Fluorimetric high-performance liquid chromatography of prostaglandins and its application to their determination in human seminal fluid.

A highly sensitive and simple high-performance liquid chromatographic method with fluorescence detection for the determination of prostaglandins is described. Prostaglandins are converted into the corresponding fluorescent derivatives by reaction with 3-bromomethyl-6,7-dimethoxy-1-methyl-2(1H)-quinoxalinone in the presence of potassium hydrogen carbonate and 18-crown-6 in acetonitrile. The derivatives are separated simultaneously within 34 min (the total run time per injection, 56 min) on a reversed-phase column (YMC Pack C8) by a stepwise elution with mixtures of acetonitrile, methanol and water and detected fluorimetrically. The detection limits are 10-15 fmol at a signal-to-noise ratio of 5 in a 10-microliter injection volume. Prostaglandins E1, E2, F1 alpha and F2 alpha in human seminal fluids are measured by this method.