A Simple, Differential Extraction Method for the Simultaneous Direct Radioimmunoassay of Androgens and Androgen Glucuronides in Human Serum

Steroids. 1996 Jul;61(7):429-32. doi: 10.1016/0039-128x(96)00063-3.

Abstract

A method is described for the differential extraction of unconjugated androgens (testosterone and dihydrotestosterone) and glucuronidated androgens (androstane-3 alpha,17 beta-diol glucuronide and androsterone glucuronide) from human serum using solid-phase, gravity-flow extraction columns. In this method, 100-microL aliquots of serum are loaded onto the normal-phase columns, unconjugated androgens are eluted with ethyl ether, and glucuronides are eluted with ethyl ether containing 2% acetic acid. Glucuronide eluates are washed with 1% aqueous acetic acid to remove cross-reacting steroid sulfates. Assays of sera for the four steroids were performed using standard radioimmunoassay methodology, except for androsterone glucuronide. This steroid was assayed with a novel radioimmunoassay method that employees a tritiated, unconjugated androsterone tracer and an anti-dehydroepiandrosterone sulfate antiserum. The new method is well suited for the assay of conjugated and unconjugated steroids in large numbers of specimens, particularly where the sample volume is limited.

MeSH terms

  • Adult
  • Aged
  • Androgens / blood*
  • Androgens / chemistry
  • Androstane-3,17-diol / analogs & derivatives
  • Androstane-3,17-diol / blood
  • Androstane-3,17-diol / chemistry
  • Androsterone / analogs & derivatives
  • Androsterone / blood
  • Androsterone / chemistry
  • Dihydrotestosterone / blood
  • Dihydrotestosterone / chemistry
  • Humans
  • Hydrogen-Ion Concentration
  • Male
  • Radioimmunoassay / methods*
  • Reference Values
  • Testosterone / blood
  • Testosterone / chemistry
  • Tritium

Substances

  • Androgens
  • Dihydrotestosterone
  • Tritium
  • androsterone glucuronide
  • Androstane-3,17-diol
  • androstane-3,17-diol glucuronide
  • Testosterone
  • Androsterone