19-08-2014, 10:17 AM
Gradient elution chromatography is a very powerful tool for the separation of components having a wide range of retentivity. It is not only used for chemical analysis, but also used in preparative and large-scale chromatography for the separation of many macromolecules, such as proteins. In gradient elution chromatography, the composition of the mobile phase is changed during the elution process.
Gradient elution chromatography is a very powerful tool for the separation of components having a wide range of retentivity. It is not only used for chemical analysis, but also used in preparative and large-scale chromatography for the separation of many macromolecules, such as proteins. In gradient elution chromatography, the composition of the mobile phase is changed during the elution process.
Therefore, it is possible to change the elution strength of mobile phase for the elution of individual sample components, which further results good resolution, selectivity, yield and purity of desired sample component in a reasonable time.
The present work comprises of developing a rigorous mathematical model to highlight the effect of gradient elution of adsorbed molecules and also to study the influence of the various parameters on the elution curve for the condition of both retaining as well as non-retaining. Model equations were solved using a new code developed in MATLAB 6.5 version. Experiments were carried out to study the effect of salt concentration on the adsorption isotherms of BSA protein on FP-DA adsorbent using different ionic strength of NaCl in 50 mM Tris buffer at pH 8.0. Later column experiments were carried out to validate the model predictions for different gradient condition.