Studies on Neurosteroids. II. Retention Behavior of Derivatized 20-Oxosteroids and Their Sulfates Using High-Performance Liquid Chromatography

Abstract The retention behavior of five 20-oxosteroids (pregnenolone, 3 β - hydroxy- 5 α -pregnan-20-one, 3 β -hydroxy- 5 β - pregnan-20-one, 3 α -hydroxy- 5 α -pregnan-20-one, and 3 α - hydroxy-5 β -pregnan-20-one) and their sulfates derivatized with 4-(N,N-dimethylaminosulfonyl)- 7-hydrazino-2,1,3-benzoxadiazole were examined using reversed-phase high-performance liquid chromatography with fluorescence detection. Inclusion chromatography using cyclodextrin as a mobile phase additive was also used for this purpose and found effective in separating the isomeric derivatized 20-oxosteroids and their sulfates.