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Derivatization of Ketosteroids for High-Performance Liquid Chromatography with Electrochemical Detection
Abstract:Abstract

A highly sensitive and selective derivatization of ketosteroids for use in high-performance liquid chromatography with electrochemical detection (HPLC/EC) is described. When the detector response for three phenylhydrazone derivatives was compared with one another using dehydroepiandrosterone, the p-nitrophenylhydrazone showed the highest sensitivity with a detection limit of 200 pg. Dehydroepiandrosterone and other principal 17-ketosteroids in human blood were quantitatively derivatized into p-nitrophenylhydrazones and efficiently separated by HPLC/EC on a μBondapak C1 8 column using 0.5%. NH4H2PO4/methanol (2:7, v/v).
Keywords:electrochemical detection  HPLC  ketosteroids  p-nitrophenylhydrazone  blood plasma
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