Abstract
The effect of the particle size of column packings on column efficiency in the reversed-phase high-performance liquid chromatographic separation of proteins was studied. Fast protein separations on non-porous octadecylsilyl (C 18) spherical silica gels of three different particle diameters (2, 5 and 20 μm) were examined, and five standard proteins were completely resolved within 20 s using a steep gradient elution with 0.1% aqueous trifluoroacetic acid—acetonitrile in all instances. The column efficiencies were approximately the same for all particle sizes. The large-particle (20 μm), non-porous C 18 column could be operated at room temperature for the fast protein separation. Rapid separations on conventional macroporous C 18 silica particles of 7 and 20 μm were also examined.
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