Morphological and chromatographic characterization of molecularly imprinted monolithic columns

The chromatographic stationary phases based on molecularly imprinted monoliths were prepared by free radical polymerization and subsequently characterized. The mixture of commonly used comonomers of ethylene dimethacrylate/methacrylic acid was polymerized in presence of various porogenic mixtures. Tosyl-L-phenylalanine was selected as a model template. Polymerization steps were carried out in the glass columns (i.d. of 3.3 mm) enabling UV initiation. Difficulties encountered with polymerization in situ, e. g. volume contraction and adhesion of polymer onto the glass inner wall, were successfully solved. The morphology of monoliths was investigated by electron microscopy, mercury porosimetry and surface area measurements. The influence of polymerization conditions on monolith morphology and subsequent chromatographic properties was evaluated. Polymers prepared by UV polymerization showed higher total porosity and the most frequent pore radius compared to the same polymers prepared thermally. The prepared monoliths by UV irradiation were also significantly more permeable for mobile phase during the chromatography tests. Morphology of monoliths prepared in dodecanol/toluene porogenic mixture markedly depended on the temperature of polymerization; the most frequent pore radius decreased with increasing temperature. From chromatographic point of view, monolithic columns were tested by separations of standard hydrophobic solutes. Height equivalent to a theoretical plate reached the values of approx. 25 mu m, Walters indices of hydrophobicity laid slightly above 3. Prepared monoliths were compared to the conventional columns, filled with the particles prepared from the polymer of the same composition and preparation. In many cases the monolithic columns showed better chromatographic parameters.