A procedure for microdetermination of testosterone in human spermatic vein blood is presented. C14-testosterone was added to the plasma to facilitate chemical studies. The plasma was extracted with ether and partitioned between petroleum ether and 70 per cent ethanol. The alcohol fraction was treated with 2,4-dinitrophenylhydrazine and the derivative was chromatographed on 2 different columns to constant specific activity. The validity of the method was tested further by changing the derivative obtained from dog plasmas to the free steroid, re-processing it, and demonstrating that the specific activity was not changed by these procedures. Data obtained from clinical material showed that the level of testosterone in human spermatic vein blood varies from 1.6 to 0.025 μg. per cc. of plasma, and decreases with aging.
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