Sensitive high-performance liquid chromatographic method for prostaglandins using a fluorescence reagent,...

High-performance liquid chromatography of prostaglandins is developed in which a fluorescence reagent, 4-bromomethyl-7-acetoxycoumarin is used to perform the high-sensitivity detection. The reagent reacts with prostaglandins and related compounds to form the ester derivatives, which are separated using a reversed-phase system. Each labeled compound eluted from the column is successively hydrolyzed to the fluorescent coumarin derivative, and this fluorophore is introduced into a flow-through fluorometer. Prostaglandins can be determined in the range of at least 1 nmol to 5 pmol, and the detection limit is about 10 fmol. This system is applied to the analysis of prostaglandins in human seminal fluid.