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MB Sample ID: SA239023

Local Sample ID:WT-22R
Subject ID:SU002485
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090

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Subject:

Subject ID:SU002485
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
WT-22RSA239023FL029779p53 +/+experimental factor(s)

Collection:

Collection ID:CO002478
Collection Summary:Mouse liver samples were collected from mice and immediately flash frozen in liquid nitrogen. Frozen tissue samples were crushed using a morter and pestle on dry ice to homogenize tissue prior to lipid extraction.
Sample Type:Liver

Treatment:

Treatment ID:TR002497
Treatment Summary:Male mice were C57BL/6J background, kept on a 12hr day/night cycle. Mice were 3-4 months old at the time of tissue isolation. Four littermate pairs were analyzed.

Sample Preparation:

Sampleprep ID:SP002491
Sampleprep Summary:Mouse liver sections were weighed into Eppendorf tubes. The liver tissue was then mashed with a spatula prior to extraction. The samples were extracted using a liquid liquid partition with water (250 µL), methanol (300 µL), and MTBE (1 mL). Avanti’s deuterated lipid mix, Equisplash, was used as an internal standard. This was spiked into the methanol at 1.5 µg/mL and used for extraction. The extracts were centrifuged at 20,000 rcf for 10 min. The top layer was removed, dried down, and reconstituted in 1 mL of IPA for analysis.

Combined analysis:

Analysis ID AN003903
Analysis type MS
Chromatography type Reversed phase
Chromatography system Waters Acquity H-Class
Column Waters Acquity BEH C18 (100 x 2.1mm,1.7um)
MS Type ESI
MS instrument type Orbitrap
MS instrument name Thermo Q Exactive HF-X Orbitrap
Ion Mode UNSPECIFIED
Units peak area

Chromatography:

Chromatography ID:CH002889
Chromatography Summary:Analysis was performed using a Thermo Q Exactive Plus coupled to a Waters Acquity H-Class LC. A 100 mm x 2.1 mm, 1.7 µm Waters BEH C18 column was used for separations. The following mobile phases were used: A- 60/40 ACN/H20 B- 90/10 IPA/ACN; both mobile phases had 10 mM Ammonium Formate and 0.1% Formic Acid. A flow rate of 0.2 mL/min was used. Starting composition was 32% B, which increased to 40% B at 1 min (held until 1.5 min) then 45% B at 4 minutes. This was increased to 50% B at 5 min, 60% B at 8 min, 70% B at 11 min, and 80% B at 14 min (held until 16 min). At 16 min the composition switched back to starting conditions (32% B) and was held for 4 min to re-equilibrate the column. Samples were analyzed in positive/negative switching ionization mode with top 5 data dependent fragmentation.
Methods Filename:Sanford_2022_Lipidomics_protocol.docx
Instrument Name:Waters Acquity H-Class
Column Name:Waters Acquity BEH C18 (100 x 2.1mm,1.7um)
Chromatography Type:Reversed phase

MS:

MS ID:MS003642
Analysis ID:AN003903
Instrument Name:Thermo Q Exactive HF-X Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:Analysis was performed using a Thermo Q Exactive Plus coupled to a Waters Acquity H-Class LC. A 100 mm x 2.1 mm, 2.1 µm Waters BEH C18 column was used for separations. The following mobile phases were used: A- 60/40 ACN/H20 B- 90/10 IPA/ACN; both mobile phases had 10 mM Ammonium Formate and 0.1% Formic Acid. A flow rate of 0.2 mL/min was used. Starting composition was 32% B, which increased to 40% B at 1 min (held until 1.5 min) then 45% B at 4 minutes. This was increased to 50% B at 5 min, 60% B at 8 min, 70% B at 11 min, and 80% B at 14 min (held until 16 min). At 16 min the composition switched back to starting conditions (32% B) and was held for 4 min to re-equilibrate the column. Samples were analyzed in positive/negative switching ionization mode with top 5 data dependent fragmentation.
Ion Mode:UNSPECIFIED
Analysis Protocol File:Sanford_2022_Lipidomics_protocol.docx
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