Simultaneous Separation of Monoacylglycerols, Free Fatty Acids, and Fatty Acid Methyl and Ethyl Esters by Reversed‐Phase HPLC

Abstract We have developed a reversed‐phase C18 high performance liquid chromatography (HPLC) method to separate molecular species of monoacylglycerols (MAG), fatty acids, fatty acid methyl esters, and fatty acid ethyl esters, simultaneously. This system also separates the regioisomers, 2‐acyl‐sn‐glycerol and 1‐acyl‐sn‐glycerol, with 2‐acyl‐sn‐glycerol eluting earlier than 1‐acyl‐sn‐glycerol. The elution order of the fatty acid and esters was ricinoleate, linolenate, linoleate, palmitate, oleate, and stearate. This method is also useful for the identification of radiolabeled metabolites using co‐chromatography.