Abstract
Columns with inner diameters of 50 and 28 ΞΌm were slurryβpacked with 4β5 ΞΌm sizeβexclusion chromatography particles (Du Pont GF 250 Zorbax Bioseries) and compared with a commercially prepared 9.4βmm i.d. column that was packed with the same particles. The columns were compared on the basis of efficiencies, peak shapes, and van Deemter coefficients obtained for three proteins used as test analytes. The microcolumns provided more symmetrical peak shapes and lower plate heights than those provided by the commercial column if the peaks were analyzed using statistical moments. For example, h~min~ for bovine serum albumin was 5.6 in the commercial column and 2.1 in the microcolumn. This difference is attributed to the quality of the packing process, based on the fact that the van Deemter A term was significantly lower for the microcolumn than for the conventional column. The van Deemter data demonstrate that the A term was the major source of band broadening in the commercial column throughout the range of velocities used, and both the A and C terms were significant for the microcolumn. A 90 cm Γ 50 ΞΌm column was prepared and showed approximately 80% of the expected increase in efficiency.