrile with 0.1% formic acid) over 10 minutes |
Methods Filename: | pass1b_hilicpos_methods.pdf |
Instrument Name: | Shimadzu Nexera X2 |
Column Name: | Waters Atlantis HILIC (150 x 2.1mm,3um) |
Column Temperature: | - |
Flow Gradient: | The column was eluted isocratically at a flow rate of 250 µL/min with 5% mobile phase A for 1 minute followed by a linear gradient to 40% mobile phase B over 10 minutes |
Flow Rate: | 250 µL/min |
Solvent A: | 100% water; 0.1% formic acid; 10 mM ammonium formate |
Solvent B: | 100% acetonitrile; 0.1% formic acid |
Chromatography Type: | HILIC |
MS:
MS ID: | MS004061 |
Analysis ID: | AN004314 |
Instrument Name: | Thermo Q Exactive Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | High resolution, accurate mass data were acquired using a system comprised of a Shimadzu Nexera X2 U-HPLC (Shimadzu Corp.; Marlborough, MA) coupled to a Q Exactive hybrid quadrupole orbitrap mass spectrometer (Thermo Fisher Scientific; Waltham, MA). MS analyses were carried out using electrospray ionization in the positive ion mode using full scan analysis over 70-800 m/z at 70,000 resolution and 3 Hz data acquisition rate. Other MS settings were: sheath gas 40, sweep gas 2, spray voltage 3.5 kV, capillary temperature 350°C, S-lens RF 40, heater temperature 300°C, microscans 1, automatic gain control target 1e6, and maximum ion time 250 ms. Raw data were processed using TraceFinder software (Thermo Fisher Scientific; Waltham, MA) and Progenesis QI (Nonlinear Dynamics; Newcastle upon Tyne, UK). The identities of 202 profiled metabolites were confirmed using reference standards. |
Ion Mode: | POSITIVE |