Various methods were tested for packing columns with silica having an average particle size of 15 μm for use in preparative medium pressure liquid chromatography (MPLC). Depending on the column type investigated, of the five methods tested, the two newly elaborated dry-packing procedures, using vacuum and nitrogen overpressure, resulted in the best packing density. Using these dry-filling techniques, the influence of various types of silica [with different average particle sizes (15 μm up to 130 μm) and particle size distributions (5-25 μm up to 63-200 μm)] on resolution and retention times was examined. Evaluation with different particle sizes showed a pronounced decrease of band broadening on using silica with an average particle size of 15 μm, which in turn resulted in a significant increase of resolution. The fluorescence of the test compounds used permitted observation of the chromatographic development. For preparative MPLC separations of the closely related test compounds a significant increase in resolution was obtained using silica with small particles and a narrow particle size distribution.
|Number of pages||18|
|Journal||Journal of Liquid Chromatography|
|Publication status||Published - szept. 1 1989|
ASJC Scopus subject areas
- Molecular Medicine