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MB Sample ID: SA035193
| Local Sample ID: | BALF_12558_15uL_C |
| Subject ID: | SU000650 |
| Subject Type: | Human | Animal |
| Subject Species: | Homo sapiens;Mus musculus |
| Taxonomy ID: | 9606;10090 |
| Genotype Strain: | Animal: C57BL/6 |
| Species Group: | Mammal |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU000650 |
| Subject Type: | Human | Animal |
| Subject Species: | Homo sapiens;Mus musculus |
| Taxonomy ID: | 9606;10090 |
| Genotype Strain: | Animal: C57BL/6 |
| Species Group: | Mammal |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| BALF_12558_15uL_C | SA035193 | FL007510 | Human | Species |
| BALF_12558_15uL_C | SA035193 | FL007510 | Smoker | SmokingStatus |
| BALF_12558_15uL_C | SA035193 | FL007510 | Healthy | Phenotype |
Collection:
| Collection ID: | CO000644 |
| Collection Summary: | Mouse BALF collection was performed using a total of 1.0 mL PBS divided into three washes and stored at -80°C until analysis. Human BALF was collected in the right middle lobe and lingual by instilling two aliquots of 40 mL and one aliquot of 50 mL of sterile saline per lobe (i.e., 130 mL per lobe, total volume = 260 mL per subject), which is withdrawn by gentle manual suction and immediately placed on ice. Aliquots for metabolomics analysis were frozen at -80 °C and stored until sample preparation and MS analysis. |
| Sample Type: | BALF |
| Collection Method: | Broncheoalveolar lavage |
Treatment:
| Treatment ID: | TR000664 |
| Treatment Summary: | None |
Sample Preparation:
| Sampleprep ID: | SP000657 |
| Sampleprep Summary: | 100uL of BALF were prepared using methanol assisted protein precipitation followed by methyl-tert butyl ether (MTBE) liquid-liquid extraction. Both liquid and aqueous fractions were preserved for analysis. The lipid fraction was reconstituted in 200µL of methanol. The lipid fraction was dried down in a SpeedVac at 45°C for reconstitution in 50 µL of 95:5 water:acetonitrile. |
Combined analysis:
| Analysis ID | AN000959 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Agilent 1290 |
| Column | Agilent Zorbax RRHD SB-C18 (100 x 2.1mm,1.8um) |
| MS Type | ESI |
| MS instrument type | QTOF |
| MS instrument name | Agilent 6510 QTOF |
| Ion Mode | POSITIVE |
| Units |
Chromatography:
| Chromatography ID: | CH000684 |
| Chromatography Summary: | The BALF samples were separated using C18 reverse phase chromatography. The separations used an Agilent Zorbax Rapid Resolution HD (RRHD) SB-C18, 1.8 micron (2.1 x 100 mm) analytical column with an Agilent Zorbax SB-C18, 1.8 micron (2.1 x 5 mm) guard column and used an Agilent 1290 series high performance liquid chromatography (HPLC) pump. Injection volumes were 4µL for mouse BALF and 15µL for human BALF. Flow rate was 0.7 mL/min using the following mobile phases: mobile phase A was water with 0.1% formic acid, and mobile phase B was 60:36:4 isopropyl alcohol:acetonitrile:water with 0.1% formic acid. Gradients were 0-0.5 minutes 30-70% B, 0.5-7.42 minutes 70-100% B, 7.42-9.9 minutes 100% B, 9.9-10.0 minutes 100-30% B, 10-14.6 minutes 30% B for the positive mode injections. For negative mode the following eluant profile was used: 0-1 minutes 30-70% B, 1-7.92 minutes 70-100% B, 7.92-10.4 minutes 100% B, 10.4-10.5 minutes 100-30% B, 10.5-15.1 minutes 30% B. |
| Instrument Name: | Agilent 1290 |
| Column Name: | Agilent Zorbax RRHD SB-C18 (100 x 2.1mm,1.8um) |
| Flow Gradient: | 0-0.5 minutes 30-70% B, 0.5-7.42 minutes 70-100% B, 7.42-9.9 minutes 100% B, 9.9-10.0 minutes 100-30% B, 10-14.6 minutes 30% B |
| Flow Rate: | 0.7 mL/min |
| Solvent A: | 100% water; 0.1% formic acid |
| Solvent B: | 60% isopropanol/36% acetonitrile/4% water; 0.1% formic acid |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS000854 |
| Analysis ID: | AN000959 |
| Instrument Name: | Agilent 6510 QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| Ion Mode: | POSITIVE |
| Capillary Voltage: | 4000V |
| Fragment Voltage: | 120 |
| Desolvation Gas Flow: | 12mL/min |
| Desolvation Temperature: | 300C |
| Nebulizer: | 30psi |
| Scan Range Moverz: | 60-1600 |
| Scanning: | 2.02 spectra/second |
| Skimmer Voltage: | 60V |
