Separation and quantitation of hydroxy and epoxy fatty acid by high-performance liquid chromatography with an evaporative light-scattering detector

A new high-performance liquid chromatography technique with an evaporative light-scattering detector (ELSD) has been developed for the separation and quantitative analysis of hydroxy and epoxy fatty acids. This method employs a gradual binary gradient (hexane/isopropanol) and ELSD detection. The minimum limit of detection is about 1 µg and ratio of mass to signal is essentially linear in the range of 10 to 200 µg. This high-performance liquid chromatography (HPLC) technique is able to separate various positional isomers of mono-hydroxy and dihydroxy fatty acids and can also discriminate between monohydroxy, epoxy, epoxyhydroxy, dihydroxy and trihydroxy fatty acids.