HPLC-RIA analysis of steroid hormone profile in a virilizing stromal tumor of the ovary

Abstract The pathological steroid biosynthesis of a virilizing ovarian tumor was examined via high performance liquid chromatography–radioimmunoassay (HPLC–RIA) determination of the intratissular steroid concentrations. Sex cord-stromal tumor of the ovary was obtained surgically from an 18-year-old female patient with extremely high androst-4-ene-3,17-dione (4-en-dione) and testosterone (Test) blood serum levels. The tissue specimen was extracted with ethyl acetate and the extract was then purified on a C 18 mini-column with methanol–water eluents. Steroids were isolated by reversed-phase HPLC on a C 18 -silica gel column with 51%, 55% and 64% v/v methanol–water eluents. Steroids in the collected eluent fractions were detected by the radioactivity of tritiated internal standards and then quantified by specific RIAs. In the tumor specimen, very high 17α-hydroxyprogesterone (17-OH-Prog; 6300 fmol/g), dehydro-epiandrosterone (2870 fmol/g), androst-4-ene-3,17-dione (3000 fmol/g), testosterone (5700 fmol/g) concentrations, and less progesterone (PROG; 320 fmol/g) and androst-5-ene-3β,17β-diol (5-en-diol; 320 fmol/g), were determined. Tissue levels of 5α-dihydrotestosterone (DHT), 5α-androstane-3α,17β-diol (3α-diol), 5α-androstane-3β,17β-diol (3β-diol), and 17β-estradiol were found to be 71, 20, 28, and 12 fmol/g, respectively. Steroid profile analysis verified a pathological steroid biosynthesis in the ovarian tumor and suggested that the 17α-hydroxylase (17α-H), 17,20-lyase (17,20-L), and 3β-hydroxysteroid dehydrogenase/Δ 5–4 -isomerase (Δ 5 -3β-HSD) activities were particularly elevated in this tumorous tissue. Present data demonstrate that the analysis of intratissular steroid profile by a HPLC–RIA method may valuably contribute to the steroidal pathophysiology of endocrine tumors.