Complete chromatographic separation of steroids, including 17α and 17β-estradiols, using a carbazole-based polymeric organic phase in both reversed and normal-phase HPLC

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Poly(2-N-carbazolylethyl acrylate) with terminal trimethoxysilyl groups was prepared as an 2 organic phase and immobilized onto silica. The retention behavior of the column packed with this carbazole-based polymer-immobilized silica (Sil-CEA) was investigated by using various estrogenic steroids and corticoids in both reversed phase and normal phase liquid chromatographies. As a result, the complete separation was confirmed for eight kinds of steroids with Sil-CEA. The most specific separation with Sil-CEA can be emphasized by the high separation factor (e.g., α = 1.39 in methanol-water (7:3, v/v) at 35 °C) for 17α- and 17β-estradioles, that are one of the most difficult isomers in chromatographic separation, while two kinds of commercially available polymeric ODSs as references shows only α = 1.01 at the same condition. Since the excellent separation and retention order with Sil-CEA was maintained even in a normal phase mobile phase such as a hexane-2-propanol mixture, it is estimated that the CEA phase has multiple interaction mechanisms through stronger interactions such as dipole-dipole, carbonyl-π and hydrogen bonding interactions than a hydrophobic effect expected in ODS.

http://www.springerlink.com/content/k4003417ug062622/

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