Analysis of Fatty Acids in Mouse Cells Using Reversed-Phase High-Performance Liquid Chromatography
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چکیده
A reversed-phase high-performance liquid chromatography (RP-HPLC) method was developed to analyze various fatty acids in recombinant mouse L cells. These fatty acids were the metabolites of oleic acid. A process was developed to extract fatty acids from the cell samples before RP-HPLC analysis. The samples were first saponified w~th 0.5 M NaOH in 96% ethanol then extracted with acidified ethyl acetate. After extraction, the sample was dried and dissolved in HPLCgrade methanol.Aher centrifugation to remove insoluble impurities, the sample was applied to a C18 RP-HPLC column using a gradient of acetonitrile (ACN)-H20. The eluted fatty acids were monitored by ultraviolet (UV) absorption at 195 n m and identified by retention time and adsorption spectrum comparison. This method successfully resolved various fatty acids and provided a tool for the elucidation of the fatty acid metabolic pathway in the cells.
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