ABSTRACT
This chapter is concerned specifically with one aspect of these developments, namely general considerations of the basic theory which underpin the successes of high-performance liquid chromatography (HPLC) in peptide protein chemistry as evident today. The separation of a mixture of polar, ionizable substances such as peptides and proteins in a chromatographic column is the result of two events that occur within the column. The time taken for a solute to completely pass through a chromatographic bed is called the retention time. Differences in the molecular characteristics and interactive behavior of peptide and protein solutes can thus be revealed in chromatographic separation, either through differences in unitary free energies of the solutes, or as changes in selectivity. Zone broadening of solutes in chromatographic systems arises from two causes, an intracolumn cause and an extra-column cause due to the instrumentation characteristics. Retention behavior will be affected by the hydrodynamic characteristics and fluid dynamic properties of the chromatographic system and the solute components.
