Chromatographic Separation of Steroid Hormones by Centrifugation Through Columns of Microparticulate Silica

Abstract

A chromatographic technique for separating steroid hormones utilizes centrifugal force to enhance the migration of a sample through columns of densely packed gel. Application of centrifugal force permits the use of ultra fine silica (150 A-wide leaflets) for preparing the adsorbent bed, thereby providing improved resolution. By the use of several solvent systems various closely related species of steroids were separated, including 1) 17-keto steroids, 2) axial-equatorial hydroxyl isomers and cis-trans isomers of pregnanediol, and 3) other combinations of steroids heretofore not readily resolved, such as cortisone and prednisone, 5β-pregnane-3,20-dione and 5α-pregnane-3,20-dione, and 17α-OH progesterone and testosterone. The method is distinctive in its simplicity, high resolving power, reproducibility, rapidity, and low cost.