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Clinical Chemistry 39: 936-941, 1993;
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Clinical Chemistry, Vol 39, 936-941, Copyright © 1993 by the American Association for Clinical Chemistry

Simultaneous Determination of Free Testosterone and Testosterone Bound to Non-Sex-Hormone-Binding Globulin by Equilibrium Dialysis

Angela Barini 1, Italiana Liberale 1, and Edoardo Menini 1

1 Institute of Biological Chemistry, Hormone Laboratory, Catholic University S.C., 00168 Rome, Italy

We describe a procedure based on equilibrium dialysis that allows the simultaneous determination of free testosterone and testorerone bound to non-sex-hormone-binding globulin (non-SHBG) in plasma. After saturating SHBG with 5agr-dihydrotestosterone (DHT) according to a technique recently described, the percentage of free testosterone in the treated and the untreated samples is measured by equilibrium dialysis with use of a semiautomated instrument that allows rigorous standardization of the experimental conditions. The present method is simpler and faster than the previously described technique in which, after the saturation of SHBG with DHT, the unbound fractions were measured by centrifugal ultrafiltration dialysis. The method is also reproducible and suited for the analysis of a large number of samples. The technique has been applied to the determination of the fractional distribution of testosterone in plasma pools from normally menstruating, pregnant, and postmenopausal women and from normal men.

Submitted on August 14, 1992
Accepted on December 15, 1992






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Copyright © 1993 by the American Association for Clinical Chemistry.