SEPARATION OF HIGHLY COMPLEX MIXTURES By TWO-DIMENSION LIQUID CHROMATOGRAPHY Principal Investigator

Abstract

ABSTRACT This report summarizes the progress made on the title project during the grant period. We developed a new classification of two-dimensional separations based on the observation that separations can be made in time or in space. Thus, two-dimensional separations can be made in time×time, space×space, space×time, or time×space. The two successive separations must use two different modes of chromatography that afford uncorrelated or weakly correlated patterns of retention factors for the components of the samples analyzed. Our attention was mainly focused on the separation of protein digests, particularly, on those of the digests of myoglobin and bovine serum albumin as model systems and extremely efficient temporal separations were developed. We also designed and constructed new instruments to carry out space×space separations (True Bidimensional Chromatography, HPLC 2 or spacial separations) and time×space separations (a new hybrid combination of a temporal and a spacial separation that we designed).