Fatty acids participate in different metabolic mechanisms, but their key role is connected with their participation in membrane phospholipid organizations. This review introduces the current knowledge of fatty acid analysis with an emphasis on the analysis of these compounds in biological samples. Therefore this manuscript is focused on various aspects of biological sample preparation methods, such as lipid extraction, lipid and phospholipid fractionation (TLC and SPE), derivatization, and on the methodologies of fatty acid analysis by gas chromatography (GC) and high-performance liquid chromatography (HPLC). The wide spectrum of columns and detectors used in these techniques for selective separation and determination of bioactive fatty acids are discussed. The abilities of using a sensitive tandem LC/MS and GC/MS assay are also shown.
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