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MB Sample ID: SA316702
Local Sample ID: | 20120522-zhxl-B3-L-1 |
Subject ID: | SU003031 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU003031 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
20120522-zhxl-B3-L-1 | SA316702 | FL037717 | infection | Treatment |
Collection:
Collection ID: | CO003024 |
Collection Summary: | Samples were counted, washed with cold PBS and then flash-frozen in liquid N2 |
Sample Type: | Spleen |
Treatment:
Treatment ID: | TR003040 |
Treatment Summary: | Spleen tissues were weighed and homogenized with the first solvent (the mixture of chloroform, methanol and water (1:2:1, v/v/v)) for 30 s at 4 0C and then centrifuged at 12,000 rpm for 10 min at 4 0C. The supernatant was collected and deposit was re-homogenized with the second solvent (methanol alone) before a second centrifugation. The two supernatants were mixed, and aliquot of sample was transferred to a GC sampling vial containing 5 μL 0.1 mg/mL ribitol (Sigma) as an analytical internal standard and then dried in a vacuum centrifuge concentrator before the subsequent derivatization. Two technical replicates were prepared for each sample. |
Sample Preparation:
Sampleprep ID: | SP003037 |
Sampleprep Summary: | Spleen tissues were weighed and homogenized with the first solvent (the mixture of chloroform, methanol and water (1:2:1, v/v/v)) for 30 s at 4 0C and then centrifuged at 12,000 rpm for 10 min at 4 0C. The supernatant was collected and deposit was re-homogenized with the second solvent (methanol alone) before a second centrifugation. The two supernatants were mixed, and aliquot of sample was transferred to a GC sampling vial containing 5 μL 0.1 mg/mL ribitol (Sigma) as an analytical internal standard and then dried in a vacuum centrifuge concentrator before the subsequent derivatization. Two technical replicates were prepared for each sample. |
Combined analysis:
Analysis ID | AN004788 |
---|---|
Analysis type | MS |
Chromatography type | GC |
Chromatography system | Thermo Scientific Trace GC Ultra with DSQ II GC/MS |
Column | Agilent DB5-MS (30m x 0.25mm, 0.25um) |
MS Type | EI |
MS instrument type | Triple quadrupole |
MS instrument name | Thermo Scientific Trace GC Ultra with DSQ II GC/MS |
Ion Mode | POSITIVE |
Units | Peak area |
Chromatography:
Chromatography ID: | CH003619 |
Chromatography Summary: | Low pH polar (LC/MS Pos early) |
Instrument Name: | Thermo Scientific Trace GC Ultra with DSQ II GC/MS |
Column Name: | Agilent DB5-MS (30m x 0.25mm, 0.25um) |
Column Temperature: | 270 °C |
Flow Gradient: | none |
Flow Rate: | 1.0 mL/min |
Solvent A: | none |
Solvent B: | none |
Chromatography Type: | GC |
MS:
MS ID: | MS004534 |
Analysis ID: | AN004788 |
Instrument Name: | Thermo Scientific Trace GC Ultra with DSQ II GC/MS |
Instrument Type: | Triple quadrupole |
MS Type: | EI |
MS Comments: | samples was derivatized and then used to firstly protect carbonyl moieties through methoximation, through a 90 min 37 ℃ reaction with 40 μL of 20 mg/mL methoxyamine hydrochloride (Sigma-Aldrich) in pyridine, followed by derivatization of acidic protons through a 30 min 37 0C reaction with the addition of 80 μL N-methyl-N-trimethylsilyltrifluoroace-tamide (MSTFA, Sigma-Aldrich). The derivatized sample of 1 μL was injected into a 30m × 250 μm i.d. × 0.25 μm DBS-MS column using splitless injection and analysis was carried out by Trace DSQ II (Thermo Scientific). The initial temperature of the GC oven was held at 85 0C for 5 min followed by an increase to 330 0C at a rate of 15 0C min-1 then held for 5 min. Helium was used as carrier gas and flow was kept constant at 1 mL min-1. The MS was operated in a range of 50-600 m/z. |
Ion Mode: | POSITIVE |