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MB Sample ID: SA260940
| Local Sample ID: | 90585016811 |
| Subject ID: | SU002716 |
| Subject Type: | Mammal |
| Subject Species: | Rattus norvegicus |
| Taxonomy ID: | 10116 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002716 |
| Subject Type: | Mammal |
| Subject Species: | Rattus norvegicus |
| Taxonomy ID: | 10116 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 90585016811 | SA260940 | FL033854 | Training | Group |
| 90585016811 | SA260940 | FL033854 | 2 weeks of training | Timepoint |
| 90585016811 | SA260940 | FL033854 | Female | Sex |
Collection:
| Collection ID: | CO002709 |
| Collection Summary: | - |
| Sample Type: | Liver |
Treatment:
| Treatment ID: | TR002726 |
| Treatment Summary: | - |
Sample Preparation:
| Sampleprep ID: | SP002722 |
| Sampleprep Summary: | 10 µl of plasma or 300 µl of tissue homogenate prepared at 100 mg/ml in 3M perchloric acid containing isotopically labeled internal standards KIC-d3, KIV-5C13, (Cambridge Isotope Laboratories), and KMV-d8 (Toronto Research Chemicals) are precipitated with 150 µl of 3M PCA. 200 µl of 25 M o-phenylenediamine (OPD) in 3M HCl is added to the supernatants and the samples are incubated at 80oC for 20 minutes. Keto acids are extracted with ethyl acetate. The extracts are dried under nitrogen and reconstituted in 200 mM ammonium acetate. |
| Sampleprep Protocol Filename: | pass1b_experimental_design_metabolomics.pdf |
Combined analysis:
| Analysis ID | AN004286 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Waters Acquity UPLC |
| Column | Waters Acquity UPLC BEH C18 (50 x 2.1 mm,1.7 um) |
| MS Type | ESI |
| MS instrument type | Triple quadrupole |
| MS instrument name | Waters Xevo TQ-S |
| Ion Mode | POSITIVE |
| Units | pmol/mg of tissue |
Chromatography:
| Chromatography ID: | CH003192 |
| Chromatography Summary: | The analytical column is used at 30°C. 10 µl of the sample are injected onto the column and eluted at a flow rate of 0.4 ml/min. The gradient begins with 45% eluent A (5 mM ammonium acetate in water) and is then programmed as follows: 0 to 2 min – gradient to 55% eluent B (methanol); 2 to 2.5 min - gradient to 95% eluent B; 2.5 to 3.2 min - hold at 95% eluent B, return to 45% A, and re-equilibrate the column at initial conditions for 1 minute. |
| Methods Filename: | pass1b_ka_methods.pdf |
| Instrument Name: | Waters Acquity UPLC |
| Column Name: | Waters Acquity UPLC BEH C18 (50 x 2.1 mm,1.7 um) |
| Column Temperature: | 30C |
| Flow Gradient: | 45% A and 55% B for 2 minutes, followed by a linear gradient to 95% B from 2 to 2.5 minutes, held at 95% B for 0.7 minutes, returned to 45% A, and finally the column was re-equilibrated at initial conditions for 1 minute. The total run time was 4.7 minutes. |
| Flow Rate: | 0.4 ml/min |
| Solvent A: | 100% water; 5 mM ammonium acetate |
| Solvent B: | 100% methanol |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS004033 |
| Analysis ID: | AN004286 |
| Instrument Name: | Waters Xevo TQ-S |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | Mass transitions of m/z 203 -> 161 (ketoleucine or KIC), 206 -> 161 (KIC-d3), 189 -> 174 (ketoisovalerate or KIV), 194 -> 178 (KIV-5C13), 203 -> 174 (3-methyl-2-oxovalerate or KMV), and 211 -> 177 (KMV-d8) are monitored in a positive ion electrospray ionization mode. |
| Ion Mode: | POSITIVE |
