THE CHARACTERIZATION OF BONDED PHASES FOR HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

The physical and chemical nature of chemically bonded phases used in high performance liquid chromatography have been studied. These bonded phases were characterized by a variety of chromotographic and non-chromatographic experiments. The non-chromatographic techniques included ¹³C NMR and batch extraction methods. The role played by the bonded phase as well as the mobile phase in determining the retention characteristics of the "stationary phase" were determined. The retention of solute molecules on bonded phases was found to be a function of the chain length of the bonded phase, the chemical nature of the bonded molecule, and the type of organic modifier used in the mobile phase. The energetics of the solute-stationary phase interactions was determined by the differential enthalpy and was found to be indicative of a partitioning process between two liquid phases. The retention process was also affected by the surface coverage of the bonded molecule. Optimum retention and separation characteristics were obtained with a hydrocarbon bonded phase of high surface coverage when used with a mobile phase containing a very polar organic modifier. The efficiency of these bonded phases was found to be independent of chain length as well as surface coverage of the bonded molecule. Some bonded phases which have specific functionalities incorporated into the bonded molecule are not true reversed phases. The selectivity of the bonded phases towards polar solute molecules was found to be affected by the type of organic modifier used in the mobile phase. The major accomplishment of this work shows that the stationary phase consists of the bonded molecule as well as trapped mobile phase. The composition of this ternary mixture is a function of the type and amount of bonded material and the type and amount of organic modifier used in the mobile phase