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Cellular Lipid Extraction for Targeted Stable Isotope Dilution Liquid Chromatography-Mass Spectrometry Analysis

Article DOI: 10.3791/3399-v • 09:26 min • November 17th, 2011
November 17th, 2011 •
1,2, 1,2, 1,2
1Centers for Cancer Pharmacology and Excellence in Environmental Toxicology, University of Pennsylvania, 2Department of Pharmacology, University of Pennsylvania

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This protocol will demonstrate the extraction and analysis of free and esterified bioactive fatty acids from cells. Fatty acids are accurately quantified using stable isotope dilution, chiral liquid chromatography, electron capture atmospheric chemical ionization multiple reaction monitoring mass spectrometry (SID-LC-ECAPCI-MRM/MS).

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Cellular Lipid Extraction Targeted Stable Isotope Dilution Liquid Chromatography-mass Spectrometry Analysis Fatty Acids Arachidonic Acid Linoleic Acid Oxidized Bioactive Lipids Stereoisomers Enzymatic Metabolism Reactive Oxygen Species Cyclooxygenase Lipoxygenase Cytochromes P450 Cell Lysate Liquid-liquid Extraction Stable Isotope Internal Standards Ion Suppression Mass Spectrometry Analysis
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