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MB Sample ID: SA020204

Local Sample ID:140404ajlsa28_1
Subject ID:SU000433
Subject Type:Human
Subject Species:Homo sapiens
Taxonomy ID:9606
Species Group:Human

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Subject:

Subject ID:SU000433
Subject Type:Human
Subject Species:Homo sapiens
Taxonomy ID:9606
Species Group:Human

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
140404ajlsa28_1SA020204FL006490fresh liverTreatment
140404ajlsa28_1SA020204FL0064902 HrCollection Time

Collection:

Collection ID:CO000427
Collection Summary:Liver biopsies were frozen in liquid nitrogen and then stored at -80. They were then pulverized and the crushed tissue from one biopsy was divided into three 0.6mL tubes, containing ±25-70mg. These are triplicates from each biopsy. Tissue was kept at cryo temperatures throughout.
Collection Protocol Filename:StudyDesign_BoteBruinsma_02.11.14.pdf
Sample Type:Tissue

Treatment:

Treatment ID:TR000447
Treatment Summary:9 livers were run for metabolites at 4 timepoints (0,1,2,3), in triplicate. Warm ischemic livers (27, 8, 34) and non-warm ischemic livers (25, 28, 35) and fresh liver samples (R). Liver 30 was a steatotic liver, for which there were 4 timepoints.
Treatment Protocol Filename:StudyDesign_BoteBruinsma_02.11.14.pdf

Sample Preparation:

Sampleprep ID:SP000440
Sampleprep Summary:1. Weigh 4 mg tissue sample ( muscle 20 mg) in to a 2.0 ml eppendorf tube. Add 1.0 mL extraction solvent to the tissue sample and using GenoGrinder homogenize samples for 45 seconds ensuring that sample resembles a powder. 2. Centrifuge the samples at 2500 rpm. for 5 minutes. Aliquot 2 X 500µl supernatant, one for analysis and one for a backup sample. Store backup aliquot in the -20°C freezer. 3. Evaporate one 500µl aliquot of the sample in the Labconco Centrivap cold trap concentrator to complete dryness 4. The dried aliquot is then re-suspended with 500l 50% acetonitrile (degassed as given) (only for liver and brain samples). 5. Centrifuge for 2 min at 14000 rcf using the centrifuge Eppendorf 5415. 6. Remove supernatant to a new Eppendorff tube. 7. Evaporate the supernatant to dryness in the the Labconco Centrivap cold trap concentrator. 8. Submit to derivatization.
Sampleprep Protocol Filename:SOP_Extraction_of_Liver_Tissue_Samples.pdf

Combined analysis:

Analysis ID AN000652
Analysis type MS
Chromatography type GC
Chromatography system Leco Pegasus III GC
Column Restek Corporation Rtx-5Sil MS
MS Type EI
MS instrument type GC-TOF
MS instrument name Leco Pegasus III GC TOF
Ion Mode POSITIVE
Units counts

Chromatography:

Chromatography ID:CH000470
Methods Filename:Data_Dictionary_Fiehn_laboratory_GCTOF_MS_primary_metabolism_10-15-2013_general.pdf
Instrument Name:Leco Pegasus III GC
Column Name:Restek Corporation Rtx-5Sil MS
Column Pressure:7.7 PSI
Column Temperature:50-330C
Flow Rate:1 ml/min
Injection Temperature:50 C ramped to 250 C by 12 C/s
Sample Injection:0.5 uL
Oven Temperature:50°C for 1 min, then ramped at 20°C/min to 330°C, held constant for 5 min
Transferline Temperature:230C
Washing Buffer:Ethyl Acetate
Sample Loop Size:30 m length x 0.25 mm internal diameter
Randomization Order:Excel generated
Chromatography Type:GC

MS:

MS ID:MS000578
Analysis ID:AN000652
Instrument Name:Leco Pegasus III GC TOF
Instrument Type:GC-TOF
MS Type:EI
Ion Mode:POSITIVE
Ion Source Temperature:250 C
Ionization Energy:70 eV
Mass Accuracy:Nominal
Source Temperature:250 C
Scan Range Moverz:85-500 Da
Scanning Cycle:17 Hz
Scanning Range:85-500 Da
Skimmer Voltage:1850 V
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