The Influence of Reversed-Phase n-Alkyl Chain Length on Protein Retention,Resolution and Recovery: Implications for Preparative HPLC

Abstract

High-performance liquid chromatography (HPLC) of proteins on reversed-phase columns of varying n-alkyl chain length (C₂ to C₂₂ was studied using a trifluoroacetic acid/2-propanol mobile phase system. Protein resolution was influenced by chain length but retention times for proteins, unlike those of small molecules, were relatively constant, independent of chain length or carbon loading. Loading capacities were found to be affected by chain length, and aspects of protein interaction with stationary phase are discussed.