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MB Sample ID: SA033367
| Local Sample ID: | Inj053_CSH_Lung_CF70.d |
| Subject ID: | SU000627 |
| Subject Type: | Animal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Gender: | Male |
| Species Group: | Mammal |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU000627 |
| Subject Type: | Animal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Gender: | Male |
| Species Group: | Mammal |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| Inj053_CSH_Lung_CF70.d | SA033367 | FL007376 | Lung | Source |
| Inj053_CSH_Lung_CF70.d | SA033367 | FL007376 | Mouse | Species |
| Inj053_CSH_Lung_CF70.d | SA033367 | FL007376 | Control Diet | Treatment |
Collection:
| Collection ID: | CO000621 |
| Collection Summary: | C57BL/6 mice, 6-7 weeks of age upon diet assignment were fed for 150 days and weighed daily. Mice were euthanized with an overdose of pentobarbital IP and lungs were flash frozen. 6 mg of lung tissue was extracted for GC-TOFMS and HILIC-QTOFMS analysis. |
| Sample Type: | Tissue |
Treatment:
| Treatment ID: | TR000641 |
| Treatment Summary: | Male C57BL/6N mice (6-7 weeks of age) were provided ad libitum access to one of three diets for 150 days: low fat (10% kcals) control (CTRL) chow, high fat (45% kcals) with sugar (HFS) chow or very high (60% kcals) fat (VHF) chow. Body weight and food intake were measured daily. |
| Treatment Doseduration: | 150 Days |
Sample Preparation:
| Sampleprep ID: | SP000634 |
| Sampleprep Summary: | 1. Weigh 50 mg tissue sample in to a 25 ml conical polypropylene centrifuge tube. 2. Add 2.5mL extraction solvent to the tissue sample and homogenize for 45 seconds ensuring that sample resembles a powder. In between samples, clean the homogenizer in solutions of methanol, acetone, water, and the extraction solvent. 3. Centrifuge the samples at 2500 rpm. for 5 minutes. Aliquot 2 X 500μl supernatant, one for analysis and one for a backup sample. Store backup aliquot in the -20°C freezer. 4. Evaporate one 500μl aliquot of the sample in the Labconco Centrivap cold trap concentrator to complete dryness 5. The dried aliquot is then re-suspended with 500l 50% acetonitrile (degassed as given) 6. Centrifuge for 2 min at 14000 rcf using the centrifuge Eppendorf 5415. 7. Remove supernatant to a new Eppendorff tube. 8. Evaporate the supernatant to dryness in the the Labconco Centrivap cold trap concentrator. 9. Submit to derivatization. |
| Sampleprep Protocol Filename: | SOP_Extraction_of_Mammalian_Tissue_Samples.pdf |
| Sampleprep Protocol Comments: | This study combined 2 different injection volumes for certain samples when processing positive mode data. The reason for this was because in certain samples the TG's were overloaded with the 1uL injections, but everything else was at a normal peak height. Therefore samples were run at a 0.1uL injection volume and took the data from 9-12.5 minutes because that's when TG's elute. The samples that were injected at 0.1uL were used for times 9 min-12.5 min and the 1uL samples were used for times 0-8.99min. |
Combined analysis:
| Analysis ID | AN000924 | AN000925 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | Reversed phase |
| Chromatography system | Agilent 6530 | Agilent 6550 |
| Column | Waters Acquity CSH C18 (100 x 2.1mm,1.7um) | Waters Acquity CSH C18 (100 x 2.1mm,1.7um) |
| MS Type | ESI | ESI |
| MS instrument type | QTOF | QTOF |
| MS instrument name | Agilent 6530 QTOF | Agilent 6550 QTOF |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | Counts | Counts |
Chromatography:
| Chromatography ID: | CH000660 |
| Chromatography Summary: | See file below. |
