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MB Sample ID: SA352095
Local Sample ID: | AWT-IR_3 |
Subject ID: | SU003341 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU003341 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
AWT-IR_3 | SA352095 | FL041093 | Heart | Sample source |
AWT-IR_3 | SA352095 | FL041093 | Aged | Age |
AWT-IR_3 | SA352095 | FL041093 | WT | Genotype |
AWT-IR_3 | SA352095 | FL041093 | I/R | Treatment |
Collection:
Collection ID: | CO003334 |
Collection Summary: | The sample collection method was: All mice groups underwent anesthesia with 2%-3% isoflurane and 100% O2. The mouse hearts were excised and rinsed in ice-cold PBS. Both atriums were removed; only the left ventricular myocardium, including the infarct area, was collected and was quickly frozen by using liquid nitrogen. Samples were preserved below -80 °C and were sent out for metabolomic and lipidomic analysis. |
Sample Type: | Heart |
Treatment:
Treatment ID: | TR003350 |
Treatment Summary: | YWT: young wild type; Sham: no treatment; MIF20: treatment with MIF20; AWT: aged wild type; MIFff: with MIF; cMIFKO: knock out MIF; I/R: "ischemia-reperfusion"; For I/R, the myocardium underwent 45 minutes long ischemic condition, then followed by 24 hours long reperfusion. MIF20 was administered 5 minutes before the onset of reperfusion by I.V. injection via the jugular vein. MIF20 was administered until the MIF20 concentration in blood reached 8nM. Procedure is : We injected 100 uL of MIF20 solution (160 nM) into a mouse by I.V. Consider the total blood volume of a mouse is about 2 mL. Therefore, the MIF20 solution was diluted 20 times, reaching 8 nM in blood. |
Sample Preparation:
Sampleprep ID: | SP003348 |
Sampleprep Summary: | The lipidomic sample preparation method was: The samples were thawed on ice. Approximately 10 mg of each sample was weighed and homogenized by ball-mill in 1mL of extraction solution (MTBE:Methanol = 3:1, V/V) with internal standards. The mixture was vortexed for 15 min. Next, the mixture was added with 200 µL of water and vortexed for 1 min and incubating at 4 °C for 10 min. After centrifugation at 12000 rpm for 10 min (4 °C), 200 µL of the upper phase was collected for complete solvent drying under 20 °C. The residue was reconstituted using 200 µL of reconstitution solution (ACN:IPA = 1:1, V/V), followed by vortex for 3 min and centrifugation at 12000 rpm for 3 min. A 120 µL of the final supernatant was used for LC-MS analysis. The data acquisition instruments consisted of Ultra Performance Liquid Chromatography (UPLC) (Nex-era LC-40) and tandem mass spectrometry (MS/MS) (Triple Quad 6500+). |
Combined analysis:
Analysis ID | AN005283 |
---|---|
Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Nexera LC-40 |
Column | Thermo Accucore C30 (150 x 2.1mm,2.6um) |
MS Type | ESI |
MS instrument type | Triple quadrupole |
MS instrument name | ABI Sciex 6500+ |
Ion Mode | UNSPECIFIED |
Units | lipid content in the sample (nmol/g) |
Chromatography:
Chromatography ID: | CH003996 |
Chromatography Summary: | Solvent A: acetonitrile /water (60/40, V/V) (0.1% formic acid added, 10 mmol/L ammonium formate) Solvent B: acetonitrile / Isopropyl alcohol (10/90, V/V) (0.1% formic acid added, 10 mmol/L ammonium formate) |
Instrument Name: | Nexera LC-40 |
Column Name: | Thermo Accucore C30 (150 x 2.1mm,2.6um) |
Column Temperature: | 45 |
Flow Gradient: | A:B; 80:20(V/V) at 0 min, 70:30(V/V) at 2 min, 40:60(V/V) at 4 min , 15:85(V/V) at 9 min, 10:90(V/V) at 14 min, 5:95(V/V) at 15.5 min, 5:95(V/V) at 17.3 min, 80:20(V/V) at 17.5 min, 80:20(V/V) at 20 min |
Flow Rate: | 0.35ml/min |
Solvent A: | 60% acetonitrile:40% water; 0.1% formic acid; 10 mmol/L ammonium formate |
Solvent B: | 10% acetonitrile:90% isopropyl alcohol; 0.1% formic acid added; 10 mmol/L ammonium formate |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS005013 |
Analysis ID: | AN005283 |
Instrument Name: | ABI Sciex 6500+ |
Instrument Type: | Triple quadrupole |
MS Type: | ESI |
MS Comments: | LIT and triple quadrupole (QQQ) scans were acquired on a triple quadrupole-linear ion trap mass spectrometer (QTRAP), QTRAP® 6500+ LC-MS/MS System, equipped with an ESI Turbo Ion-Spray interface, operating in positive and negative ion mode and controlled by Analyst 1.6.3 software (Sciex). The ESI source operation parameters were as follows: ion source, turbo spray; source temperature 500 ℃; ion spray voltage (IS) 5500 V(Positive),-4500 V(Neagtive); Ion source gas 1 (GS1), gas 2 (GS2), curtain gas (CUR) were set at 45, 55, and 35 psi, respectively. Instrument tuning and mass calibration were performed with 10 and 100 μmol/L polypropylene glycol solutions in QQQ and LIT modes, respectively. QQQ scans were acquired as MRM experiments with collision gas (nitrogen) set to 5 psi. DP and CE for individual MRM transitions was done with further DP and CE optimization. A specific set of MRM transitions were monitored for each period according to the lipids eluted within this period. |
Ion Mode: | UNSPECIFIED |