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THE SYNTHESIS AND CHARACTERIZATION OF BONDED PHASE CHROMATOGRAPHIC ADSORBENTS
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THE SYNTHESIS AND CHARACTERIZATION OF BONDED PHASE CHROMATOGRAPHIC ADSORBENTS
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Dissertation
THE SYNTHESIS AND CHARACTERIZATION OF BONDED PHASE CHROMATOGRAPHIC ADSORBENTS
1982
Overview
Several phenyl alkyl bonded phases for liquid chromatography were synthesized and characterized by liquid chromatography, gas chromatography and ('13)C nuclear magnetic resonance (NMR) spectroscopy. The chromatographic physical parameters investigated include a quantitative determination of the mobile phase volume and the stationary phase volume. The stationary phase volume was determined to be a function of the bonded moiety chain length and the chromatographic solvent system employed. The interpretation of the stationary phase volume is discussed in terms of the porous nature of the silica gel support. The chemical parameters determined include a quantitative determination of the mobile phase and stationary phase composition (which were different from each other). The selectivity of the chromatographic separations was dependent on the chemical composition and the volume of both the stationary and mobile phases. Carbon 13 NMR spectroscopy provided information about the environment of the bonded moiety in the stationary phase. The liquid-like nature of the bonded moiety was influenced by the chain length of the attached species, the choice of organic modifier, and the chemical composition of the solvents. Temperature did not appear to play a role in the line widths under the experimental conditions examined. The separation of several peptide diastereoisomers on different commercially available hydrocarbon bonded adsorbents is also reported. Select diastereoisomers of arginine vasopressin and oxytocin are extremely sensitive to differences in the composition of the stationary phase. The selectivity and elution order were dependent upon the choice of adsorbent and solvent system employed. The addition of a second organic modifier provided a method for the dynamic modification of the stationary phase. The ability to dynamically modify the stationary phase can enhance the selectivity for the separation of selected peptide diastereoisomers.
