SEPARATION OF ARACHIDONIC-ACID METABOLITES BY ONLINE EXTRACTION AND REVERSED-PHASE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY OPTIMIZED BY COMPUTER-SIMULATION

A complex mixture of arachidonic acid metabolites was separated by rev ersed-phase HPLC using a multi-step gradient, which was modelled by co mputer-assisted HPLC method development. The metabolites were extracte d on-line on a precolumn connected to the analytical column in the sam e HPLC system. The predictions of the resolution and also the retentio n times calculated by computer simulation were very accurate when comp ared with the corresponding experimental run (maximum deviation 0.86%) . An appropriate HPLC method and additionally an on-line extraction pr ocedure could be developed with just three experimental HPLC runs. Thi s method could be useful for evaluating the concentrations of arachido nic acid metabolites involved in inflammatory diseases.