6533b86efe1ef96bd12cbd03

RESEARCH PRODUCT

Evaluation of advanced silica packings for the separation of biopolymers by high-perforamnce liquid chromatography

subject

description

Non-porous monodisperse 1.5-μm silicas were allowed to react with (A) and (B) N-acetylaminopropyltriethoxysilane to generate bonded phases useful in high-performance hydrophobic-interaction chromatography (HIC). Differences in the selectivity were observed between he amide and the ether phase. Peak capacities between 10 and 30 were achieved for several proteins with the amide and ether phase packed into columns of 36 × 8 mm I.D. and elution of the proteins under chromatographic conditions in which the gradient volume, VG, was held constant by varying the gradient time between 20 and 2.5 min and the flow-rate between 0.5 and 4.0 ml/min. The S values derived from the dependences of log k′ on the volume fraction of the low ionic strength buffer, ϕb, were of the same magnitude as reported for porous HIC silicas and showed a dependence on the molecular weight of the protein. Using these HIC stationary phases based on non-porous 1.5-μm supports, fast separations (<5 min)_could be carried out with high biological recoveries.