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MB Sample ID: SA077856
| Local Sample ID: | IC43_64 |
| Subject ID: | SU001182 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001182 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| IC43_64 | SA077856 | FL011851 | Interstitial cystitis | Condition |
Collection:
| Collection ID: | CO001176 |
| 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: | TR001197 |
| Treatment Summary: | Patients were not receiving any treatment at the time the urine was collected. |
Sample Preparation:
| Sampleprep ID: | SP001190 |
| 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 lipidomics analysis, samples were resuspened in 110 µL of 9:1 methanol:toluene with 50 nM of CUDA 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. 100 µL of the supernatant was removed and submited to lipidomics analysis. |
Combined analysis:
| Analysis ID | AN001846 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Thermo Vanquish |
| Column | Waters Acquity CSH C18 (100 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: | CH001323 |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Waters Acquity CSH C18 (100 x 2.1mm,1.7um) |
| Column Pressure: | 600 bar - 900 bar |
| Column Temperature: | 65°C |
| Flow Gradient: | 0 min 15% B; 0−2 min 30% B; 2−2.5 min 48% B; 2.5−11 min 82% B; 11−11.5 min 99% B; 11.5−12 min 99% B; 12−12.1 min 15% B; 12.1−15 min 15% B. |
| Flow Rate: | 600 µL/min |
| Internal Standard: | 50 nM CUDA |
| Sample Injection: | 1 µL |
| Solvent A: | 40% water/60% acetonitrile; 0.1% formic acid; 10 mM ammonium formate |
| Solvent B: | 90% isopropanol/10% acetonitrile; 0.1% formic acid; 10 mM ammonium formate |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS001707 |
| Analysis ID: | AN001846 |
| 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 |
