STUDIES ON PYROGENIC STEROIDS. I. SEPARATION, IDENTIFICATION, AND MEASUREMENT OF UNCONJUGATED DEHYDROEPIANDROSTERONE, ETIOCHOLANOLONE, AND ANDROSTERONE IN HUMAN PLASMA*

Abstract

A method is presented for the separation of unconjugated plasma 17-ketosteroids and micromeasurement of their 2,4-dinitrophenylhydrazone derivatives. An isotopic radiochemical dilution technique with pure tritium-labeled etiocholanolone, dehydroepiandrosterone, and androsterone is applied to correct for losses during the procedure. The normal plasma ranges are: etiocholanolone < 0.5 to 1.2 ug per 100 ml; androsterone, <0.5 to 0.8 [mu]g per 100 ml; dehydroepiandrosterone, < 0.5 to 1.2 [mu]g per 100 ml. The total mean [plus or minus] SD isotopic and chemical recoveries are 72 [plus or minus] 14 and 67 [plus or minus] 21 per cent, respectively. Each of the measured 2,4-dinitrophenylhydrazone derivatives of unconjugated etiocholanolone, dehydroepiandrosterone, and androsterone was identified by characteristic elution mobility and maintenance of constant specific activity after four separate column chromatographic procedures. Elevation of unconjugated plasma etiocholanolone (a known pyrogenic steroid) and dehydroepiandrosterone was demonstrated in five patients during attack of etiocholanolone fever. In addition, three of the five patients had elevated unconjugated plasma androsterone levels.