Probing topography and tailing for commercial stationary phases using AFM, FT-IR, and HPLC.

Atomic force microscopy was used to study surface characteristics of three chromatographic silica products: Agilent Zorbax SB300, Waters Symmetry 300, and Merck Chromolith. Each is modified with a monomeric C18 monolayer. Both topographic and adhesive force measurements were made for each product. Topographical images revealed that all three materials are as smooth as glass on the scale of 100 nm and below. Adhesive forces for all three materials were much lower and much more uniform than for chemically modified fused silica. FT-IR spectra for all three materials showed a low abundance of isolated silanols, thus explaining the low adhesion. Chromatograms of a cationic dye, 1,1'-didodecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI), ranging in concentration from 1 to 300 microM were obtained for each column. All three materials exhibited classic nonlinear tailing; the Zorbax exhibited fronting as well. Chromatographic simulations were performed for the Symmetry and Chromolith products to determine the number of strong adsorption sites. The AFM, FT-IR, and HPLC were all consistent in indicating that the Chromolith material had half as many strong adsorption sites as the Symmetry material. The Zorbax material exhibited a number of isolated silanols that was comparable to the other materials, yet its adhesive force suggested a less adsorptive material, and its chromatographic performance suggested a more adsorptive material. Its topography is discussed as a possible reason for its anomalous chromatographic behavior.