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MB Sample ID: SA212401
| Local Sample ID: | 37904_sub |
| Subject ID: | SU002314 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | C57BL6 and FVB mixed strain |
| Age Or Age Range: | 54-59 weeks old |
| Gender: | Male and female |
| Animal Feed: | Specialty Feeds Irradiated Rat and Mouse Standard Chow Diet |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002314 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | C57BL6 and FVB mixed strain |
| Age Or Age Range: | 54-59 weeks old |
| Gender: | Male and female |
| Animal Feed: | Specialty Feeds Irradiated Rat and Mouse Standard Chow Diet |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 37904_sub | SA212401 | FL025925 | Male | Sex |
| 37904_sub | SA212401 | FL025925 | ERalpha-knockout | Genotype |
Collection:
| Collection ID: | CO002307 |
| Collection Summary: | Ventricles were dissected from mice such that each sample will include left and right ventricular tissue.Tissues were snap frozen in liquid nitrogen and stored in -80 freezer until tissues were processed for metabolomics. |
| Sample Type: | Ventricles, Skeletal Muscles, Subcutaneous fat |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR002326 |
| Treatment Summary: | Mice did not undergo specific treatment, as this was a basal phenotyping study. Mice were fasted for 6 hours before dissections, and a lethal dose of anesthesia was delivered via intraperitoneal injection before tissue collection. |
| Animal Anesthesia: | Pentobarbitone |
| Animal Fasting: | 6 hours |
Sample Preparation:
| Sampleprep ID: | SP002320 |
| Sampleprep Summary: | 600 µL of 3:1 methanol/water (containing 13C6 Sorbitol & 13C5, 15N Valine as internal standards) was added to the samples and cryomilled for 45 sec x 3 at 6800 rpm. 480 µL of the homogenate was transferred into a fresh tube and centrifuged for 10 min at 12700 rpm. 100 µL of the supernatant of each sample was pooled to make up the pooled biological quality controls (PBQCs). 100 µL of each sample (study samples & PBQCs) was dried using a SpeedVac vacuum concentrator prior to derivatization. The samples and PBQCs were methoximated (methoxyamine hydrochloride - 25 µL) and trimethylsilylated (BSTFA - 25 µL) to enable gas phase analysis. |
Combined analysis:
| Analysis ID | AN003637 |
|---|---|
| Analysis type | MS |
| Chromatography type | GC |
| Chromatography system | Shimadzu Nexis GC-2030 |
| Column | Agilent DB5-MS (30m x 0.25mm, 0.25um) |
| MS Type | EI |
| MS instrument type | Triple quadrupole |
| MS instrument name | Shimadzu TQ8050 NX |
| Ion Mode | POSITIVE |
| Units | abundance |
Chromatography:
| Chromatography ID: | CH002692 |
| Chromatography Summary: | The GC-MS system used comprised of an AOC6000 autosampler, a 2030 Shimadzu gas chromatograph and a TQ8050NX triple quadrupole mass spectrometer (Shimadzu, Japan). The mass spectrometer was tuned according to the manufacturer’s recommendations using tris-(perfluorobutyl)-amine (CF43). GC-MS was performed on a 30m Agilent DB-5 column with 0.25mm internal diameter column and 1µm film thickness. The injection temperature (inlet) was set at 280°C, the MS transfer line at 280°C and the ion source adjusted to 200°C. Helium was used as the carrier gas at a flow rate of 1 mL/min and argon gas was used in the collision cell to generate the MRM product ion. The analysis of TMS samples was performed under the following oven temperature program; 100°C start temperature, hold for 4 minutes, followed by a 10°C min-1 oven temperature ramp to 320°C with a following final hold for 11 minutes. Approximately 520 targets were collected using the Shimadzu Smart Metabolite Database, where each target comprised a quantifier MRM along with a qualifier MRM, which covers approximately 350 endogenous metabolites and multiple stable isotopically labelled internal standards. Resultant data was processed using Shimadzu LabSolutions Insight software, where peak integrations were visually validated and manually corrected where required. |
| Instrument Name: | Shimadzu Nexis GC-2030 |
| Column Name: | Agilent DB5-MS (30m x 0.25mm, 0.25um) |
| Chromatography Type: | GC |
MS:
| MS ID: | MS003388 |
| Analysis ID: | AN003637 |
| Instrument Name: | Shimadzu TQ8050 NX |
| Instrument Type: | Triple quadrupole |
| MS Type: | EI |
| MS Comments: | The GC-MS system used comprised of an AOC6000 autosampler, a 2030 Shimadzu gas chromatograph and a TQ8050NX triple quadrupole mass spectrometer (Shimadzu, Japan). The mass spectrometer was tuned according to the manufacturer’s recommendations using tris-(perfluorobutyl)-amine (CF43). GC-MS was performed on a 30m Agilent DB-5 column with 0.25mm internal diameter column and 1µm film thickness. The injection temperature (inlet) was set at 280°C, the MS transfer line at 280°C and the ion source adjusted to 200°C. Helium was used as the carrier gas at a flow rate of 1 mL/min and argon gas was used in the collision cell to generate the MRM product ion. The analysis of TMS samples was performed under the following oven temperature program; 100°C start temperature, hold for 4 minutes, followed by a 10°C min-1 oven temperature ramp to 320°C with a following final hold for 11 minutes. Approximately 520 targets were collected using the Shimadzu Smart Metabolite Database, where each target comprised a quantifier MRM along with a qualifier MRM, which covers approximately 350 endogenous metabolites and multiple stable isotopically labelled internal standards. Resultant data was processed using Shimadzu LabSolutions Insight software, where peak integrations were visually validated and manually corrected where required. |
| Ion Mode: | POSITIVE |
