Summary of Study ST001286

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 PR000868. The data can be accessed directly via it's Project DOI: 10.21228/M8GT28 This work is supported by NIH grant, U2C- DK119886.

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Study IDST001286
Study TitleLipid composition of isolated lipid droplets from the functional bovine corpus luteum
Study TypeLipidomics
Study SummaryEstablishment and maintenance of pregnancy is dependent on progesterone synthesized by the corpus luteum (CL). The CL is known for the prominent presence of intracellular lipid droplets (LDs). However relatively little is known about the composition and function of these luteal LDs. Our objective was to identify the lipid composition of LDs from fully functional bovine CLs. Luteal LDs were isolated by flotation through a discontinuous sucrose gradient, lipids were then extracted using a standard Bligh and Dyer protocol, dried, and sent to Avanti Polar Lipids for lipidomics analysis. The samples were provided for lipidomic profiling of free sterols, cholesteryl esters, triglycerides, diacylglycerols, phospholipids, and sphingolipids. Molecular species were resolved by reversed-phase liquid chromatography in the presence of class and sub-class specific internal standard compounds added to each sample. The compounds were detected by tandem mass spectrometry (MS/MS) with scheduled multiple reaction monitoring (MRM) for mass-specific fragment ions according to the lipid class and molecular weight of the compound. Quantification of cholesterol, cholesteryl esters, triglycerides, and diglycerides were directly calculated with standards and internal standards from calibration response curves. The remaining lipid species were semi-quantization using the integrated area of each analyte’s MRM peak, divided by the appropriate internal standard peak area, and multiplied by the standard’s known concentration. Lipid concentrations were normalized to the corresponding protein concentration of each sample and as a mol % relative to total lipids or within each lipid class. Isolated luteal LDs were composed primarily of triglyceride (88%, mol% of lipid class to total lipids). Other neutral lipids included diacylglycerol, 2.9%; and cholesteryl esters, 1.5%. Polar lipids were primarily composed of phosphatidylcholine (3.1%), sphingomyelin (1.5%), phosphatidylinositol (0.9%), phosphatidylethanolamine (0.8%) and phosphatidylserine (0.4%). A number of other minor lipids representing less than 0.32% of the total lipid pool were also detected including phosphatidylglycerol, lysophospholipids, ceramides, and glycosylated ceramides. Lipid composition of bovine luteal LDs are distinct from LDs isolated from other tissues and in other species.
Institute
University of Nebraska Medical Center
DepartmentObstetrics and Gynecology
LaboratoryJohn S. Davis
Last NameDavis
First NameJohn
Address983255 Nebraska Medical Center Omaha, NE 68198-3255
Emailjsdavis@unmc.edu
Phone402-559-9079
Submit Date2019-11-18
Num Groups1
Total Subjects3
Num Females3
Analysis Type DetailLC-MS
Release Date2020-05-18
Release Version1
John Davis John Davis
https://dx.doi.org/10.21228/M8GT28
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Combined analysis:

Analysis ID AN002130 AN002131 AN002132 AN002133 AN002134 AN002135 AN002136 AN002137 AN002138
Analysis type MS MS MS MS MS MS MS MS MS
Chromatography type Reversed phase Reversed phase Reversed phase Reversed phase Reversed phase Reversed phase Reversed phase Reversed phase Reversed phase
Chromatography system Waters Acquity Waters Acquity Waters Acquity Waters Acquity Waters Acquity Waters Acquity Waters Acquity Waters Acquity Waters Acquity
Column Waters Acquity BEH C18 (50 x 1.2mm,1.7um) Agilent Eclipse XBD C8 (50 x 4.6mm, 1.8um) Agilent Eclipse XBD C8 (50 x 4.6mm, 1.8um) Agilent Eclipse XBD C8 (50 x 4.6mm, 1.8um) Agilent Eclipse XBD C8 (50 x 4.6mm, 1.8um) Agilent Eclipse XBD C8 (50 x 4.6mm, 1.8um) Agilent Eclipse XBD C8 (50 x 4.6mm, 1.8um) Thermo Hypersil Gold C18 (50 x 1.2mm,1.7um) Agilent Eclipse XDB-PLUS C18 (50 x 1.2mm, 1.8um)
MS Type ESI ESI ESI ESI ESI ESI ESI ESI ESI
MS instrument type Triple quadrupole Triple quadrupole Triple quadrupole Triple quadrupole Triple quadrupole Triple quadrupole Triple quadrupole Triple quadrupole Triple quadrupole
MS instrument name ABI Sciex 5500 QTrap ABI Sciex 5500 QTrap ABI Sciex 5500 QTrap ABI Sciex 5500 QTrap ABI Sciex 5500 QTrap ABI Sciex 5500 QTrap ABI Sciex 5500 QTrap ABI Sciex 5500 QTrap ABI Sciex 5500 QTrap
Ion Mode POSITIVE POSITIVE POSITIVE NEGATIVE POSITIVE NEGATIVE POSITIVE POSITIVE POSITIVE
Units nM nM nM nM nM nM nM nM nM
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