Analysis of applying different solvents for the mobile phase and for sample injection

Overloading a chromatographic column with a compound possessing low solubility in the mobile phase has been investigated. In order to increase the concentration of injection a strong solvent for dissolving the feed was used. The injection of such concentrated samples brings the risk of triggering undesired crystallisation processes. A model system has been investigated with ethanol-water as the mobile phase and dl-threonine as the sample dissolved in pure water. Under extreme overloaded conditions band splitting was observed. Measurements of the adsorption isotherms and systematic solubility studies were carried out. For the process analysis a simplified mathematical model was applied. The simulations of the band profiles were compared with the experimental data.