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

M. Szécsi, I. Tóth, J. Gardi, T. Nyári, J. Julesz

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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 C18 mini-column with methanol-water eluents. Steroids were isolated by reversed-phase HPLC on a C18-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/Δ5-4-isomerase (Δ5-3β-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.

Original languageEnglish
Pages (from-to)47-56
Number of pages10
JournalJournal of biochemical and biophysical methods
Issue number1-2 SPEC. ISS.
Publication statusPublished - Oct 29 2004



  • High-performance liquid chromatography
  • Ovarian neoplasms
  • Radioimmunoassay
  • Sex cord-stromal tumor
  • Steroid hormone biosynthesis
  • Steroid isolation and purification
  • Virilism

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry

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