Abstract
A column switching high-performance liquid chromatographic method with electrochemical detection was developed for the simultaneous determination of two modified steroidal antiandrogen anthrasteroids: 3aβ, 6-dimethyl- 2, 3, 3a, 4, 5, 8, 9, 10, 10aβ, 11, 11aβ, 11bα-dodecahydro-1H-cyclopenta[α]anthracene-3, 8-dione (I) and its 3-hydroxy derivative (II) in human serum. After solid phase extraction from serum, the anthrasteroids were derivatized with 2, 5-dihydroxybenzohydrazide during the evaporation process of the reaction mixture at 20°C for 25min. The derivatives were detected electrochemically at +0.45V vs. Ag/AgCl. The detection limits (S/N=3) for compounds I and II in 0.5ml of human serum were 0.5 and 1ng ml-1, respectively. Calibration curves for both compounds were linear over concentration ranges of 2-20ng ml-1 and 20-200 ng ml-1, respectively.
