High-performance liquid chromatography and post-column derivatization with diphenyl-1-pyrenylphosphine for fluorimetric determination of triacylglycerol hydroperoxides

Abstract Triacylglycerol monohydroperoxides (TG-mHPO) were selectively detected at the picomole levels after post-column reaction with diphenyl-1-pyrenylphosphine (DPPP). TG-mHPO were separated on two types of reserved-phase columns, an ODS column and a phenylated silica gel, column which were useful for determining TG-mHPO at their molecular species levels and their class levels, respectively. After the separation, DPPP solution was mixed with the eluent followed by reaction in a stainless-steel coil 20 m × 0.5 mm I.D. at 80°C, then the fluorescence intensity of DPPP oxide was measured (λ ex. 352 nm, λ em. 380 nm). Using these systems, TG-mHPO were determined in the range 2–1000 pmol. The relative standard deviations were 2.3–2.8%.