Presence of fatty acid esters of pregnenolone in follicular fluid from women undergoing follicle stimulation.

In the following investigation the presence of lipoidal pregnenolone derivatives in the preovulatory follicular fluid obtained from women undergoing in vitro fertilization was established. Concentrations of lipoidal pregnenolone proved to be at least twofold greater than those of the unconjugated counterpart. Indirect identification of these lipoidal pregnenolone derivatives was achieved by comparing the C-18 column, thin-layer silica gel (TLC), high performance liquid chromatography (HPLC), and gas chromatography/mass spectrometry (GC/MS) chromatographic properties of the endogenous lipoidal pregnenolone derivatives in follicular fluid with those of synthetic acyl pregnenolone esters. Lipoidal pregnenolone derivatives recovered after HPLC subfractionation were treated with alkali to hydrolyze the acyl group thus liberating nonconjugated pregnenolone. Concentrations of this steroid were then measured using radioimmunoassay upon which analysis of HPCL and gas chromatograms permitted the calculation of the individual pregnenolone ester contributions within the samples. Five lipoidal pregnenolone derivatives constituted more than 90% of the total lipoidal pregnenolone concentration observed, these derivatives being: pregnenolone oleate (30.7%), linoleate (20.7%), palmitate (20.1%), linolenate (14.8%), and palmitoleate (7.1%).