Summary of study ST001323

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR000900. The data can be accessed directly via it's Project DOI: 10.21228/M8BX09 This work is supported by NIH grant, U2C- DK119886.

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Study IDST001323
Study TitleEffect of high-fat diet on serum lipidome in mice
Study SummaryWe analyzed mouse serum samples from a mouse dietary intervention experiment. Briefly, C57BL/6 mice (n=44) were divided into 4 groups (n=11 per group) and fed High-fat diet (HFD), 1% deoxycholic acid (DCA) in drinking water, both, or left as control for 9 months. For quality control, 12 TQC samples and 2 blanks were also included in the analysis (total 58 samples and 6 groups). The two treatments were selected to demonstrate the ability of lipidomics to detect gross changes induced by HFD in the serum lipidome, as well as specific/minor changes induced by the secondary bile acid (DCA) through regulation of liver lipid metabolism.
Institute
QIMR Berghofer Medical Research Institute
Last NameMohamed
First NameAhmed
Address300 Herston Road
Emailahmed.mohamed@qimrberghofer.edu.au
Phone+61738453992
Submit Date2020-03-02
Raw Data AvailableYes
Raw Data File Type(s).d
Analysis Type DetailLC-MS
Release Date2020-03-20
Release Version1
Ahmed Mohamed Ahmed Mohamed
https://dx.doi.org/10.21228/M8BX09
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Sample Preparation:

Sampleprep ID:SP001405
Sampleprep Summary:Mouse serum lipids were extracted using the lipid extraction method by Matyash et al. Mouse serum (30 µL) was added to 215 µL of ice-cold methanol containing 50 µg/mL butylated hydroxytoluene (BHT). Samples were homogenized by three rounds of vortex mixing for 30 seconds, freezing in liquid nitrogen for one minute, thawing for two minutes and sonicating for 10 minutes at 15°C, power 100% in a Grant XUB18 bath sonicator. SPLASH LipidoMix Mass Spec Standard (10 µL, undiluted) and Cer/Sph mixture II (10 µL, undiluted) internal standards mixes from Avanti Polar Lipids were then added to each sample. After overnight incubation at -30°C, 750 µL MTBE was added and each tube was vortex mixed for 10 seconds and shaken for 10 minutes on a tube rotator (4°C). MilliQ water (188 µL) was then added, and the tube was vortex mixed for 30 seconds to form a biphasic separation. After centrifuging for 15 minutes at 15,000 ×g, the clear upper phase containing lipids in MTBE was transferred to another tube and dried down using a gentle stream of nitrogen. Prior to LC-MS/MS analysis, the samples were resuspended in a mixture of 50 µL methanol (containing 50 µg/mL BHT)/toluene (90/10%, v/v) and 4µL was used as the injection volume.
Processing Storage Conditions:On ice
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