Direct observation of the separation process by the chromato-videoscope

Abstract

Separation processes in a liquid chromatographic column were visualized and analyzed by a developed chromatovideoscope. The migration aspects were evaluated with successively obtained densitograms. In reversed-phase chromatography, the band widths of each solute band were almost equal fore both weakly and strongly retained solutes when compared at the same column position (the same migration distance in the column). In the gradient elution mode, the position of the solute band showed that the migration velocity of the solute band changed gradually according to changes in solvent composition. The drug trapping process to BSA- coated ODS packings for direct injection of biological fluid was also observed. In the absence of BSA from the sample solution, the drug molecules were trapped in a narrow band. However, at higher BSA concentrations in the sample solution, a broader band shape was observed. This band broadening shows how the drug molecules were retained on the protein.