| Methods Filename: | Data_Dictionary_Fiehn_laboratory_CSH_QTOF_lipidomics_05-29-2014.pdf |
| Instrument Name: | Agilent 6530 |
| Column Name: | Waters Acquity CSH C18 (100 x 2.1mm,1.7um) |
| Column Pressure: | 450-850 bar |
| Column Temperature: | 65 C |
| Flow Gradient: | 15% B to 99% B |
| Flow Rate: | 0.6 mL/min |
| Injection Temperature: | 4 C |
| Internal Standard: | See data dictionary |
| Retention Time: | See data dictionary |
| Sample Injection: | 1.67 uL |
| Solvent A: | 60% acetonitrile/40% water; 10mM formic acid; 10mM ammonium formate |
| Solvent B: | 90% isopropanol/10% acetonitrile; 10mM formic acid; 10mM ammonium formate |
| Analytical Time: | 13 min |
| Capillary Voltage: | 3500 |
| Time Program: | 15 min |
| Weak Wash Solvent Name: | Isopropanol |
| Strong Wash Solvent Name: | Isopropanol |
| Target Sample Temperature: | Autosampler temp 4 C |
| Randomization Order: | Excel |
| Chromatography Type: | Reversed phase |
| Chromatography ID: | CH000661 |
| Chromatography Summary: | See file below |
| Methods Filename: | Data_Dictionary_Fiehn_laboratory_CSH_QTOF_lipidomics_05-29-2014.pdf |
| Instrument Name: | Agilent 6550 |
| Column Name: | Waters Acquity CSH C18 (100 x 2.1mm,1.7um) |
| Column Pressure: | 450-850 bar |
| Column Temperature: | 65 C |
| Flow Gradient: | 15% B to 99% B |
| Flow Rate: | 0.6 mL/min |
| Injection Temperature: | 4 C |
| Internal Standard: | See data dictionary |
| Retention Time: | See data dictionary |
| Sample Injection: | 5.0 uL |
| Solvent A: | 60% acetonitrile/40% water; 10 mM ammonium acetate; 10mM acetic acid formate; 10mM formic acid |
| Solvent B: | 90% isopropanol/10% acetonitrile; 10mM acetic acid; 10mM ammonium acetate |
| Analytical Time: | 13 min |
| Capillary Voltage: | 3500 |
| Time Program: | 15 min |
| Weak Wash Solvent Name: | Isopropanol |
| Strong Wash Solvent Name: | Isopropanol |
| Target Sample Temperature: | Autosampler temp 4 C |
| Randomization Order: | Excel |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS000822 |
| Analysis ID: | AN000924 |
| Instrument Name: | Agilent 6530 QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| Ion Mode: | POSITIVE |
| Capillary Voltage: | 3500 |
| Collision Energy: | 25 eV |
| Collision Gas: | Nitrogen |
| Dry Gas Flow: | 8L/min |
| Dry Gas Temp: | 325 C |
| Fragment Voltage: | 120 |
| Fragmentation Method: | Auto MSMS |
| Ion Source Temperature: | 325 C |
| Ion Spray Voltage: | 1000 |
| Ionization: | Pos |
| Precursor Type: | Intact Molecule |
| Reagent Gas: | Nitrogen |
| Source Temperature: | 325 C |
| Dataformat: | .d |
| Desolvation Gas Flow: | 11 L/min |
| Desolvation Temperature: | 350 C |
| Nebulizer: | 35 psig |
| Octpole Voltage: | 750 |
| Resolution Setting: | Exteded Dyamic Range |
| Scan Range Moverz: | 60-1700 Da |
| Scanning Cycle: | 2 Hz |
| Scanning Range: | 60-1700 Da |
| Skimmer Voltage: | 65 |
| MS ID: | MS000823 |
| Analysis ID: | AN000925 |
| Instrument Name: | Agilent 6550 QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| Ion Mode: | NEGATIVE |
| Capillary Voltage: | 3500 |
| Collision Energy: | 25 eV |
| Collision Gas: | Nitrogen |
| Dry Gas Flow: | 13L/min |
| Dry Gas Temp: | 200 C |
| Fragment Voltage: | 175 |
| Fragmentation Method: | Auto MSMS |
| Ion Source Temperature: | 325 C |
| Ion Spray Voltage: | 1000 |
| Ionization: | Neg |
| Precursor Type: | Intact Molecule |
| Reagent Gas: | Nitrogen |
| Source Temperature: | 325 C |
| Dataformat: | .d |
| Desolvation Gas Flow: | 11 L/min |
| Desolvation Temperature: | 350 C |
| Nebulizer: | 35 psig |
| Octpole Voltage: | 750 |
| Resolution Setting: | Exteded Dyamic Range |
| Scan Range Moverz: | 60-1700 Da |
| Scanning Cycle: | 2 Hz |
| Scanning Range: | 60-1700 Da |
| Skimmer Voltage: | 65 |
