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MB Sample ID: SA052717
Local Sample ID: | M35 |
Subject ID: | SU000933 |
Subject Type: | Animal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Species Group: | Mammals |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU000933 |
Subject Type: | Animal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Species Group: | Mammals |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
M35 | SA052717 | FL009996 | PBS Ctl | Treatment |
Collection:
Collection ID: | CO000927 |
Collection Summary: | Skeletal muscle tissue was harvested and immediately flash frozen in a cryo tube in iquid nitrogen. Skeletal muscle tissue was harvested and immediately flash frozen in a cryo tube in iquid nitrogen. |
Sample Type: | Muscle |
Treatment:
Treatment ID: | TR000947 |
Treatment Summary: | Fraction of muscle tissue weighed (~25-50 mg wet weight), then the finely cut up tissue quickly added to fresh pre-made buffer (50 % acetyl-nitrile, 50 % water, 0.3% formic acid) at a standard concentration of 25 mg/475 mcl buffer then fully homogenized on ice for ~20 s and placed on dry ice/stored at - 80C. Fraction of muscle tissue weighed (~25-50 mg wet weight), then the finely cut up tissue quickly added to fresh pre-made buffer (50 % acetyl-nitrile, 50 % water, 0.3% formic acid) at a standard concentration of 25 mg/475 mcl buffer then fully homogenized on ice for ~20 s and placed on dry ice/stored at - 80C. |
Sample Preparation:
Sampleprep ID: | SP000940 |
Sampleprep Summary: | Fraction of muscle tissue weighed (~25-50 mg wet weight), then the finely cut up tissue quickly added to fresh pre-made buffer (50 % acetyl-nitrile, 50 % water, 0.3% formic acid) at a standard concentration of 25 mg/475 mcl buffer then fully homogenized on ice for ~20 s and placed on dry ice/stored at - 80C. Fraction of muscle tissue weighed (~25-50 mg wet weight), then the finely cut up tissue quickly added to fresh pre-made buffer (50 % acetyl-nitrile, 50 % water, 0.3% formic acid) at a standard concentration of 25 mg/475 mcl buffer then fully homogenized on ice for ~20 s and placed on dry ice/stored at - 80C. The samples were crash deprotonized by methanol precipitation and spiked with D27-deuterated myristic acid (D27-C14:0) as an internal standard for retention-time locking and dried. The trimethylsilyl-D27-C14:0 standard retention time (RT) was set at 16.727 min. Reactive carbonyls were stabilized at 50C with methoxyamine hydrochloride in dry pyridine. Metabolites were made volatile with TMS groups using N-methyl-N-(trimethylsilyl) trifluoroacetamide or MSTFA with catalytic trimethylchlorosilane at 50C. The samples were crash deprotonized by methanol precipitation and spiked with D27-deuterated myristic acid (D27-C14:0) as an internal standard for retention-time locking and dried. The trimethylsilyl-D27-C14:0 standard retention time (RT) was set at 16.727 min. Reactive carbonyls were stabilized at 50C with methoxyamine hydrochloride in dry pyridine. Metabolites were made volatile with TMS groups using N-methyl-N-(trimethylsilyl) trifluoroacetamide or MSTFA with catalytic trimethylchlorosilane at 50C. |
Combined analysis:
Analysis ID | AN001458 |
---|---|
Analysis type | MS |
Chromatography type | GC |
Chromatography system | Agilent 6890N |
Column | Agilent DB5-MS (15m x 0.25mm,0.25um) |
MS Type | EI |
MS instrument type | Single quadrupole |
MS instrument name | Agilent 5975 |
Ion Mode | POSITIVE |
Units | Peak values (Log transformed) |
Chromatography:
Chromatography ID: | CH001025 |
Chromatography Summary: | GC/MS methods follow previous studies using a 6890 N GC connected to a 5975 Inert single quadrupole MS (Agilent Technologies, Santa Clara, CA) (Bonikos et al. 1975; Fiehn 2008; Kind et al. 2009). The two wall-coated, open-tubular GC columns connected in series are both from J&W/Agilent, DB5-MS, 15 meters in length, 0.25 mm in diameter, with an 0.25-l m luminal film. Positive ions generated with conventional electron-ionization at 70 eV are scanned broadly from 600 to 50 m/z in the detector throughout the 45 min cycle time. |
Instrument Name: | Agilent 6890N |
Column Name: | Agilent DB5-MS (15m x 0.25mm,0.25um) |
Chromatography Type: | GC |
MS:
MS ID: | MS001346 |
Analysis ID: | AN001458 |
Instrument Name: | Agilent 5975 |
Instrument Type: | Single quadrupole |
MS Type: | EI |
Ion Mode: | POSITIVE |