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MB Sample ID: SA212877

Local Sample ID:batch1_MT_20210726_099
Subject ID:SU002319
Subject Type:Human
Subject Species:Homo sapiens
Taxonomy ID:9606

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Subject:

Subject ID:SU002319
Subject Type:Human
Subject Species:Homo sapiens
Taxonomy ID:9606

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
batch1_MT_20210726_099SA212877FL025967PooledLiLabSampleType

Collection:

Collection ID:CO002312
Collection Summary:Two different deidentified human plasma pooled samples were used for this study: a) commercial Qstd plasma pool ; b) biosamples (from 1R01AI149746 (NIH-NIAID) samples) plasma pool
Sample Type:Blood (plasma)
Storage Conditions:-80℃

Treatment:

Treatment ID:TR002331
Treatment Summary:No treatment

Sample Preparation:

Sampleprep ID:SP002325
Sampleprep Summary:The plasma samples were thawed in ice. Metabolites extraction were carried out by protein precipitation using extraction solvent, acetonitrile:methanol (8:1, v/v) containing 0.1% formic acid and isotope labelled Trimethyl-13C3]-caffeine, [13C5]-L-glutamic acid, [15N2]-Uracil, [15N,13C5]-L-methionine, [13C6]-D-glucose and [15N]-L-tyrosin as spike-in controls. 30 μl of plasma was taken and 60 μl of extraction solvent was added. All samples were vortexed and incubated with shaking at 1000 rpm for 10 min at 4°C followed by centrifugation at 4°C for 15 min at 15,000 rpm. The supernatant was transferred into mass spec vials and 2 μl injected into UHPLC-MS.
Extraction Method:Protein precipitation
Extract Storage:-80℃

Combined analysis:

Analysis ID AN003643
Analysis type MS
Chromatography type HILIC
Chromatography system Thermo Scientific Vanquish Horizon UHPLC system
Column HILIC column, AccucoreTM-150-Amide (50 x 2.1mm,2.6um)
MS Type ESI
MS instrument type Orbitrap
MS instrument name Thermo Orbitrap ID-X Tribrid
Ion Mode POSITIVE
Units m/z values

Chromatography:

Chromatography ID:CH002697
Chromatography Summary:Mobile phase: 10 mM ammonium acetate in 95% acetonitrile containing 0.1% acetic acid as mobile phase A and 10 mM ammonium acetate in 50% acetonitrile containing 0.1% acetic acid as mobile phase B were used for HILIC method. Mobile phase gradient: For HILIC acquisition, following gradient was applied at a flow rate of 0.55 ml/min: 0-0.1 min: 0% B, 0.10-5.0 min: 98% B, 5.00-5.50 min: 0% B and 4.5 min for cleaning and equilibration of column.For RP column, following gradient was applied at a flow rate of 0.4 ml/min: 0-0.1 min: 0% B, 0.10-1.9 min: 60% B, 1.9-5.0 min: 98% B, 5.00-5.10 min: 0% B and 4.9 min cleaning and column equilibration.
Instrument Name:Thermo Scientific Vanquish Horizon UHPLC system
Column Name:HILIC column, AccucoreTM-150-Amide (50 x 2.1mm,2.6um)
Column Temperature:45
Flow Gradient:0-0.1 min: 0% B, 0.10-5.0 min: 98% B, 5.00-5.50 min: 0% B and 4.5 min for cleaning and equilibration of column.
Flow Rate:0.55 ml/min
Solvent A:95% acetonitrile/5% water; 0.1% acetic acid; 10 mM ammonium acetate
Solvent B:50% acetonitrile/50% water; 0.1% acetic acid; 10 mM ammonium acetate
Chromatography Type:HILIC

MS:

MS ID:MS003394
Analysis ID:AN003643
Instrument Name:Thermo Orbitrap ID-X Tribrid
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:None
Ion Mode:POSITIVE
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