Chromatographic behavior of epirubicin and its analogues on high-purity silica in hydrophilic interaction chromatography

被引:84
作者
Li, RP [1 ]
Huang, JX [1 ]
机构
[1] Chinese Acad Sci, Ecoenvironm Sci Res Ctr, Beijing 100085, Peoples R China
关键词
hydrophilic interaction chromatography; mobile phase composition; adsorption isotherms; epirubicin; antibiotics;
D O I
10.1016/j.chroma.2004.04.033
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Hydrophilic interaction chromatography has been applied for the separation of epirubicin and its analogues using high-purity silica column with aqueous-organic mobile phase. Parameters affecting the chromatographic behavior of the solutes such as organic modifier, buffer pH, ionic strength and sample size, have been investigated. Of utmost importance for successful separation of these analogues is the choice of organic modifier, since it impacts both the solvent selectivity and the ionization of silica silanols as well as buffer solution, and consequently the retention behavior of solutes. Acetonitrile was shown to offer superior separation of these analogues to methanol, isopropanol or tetrahydrofuran. Results of the effects of organic modifier, buffer pH and ion strength indicate that the retention mechanism is a mixed-mode of adsorption and ion exchange. In addition, an irreversible adsorption of these compounds was found on silica in the weakly acidic or neutral mobile phases, and the effect of various factors on irreversible adsorption was also preliminarily discussed. More significantly, these basic compounds have exhibited peaks with a slanted front and a sharp tail, a typical overloading peak profile belonging to the behavior of competitive anti-Langmuir isotherm by increasing the sample size at the experimental conditions. (C) 2004 Elsevier B.V. All rights reserved.
引用
收藏
页码:163 / 169
页数:7
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