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Bound fatty acids When fatty acids are combined in more complex molecules such as acylglycerols, cholesterol esters, waxes and glycosphingolipids, they can be obtained free by saponification (inorganic or organic basic solution) or acidic hydrolysis and then derivatized. FAME may be also obtained plump hardcore directly by transesterification (alcoholysis or methanolysis) of the fatty acid-containing lipids. The extraction and methylation may also be combined in a one-step procedure, this is particularly recommended for very small samples in order to prevent any loss of fatty acids during the plump hardcore classical procedures. Saponification When fatty acids are required in free form for plump hardcore further analysis, lipids (present as glycerides, glycerophosphatides, glycosyldiglycerides, sterol esters or waxes) are first hydrolyzed in alkaline medium allowing to extract also the unsaponifiable material if present in the crude lipid mixture (sterols, alcohols, hydrocarbons, pigments, vitamins...).
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