Isolation and Identification of Progesterone and 20α-Hydroxypregn-4-en-3-one in Ovarian Venous Blood of Rabbits

Abstract

Pregn-4-ene-3,20-dione (progesterone) and 20a-hydroxy-pregn-4-en-3-one (Δ4-3-ketopregnen-20α-ol) were identified in the ovarian venous blood of mature New Zealand white rabbits. Pregn-4-ene-3,20-dione was identified by chromatographic mobilities in 4 different paper chromatographic systems, by refractiveness to acetylation, by UVabsorption spectra in methanol and in concentrated sulfuric acid, and finally by an infrared absorption spectrum. The criteria of the identity of 20α-hydroxy-pregn-4-en-3-one were the following: R_T values in 4 different systems, the oxidation product, the acetate, the Zimmermann reaction and the KI-I₂ reaction on paper, the UV spectrum in methanol, the UV and visible spectrum in concentrated sulfuric acid and the infrared spectrum in chloroform.