A non-chromatographic radioimmunoassay of testosterone in serum

Convenient methodology based on separation of testosterone from non-alcoholic neutral steroids by means of a sulfation procedure has been developed for the radioimmunoassay (RIA) of testosterone in male and in female serum. When coordinated with our previously published non-chromatographic procedure (1) for the RIA of estrone and 17 beta-estradiol in serum all 3 steroids can be determined in the same specimen. With only minor modification (14) progesterone also can be determined. Validation of the procedure was based on: 1. agreement between results obtained using TLC and sulfation to fractionate testosterone (r=0.99; b=1.03), 2. accurate recovery of different quatities of testosterone added to serum, 3. independence of the concentration of testosterone and volume of serum used for assay, 4. low procedural blanks (1.1+/-0.7 pg), 5. low intra-assay (4.7-4.8%) and interassay (4.8-8.8%) variability and 6. correspondence of observed values for testosterone in male serum (7.00+/-2.03 ng/ml) and in femal serum (420+/-115 pg/ml) with those reported previously by others.