Specific estimation of fifteen unconjugated, non-metabolized steroid hormones in human urine

Abstract

A method for the specific determination of 15 free, non-metabolized steroid hormones in human urine is described. The steroids progesterone (P), androstenedione (AD), pregnenolone (PL), 5α-dihydrotestosterone (DHT), dehydroepiandrosterone (DHEA), testosterone (T), 11-deoxycorticosterone (DOC), 17-OH-progesterone (17-OHP), 17-OH-pregnenolone (17-PL), 11-deoxycortisol (S), 18-OH-11-deoxycorticosterone (18-OH-DOC), corticosterone (B), aldosterone (Aldo), cortisol (F), and 18-OH-corticosterone (18-OH-B) were extracted from 2 ml urine samples by a solid-phase technique, subjected to automatic high performance liquid chromatography (HPLC) and finally quantitated by radioimmunoassay (RIA). The combination of HPLC and RIA provides a high specificity of steroid estimates. The automatic mode of HPLC renders the method quite suitable for series analyses in research or for routine purposes. Precision and accuracy are satisfactory, and comparable with the level commonly achieved in RIA techniques. The mean values (nmol/24 h) of reference ranges established from 32 normal males were as follows: P: 0.36; AD: 9.76; PL: 0.90; DHT: 0.61; DHEA: 6.76; T: 1.43; DOC: 0.35; 17-OHP: 1.03; 17-PL: 0.20; S: 0.24; 18-OH-DOC: 2.11; B: 1.49; Aldo: 0.46; F: 68.3; 18-OH-B: 5.41. This highly practicable method may be particularly useful for the further investigation of the physiological or diagnostic significance of the non-conjugated, non-metabolized fraction of steroid hormones in urine.