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MB Sample ID: SA076272

Local Sample ID:HC115_016
Subject ID:SU001164
Subject Type:Human
Subject Species:Homo sapiens
Taxonomy ID:9606

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Subject:

Subject ID:SU001164
Subject Type:Human
Subject Species:Homo sapiens
Taxonomy ID:9606

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
HC115_016SA076272FL011692ControlTreatment Group

Collection:

Collection ID:CO001158
Collection Summary:Plasma samples from patients with WD and healthy subjects were obtained from the Institute of Neurology and Psychiatry in Warsaw. Subjects fasted for 8 hours prior to sampling. Plasma samples were de-identified, shipped to the University of California, Davis and stored at -80°C until further analysis.
Sample Type:Blood (plasma)
Storage Conditions:-80℃

Treatment:

Treatment ID:TR001178
Treatment Summary:NA

Sample Preparation:

Sampleprep ID:SP001171
Sampleprep Summary:All samples were removed from -80°C and allowed to thaw on ice. Of the 100-250 uL of plasma shipped to the facility for analysis, 30 µL of plasma was pipetted into a new 1.5 mL Eppendorf tube. 1 mL of 3:3:2 acetonitrile:isopropanol:water was added to each sample. Samples were vortexed for 10 seconds using a MiniVortexer. Samples were then shaken on an Orbital Mixing Chilling/Heating Plate for 5 minutes at 4°C. Samples were centrifuged for 2 minutes at 14,000 rcf used the centrifuge Eppendorf 5415 D. Two 475 µL aliquots were removed from the supernatant. Both samples were dried to complete using Labconco Centrivap cold trap. 500 µL of 50:50 acetonitrile was added to each aliquot of all samples. Samples were vortexed for 10 seconds using the MiniVortexer. Samples were centrifuged for 2 minutes at 14,000 rcf using the centrifuge Eppendorf 5415 D. 475 µL of supernatant was removed and evaporated to dryness using the Labconco Centrivap cold trap. One sample is submitted to derivitization, the other sample is saved for LC-MS analysis.
Sample Derivatization:To the dried down samples, 10 µL of a prepared methoxyamine solution was added to each sample. Samples were shaken on an Orbital Mixing Chilling/Heating Plate at 30°C for 1.5 hours. 91 µL of a N-methyl-N-(trimethylsilyl)-trifluoroacetamide (MSTFA) and fatty-acid methyl-esters (FAMES) solution was added to each sample. Samples were shaken on an Orbital Mixing Chilling/Heating Plate at 37°C for 30 minutes. Contents were transferred to individual glass vials with micro-inserts and capped immediately. Submitted to GC-MS analysis.

Combined analysis:

Analysis ID AN001817
Analysis type MS
Chromatography type GC
Chromatography system Agilent 6890N
Column Restek Rtx-5Sil (30m x 0.25mm,0.25um)
MS Type EI
MS instrument type GC-TOF
MS instrument name Leco Pegasus III GC TOF
Ion Mode POSITIVE
Units Peak Height

Chromatography:

Chromatography ID:CH001283
Instrument Name:Agilent 6890N
Column Name:Restek Rtx-5Sil (30m x 0.25mm,0.25um)
Chromatography Type:GC

MS:

MS ID:MS001676
Analysis ID:AN001817
Instrument Name:Leco Pegasus III GC TOF
Instrument Type:GC-TOF
MS Type:EI
Ion Mode:POSITIVE
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