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MB Sample ID: SA015154
Local Sample ID: | Inj09_Kid Met_3 |
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 |
---|---|---|---|---|
Inj09_Kid Met_3 | SA015154 | FL003371 | Metformin | Treatment |
Inj09_Kid Met_3 | SA015154 | FL003371 | Kidney | 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 |