WHOLE-COLUMN DETECTION CHROMATOGRAPHY FOR HPLC

A NEW MULTIDIMENSIONAL TECHNIQUE FOR HIGH-PERFORMANCE LIQUIDCHROMATOGRAPHY (HPLC) WILL BE DEVELOPED FOR THE ANALYSIS OF BIOLOGICAL SAMPLES. IN WHOLE-COLUMN DETECTION CHROMATOGRAPHY (WCDC), COMPONENTS OF A COMPLEX MIXTURE ARE DETECTED SIMULTANEOUSLY AT MULTIPLE DETECTION ZONES ALONG THE ENTIRE LENGTH OF A LIQUID CHROMATOGRAPHIC COLUMN. IT IS ANTICIPATED THAT THIS TECHNIQUE WILL OFFER MANY ADVANTAGES COMPARED WITH PRESENT CHROMATOGRAPHIC AND DETECTION METHODS DUE TO THE ABILITY TO CONSTANTLY MONITOR THE SEPARATION PROCESS, ALTER CONDITIONS DURING THE SEPARATION TO ACHIEVE OPTIMUM RESOLUTION OF THE COMPONENTS, AND THE POSSIBLE INCREASE IN SENSITIVITY THAT COULD BE OBTAINED BY THE MEASUREMENT OF THE ANALYTE SIGNAL THROUGHOUTTHE SEPARATION PROCESS VERSUS POSTCHROMATOGRAPHIC DETECTION. DURING PHASE I, THE TECHNIQUE WILL BE DEVELOPED FOR USE WITHISOCRATIC HPLC USING UV DETECTION. THIS RESEARCH SHOULD LEAD TO A BETTER UNDERSTANDING OF THE MECHANISMS OF SEPARATION USING DIFFERENT MOBILE PHASES OR SOLVENT PLUGS DURING A SINGLE CHROMATOGRAPHIC ANALYSIS. PHASE II RESEARCHWILL EXTEND THIS TECHNIQUE TO ENABLE AN EXPERT SYSTEM SEPARATION IN REAL TIME AND TO DEVELOP WCDC ANALYSES FOR MEASUREMENT OF BIOLOGICALLY IMPORTANT COMPOUNDS THAT CANNOT BE ACHIEVED BY CURRENT HPLC TECHNIQUES.