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MB Sample ID: SA210450

Local Sample ID:101_sp_4w_pos
Subject ID:SU002281
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090

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Subject:

Subject ID:SU002281
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
101_sp_4w_posSA210450FL025708SpadeGenotype
101_sp_4w_posSA210450FL025708Sp_4wBatch

Collection:

Collection ID:CO002274
Collection Summary:Mice were euthanized. Then, ears were collected and stored at -80˚C after rapid freezing in liquid nitrogen.
Sample Type:skin

Treatment:

Treatment ID:TR002293
Treatment Summary:Mice were bred and maintained in the Keio specific pathogen-free facility, with a 12 h light/12 h dark cycle and food (CLEA Rodent Diet CE-2, CLEA Japan, Tokyo, Japan) and water available ad libitum.

Sample Preparation:

Sampleprep ID:SP002287
Sampleprep Summary:Frozen ear was pulverized by a metal cone using MULTI-BEADS SHOCKER MB1200 (YASUI KIKAI, Osaka, Japan), and dissolved in 1 ml methanol. The resulting solvents were transferred to a 2 ml glass tube, and 500 µl CHCl3 in internal standard (1 µM d9-EOS d18:1/32:0/18:2 (Cayman, Ann Arbor, MI, USA)) was added followed by incubation at -30˚C for 16 h. After centrifugation (2,000 g, 4˚C, 10 min), A total of 200 µl supernatants were dissolved in 100 µl methanol, and 20 μl MilliQ was added. After 15 min of incubation at room temperature, the tubes were centrifuged (2,000 g, 20˚C, 10 min). The supernatants were subjected to untargeted LC-MS/MS analysis.

Combined analysis:

Analysis ID AN003593 AN003594
Analysis type MS MS
Chromatography type Reversed phase Reversed phase
Chromatography system Waters Acquity UPLC Waters Acquity UPLC
Column Waters Acquity BEH C18 (50 x 2.1mm,1.7um) Waters Acquity BEH C18 (50 x 2.1mm,1.7um)
MS Type ESI ESI
MS instrument type Triple TOF Triple TOF
MS instrument name ABI Sciex 6600 TripleTOF ABI Sciex 6600 TripleTOF
Ion Mode NEGATIVE POSITIVE
Units CPM/mg CPM/mg

Chromatography:

Chromatography ID:CH002655
Chromatography Summary:LC gradient consisted of holding solvent (A/B: 100/0) for 1 min, then linearly converting to solvent (A/B: 60/40) for 4 min, linearly converting to solvent (A/B:36/64) for 2.5 min and holding for 4.5 min, then linearly converting to solvent (A/B: 17.5/82.5) for 0.5 min, linearly converting to solvent (A/B: 15/85) for 8.5 min, and linearly converting to solvent (A/B: 5/95) for 1 min followed by returning to solvent (A/B: 100/0) and holding for 5 min for re-equilibration.
Instrument Name:Waters Acquity UPLC
Column Name:Waters Acquity BEH C18 (50 x 2.1mm,1.7um)
Column Temperature:45
Flow Gradient:LC gradient consisted of holding solvent (A/B: 100/0) for 1 min, then linearly converting to solvent (A/B: 60/40) for 4 min, linearly converting to solvent (A/B:36/64) for 2.5 min and holding for 4.5 min, then linearly converting to solvent (A/B: 17.5/82.5) for 0.5 min, linearly converting to solvent (A/B: 15/85) for 8.5 min, and linearly converting to solvent (A/B: 5/95) for 1 min followed by returning to solvent (A/B: 100/0) and holding for 5 min for re-equilibration.
Flow Rate:0.30mL/min
Solvent A:20% methanol/20% acetonitrile/60% water; 5 mM ammonium acetate; 10 nM EDTA
Solvent B:100% isopropanol; 5 mM ammonium acetate; 10 nM EDTA
Chromatography Type:Reversed phase

MS:

MS ID:MS003348
Analysis ID:AN003593
Instrument Name:ABI Sciex 6600 TripleTOF
Instrument Type:Triple TOF
MS Type:ESI
MS Comments:MS-DIAL was used for peak picking and alignment
Ion Mode:NEGATIVE
  
MS ID:MS003349
Analysis ID:AN003594
Instrument Name:ABI Sciex 6600 TripleTOF
Instrument Type:Triple TOF
MS Type:ESI
MS Comments:MS-DIAL was used for peak picking and alignment
Ion Mode:POSITIVE
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