CHARACTERIZATION OF WIDE-PORE REVERSED-PHASE COLUMNS FOR BIOPOLYMER SEPARATIONS .1. SINGLE-PARAMETRIC EVALUATION

Nine prepacked wide-pore columns recommended for protein separation we re studied. The stationary phases investigated were widely different w ith respect to the type of ligands. In order to characterize the colum ns, measurements were made with six different test mixtures containing two different homologous series, substituted benzenes and amino acids , peptides and proteins. The isopotential eluent compositions obtained for the homologous series revealed the polar-apolar nature of the sta tionary phases, The relative hydrophobicity calculated for the individ ual test compounds provided more thorough but still partial characteri zation. The involvement of substituted benzenes resulted in the most c omplete description. The characterization with amino acids, peptides a nd proteins furnished results similar to those obtained for smaller te st compounds having low or intermediate polarity. The examination of t he behaviour of proteins revealed that the retention is determined by the hydrophobicity of the stationary phases and the other characterist ics affect the selectivity.