Can the theory of gradient liquid chromatography be useful in solving practical problems?
Advances in the theory of gradient liquid chromatography and their practical impacts are reviewed. Theoretical models describing retention in reversed-phase, normal-phase and ion-exchange modes are compared. Main attention is focused on practically useful models described by two- or three-parameter equations fitting the experimental data in the range of mobile phase composition utilized for sample migration during gradient elution. The applications of theory for gradient method development, optimization and transfer are addressed. The origins and possibilities for overcoming possible pitfalls are discussed, including the effects of the instrumental dwell volume, uptake of mobile phase components on the column and size of the sample molecules. Special attention is focused on gradient separations of large molecules.
Jandera
Pavel
P
Department of Analytical Chemistry, Faculty of Chemical Technology, University of Pardubice, Nám. Cs. Legií 565, CZ-53210 Pardubice, Czech Republic. pavel.jandera@upce.cz
D001704:Biopolymers / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange; D002853:Chromatography, Liquid; D008962:Models, Theoretical; D012997:Solvents
D001704:Biopolymers; D012997:Solvents