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Updated: Jun 28, 2025

Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns
Published on: April 26, 2016
Elution behavior of drugs in high-speed counter-current chromatography using on-column complexation with metal ions
Yukiko Moriiwa1, Atsushi Shoji2, Yoichi Shibusawa2
1Department of Biomedical Analysis, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, Tokyo, 192-0392, Japan. moriiwa@toyaku.ac.jp.
Abstract:
In this study, determination of (nitrogen containing) drugs by on-column complexation with metal ions in high-speed counter-current chromatography (HSCCC) was investigated. Bromazepam (BMP) was strongly retained in the organic upper stationary phase (UP) of the two-phase solvent system composed of tert-butyl methyl ether-acetonitrile-water (2:2:3, v/v/v) by eluting the aqueous lower mobile phase (LP) at a flow rate of 2 mL min-1. On the other hand, BMP (200 µg mL-1) was eluted faster without retention to the organic UP with the two-phase system containing 100 μg mL-1 of copper ions (CuCl2) because a very polar BMP-Cu2+ complex was immediately formed in the aqueous LP. The dramatic change in the retention behavior of BMP resulted from on-column complexation. The on-column complexation in HSCCC was further investigated for five (nitrogen containing) drugs and seven metal ions. In the result, tizanidine and phentolamine formed complexes with Al3+, Fe2+, Co2+, Ni2+, Cu2+, and Zn2+, ambroxol formed complexes with Al3+, Fe2+, and Cu2+, but voriconazole formed no complexes with all metal ions tested.
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