Simple method for determining the amount of ion-exchange groups on chromatographic supports

被引：38

作者：

Lendero, N ^{[1
]}

Vidic, J ^{[1
]}

Brne, P ^{[1
]}

Podgornik, A ^{[1
]}

Strancar, A ^{[1
]}

机构：

[1] BIA Separat DOO, SI-1000 Ljubljana, Slovenia

来源：

JOURNAL OF CHROMATOGRAPHY A | 2005年 / 1065卷 / 01期

关键词：

ion exchangers; total ionic capacity; pH transient; monolithic columns; convective interaction media (CIM); resin characterization;

D O I：

10.1016/j.chroma.2004.10.072

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The objective of this study was to develop a fast, simple, non-destructive, non-toxic and low-priced method for determining the amount of ionic groups on resins, since the conventional titration method fails to give proper results on methacrylate monoliths. After the column had been pre-saturated with a high concentration buffer solution, a low concentration buffer solution of the same pH value was pumped through the column. Measuring pH and absorbance, the profiles with a shape of typical break-through curve were obtained. It was shown that the time of the pH transient, which appeared under such conditions, could be used as a measure of the total ionic capacity of ion-exchange monolithic columns. The effect of the column length, linear velocity and varying concentrations of buffer solutions on the time of the pH transient was examined. The method was shown to be suitable for determining the amount of ionic groups on both anion and cation monolithic columns. In addition, it could also be applied to particle bed columns. The time of the pH transient and the protein dynamic binding capacity were also compared and it was concluded that for a given monolith the protein capacity can be derived from the data obtained by the new method. (C) 2004 Elsevier B.V. All rights reserved.

引用

页码：29 / 38

页数：10

共 6 条

[1] High-performance chromatofocusing using linear and concave pH gradients formed with simple buffer mixtures I. Effect of buffer composition on the gradient shape [J].