Rapid method for the analysis of red blood cell fatty acids by reversed-phase high-performance liquid chromatography

A high-performance liquid chromatographic method was developed for a rapid qualitative and quantitative analysis of p-bromophenacyl esters of red blood cell fatty acids in humans. Both free and bound fatty acids, extracted with hexane-2-propanol (3:2) from packed red blood cells were derivatized with p-bromophenacyl bromide and analysed. Ten identical samples taken from a mixed pool of packed red blood cells from healthy subjects were analysed on two different columns. The fatty acid p-bromophenacyl esters were analysed on a 10 RP-18 column with methanol-acetonitrile-0.01 M ammonium formate as mobile phase and also on a 10 RP-8 column with acetonitrile-0.01 M ammonium formate as mobile phase. The two methods gave analogous results except in total analysis time: that on a 10 RP-8 column is ca. 40% shorter. Furthermore, a quantitative analysis of a standard solution to evaluate the extraction procedure in the absence or in the presence of the red blood cell core indicated a significant difference when the core is present.