Liquid Chromatographic Studies of Silica-Immobilized Bovine Serum Albumin Under Normal-Phase Conditions: Separation of Phenolic Solutes Using Ternary Mixtures of Hydrogen Chloride-Diethyl Ether-Hexane as Eluents

The capacity factors, k', for multifunctional solutes with hydroxyl and carboxyl groups are measured on silica-immobilized bovine serum albumin (BSA) using various compositions of hydrogen chloride saturated diethyl ether, hexane, and neat diethyl ether. The values of k' are smaller at higher concentrations of free ether and larger as more of the ether is complexed with hydrogen chloride. Additionally, solute retention increases with increasing hydroxyl and carboxyl substituents in the molecule. The data indicate that solute retention is consistent with normal-phase behavior, which, to our knowledge, makes this work the first successful normal-phase application of silica-immobilized BSA to be reported.