Summary of Study ST001336

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 IDST001336
Study TitleEffect of high-fat diet and bile acid treatment on serum and tissue lipidomes in mice
Study SummaryWe analyzed mouse serum and esophageal tissue 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), 0.2% deoxycholic acid (DCA) in drinking water, both, or left as control for 9 months. For quality control,TQC samples and blanks were also included in the analysis. The two treatments were selected to demonstrate the ability of lipidomics to detect gross changes induced by HFD in the serum and tissue lipidomes, as well as specific/minor changes induced by the secondary bile acid (DCA) through regulation of liver lipid metabolism. The serum and tissue samples were analyzed using targeted and untargeted lipidomics methods. The targeted serum lipidomics data has previously been uploaded as part of study ST001323.
Institute
QIMR Berghofer Medical Research Institute
Last NameMolendijk
First NameJeffrey
Address300 Herston Road, Herston, QLD, 4006, Australia
Emailjeffrey.molendijk@qimrberghofer.edu.au
Phone+61738453992
Submit Date2020-03-26
Raw Data AvailableYes
Raw Data File Type(s)d
Analysis Type DetailLC-MS
Release Date2020-04-27
Release Version1
Jeffrey Molendijk Jeffrey Molendijk
https://dx.doi.org/10.21228/M8BX09
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Sample Preparation:

Sampleprep ID:SP001418
Sampleprep Summary: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. Tissue wet weight was determined using a Mettler-Toledo XS105 balance (Mettler-Toledo, Melbourne, Australia). Biopsies were transferred to Eppendorf tubes containing 500 µL ice-cold methanol, 50 µg/ml BHT and one steel bead and homogenized in a TissueLyzer LT (Qiagen, Melbourne, Australia) for six minutes at 50 Hz. Homogenate was transferred to new tubes and the original tube was washed with 400 µL methanol and transferred. Samples were dried down under nitrogen flow and resuspended in 20 µL water and 200 µL methanol (50 µg/ml 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) and Cer/Sph mixture II (10 µL) 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, 700 µL of the clear upper phase containing lipids in MTBE was transferred to another tube and dried down using a gentle stream of nitrogen. After drying down of lipid extracts were resuspended in 50 µL methanol (containing 50 µg/mL BHT)/toluene (90/10%, v/v).
Processing Storage Conditions:On ice
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