Separation of enantiomers on chiral stationary phase based on chicken α1-acid glycoprotein: Effect of silica particle diameters on column performance

The effects of silica particle diameters on performances of chicken alpha(1)-acid glycoprotein (c-AGP)-immobilized silica particle columns were investigated. c-AGP was immobilized onto aminopropyl silica particles, whose nominal particle diameters were 5, 3 and 2.1 mu m, activated with N,N'-disuccinimidyl carbonate. The retention factor (k), enantioseparation factor (alpha), resolution (R-s) and height equivalent to a theoretical plate (H) of solutes on three c-AGP columns were evaluated using a mixture of phosphate buffer and organic modifier as a mobile phase in LC. There were not so much differences in their k and a values among three c-AGP columns, while their R-s values were in the order of 2.1 mu m >3 mu m > 5 mu m silica particles and their H values were in the reversed order. Since three c-AGP columns gave almost the same enantioseparation factors for solutes, their highest R-s and lowest H values on a c-AGP-immobilized column prepared with 2.1-mu m silica particles came from its highest column efficiency among there c-AGP columns. These results suggest that 2.1-mu m silica particles could be useful for the preparation of c-AGP or protein-based CSPs. (C) 2014 Elsevier B.V. All rights reserved.