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MB Sample ID: SA035143
Local Sample ID: | MBAL_MET106 |
Subject ID: | SU000648 |
Subject Type: | Human | Animal |
Subject Species: | Homo sapiens;Mus musculus |
Taxonomy ID: | 9606;10090 |
Genotype Strain: | Animal: C57BL/6 |
Species Group: | Mammal |
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Subject:
Subject ID: | SU000648 |
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 |
---|---|---|---|---|
MBAL_MET106 | SA035143 | FL007503 | Mouse | Species |
MBAL_MET106 | SA035143 | FL007503 | Smoker | SmokingStatus |
MBAL_MET106 | SA035143 | FL007503 | Emphysema | Phenotype |
Collection:
Collection ID: | CO000642 |
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one aliquot of 50 mL of sterile saline per lobe (i.e., 130 mL per lobe, total volume = 260 mL per subject), which is withdrawn by gentle manual suction and immediately placed on ice.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: | TR000662 |
Treatment Summary: | None |
Sample Preparation:
Sampleprep ID: | SP000655 |
Sampleprep Summary: | 200uL of BALF was prepared using methanol assisted protein precipitation followed by methyl-tert butyl ether (MTBE) liquid-liquid extraction. The aqueous fraction was dried down in a SpeedVac at 45°C, then reconstituted in 50 µL of 95:5 water:acetonitrile. |
Combined analysis:
Analysis ID | AN000957 |
---|---|
Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Agilent 1200 |
Column | Agilent Zorbax Narrow Bore RRHT RRHD SB-AQ (100 x 2.1mm,1.8um) |
MS Type | ESI |
MS instrument type | QTOF |
MS instrument name | Agilent 6520 QTOF |
Ion Mode | POSITIVE |
Units | peak intensity |
Chromatography:
Chromatography ID: | CH000682 |
Chromatography Summary: | Analysis of the aqueous fraction was by reversed-phase chromatography using an Agilent 1200 series pump with an Agilent Zorbax Narrow Bore RRHT SB-AQ (1.8 micron, 2.1 x 100 mm, 80Å) analytical column and an Agilent Zorbax SB-AQ (5 micron, 2.1 x 12.5 mm) guard column. Sample injection volumes were 10 µL. A flow rate of 0.3ml/min was used with the following mobile phases: mobile phase A was water with 0.1% formic acid, and mobile phase B was 90:10 acetonitrile:water with 0.1% formic acid. Gradient elution was as follows: 0-3 minutes 2% B, 3-5 minutes 2-40% B, 5-20 minutes 40-100% B, 20-30 minutes 100% B, 30-30.01 100-2% B, 30.01-40 minutes 2% B. |
Instrument Name: | Agilent 1200 |
Column Name: | Agilent Zorbax Narrow Bore RRHT RRHD SB-AQ (100 x 2.1mm,1.8um) |
Flow Gradient: | 0-3 minutes 2% B, 3-5 minutes 2-40% B, 5-20 minutes 40-100% B, 20-30 minutes 100% B, 30-30.01 100-2% B, 30.01-40 minutes 2% B |
Flow Rate: | 0.3ml/min |
Solvent A: | 100% water; 0.1% formic acid |
Solvent B: | 90% acetonitrile/10% water; 0.1% formic acid |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS000852 |
Analysis ID: | AN000957 |
Instrument Name: | Agilent 6520 QTOF |
Instrument Type: | QTOF |
MS Type: | ESI |
Ion Mode: | POSITIVE |
Capillary Voltage: | 4000V |
Desolvation Gas Flow: | 10L/min |
Desolvation Temperature: | 300C |