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MB Sample ID: SA289477
Local Sample ID: | RP_experiment_set_2_8 |
Subject ID: | SU002859 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU002859 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
RP_experiment_set_2_8 | SA289477 | FL035711 | mouse liver | Input_metabolites |
Collection:
Collection ID: | CO002852 |
Collection Summary: | Mouse metabolites were collected from the liver of female mice using methanol extraction. After euthanizing a mouse, the liver was immediately frozen in liquid nitrogen. We then used cold 80% methanol to extract metabolites. First, 1 ml of 80% methanol was added to the liver and incubated for 10 min at -20oC. Glass beads were added to the liver and then the liver was lysed by bead-beating for 45 s using a Tissuelyser cell disrupter (Qiagen). The lysate was incubated for 10 min at -20oC and centrifuged (13200 rpm, 5 min) to separate metabolites from macromolecules. The supernatant was collected and 200 µl of 80% methanol was added to the pellet. The incubation, shaking and centrifugation steps were repeated twice to extract more metabolites from the pellet. The three supernatants were combined and centrifuged (14000 rpm, 10 min) to separate any remaining macromolecules from the metabolites. The combined supernatants were dried using a SpeedVac Concentrator (Savant, SPD131DDA) at 25oC and the dried metabolite samples were stored at -80oC. |
Sample Type: | Liver |
Collection Method: | 80% methanol |
Storage Conditions: | -80℃ |
Treatment:
Treatment ID: | TR002868 |
Treatment Summary: | Metabolites were extracted from mouse livers as discussed in collection. No special treatment was performed on the mice. |
Sample Preparation:
Sampleprep ID: | SP002865 |
Sampleprep Summary: | Dried-down extracts were reconstituted in 150 µl 70% acetonitrile, at a relative protein concentration of ~ 2 µg/µl. These were were injected (4 µl) for LC/MS-based targeted metabolite profiling. |
Processing Storage Conditions: | -80℃ |
Extract Storage: | -80℃ |
Combined analysis:
Analysis ID | AN004465 | AN004466 |
---|---|---|
Analysis type | MS | MS |
Chromatography type | Reversed phase | Reversed phase |
Chromatography system | Agilent Model 1290 Infinity II liquid chromatography system | Agilent Model 1290 Infinity II liquid chromatography system |
Column | Cogent Diamond Hydride (150 × 2.1 mm, 4um) | Cogent Diamond Hydride (150 × 2.1 mm, 4um) |
MS Type | ESI | ESI |
MS instrument type | QTOF | QTOF |
MS instrument name | Bruker Impact HD | Bruker Impact HD |
Ion Mode | POSITIVE | NEGATIVE |
Units | Ion abundance (max peak height) | Ion abundance (max peak height) |
Chromatography:
Chromatography ID: | CH003351 |
Chromatography Summary: | Chromatography of metabolites utilized reversed phase chromatography on a Agilent ZORBAX Eclipse Plus C18, 100 × 2.1 mm, 1.8 μm. Mobile phases consisted of (A) 10 mM ammonium formate with 5 μM Agilent deactivator additive in 5:3:2 water:acetonitrile:2-propanol and (B) 10 mM ammonium formate in 1:9:90 water:acetonitrile:2-propanol. Column temperature was set at 60°C and autosampler temperature was at 20°C. The flow rate was 0.4 mL/min. The following gradient was applied: 0 min, 15% B; 0-2.5 min, to 50% B; 2.5-2.6 min, to 57%, 2.6-9 min, to 70% B; 9-9.1 min, to 93% B; 9.1-11.1 min, to 96%; 11.1- 15min, 100% B; 15-20 min, 15% B. |
Instrument Name: | Agilent Model 1290 Infinity II liquid chromatography system |
Column Name: | Cogent Diamond Hydride (150 × 2.1 mm, 4um) |
Column Temperature: | 60 |
Flow Gradient: | 0 min, 15% B; 0-2.5 min, to 50% B; 2.5-2.6 min, to 57%, 2.6-9 min, to 70% B; 9-9.1 min, to 93% B; 9.1-11.1 min, to 96%; 11.1- 15min, 100% B; 15-20 min, 15% B. |
Flow Rate: | 0.4 mL/min |
Solvent A: | 50% water/30% acetonitrile/20% isopropanol;10 mM ammonium formate with 5 µM Agilent deactivator additive |
Solvent B: | 1% water/9% acetonitrile/90% isopropanol;10 mM ammonium formate |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS004212 |
Analysis ID: | AN004465 |
Instrument Name: | Bruker Impact HD |
Instrument Type: | QTOF |
MS Type: | ESI |
MS Comments: | The Bruker Impact II QTOF was equipped with a vacuum insulated probe heated electrospray ionization source (VIP-HESI) (Bruker Daltonics, Billerica, USA) to identify representative lipid structures using auto-MS/MS with and without scheduled precursor list fragmentation. Fragments were compared with those deposited in LIPID MAPS, HMDB and MassBank |
Ion Mode: | POSITIVE |
MS ID: | MS004213 |
Analysis ID: | AN004466 |
Instrument Name: | Bruker Impact HD |
Instrument Type: | QTOF |
MS Type: | ESI |
MS Comments: | The Bruker Impact II QTOF was equipped with a vacuum insulated probe heated electrospray ionization source (VIP-HESI) (Bruker Daltonics, Billerica, USA) to identify representative lipid structures using auto-MS/MS with and without scheduled precursor list fragmentation. Fragments were compared with those deposited in LIPID MAPS, HMDB and MassBank. |
Ion Mode: | NEGATIVE |