DETERMINATION OF TRIACYLGLYCEROL AND CHOLESTEROL ESTER HYDROPEROXIDES IN HUMAN PLASMA BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY WITH FLUOROMETRIC POSTCOLUMN DETECTION

Cholesterol ester (ChE) and triaclglycerol (TG) hydroperoxides in human plasma were determined by high-performance liquid chromatography with postcolumn detection with diphenyl-1-pyrenylphosphine. Human plasma was extracted once with n-hexane. 2,6-Di-tert.-butyl-4-methylphenol and N-stearylcinnamide were added to human plasma before extraction as an antioxidation agent and an internal standard, respectively. The detection limits of both ChE and TG hydroperoxides were 1 pmol. The sample size was minimized to 250 mul for each run. The recoveries of ChE and TG hydroperoxides from fresh plasma were ca. 90 and 80%, respecively. The relative standard deviations (n = 8) of their values in frozen human plasma 5.4% (ChE hydroperoxides, 298 nM) and 5.7% (TG hydroperoxides, 267 nM). No TG hydroperoxides and 24.5 +/- 9.6 nM (n = 15) ChE hydroperoxides were detected in fresh human plasma. The relative standard deviation (n = 8) of ChE hydroperoxides values in fresh plasma was 5.8% (27.1 nM).