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Cellular Lipid Extraction for Targeted Stable Isotope Dilution Liquid Chromatography-Mass Spectrometry Analysis
Published on: November 17, 2011
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An optimized method for measuring fatty acids and cholesterol in stable isotope-labeled cells
Joseph P Argus1, Amy K Yu1, Eric S Wang1
1Departments of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095.
Journal of Lipid Research
|December 16, 2016
Summary
This study presents a new method for analyzing fatty acids and cholesterol in cells using stable isotope labeling. The optimized technique allows accurate lipid analysis from small cell samples, aiding metabolic research.
Area of Science:
- Biochemistry
- Cell Biology
- Metabolomics
Background:
- Stable isotope labeling is crucial for studying lipid metabolism in normal and cancerous cells.
- Existing methods for fatty acid and cholesterol analysis have limitations for in vitro stable isotope-labeling studies.
Purpose of the Study:
- To develop an optimized method for measuring both fatty acids and cholesterol from small numbers of stable isotope-labeled cultured cells.
- To enable simultaneous quantitation of cholesterol and fatty acids from the same sample.
Main Methods:
- Quantitative derivatization and extraction of fatty acids and cholesterol.
- Application of stable isotope labeling for lipid analysis.
- Analysis of mass isotopomer distributions.
Main Results:
- The method allows quantitative recovery of fatty acids from diverse lipid classes.
- Cholesterol is recovered with modest yield, enabling simultaneous analysis with fatty acids.
- Accurate measurement of mass isotopomer distributions for cholesterol and many fatty acids from small cell numbers is achievable.
- Background contamination can affect quantitation of low-abundance fatty acids.
Conclusions:
- This optimized method facilitates the acquisition of lipid parameters for flux quantification.
- The technique enhances understanding of how lipid metabolism impacts cellular function.
- It is suitable for analyzing lipid metabolic parameters in both normal and neoplastic cells.
