Measurement and use of retention data from high-performance gradient elution

摘要: Under “ideal” conditions it is possible to model retention in gradient elution so as be able calculate times, tg, a function of isocratic corresponding liquid chromatographic systems. In this paper we consider various “non-ideal” processes that lead errors calculated values tg. The more important these are solvent demixing due uptake one mobile phase component by the column packing, non-linear plots log k′ vs. time or composition and changes dead-time, t0, flow-rate. Expressions derived correct for effects, including equipment limitations discussed preceding paper. Calculated tg reversed-phase systems then agree with experimental within ± 1 % (1 standard deviation) total time, tG. These results should prove useful (a) improving precision (which comparable elution), (b) using efficient method development procedures (c) better understanding separations macromolecules such proteins.