High-performance liquid chromatographic methods for monitoring of isomers of 17-hydroxy-16-hydroxymethyl-3-methoxyestra-1,3,5(10)-triene

Claisen condensation and consecutive reduction of 3-methoxyestra-1,3,5(10)- trien-17-one theoretically leads to four diastereomers of 17-hydroxy-16-hyd roxymethyl-3-methoxyestra-1,3,5(10)-triene and their further transformation s give different compounds with different biological activities. High-perfo rmance liquid chromatographic methods were developed for separation of the four isomers of 17-hydroxy-16-hydroxymethyl-3-methoxyestra-1,3,5(10)-triene : reversed-phase separation on a Nucleosil ODS C-18 column with water-metha nol as mobile phase; and normal-phase separation on an APEX Silica column w ith hexane-dichloromethane-2-propanol as mobile phase; The effects of eluen t composition and flow-rate on the separation were investigated. This is th e first chromatographic evidence for the formation of the 16 alpha,17 alpha isomer in the reduction of 16-hydroxymethylene-3-methoxyestra-1,3,5(10)-tr ien-17-one. (C) 1999 Elsevier Science B.V. All rights reserved.