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MB Sample ID: SA351623
Local Sample ID: | 2738 |
Subject ID: | SU003330 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU003330 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
2738 | SA351623 | FL041042 | cardiac blood | Sample source |
2738 | SA351623 | FL041042 | WT | Genotype |
Collection:
Collection ID: | CO003323 |
Collection Summary: | Fetuses of mice embryo at E17.5 developmental stage were isolated from pregnant females and decapitated on ice. ~50ul blood was collected from each fetus using a pipette and transferred into tubes containing 2ul 0.5M EDTA. collected by centrifugation and frozen at -80C before metabolic analysis. Cardiac blood and serum samples were also collected from the mothers of each litter using the same approach. |
Sample Type: | fetus blood or cardiac blood |
Treatment:
Treatment ID: | TR003339 |
Treatment Summary: | N/A |
Sample Preparation:
Sampleprep ID: | SP003337 |
Sampleprep Summary: | A monophasic extraction protocol was used to extract the metabolites from the serum. To 20 µL of serum, 140 µL of chilled 6:1 MeOH/Milli-Q water containing 0.8 nmol of 13C515N valine and 0.8 nmol of 13C6 sorbitol was added. Each sample was vortexed and then incubated at 4°C for 10 min with continuous agitation (950 rpm) using an Eppendorf Thermomixer C. The samples were centrifuged at 4°C for 10 min at 12700 rpm using an Eppendorf centrifuge 5430 R. The supernatant was transferred into a fresh 1.5 mL Eppendorf tube and the cell debris was discarded. A 16 µL aliquot of each sample was pooled to create the pooled biological quality control (PBQC). Sixteen µL of each study sample and the PBQC were transferred into HPLC inserts and evaporated at 30 °C to complete dryness, using a CHRIST RVC 2-33 CD plus speed vacuum. To limit the amount of moisture present in the insert, 20 µL 100% methanol (LCMS grade) was added to each insert and evaporated using a speed vacuum. |
Combined analysis:
Analysis ID | AN005266 |
---|---|
Analysis type | MS |
Chromatography type | GC |
Chromatography system | Shimadzu GC-2030 |
Column | Agilent DB5-MS (30m x 0.25mm, 0.25um) |
MS Type | EI |
MS instrument type | Triple quadrupole |
MS instrument name | Shimadzu TQ8050NX |
Ion Mode | POSITIVE |
Units | Relative abundance |
Chromatography:
Chromatography ID: | CH003985 |
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)with an electron ionisation source(-70eV). 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 the derivatised samples was performed under the following oven temperature program; 100°C start temperature, hold for 4 minutes, followed by a 10°C/min oven temperature ramp to 320°C with a following final hold for 11 minutes. |
Instrument Name: | Shimadzu GC-2030 |
Column Name: | Agilent DB5-MS (30m x 0.25mm, 0.25um) |
Column Temperature: | 100 - 320 |
Flow Gradient: | N/A |
Flow Rate: | 1 mL/min |
Solvent A: | N/A |
Solvent B: | N/A |
Chromatography Type: | GC |
MS:
MS ID: | MS004997 |
Analysis ID: | AN005266 |
Instrument Name: | Shimadzu TQ8050NX |
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)with an electron ionisation source(-70eV). 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 the derivatised samples was performed under the following oven temperature program; 100°C start temperature, hold for 4 minutes, followed by a 10°C/min oven temperature ramp to 320°C with a following final hold for 11 minutes. |
Ion Mode: | POSITIVE |