FAST PROTEIN SEPARATION BY REVERSED-PHASE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY ON OCTADECYLSILYL-BONDED NONPOROUS SILICA-GEL - EFFECT OF PARTICLE-SIZE OF COLUMN PACKING ON COLUMN EFFICIENCY

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 (C18) spherical silica gels of three different particle diameters (2, 5 and 20-mu-m) 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-mu-m), non-porous C18 column could be operated at room temperature for the fast protein separation. Rapid separations on conventional macroporous C18 silica particles of 7 and 20-mu-m were also examined.