Prostaglandins E and F, and 19-hydroxylated E and F (series I and II) in semen of fertile men. Gas and liquid chromatographic separation with selected ion detection.

Low concentrations of prostaglandins (PG) could be related to male clinical infertility althought relevant experimental data are scarce. The aim of this work is to establish reliable seminal PG levels in fertile men by rigorous sample control, to prevent degradation, and by rapid and simple extraction and assay procedures. Single semen samples from healthy fertile men were immediately centrifuged (within 30 min of ejaculation) adding PGF2 alpha D4 to the seminal plasma as internal standard. The samples were next ultrafiltered and the PGs in the ultrafiltrate were derivatized with a mixture of N,O-bis(trimethylsilyl)trifluoroacetamide and piperidine (1:1) at 60 degrees for 30 min. Optimum gas-liquid chromatographic separation of all of the peaks of interest was achieved on 4 m x 1/4 in. I.D. Dexsil 300 packed columns at 280 degrees. The detection and quantitation of all the peaks of interest depends on the selected ion monitoring of specific masses. The values obtained (in micrograms/ml, range in parentheses) were: PGEs, 63.5 +/- 49.3 (9--164); PGFs, 2.6 +/- 1.92 (0.95--6.63); 19-OH PGEs, 592.6 +/- 312.5 (142.1--1047); and 19-OH PGFs, 12.66 +/- 5.21 (4--19). Individual values for members of both series I and II are also presented. The sample collection and extraction procedures were further checked by high-performance liquid chromatography on a muPorasil column, with individual isolation and collection of all of the PGs, including the 19-OH PGs not previously separted by liquid chromatography.