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MB Sample ID: SA015156
| Local Sample ID: | Inj06_Liv Met_2 |
| Subject ID: | SU000360 |
| Subject Type: | Animal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Species Group: | Mammal |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU000360 |
| Subject Type: | Animal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Species Group: | Mammal |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| Inj06_Liv Met_2 | SA015156 | FL003372 | Metformin | Treatment |
| Inj06_Liv Met_2 | SA015156 | FL003372 | Liver | Organ |
Collection:
| Collection ID: | CO000354 |
| Collection Summary: | Liver tissues were isolated and frozen down in liquid nitrogen. |
| Collection Protocol Filename: | SookWahYee_StudyDesign_5.13.14.pdf |
| Sample Type: | Liver and Kidney tissue |
Treatment:
| Treatment ID: | TR000374 |
| Treatment Summary: | 5 mice were treated with biguanide (metformin) and 5 mice were treated with saline. Liver tissues were obtained 7 days after treatment. |
| Treatment Protocol Filename: | SookWahYee_StudyDesign_5.13.14.pdf |
Sample Preparation:
| Sampleprep ID: | SP000367 |
| Sampleprep Summary: | 1. Weigh 4mg tissue sample in to a 2mL Eppendorf tube. 2. Add 1mL 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 in the order listed. 3. Vortex samples for 10 seconds, then 5 minutes on 4°C shaker. 4. Centrifuge the samples for 2 minutes at 14,000 rcf. Aliquot 500μL supernatant for analysis, and 500μL for a backup. Store backup aliquots in the -20°C freezer. 5. Evaporate one 500μl analysis aliquot in the Labconco Centrivap cold trap concentrator to complete dryness (typically overnight). 6. The dried aliquot is then re-suspended with 500l 50% acetonitrile (degassed as given) 7. Centrifuge for 2 minutes at 14,000 rcf using the centrifuge Eppendorf 5415. 8. Remove supernatant to a new Eppendorf tube. 9. Evaporate the supernatant to dryness in the the Labconco Centrivap cold trap concentrator. 10. Submit to derivatization. |
| Sampleprep Protocol Filename: | SOP Extraction of Liver Tissue Samples.pdf |
Combined analysis:
| Analysis ID | AN000552 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | Unspecified |
| Column | Waters Acquity BEH Amide (150 x 2.1mm,1.7um) |
| MS Type | ESI |
| MS instrument type | QTOF |
| MS instrument name | Agilent 6530 QTOF |
| Ion Mode | POSITIVE |
| Units | Peak height |
Chromatography:
| Chromatography ID: | CH000389 |
| Methods Filename: | Data_Dictionary_Fiehn_laboratory_HILIC_QTOF_MS.pdf |
| Instrument Name: | Unspecified |
| Column Name: | Waters Acquity BEH Amide (150 x 2.1mm,1.7um) |
| Column Pressure: | 200-700 bar |
| Column Temperature: | 45 C |
| Flow Gradient: | 100% B to 30% B |
| Flow Rate: | 0.4 mL/min |
| Injection Temperature: | 4 C |
| Internal Standard: | See data dictionary |
| Sample Injection: | 3 μL |
| Analytical Time: | 14 min |
| Capillary Voltage: | 4500 |
| Time Program: | 16.75 min |
| Weak Wash Solvent Name: | 1:1 ACN:H2O |
| Weak Wash Volume: | 5 seconds |
| Target Sample Temperature: | Autosampler temp 4 C |
| Randomization Order: | Excel |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS000488 |
| Analysis ID: | AN000552 |
| Instrument Name: | Agilent 6530 QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| Ion Mode: | POSITIVE |
| Capillary Voltage: | 3500 |
| Collision Gas: | Nitrogen |
| Dry Gas Flow: | 8 l/min |
| Dry Gas Temp: | 325 |
| Fragment Voltage: | 120 |
| Fragmentation Method: | Auto MS/MS |
| Ion Source Temperature: | 325 |
| 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: | Extended Dynamic Range |
| Scan Range Moverz: | 60-1700 Da |
| Scanning Cycle: | 2 Hz |
| Scanning Range: | 60-1700 Da |
| Skimmer Voltage: | 65 |
