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MB Sample ID: SA077902

Local Sample ID:IC23_44
Subject ID:SU001183
Subject Type:Human
Subject Species:Homo sapiens
Taxonomy ID:9606

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Subject:

Subject ID:SU001183
Subject Type:Human
Subject Species:Homo sapiens
Taxonomy ID:9606

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
IC23_44SA077902FL011852Interstitial cystitisCondition

Collection:

Collection ID:CO001177
Collection Summary:Urine samples were collected from 43 patients with interstial cystitis. Urine was frozen to -80°C and shipped to the WCMC on dry ice, where samples were stored at -80°C until analysis.
Sample Type:Urine
Storage Conditions:-80℃

Treatment:

Treatment ID:TR001198
Treatment Summary:Patients were not receiving any treatment at the time the urine was collected.

Sample Preparation:

Sampleprep ID:SP001191
Sampleprep Summary:Samples were removed from the -80°C freezer and allowed to thaw on ice. 225 µL of MeOH with lipid internal standards was added to each sample. Samples were vortexed for 10 seconds. 750 µL of MTBE plus quality controls was added to each sample. Samples were vortexed for 10 seconds. 188 µL of LC-MS grade water was added to each sample. Samples were shaken at 4°C on an orbital mixing plate for 6 minutes. Samples were centrifuged for 2 minutes at 14,000 rcf. 2 aliquots of 350 µL were removed from the top, organic phase. One aliquot was stored at -20°C for backup and the other was dried using a Labconco centrivap. 2 aliquots of 125 µL of the bottom polar phase were removed. One aliquot was stored at -20°C for backup and the other was dried using a Labconco centrivap.
Sample Resuspension:For HILIC analysis, samples were resuspened in 100 µL of 9:1 methanol:toluene with deuterated compounds as an internal standard. Samples were vortexed for 10 seconds and then sonicated in room temperature water for 5 minutes. Samples were run in a centrifuge for 2 minutes at 16,100 rcf. 90 µL of the supernatant was removed and submitted to HILIC analysis.

Combined analysis:

Analysis ID AN001847
Analysis type MS
Chromatography type HILIC
Chromatography system Thermo Vanquish
Column Waters Acquity BEH Amide (150 x 2.1mm,1.7um)
MS Type ESI
MS instrument type Orbitrap
MS instrument name Thermo Q Exactive HF hybrid Orbitrap
Ion Mode POSITIVE
Units Peak Height

Chromatography:

Chromatography ID:CH001324
Instrument Name:Thermo Vanquish
Column Name:Waters Acquity BEH Amide (150 x 2.1mm,1.7um)
Column Pressure:200 bar - 600 bar
Column Temperature:45°C
Flow Gradient:0 min 100% B; 0−2 min 100% B; 2−7.7 min 70% B; 7.7−9.5 min 40% B; 9.5−10.25 min 30% B; 10.25−12.75 min 100% B; 12.75−17 min 100% B
Flow Rate:400 µL/min
Sample Injection:1 µL
Solvent A:100% water; 0.125% formic acid; 10 mM ammonium formate
Solvent B:95% acetonitrile/5% water; 0.125% formic acid; 10 mM ammonium formate
Chromatography Type:HILIC

MS:

MS ID:MS001708
Analysis ID:AN001847
Instrument Name:Thermo Q Exactive HF hybrid Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:Data was processing using X-Calibur Qual Browser, MS-Dial, and MS-Flo following the data processing procedure outlined by the WCMC.
Ion Mode:POSITIVE
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