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MB Sample ID: SA258272

Local Sample ID:5980
Subject ID:SU002670
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090
Gender:Male
Species Group:Mammals

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Subject:

Subject ID:SU002670
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090
Gender:Male
Species Group:Mammals

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
5980SA258272FL033145High Fat dietDiet
5980SA258272FL033145FastedNutritional status
5980SA258272FL033145VLCAD -/-Genotype

Collection:

Collection ID:CO002663
Collection Summary:Mice were sacrificed under anesthesia using ketamine (100mg/ml) and xylazine (20mg/ml). Immediately after mouse sacrifice, hearts were rapidly excised, snap-frozen with liquid nitrogen, and then stored at -80°C.
Sample Type:Heart
Storage Conditions:-80℃
Collection Tube Temp:-80

Treatment:

Treatment ID:TR002682
Treatment Summary:VLCAD-/- males (129svJ X C57BL/6J genetic background) were crossed with C57Bl/6J females to generate heterozygous VLCAD+/- mice. Coupling of heterozygous mice then produced VLCAD-/- mice and control littermate counterparts (VLCAD+/+). All animals were genotyped as previously described. Mice were housed in a specific pathogen-free facility under a 12-h light/dark cycle at constant temperature and with access to water and food ad libitum. At 7-month of age, mice were fed for 2 weeks with either a chow diet (Harlan no. 2018; 3.1 kcal/g) or a high-fat diet (Harlan Teklab no. 3584; 5.4kcal/g) ad libitum. Mice were sacrificed in fed state for the CD group (CD/fed condition) or after fasting for 24 hours for the HFD group (HFD/fasted condition. Percent calories from carbohydrates, lipids and proteins were 58/18/24 for the CD and 26.6/58.4/15 for the HFD.

Sample Preparation:

Sampleprep ID:SP002676
Sampleprep Summary:Total lipids were extracted from 40mg of pulverized hearts under liquid nitrogen using in methyl tert-butyl ether (MTBE) and ethyl acetate after spiking with the following internal standards lysophosphatidylcholine (LPC) 13:0, phosphatidylcholine (PC) 19:0/19:0, PC14:0/14:0, phosphatidylserine (PS) 12:0/12:0, phosphatidylglycerol (PG) 15:0/15:0 and phosphatidylethanolamine (PE) 17:0/17:0). Supernatants were evaporated using a Speed Vacuum concentrator overnight, dissolved in methanol/chloroform (2:1) and aliquots (50 µL) were stored at -80ºC until use.
Sampleprep Protocol Filename:Sample preparation protocol of untargeted lipidomic analysis
Processing Storage Conditions:-80℃

Combined analysis:

Analysis ID AN004233
Analysis type MS
Chromatography type Reversed phase
Chromatography system Agilent 1290 Infinity
Column Agilent ZORBAX Eclipse Plus C18 (100 x 2.1mm,1.8um)
MS Type ESI
MS instrument type QTOF
MS instrument name Agilent 6530 QTOF
Ion Mode POSITIVE
Units Counts

Chromatography:

Chromatography ID:CH003141
Methods Filename:LC-MS and data processing protocol of untargeted lipidomic analysis
Instrument Name:Agilent 1290 Infinity
Column Name:Agilent ZORBAX Eclipse Plus C18 (100 x 2.1mm,1.8um)
Column Temperature:40
Flow Gradient:0–2 min, 50% B; 2–11 min, 50–85% B; 11–33 min, 85–94% B; 33–35 min, 94–98% B, 35–38 min; 98–99.5% B; 38–82 min, 98–99.5% B; 82–83 min, 99.5–50% B, plus an equilibration of 8 min
Flow Rate:0.45 ml/min
Solvent A:100% water; 0.2% formic acid; 10 mM ammonium formate
Solvent B:55% ethanol,/35% acetonitrile/10% MTBE, 55:35:10; 0.2% formic acid; 10 mM ammonium formate
Chromatography Type:Reversed phase

MS:

MS ID:MS003980
Analysis ID:AN004233
Instrument Name:Agilent 6530 QTOF
Instrument Type:QTOF
MS Type:ESI
MS Comments:MS data processing was achieved using the Mass Hunter Qualitative Analysis software package (version B.07) for peak picking and a bioinformatic script that we have developed and encoded in both Perl and R languages to enable optimal MS data alignment, filtering of presence, normalisation, batch effect correction, and missing data imputation. The resulting final dataset list reproducible high quality MS signals referred as features with their mass-over-charge (m/z), corrected signal intensity, and retention time (RT). Feature identity (ID) was as following: ionization mode:mass@RT.
Ion Mode:POSITIVE
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